Natural lipid particle formulations for agricultural applications

Abstract

Disclosed herein are compositions comprising a plurality of naturally-derived lipid particles comprising a phospholipid, a non-polar lipid, and a surface modifier, and further comprising a bioactive material, for use in various agricultural methods.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 444,331, filed February 9, 2023, and U.S. Provisional Patent Application No. 63 / 447,834, filed February 23, 2023, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] The effectiveness of agricultural chemicals, such as pesticides, that are applied to soil can be limited by the degree to which the chemical interacts with soil.High affinity to soil negatively affects the ability of chemicals to reach their intended targets, such as plants or plant pests.Similarly, agricultural chemicals that have very little affinity to soil are quickly washed away into groundwater, thereby limiting their ability to interact with their intended targets.Therefore, there is a need in the art for compositions and methods that can change the interaction between agricultural chemicals and soil, thereby affecting their mobility in soil and their ability to reach their intended targets.

[0003] Furthermore, the effectiveness of heterofunctional agricultural agents, such as pesticides, can be limited by the extent to which the agent is taken up by the plant. Thus, there is a need in the art for compositions and methods that can improve the interaction of an agent with a plant or plant part, for example, by enhancing the uptake of the agent by the plant or plant part. Summary of the Invention [Means for solving the problem]

[0004] In one aspect, the present disclosure provides an agricultural composition comprising a plurality of naturally occurring lipid particles (NLPs), each comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, wherein the NLPs comprise a hydrophobic core. In some embodiments, the at least one surface modifier modifies the mobility of the agricultural composition through soil compared to a composition without the surface modifier. In some embodiments, the hydrophobic core is composed of lipid. In some embodiments, an agricultural agent is provided in the hydrophobic core.

[0005] In some embodiments, the at least one phospholipid is selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid, phosphatidylserine, and 1,2-dimyristoyl-sn-glycero-3-phosphate. In some embodiments, the at least one phospholipid is derived from lecithin. In some embodiments, the lecithin is soybean lecithin or sunflower lecithin.

[0006] In some embodiments, the at least one non-polar lipid comprises at least one fatty acid chain selected from the group consisting of a polyunsaturated fatty acid chain, a monounsaturated fatty acid chain, and a saturated fatty acid chain.

[0007] In some embodiments, the NLP comprises at least one phospholipid layer. In some embodiments, the NLP phospholipid layer is a phospholipid bilayer. In some embodiments, the NLP has a micellar structure. In some embodiments, the NLP comprises a hydrophobic core. In some embodiments, the hydrophobic core comprises at least one non-polar lipid.

[0008] In some embodiments, the hydrophobic core is a solid.

[0009] In some embodiments, a surface modifier is incorporated into the phospholipid layer. In some embodiments, the surface modifier is selected from the group consisting of glycolipids, polysaccharides, fatty acid ethoxylates, linear alcohol ethoxylates, cetyl trimethyl esters, linear isopropylamine dodecylbenzene sulfonates, tristyrylphenol ethoxylate phosphate esters, modified styrene-acrylic copolymers, hydrophobically modified polycarboxylate polymers, anionic polymers, nonionic acrylic copolymers, nonionic combination polymers, tristyrylphenol polyalkylene oxide block copolymers, or headgroup-modified PEG lipids. In some embodiments, the headgroup-modified PEG lipid is PEG2000-C18 or PEG5000-C18. In some embodiments, the glycolipid is a rhamnolipid or sophorolipid. In some embodiments, the anionic polymer is Atlox 500L, Atlox 4917, or Atlox CS100B. In some embodiments, the polysaccharide is a C8-C10 alkyl polysaccharide.

[0010] In some embodiments, the surface modifier stabilizes the integrity of the NLP. In some embodiments, the surface modifier affects the binding of the NLP to one or more components present in the soil. In some embodiments, the surface modifier affects the affinity of the NLP for one or more components present in the soil.

[0011] In some embodiments, the surface modifier affects the surface charge of the NLPs. In some embodiments, the NLPs exhibit a negative surface charge as evidenced by a negative zeta potential. In some embodiments, the negative zeta potential is in the range of -10 to -100 mV. In some embodiments, the negative zeta potential increases the mobility of the NLPs through soil.

[0012] In some embodiments, the agricultural composition further comprises a co-solvent, in some embodiments, the co-solvent is selected from the group consisting of fatty acid methyl esters, non-ionic emulsifiers, propylene glycol, ethyl lactate, non-ionic block copolymer surfactants or non-ionic polyalkylene glycol ethers, dichloromethane, and isopropyl myristate.

[0013] In some embodiments, the NLP further comprises at least one heterologous agricultural agent. In some embodiments, the at least one heterologous agricultural agent is selected from the group consisting of pesticides, fertilizers, herbicides, plant modifiers, insect attractants, plant growth promoters, biostimulants, and plant immunity inducers. In some embodiments, the pesticide is selected from the group consisting of antifungals, antioomycetes, antibacterial agents, insecticides, molluscicides, nematicides, herbicides, and viricides. In some embodiments, the NLP comprises a combination of two or more heterologous agricultural agents. In some embodiments, the NLP comprises two or more agricultural agents independently selected from the group consisting of antifungals, antioomycetes, antibacterial agents, insecticides, molluscicides, nematicides, herbicides, and viricides. In some embodiments, the NLP comprises at least two insecticides with different modes of action. In some embodiments, the NLP comprises at least two antifungals with different modes of action. In some embodiments, the NLP comprises at least two antioomycotic agents with different modes of action. In some embodiments, the NLP comprises at least two antibacterial agents with different modes of action. In some embodiments, the NLP comprises at least two molluscicides with different modes of action. In some embodiments, the NLP comprises at least two nematicides with different modes of action. In some embodiments, the NLP comprises at least two herbicides with different modes of action. In some embodiments, the NLP comprises at least two virucides with different modes of action.

[0014] In some embodiments, (a) the antifungal agent is azoxystrobin, mancozeb, prothioconazole, folpet, tebuconazole, difenoconazole, captan, bupirimate, fosetyl-AI, strobilurin, dimoxystrobin, enestrobrin, fluoxastrobin, kresoxim-methyl, metominostrobin, picoxystrobin, pyraclostrobin, trifloxystrobin, oryzastrobin, carboxamide, carboxanilide, benalaxyl, benalaxyl-M, benodanil, carboxin, mebenil, mepronil, Fenfuram, fenhexamid, flutolanil, furalaxyl, flucarbanil, furametpyr, metalaxyl, metalaxyl-M, metofuroxam, metsulfovax, ofrace, oxadixyl, oxycarboxin, penthiopyrad, pyracarbollide, salicylanilide, tecloftalam, thifluzamide, tiadinil, N-biphenylamide, bixafen, boscalid, carboxylic acid morpholide, dimethomorph, flumorph, benzamide, flumetobir, fluopicolide, zoxamide, carboxamide, carpropamid, diclocymet, man Dipropamide, silthiofam, azoles, triazoles, bitertanol, bromuconazole, cyproconazole, diniconazole, enilconazole, epoxiconazole, fenbuconazole, flusilazole, fluquinconazole, flutriafol, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propiconazole, prothioconazole, simeconazole, tetraconazole, triadimenol, triadimefon, triticonazole, imidazole, cyazofamid, imazalil , pefurazoate, prochloraz, triflumizole, benzimidazole, benomyl, carbendazim, fuberidazole, thiabendazole, ethaboxam, etridiazole, hymexazole, pyridine, faizinam, pyrifenox, pyrimidine, cyprodinil, ferimzone, fenarimol, mepanipyrim, nuarimol, pyrimethanil, piperazine, triforine, pyrrole, fludioxonil, fenpiclonil, morpholine, aldimorph, dodemorph, fenpropimorph, tridemorph, dicarboximide, iprodione, procymidone,Vinclozolin, acibenzolar-S-methyl, anilazine, captafol, dazomet, diclomezine, fenoxanil, folpet, fenpropizine, famoxadone, fenamidone, octilinone, probenazole, proquinazide, pyroquilon, quinoxyfen, tricyclazole, carbamate, dithiocarbamate, ferbam, maneb, metiram, metam, propineb, thiram, zineb, ziram, diethofencarb, flubenthiavalicarb, iprovalicarb, propamocarb, guanidine, dodine, iminoc Tadine, guazatine, kasugamycin, polyoxins, streptomycin, validamycin A, fentin salts, sulfur-containing heterocyclyl compounds, isoprothiolane, dithianon, organophosphorus compounds, edifenphos, fosetyl, fosetyl-aluminum, iprobenfos, pyrazophos, tolclofos-methyl, organochlorine compounds, thiophanate-methyl, chlorothalonil, dichlofluanid, tolylfluanid, flusulfamide, phthalide, hexachlorobenzene, pencycuron, quintozene, nitrophenyl derivatives, binapac methylpyrazole-4-carboxamide, 3-[5-(4-chlorophenyl)-2,3-dimethylisoxazolidin-3-yl]-pyridine, N-(3',4'-dichloro-4-fluorobiphenyl-2-yl)-3-difluoromethyl-1- ... Chilpyrazole-4-carboxamide, 5-chloro-7-(4-methylpiperidin-1-yl)-6-(2,4,6-trifluorophenyl)-[1,2,4]triazolo[1,5-a]pyrimidine, 2-butoxy-6-iodo-3-propylchromen-4-one, N,N-dimethyl-3-(3-bromo-6-fluoro-2-methylindole-1-sulfonyl)-[1,2,4]triazole-1-sulfonamide, methyl-(2-chloro-5-[1-(3-methylbenzyloxyimino)-ethyl]benzyl)carbamate,Methyl-(2-chloro-5-[1-(6-methylpyridin-2-ylmethoxy-imino)ethyl]benzyl)carbamate, methyl 3-(4-chlorophenyl)-3-(2-isopropoxycarbonylamino-3-methylbutyryl-amino)propionate, 4-fluorophenyl N-(1-(1-(4-cyanophenyl)ethanesulfonyl)but-2-yl)carbamate, N-(2-(4-[3-(4-chlorophenyl)prop-2-ynyloxy]-3-methoxyphenyl)ethyl)-2-methanesulfonylamino-3-methylbutyramide, N-(2-(4-[3-(4-chlorophenyl)prop-2-ynyloxy]-3-methoxyphenyl)ethyl) )ethyl)-2-ethanesulfonylamino-3-methylbutyramide, N-(4'-bromobiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, N-(4'-trifluoromethylbiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, N-(4'-chloro-3'-fluorobiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, methyl 2-(ortho-((2,5-dimethylphenyloxy-methylene)phenyl)-3-methoxyacrylate, oxathiapiproline, and esters and salts thereof; (b) Antibacterial agents include hypochlorite, sodium hypochlorite, chloramine, dichloroisocyanurate, trichloroisocyanurate, wet chlorine, chlorine dioxide, peroxide, peracetic acid, potassium persulfate, sodium perborate, sodium percarbonate, urea hydrogen peroxide, iodine, iodopovidone, ethanol, 1-propanol, 2-propanol, 2-phenoxyethanol, phenol, cresol, halogenated phenols, hexachlorophene, triclosan, trichlorophenol, tribromophenol, pentachlorophenol, cationic surfactants, benzalkonium chloride, cetyltrimethylammonium bromide, cetyltrimethylammonium chloride, didecyldimethylammonium chloride, cetylpyridinium chloride, benzethonium chloride, chlorhexidine, glucoprotamine, octenidine dihydrochloride, ozone solution, colloidal silver, silver nitrate, mercury chloride, phenylalanine hydrochloride, silver salts, copper sulfate, copper oxide-chloride, copper hydroxide, copper octanoate, copper oxychloride sulfate, copper sulfate pentahydrate, phosphoric acid, nitric acid, sulfuric acid, amidosulfonic acid, toluenesulfonic acid, sodium hydroxide, potassium hydroxide, calcium hydroxide, sorbic acid, benzoic acid, lactic acid, salicylic acid, penicillin, cephalosporin, vancomycin, polymyxin, rifamycin, lipiamycin, quinolone, sulfonamide, aminoglycoside, kasugamycin, macrolide, lincosamide, tetracycline, cyclic lipopeptide, daptomycin, glycylcycline, tigecycline, oxazolidinone, linezolid, fidaxomicin, rifampicin, ciprofloxacin, doxycycline, ampicillin, polymyxin B, gramicidin, isoniazid, pyrazinamide, ethambutol, myanbutol, streptomycin, and esters and salts thereof; (c) Insecticides include chloronicotinyls, neonicotinoids, acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, nithiazine, thiacloprid, thiamethoxam, imidaclothiz, (2E)-1-[(2-chloro-1,3-thiazol-5-yl)methyl]-3,5-dimethyl-N-nitro-1,3,5-triazinan-2-imine, acetylcholinesterase (AChE) inhibitors, carbamates, alanycarb, aldicarb, aldoxicarb, alixicarb, aminocarb, bendiocarb, benfuracarb, and bupropion. Fencarb, butacarb, butocarboxim, butoxycarboxim, carbaryl, carbofuran, carbosulfan, cloethocarb, dimethylan, ethiofencarb, fenobucarb, fenothiocarb, formetanate, furathiocarb, isoprocarb, metam-sodium, methiocarb, methomyl, metolcarb, oxamyl, phosphocarb, pirimicarb, promecarb, propoxur, thiodicarb, thiofanox, triazamate, trimethacarb, XMC, xylylcarb, organophosphate esters, acephate, azamethiphos azinphos (-methyl, -ethyl), bromophos-ethyl, bromfenvinphos (-methyl), butathiophos, cadusafos, carbophenothion, chlorethoxyphos, chlorfenvinphos, chlormephos, chlorpyrifos (-methyl / -ethyl), coumaphos, cyanofenphos, cyanophos, demeton-S-methyl, demeton-S-methyl sulfone, dialifos, diazinon, diclofenthion, dichlorvos / DDVP, dicrotophos, dimethoate, dimethylvinphos, dioxabenzophos, disulfoton, EPN, ethion, ethoproxen Phos, etrimphos, fanfur, fenamiphos, fenitrothion, fensulfothion, fenthion, flupyrazophos, fonofos, formothion, fosmetilan, fosthiazate, heptenophos, iodofenphos, iprobenfos, isazophos, isofenphos, isopropyl O-salicylate, isoxathion, malathion, mecarbam, methacrifos, methamidophos, methidathion, mevinphos, monocrotophos, naled, omethoate, oxydemeton-methyl, parathion (-methyl / -ethyl), phenthoate, phorate, phosalone,Phosmet, phosphamidon, phosphocarb, phoxim, pirimiphos (-methyl / -ethyl), profenofos, propafos, propetamphos, prothiofos, prothoate, pyraclofos, pyridaphenthion, pyridathion, quinalphos, cebufos, sulfotep, sulprofos, tebupirimfos, temephos, terbufos, tetrachlorvinphos, thiometon, triazophos, trichlorfon, vamidothion, pyrethroids, acrinathrin, allethrin (d-cis-trans, d-trans), cypermethrin (α-, β-, θ-, ζ-), permethrin (cis-, trans-), β-cyfluthrin, bifenthrin, bioallethrin, bioallethrin-S-cyclopentyl isomer, bioethanomethrin, biopermethrin , bioresmethrin, clovaportrin, cis-cypermethrin, cis-resmethrin, cis-permethrin, clocithrin, cycloprothrin, cyfluthrin, cyhalothrin, cyphenothrin, DDT, deltamethrin, empenthrin (1R-isomer), esfenvalerate, etofenprox, fenfluthrin, fenpropathrin, fenpyrithrin, fenvalerate, flubrocythrinate, flucythrinate, flufenprox, flumethrin, fluvalinate, fubufenprox, γ-cyhalothrin, imiprothrin, kadethrin, λ, metofluthrin, fenothrin (1R-trans isomer), prallethrin, profluthrin, protrifenbut, pyresmethrin, resmethrin, RU 15525, silafluofen, tau-fluvalinate, tefluthrin, telalethrin, tetramethrin (1R-isomer), tralocitrin, tralomethrin, transfluthrin, ZXI 8901, pyrethrins, pyrethrum, oxadiazine, indoxacarb, acetylcholine receptor modulators, spinosyn, spinosad, cyclodiene, camphechlor, chlordane, endosulfan, gamma-HCH, HCH, heptachlor, organochlorines, lindane, methoxychlor, fiprole, acetoprole, ethiprole, vaniliprole, fipronil, mectin, abamectin, avermectin, emamectin, emamectin benzoate, fenoxycarb, hydroprene, kinoprene, methoprene, ivermectin, lepimectinEpofenonane, pyriproxyfen, milbemectin, milbemycin, triplen, diacylhydrazine, chromafenozide, halofenozide, methoxyfenozide, tebufenozide, benzoylurea, bistrifluron, chlorfluazuron, diflubenzuron, fluazuron, flucycloxuron, flufenoxuron, hexaflumuron, lufenuron, novaluron, noviflumuron, penfluoron, teflubenzuron, triflurane Lumulon, organotins, azocyclotin, cyhexatin, fenbutatin oxide, pyrrole, chlorfenapyr, dinitrophenol, pinapacryl, dinobuton, dinocap, DNOC, METI, fenazaquin, fenpyroximate, pyrimidifen, pyridaben, tebufenpyrad, tolfenpyrad, rotenone, acequinocyl, fluacrypyrim, microbial disruptors of insect gut membranes, Bacillus thuringiensis (Bacillus thuringiensis strains, lipid synthesis inhibitors, tetronic acid, tetramic acid, spirodiclofen, spiromesifen, spirotetramat, cis-3-(2,5-dimethylphenyl)-8-methoxy-2-oxo-1-azaspiro[4.5]dec-3-en-4-ylethyl carbonate, carboxamide, flonicamide, octopamine agonist, amitraz, inhibitor of magnesium-stimulated ATPase, propargite, ryanodine receptor agonist, phthalamide, linaxapyr, N2-[1,1-dimethyl-2-(methylsulfonyl)ethyl]-3-iodo-N1-[2-methyl-4-[1,2,2,2-tetrafluoro-1-(trifluoromethyl) Ethyl]phenyl]-1,2-benzenedi-carboxamide, spidoxamato, nicofluprole, tetraniliprole, thioxazafen, flupyradifurone, fluopyram, flubendiamide, deltamethrin, permethrin, ginpropylidaz, brofuranilide, afidopiropen, fluopyram, fluazaindolizine, triflumezopyrim, sulfoxaflor, spinetoram, chlorpyrifos, spinosad, cyantraniliprole, chlorantraniliprole, cypermethrin, prinazoline, cyclobutrifluram, spiropydione, fluensulfone, pymetrozine, thiamethoxam, lambda cyhalothrin, oxazosulfil, benzpyrimoxane,at least one of dichloromezothiaz, flupentiofenox, fluhexafon, fluxamethamide, flupirimine, cyhalodiamide, acinonapyr, cyclaniliprole, cetopyrafen, ciprofuranilide, tetrachlorantraniliprole, isocycloceram, brofuranilide, spiropydione, and esters and salts thereof; (d) the molluscicide comprises at least one of a metal salt, iron phosphate, aluminum sulfate, sodium ferric EDTA, metaldehyde, methiocarb, and an acetylcholinesterase inhibitor; (e) the nematicide comprises at least one of a fumigant, DD, 1,3-dichloropropene, ethylene dibromide, 1,2-dibromo-3-chloropropane, methyl bromide, chloropicrin, metam sodium, dazomet, methyl isothiocyanate (MITC), sodium tetrathiocarbonate, carbamate, aldicarb, aldoxycarb, carbofuran, oxamyl, creotocarb, organophosphate ester, ethoprophos, fenamiphos, cadusafos, fosthiazate, fensulfothion, thionazine, isazophos, and a biochemical; and (f) Herbicides include glufosinate, propaquizafop, metamitron, metazachlor, pendimethalin, flufenacet, diflufenican, clomazone, nicosulfuron, mesotrione, pinoxaden, sulcotrione, prosulfocarb, sulfentrazone, bifenox, quinmerac, triallate, terbuthylazine, atrazine, oxyfluorfen, diuron, trifluralin, chlorotoluron, benzoic acid herbicides, dicamba, phenoxyalkanoic acid herbicides, 2,4-D, MCPA, 2,4-DB esters, and aryloxyphenoxy. Cypropionic acid herbicides, clodinafop, cyhalofop, fenoxaprop, fluazifop, haloxyfop, quizalofop esters, pyridine carboxylic acid herbicides, aminopyralid, picloram, clopyralid esters, pyrimidine carboxylic acid herbicides, aminocyclopyrachlor esters, pyridyloxyalkanoic acid herbicides, fluoroxypyr, triclopyr, hydroxybenzonitrile herbicides, bromoxynil, ioxynil, arylpyridine carboxylic acids, arylpyrimidine carboxylic acids, acetochlor, acifluorfen, alachlor, alachlor Metrin, amitrole, asuram, azafenidin, benefin, bensulfuron, bensulide, bentazon, bromacil, butyrate, carfentrazone, chloramben, chlorimuron, chlorpropham, chlorsulfuron, clethodim, clopyralid, cloransulam, cyanazine, cycloate, DCPA, desmedipham, dichlobenil, diclofop, diclosulam, diethathyl, difenzoquat, diflufenzopyr, dimethenamid-p, diquat, DSMA, endosole, EPTC, ethalfluralin, ethametsulfuron , ethofumesate, fluazifop-P, flucarbazone, flumetsulam, flumiclorac, flumioxazin, fluometuron, fluroxypyr, fluthiaset, fomesafen, foramsulfuron, glyphosate, halosulfuron, haloxyfop, hexazinone, imazamethabenz, imazamox, imazapic, imazaquin, imazethapyr, isoxaben, isoxaflutole, lactofen, linuron, MCPB, methazole, metolachlor-s, metribuzin, metsulfuron, molinate, MSMA, napropamide, naptalam,norflurazon, oryzalin, oxadiazon, oxasulfuron, oxyfluorfen, paraquat, pebulate, pelargonic acid, pendimethalin, phenmedipham, primisulfuron, prodiamine, prometryn, pronamide, propachlor, propanil, prosulfuron, pyrazone, pyridate, pyrithiobac, quinclorac, quizalofop, rimsulfuron, sethoxydim, siduron, simazine, sulfometuron, sulfosulfuron, tebuthiuron, terbacil, thiazopyr, thifensulfuron, thiobencarb, tralkoxydim, triallate, triasulfuron, tribenuron, triflusulfuron, vernolate, and at least one of their esters and salts.

[0015] In some embodiments, the pyrethroid is deltamethrin.

[0016] In some embodiments, the heterofunctional agent comprises a plant modifier. In some embodiments, the heterofunctional agent comprises an insect modifier.

[0017] In some embodiments, the composition is formulated for application to soil. In some embodiments, the composition is formulated for delivery to a plant, plant part, or plant pest. In some embodiments, the mobility of the heterofunctional agent in soil is increased. In some embodiments, the mobility of the heterofunctional agent in soil is decreased.

[0018] In some embodiments, the agricultural composition is formulated for delivery to a plant, plant part, or plant pest. In some embodiments, the plant part is a plant seed. In some embodiments, the NLP is detected in a germinated seed. In some embodiments, the heterofunctional agent is a volatile agent. In some embodiments, the volatile agent is a fumigant, a pheromone, or an essential oil. In some embodiments, the at least one surface modifier enhances uptake of the agricultural composition by the plant or plant part compared to a composition that does not include the surface modifier. In some embodiments, the at least one surface modifier enhances biodistribution of the agricultural composition by the plant or plant part compared to a composition that does not include the surface modifier. In some embodiments, the NLP targets the meristematic region.

[0019] In some embodiments, the encapsulated heterologous functional agent is protected from ultraviolet light.

[0020] In some aspects, agricultural compositions are provided that comprise a mixture of a) a first plurality of NLPs, the first plurality of NLPs comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, and a first heterofunctional agent; and b) a second plurality of NLPs, the second plurality of NLPs comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, and a second heterofunctional agent, wherein the first plurality of NLPs comprises a hydrophobic core.

[0021] In some aspects, agricultural compositions are provided that comprise a mixture of a) a plurality of NLPs, the plurality of NLPs comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier and a first heterologous functional agent, and b) an unencapsulated agent, wherein the plurality of NLPs comprise a hydrophobic core.

[0022] In some aspects, agricultural compositions are provided comprising a plurality of NLPs, each comprising a heterogeneous functional agent, wherein the NLPs are produced by a process of applying energy to a solution comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, the heterogeneous functional agent, and an aqueous solution, and wherein the plurality of NLPs comprise a hydrophobic core.

[0023] In some embodiments, methods are provided for making an agricultural composition comprising a plurality of NLPs, the method comprising applying energy to a solution comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, a heterofunctional agent, and an aqueous solution, thereby forming NLPs, wherein the NLPs comprise a hydrophobic core.

[0024] In some embodiments, methods are provided for modifying binding of a heterofunctional agent to at least one component in soil, comprising applying to the soil a composition comprising the heterofunctional agent encapsulated in an NLP comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, wherein binding of the encapsulated heterofunctional agent to the soil differs from binding of an unencapsulated heterofunctional agent to the soil.

[0025] In some embodiments, methods are provided for modifying the mobility of a heterologous functional agent in soil, comprising applying to soil a composition comprising the heterologous functional agent encapsulated in an NLP comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, wherein the mobility of the encapsulated heterologous functional agent in the soil differs from the mobility of the unencapsulated heterologous functional agent in the soil.

[0026] In some aspects, methods are provided for reducing rootworm survival, comprising applying to soil containing rootworms a composition comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and a heterofunctional agent, wherein the heterofunctional agent contacts the rootworms, thereby reducing rootworm survival. In some embodiments, the NLP is applied to the soil as a soil drench. In some embodiments, the NLP is applied to the soil in a planting furrow. In some embodiments, the rootworm is a member of the genus Diabrotica. In some embodiments, the rootworm is the western corn rootworm (Diabrotica virgifera virgifera).

[0027] In some aspects, methods for reducing viability or fungal growth are provided, comprising applying to soil containing fungi a composition comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and a heterofunctional agent, wherein the heterofunctional agent contacts the fungus, thereby reducing fungal viability in the soil. In some embodiments, the fungus belongs to a family selected from the group consisting of Sciaridae, Diadocidiidae, Ditomyiidae, Keroplatidae, Bolitophilidae, and Mycetophilidae, and Sclerotiniaceae. In some embodiments, the fungus is Botrytis cinerea.

[0028] In some aspects, methods are provided for preventing plants from developing diseases caused by plant pests, comprising applying to soil a plurality of NLPs, each comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and a heterofunctional agent, wherein the heterofunctional agent contacts the plant pest, thereby killing the pest and thereby preventing the plant from developing the disease. In some embodiments, the NLP is applied to the soil as a soil drench. In some embodiments, the NLP is applied to the soil in a planting furrow. In some embodiments, the plant pest is a member of the order Coleoptera or Hemipteran.

[0029] In one aspect, a method is provided for increasing uptake of a heterologous functional agent by a plant or plant part, the method comprising contacting the plant or plant part with a composition comprising the heterologous functional agent encapsulated in an NLP, wherein the NLP comprises at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, and wherein the NLP comprises a hydrophobic core, and wherein uptake of the encapsulated heterologous functional agent by the plant or plant part is greater than uptake of an unencapsulated heterologous functional agent by the plant or plant part.

[0030] In one aspect, there is provided a method of delivering a heterologous functional agent to a plant or plant part, comprising contacting the plant or plant part with a composition comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and the heterologous functional agent, wherein the NLP comprises a hydrophobic core, thereby delivering the heterologous functional agent to the plant.

[0031] In some embodiments, the plant part is a plant seed.

[0032] In one aspect, a method is provided for delivering a heterologous functional agent to a meristem, the method comprising contacting a plant or plant part with a composition comprising the heterologous functional agent encapsulated in an NLP, wherein the NLP comprises at least one phospholipid, at least one non-polar lipid, and at least one surface modifier.

[0033] In one aspect, there is provided a method of distributing a heterologous functional agent in soil, the method comprising contacting a plant seed with a composition comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and a heterologous functional agent, wherein the NLP comprises a hydrophobic core, and incubating the plant seed in the soil, thereby distributing the heterologous functional agent in the soil.

[0034] In one aspect, there is provided a method of distributing a heterologous functional agent in a plant, the method comprising contacting a plant seed with a composition comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and a heterologous functional agent, wherein the NLP comprises a hydrophobic core, and incubating the plant seed under conditions that cause germination, thereby distributing the heterologous functional agent in the plant.

[0035] In some embodiments, the contacting is by injecting the composition into the plant or plant part, hi some embodiments, the composition is injected into one or more leaves.

[0036] In some embodiments, the composition is injected into the tree. In some embodiments, the composition is injected into the tree at several locations.

[0037] In one aspect, a method of treating a plant disease is provided, comprising contacting a plant or plant part with a composition comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and a heterologous functional agent, wherein the NLP comprises a hydrophobic core, thereby treating the plant disease.

[0038] In some embodiments, the disease is caused by Candidatus Liberibacter asiaticus (CLas). In some embodiments, the disease is citrus greening. In some embodiments, the disease is caused by Xylella.

[0039] In one aspect, a method for preventing a plant from developing a disease comprises contacting the plant or plant part with a composition comprising at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and a heterologous functional agent, wherein the NLP comprises a hydrophobic core, thereby preventing the plant from developing the disease.

[0040] In one aspect, a method of modifying the volatility of a heterofunctional agent is provided, comprising encapsulating a volatile heterofunctional agent in a composition comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, wherein the NLP comprises a hydrophobic core.

[0041] In one aspect, a method for sequestering a volatile heterofunctional agent is provided, comprising encapsulating the heterofunctional agent in a composition comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, wherein the NLP comprises a hydrophobic core.

[0042] In one aspect, a method is provided for the controlled release of a volatile heterologous agent into an environment, comprising encapsulating the heterologous functional agent in a composition comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, wherein the NLP comprises a hydrophobic core.

[0043] In some embodiments, the volatile heterofunctional agent is a herbicide, a fumigant, a pheromone, or an essential oil. In some aspects, a kit is provided that includes an agricultural composition, the composition comprising a plurality of naturally occurring lipid particles (NLPs), each comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, wherein the NLPs comprise a hydrophobic core, and the at least one surface modifier modifies the mobility of the agricultural composition through soil compared to a composition that does not include the surface modifier. [Brief explanation of the drawings]

[0044] [Figures 1A-1C] Comparison of the lipid profile (Figure 1C) of extracted crude lipids, and the phospholipid profile (Figure 1A), and non-polar lipid profile (Figure 1B) in the concentrated fraction from freeze-dried lemons analyzed by HPLC with an evaporative light scattering detector (ELSD). [Figures 2A-2C] Lipid analysis of the enriched fractions by comparison with soybean lipid standards is shown in Figure 2A: Enriched phospholipid fraction derived from lemon (LM) lipids; Figure 2B: Soybean phospholipid standards containing phosphatidic acid (PA), phosphatidylethanolamine (PE), phosphatidylcholine (PC), and phosphatidylinositol (PI); Figure 2C: Experimental sample spiked with soybean phospholipid standards. [Figure 3A-3C]Figure 3A shows the screening of a total of 127 NLP formulations, including 80 μg deltamethrin per ml of NLP suspension and 1 μg Exalite 594 (hereafter referred to as Exalite) per ml of NLP suspension, for soil binding. The compositions of the evaluated formulations are shown in Table 15. Figure 3A shows results in the soil retention assay. The percentage of sample remaining in the supernatant (unbound, leached from the soil) is shown. Twenty-seven of the 127 NLPs tested had soil detachment greater than 10%, and are shown in the figure along with three immobile NLPs (NLP518, NLP519, and NLP536). Unformulated deltamethrin was immobile in soil. Data points represent the average of experiments performed in triplicate. Two experiments were performed for each formulation, and the bars represent the average of the two. Figure 3B shows the screening of 27 formulations that showed reduced affinity for soil in the affinity assay in the soil mobility assay. The sample was loaded onto a soil column and eluted using simulated rain as the mobile phase. The percentage of the total applied sample leaching from the column is shown. (Figure 3C) Comparison of NLP binding in soil retention and soil mobility assays. [Figure 4] Evaluation of NLP formulations and soil leachates of NLP formulations against Western corn rootworm (WCRW) before soil exposure is shown. All NLP-formulated deltamethrin (DE) soil leachates have significantly higher mean efficacy (p<0.05) than the commercial deltamethrin formulation soil leachate (Suspend SC, last bar in the bar graph). [Figures 5A-5E]Figure 5A shows the activity of NLP580 (NLP580-DE) produced by the DCM method containing 400 μg deltamethrin per ml of suspension against WCRW in a soil drench assay. NLP containing DE can improve plant mass of corn seedlings by preventing damage from corn rootworm larvae. Water treatment (Figure 5A) did not rescue seedlings from corn rootworm damage. NLP580-DE (Figure 5D) matches the level of control achieved by Bifenture® (Figure 5B), a commercially available product for soil treatment. Deltamethrin formulated into a soluble liquid using an organic solvent (Figure 5C) did not demonstrate the same level of efficacy as NLP containing deltamethrin (Figure 5D). Quantitative plant mass data are shown in Figure 5E. P values ​​indicating the significance of the relevant comparisons are indicated at the top of the figure. [Figures 6A-6E] Figure 6 shows the activity of NLP580 (NLP580-DE) produced by the DCM method, containing 400 μg deltamethrin per ml of suspension, against WCRW in an in-furrow soil application assay. Figure 6A shows uninfested plants showing expected growth; Figure 6B shows water-treated plants infested with WCRW. Water treatment did not rescue seedlings from corn rootworm damage; Figure 6C shows Bifenture®-treated plants infested with WCRW larvae; Figure 6D shows NLP580-DE-treated plants infested with WCRW larvae. The efficacy of NLP580-DE treatment is consistent with that of Bifenture®. Quantitative plant mass data are shown in Figure 6E. P values ​​indicating the significance of the relevant comparisons are indicated at the top of the figures. [Figures 7A-7B]Figure 7 shows the delivery of a hydrophobic, water-insoluble dye (Exalite) when formulated in NLP580. NLP580 was produced by the DCM method, containing 1 μg of Exalite per ml of suspension. Delivery to green bean seeds was assessed. The positive control was Acid Red 52 (12 parts per million, ppm), a water-soluble fluorescent dye that is readily taken up by seeds. Exalite is a hydrophobic, water-insoluble dye that cannot enter seeds. However, NLP580 with Exalite entered the seedlings, indicating that the NLP enabled hydrophobic molecules to enter the seeds. Figure 7A: Visualization of fluorescence in green bean plants exposed to NLP with Exalite; Figure 7B: Quantification of the fluorescent signal. [Figure 8A-8B] Figure 8A shows the delivery of poorly water-soluble emamectin benzoate (EM) to leaves (sprouts) of 5-day-old corn seedlings when formulated in lemon NLPs. Controls are free EM and no EM (water only). Figure 8A: When EM is formulated in NLPs (lemon) at a final concentration of 32 μg per ml of suspension, significantly higher EM concentrations are achieved in corn leaves compared to unformulated EM (in water). Figure 8B: The ratio of EM concentration in leaves normalized to EM concentration in seeds was higher in treatments with NLPs containing EM than in free EM treatments. [Figure 9A-9B] Figure 9 shows the efficacy of NLP580 containing 400 μg deltamethrin per ml suspension (NLP580-DE (400 μg per ml)) against western corn rootworm (WCRW) in a corn seed treatment phytagel assay. Figure 9A shows that NLP580-DE treated seeds reached a higher fresh corn seedling weight than seeds treated with unformulated deltamethrin (DE only). Figure 9B shows that NLP580-DE treated seeds demonstrated a higher percentage of WCRW control than those treated with unformulated deltamethrin (DE only). [Figures 10A-10R]Figures 10A-10D and 10I-10M show confocal microscopy images of 6-day-old Arabidopsis roots incubated with NLP580, NLP487, and NLP544 formulations. Figures 10A-10D and 10I-10M, differentiation zone; Figures 10E-10H and 10N10R, root meristem. Figures 10A, 10E, 10J, and 10N, 1 μg / ml Exalite; Figures 10B, 10F, 10K, and 10P, NLP580; Figures 10C, 10G, 10L, and 10Q, NLP487; Figures 10D, 10H, 10M, and 10R, NLP544; and Figures 10J and 10O, NLP580 without bioactive agent (with EX but without DE). Roots were incubated for 30 min (Figures 10A-10H) or 24 h (Figures 10I-10R). The accumulation of different dyes in different cellular compartments is visualized for different NLP compositions. The insets show endomembrane localization. Arrows point to NLP580 with or without accumulation of bioactive substance in the root vasculature (v), whereas NLP487 and NLP544 preferentially accumulate in the epidermis (ep) of the root differentiation zone. [Figures 11A-11D] Confocal microscopy images of NLP580 (NLP580) produced by the DCM method, containing 1 μg of Exalite and 80 μg of deltamethrin per ml of suspension, in the epidermal cells of tomato seedlings are shown. When treated in solution for 24 hours, NLP580 was taken up by tomato roots (triplicate, Figures 11B-11D), while the dye control with Exalite showed no fluorescence in root epidermal cells (Figure 11A). Arrows point to NLP with Exalite signal. [Figures 12A-12J]Confocal microscopy images of NLP580, NLP487, and NLP544, each produced by the DCM method and containing 1 μg of Exalite and 80 μg of DE per ml of suspension in Arabidopsis organs, are shown. Figures 12A and 12F, no treatment; Figures 12B and 12G, 1 μg / ml Exalite treatment only ( ); Figures 12C and 12H, NLP580; Figures 12D and 12J, NLP487; Figures 12E and 12J, NLP544. Figures 12A-12E: cotyledons; Figures 12F-12J: hypocotyls. Fluorescence can be detected in cotyledons (Figures 12C-12E) and hypocotyls (Figures 12H-12J) when plant roots are exposed to the NLP formulations. Exalite alone did not produce fluorescence in either the roots or the aboveground plant tissues analyzed. [Figures 13A-13H] Confocal microscopy images of NLP580, NLP487, and NLP544, each produced using the DCM method and containing 1 μg of Exalite and 80 μg of deltamethrin per ml of suspension, are shown in Arabidopsis with and without Brefeldin A (BFA) treatment: Figures 13A and 13E, Exalite only; Figures 13B and 13F, NLP580; Figures 13C and 13G, NLP487; and Figures 13D and 13H, NLP544. NLP and BFA treatment (Figures 13B-13D) with counterstaining with calcofluor white (1 μg / ml) (Figures 13F-13H) showed that NLP580 (Figure 13B) and NLP487 (Figure 13C) aggregated in the intima (arrows point to the intima where they aggregate in BFA bodies), while NLP544 (Figure 13D) was insensitive to BFA. Control experiments using 1 μg / ml Exalite and BFA showed low fluorescence in the outer tissue and no fluorescence in the cytoplasm or intima (Figures 13A and 13E). [Figures 14A-14G]Confocal microscopy images of Nicotiana benthamiana leaves injected with NLPs containing 1 μg / ml Exalite and 80 μg / ml deltamethrin, or 1 μg / ml Exalite alone in 0.5x MS, or uninfiltrated leaves as controls are shown. NLPs were infiltrated and visualized 24 hours later. NLP487 (Figure 14A) localizes to the plasma membrane, while NLP530 (Figure 14E) localizes primarily to membrane-adjacent aggregates. NLP580 (Figure 14B), NLP533 (Figure 14C), NLP527 (Figure 14D), NLP646 (Figure 14F), NLP655 (Figure 14G), and NLP649 (Figure 14H) show both plasma membrane and membrane-adjacent aggregate localization. In the case of NLP649 (Figure 14H), cell details show the nuclear membrane and cytosol (arrows). In control experiments using 1 μg / ml Exalite (Figure 14I) and untreated plants (Figure 14J), Exalite alone had no fluorescence in either the roots or the aboveground plant tissues analyzed. [Figures 15A-15F] These are stereoscopic images of sections of melon plants injected with NLP487, NLP544, and NLP580, each containing 1 μg of Exalite and 80 μg of deltamethrin per ml, produced using the DCM method. From left to right, NLP signal detection 24 hours after injection for NLP487 (Figure 15A), NLP544 (Figure 15B), and NLP580 (Figure 15C). All analyzed NLPs were localized to the pith, and NLP580 was also detected in the vascular bundles (white arrows). Control experiments using 1 μg / ml Exalite alone (Figure 15D) and untreated plants (Figure 15E) show that Exalite alone is not detected in any of the analyzed root or aboveground plant tissues. Figure 15F shows a slide image of the stem under bright field. [Figures 16A-16B] Figure 16B shows confocal microscopy images of NLP580 produced by the DCM method, containing 1 μg of Exalite and 80 μg of deltamethrin per ml of suspension, in melon leaves after stem injection. Results show detection of NLP580 signal in the leaf vasculature 24 hours after injection (Figure 16B). Arrows point to the leaf vasculature. Figure 16A, control experiment using only 1 μg / ml of Exalite. [Figures 17A-17B] Tomato seedling root uptake assay followed by infestation by Colorado potato beetles (CPB) is shown. Various NLPs containing deltamethrin added to the roots of tomato seedlings cause mortality of CPB, a foliar-chewing Coleopteran insect. In Figure 17A, the numbers below the petri dishes indicate the following treatments: 1) water, 2) NLP018 without deltamethrin (blank NLP018), 3) NLP018 with 160 μg deltamethrin per ml suspension (NLP018 [160 μg / ml DE]), 4) Treatment #1 diluted 10-fold to a deltamethrin concentration of 16 μg per ml suspension (NLP018 [16 μg / ml DE]), and 5) Treatment #2 diluted 10-fold to a deltamethrin concentration of 16 μg per ml suspension (NLP018 [16 μg / ml DE]). Treatment #1 (NLP018 [1.6 μg / ml DE]), diluted 100-fold to 1.6 μg per ml, 6) NLP472 containing 160 μg deltamethrin per ml of suspension (NLP472 [160 μg / ml DE]), 7) NLP495 containing 160 μg deltamethrin per ml of suspension (NLP495 [160 μg / ml DE]), 8) 160 μg / ml deltamethrin in DCM (unformulated), 9) 100 μg / ml clothianidin. Unformulated deltamethrin caused phytotoxicity to tomato seedlings, making it unsuitable for feeding assays (number 8 in Figure 17A); Figure 17B, % larval control demonstrated by the indicated NLP formulations. All NLPs were produced using the DCM method. [Figure 18] Figure 1 shows tomato hornworm (THW) control in a tomato seedling root uptake assay, followed by infestation by THW. NLP580 produced by the DCM method, containing 400 μg deltamethrin per ml of suspension (NLP580 [400 μg / ml]), added to tomato seedlings, causes mortality of THW, a foliar-chewing lepidopteran insect. Percent larval control demonstrated by water, 50 μg / ml, 25 μg / ml, 12.5 μg / ml, and 6.25 μg / ml clothianidin, and NLP580 [400 μg / ml] is shown. [Figure 19]Results from ICP-MS analysis of gadolinium (Gd) in tomato organs after exposing the roots of tomato seedlings to various treatments are shown. The treatments were: 1) free Gd, 2) NLP527 containing DTPE-conjugated Gd (Gd-DTPE), 3) NLP529 containing Gd-DTPE, 4) NLP580 containing Gd-DTPE, 5) nanoparticles made from HSPC lipid NLP containing Gd-DTPE, and 6) DOTA-conjugated Gd. Plants whose roots were exposed to Gd-containing NLPs showed significantly better uptake and translocation into various plant organs compared to plants treated with free Gd or DOTA-conjugated Gd. NLP580 functioned similarly to HSPC(Gd), and was taken up and distributed better than free Gd or DOTA-conjugated Gd, but less well than NLP527 and NLP529. [Figure 20] The experimental setup for the tomato seedling root uptake experiment and Pseudomonas syringae infection, as well as bacterial growth readouts, are shown. [Figures 21A-21I] Shown are the results of UV stability assays testing lemon NLP (Figure 21B), carrot NLP (Figure 21C), algal autotrophic NLP (Figure 21F), and algal mixotrophic NLP (Figure 21G) containing deltamethrin, as well as unformulated deltamethrin (deltamethrin in methanol in Figure 21A), which were exposed to UV irradiation for 1, 2, 3, or 6 days. Four NLPs were also mixed with 3% lignosulfonate (Figures 21D, 21E, 21H, 21I) and tested in the same UV stability assay. After 6 days of UV exposure, the four NLPs protected deltamethrin better than deltamethrin in methanol with 55-62% deltamethrin (active ingredient, AI) retention (actual deltamethrin / input deltamethrin) (Figures 21B, 21C, 21F, 21G), compared to 15% deltamethrin retention from deltamethrin in the methanol treatment (Figure 21A). The four NLPs mixed with 3% lignosulfonate (Figures 21D, 21E, 21H, 21I) had even higher deltamethrin retention of 71-84%. [Figures 22A-22P]Confocal microscopy images showing NLP transport from roots to aboveground plant organs are shown. Figures 22A-22D and 22L: cotyledons; Figures 22E-22H and 22M-22P: hypocotyls. Figures 22A and 22E: water treatment; Figures 22B and 22F: 1 μg / ml Exalite only; Figures 22C and 22G: NLP533; Figures 22D and 22H: NLP578; Figures 22I and 22M: NLP551; Figures 22J and 22N: NLP544; Figures 22K and 22O: NLP600; Figures 22L and 22P: NLP620. Arrows indicate vascular tissues. Each NLP suspension was produced by the DCM method and contained 80 μg / ml DE and 1 μg / ml Exalite. [Figures 23A-23R] Confocal microscopy images of 5-day-old Arabidopsis primary roots showing NLP uptake in the plant differentiation zone (Figures 23A-23D and 23I-23M) and root meristem (Figures 23E-23H and Figures 23N-23R). Uptake and transport are monitored after 30 min of incubation with the indicated NLP suspensions. Differential accumulation of different NLP compositions within different cell types and cellular compartments is visualized: Figures 23A-23D and 23I-23M, differentiation zone; Figures 23E-23H and 23N-23R: root meristem. Figures 23A, 23E, 1 μg / ml Exalite only; Figures 23B, 23F, NLP551; Figures 23C, 23G, NLP533; Figures 23D, 23H, NLP578; Figures 23I, 23N, NLP600; Figures 23J, 23O, NLP620; Figures 23K, 23P, NLP699; Figures 23L, 23Q, NLP700; Figures 23M, 23R, NLP544. Each NLP suspension was produced by the DCM method and contained 80 μg / ml DE and 1 μg / ml Exalite. [Figures 24A-24R]Confocal microscopy images of Arabidopsis primary roots showing NLP uptake by the plant are shown. After 24 h of incubation with the indicated NLP suspensions, uptake and transport along Arabidopsis root tissues are monitored: Figures 24A-24D and 24I-24M, differentiation zone; Figures 24E-24H and 24N-24R: root meristem. Figures 24A, 24E, 1 μg / ml Exalite only; Figures 24B, 24F, NLP551; Figures 24C, 24G, NLP533; Figures 24D, 24H, NLP578; Figures 24I, 24N, NLP600; Figures 24J, 24O, NLP620; Figures 24K, 24P, NLP699; Figures 24L, 24Q, NLP700; Figures 24M, 24R, NLP544. Insets show intimal regions and localization (n>15). Each NLP suspension was generated by the DCM method and contained 80 μg / ml DE and 1 μg / ml Exalite. [Figures 25A-25H] Confocal microscopy images are shown demonstrating that NLPs can be transported to the shoot apical meristem. Uptake was assessed after 24 hours of root treatment with NLP formulations. Figure 25A, water only; Figure 25B, 1 μg / ml Exalite only; Figure 25C, NL487; Figure 25D, NLP580; Figure 25E, NLP578; Figure 25F, NLP533; Figure 25G, NLP544; Figure 25H, NLP551. Each NLP suspension was produced by the DCM method and contained 80 μg / ml DE and 1 μg / ml Exalite. Arrows indicate the shoot apical meristem. [Figures 26A-26T]Confocal microscopy images showing the subcellular localization of NLPs are shown. Panels showing Exalite fluorescence from labeled NLPs (panels in rows 1 and 3) and calcofluor counterstain fluorescence (1 μg / ml) (panels in rows 2 and 4). BFA promotes aggregation of the inner membrane into larger compartments (called BFA bodies), and calcofluor white stains cellulose in the cell wall. Figures 26A, 26B, 1 μg / ml Exalite only; Figures 26C, 26D, NLP578; Figures 26E, 26F, NLP544; Figures 26G, 26H, NLP551; Figures 26I, 26J, NLP533; Figures 26K, 26L, NLP600; Figures 26M, 26N, NLP580; Figures 26O, 26P, NLP620; Figures 26Q, 26R, NLP699; Figures 26S, 26T, NLP700. Arrows point to intimal membranes aggregated in BFA bodies. Each NLP suspension was generated by the DCM method and contained 80 μg / ml DE and 1 μg / ml Exalite. Arrows point to intimal membranes aggregated in BFA bodies. [Figures 27A-27G] Confocal microscopy images showing NLP localization near the plasma membrane of Nicotiana benthamiana leaves are shown. 200 μl aliquots of the indicated NLP formulations were infiltrated using a 1 ml needleless syringe, followed by fluorescence visualization using a confocal microscope 24 hours later. Figure 27A, NLP544; Figure 27B, NLP551; Figure 27C, NLP578; Figure 27D, NLP600; Figure 27E, NLP620; Figure 27F, 1 μg / ml Exalite only; Figure 27G, control, uninfiltrated. White arrows, nuclear membrane. Each NLP suspension was generated by the DCM method and contained 80 μg / ml DE and 1 μg / ml Exalite. [Figures 28A-28D]Confocal microscopy images are shown demonstrating that injected NLPs are localized to the vascular system of melon plants. Five-week-old plants were injected with 200 μl of the indicated NLP formulation using a 1 ml syringe attached to an Invaio 3 mm Trecise™ injector, followed by visualization using confocal microscopy 24 hours later. Figure 28A, NLP533; Figure 28B, NLP551; Figure 28C, water (control); Figure 28D, Exalite only at 1 μg / ml. White arrows, vascular bundles. Each NLP suspension was produced by the DCM method and contained 80 μg / ml DE and 1 μg / ml Exalite. [Figures 29A-29N] Confocal microscopy images are shown to assess NLP maize seed uptake. Figures 29A-29F, Pre-germination assay: Figure 29A, water only (24 h); Figure 29B, NL620 (24 h); Figure 29C, water only (24 h); Figure 29D, NLP600 (24 h); Figure 29E, water only (48 h); Figure 29F, NLP580 (48 h). Images show seed cross sections after incubation with the indicated treatments. Figures 29G-29J, Germination in NLP solution: Figure 29G, water only (4 days); Figure 29H, NL580 (4 days); Figure 29I, water only (5 days); Figure 29J, NLP580 (5 days). Images show cut seeds after incubation with the indicated treatments. Figures 29K-29N, 24-hour imbibition in the indicated NLP suspensions followed by germination in water (5 days): Figure 29K, water control (5 days, imaged under white light); Figure 29L, water control (5 days, imaged under fluorescent light); Figure 29M, NLP580 (5 days, imaged under white light); Figure 29N, NLP580 (5 days, imaged under fluorescent light). Each NLP suspension was produced by the DCM method and contained 80 μg / ml DE and 1 μg / ml Exalite. [Figures 30A-30G]Confocal microscopy images show that NLPs injected into corn plants are localized to the vascular system. Five-week-old plants were injected with 200 μl of the indicated NLP formulation using a 1 ml syringe attached to an Invaio 3 mm Trecise™ injector, and fluorescence was visualized 24 hours later by confocal microscopy. Figures 30A and 30D: water only; Figures 30B and 30F: NLP487; Figures 30C and 30G: NLP646; Figure 30D: NLP544. Figures 30A-30D: corn stem cross-section; Figures 30E-30G: corn leaves. Each NLP suspension was produced by the DCM method and contained 80 μg / ml DE and 1 μg / ml Nile Red. [Figure 31A-31B] Figure 31 shows the efficacy of nine NLP formulations against Western Corn Rootworm (WCRW) at a small scale in an in-furrow soil application assay. Results are shown for the efficacy of testing nine NLPs (NLP533, NLP551, NLP487, NLP646, NLP644, NLP647, NLP659, NLP660, and NLP654, each produced by the HHPH method and containing DE at 2 mg / ml) against WCRW in an in-furrow soil application assay conducted in a greenhouse. Both fresh total seedling weight (Figure 31A) and root mass (Figure 31B) demonstrate the efficacy of treatment against WCRW for all nine formulations tested. Non-infested plants show normal growth, as expected. Water treatment (water-treated WCRW-infested plants) did not rescue seedlings (or roots) from WCRW damage. Bifenture treatment served as a positive control. The efficacy of the nine NLPs (including the deltamethrin treatment) is consistent with that of the Bifenture® treatment. Unformulated deltamethrin at 2 mg / ml was less effective at protecting total plant mass (Figure 31A) or root mass (Figure 31B) from WCRW infestation than any of the nine NLPs containing the same concentration (2 mg / ml) of deltamethrin. Data shown are the average of two independent experiments, each with 9-10 replicates (plants) per treatment. All NLP suspensions were produced using the HHPH method. [Figure 32A-32B]Figure 32 shows the activity of two NLP compositions (NLP644 and NLP647) containing various concentrations of deltamethrin (80, 400, and 2000 μg / ml suspension) against western corn rootworm (WCRW) in a greenhouse in-furrow soil application assay measuring protective effects on total plant mass (Figure 32A) and root mass (Figure 32B). *, P<0.05 compared to the corresponding concentration of unformulated DE. The NLP compositions were produced using the HHPH method. [Figure 33] Figure 1 shows the efficacy of NLP644 containing 2 mg / ml deltamethrin against western corn rootworm (WCRW) in an in-furrow soil application assay in an open field test. The NLP composition was produced using the HHPH method. Data shown are the average of 20 separate replicates. [Figure 34] Figure 1 shows the antifungal efficacy of azoxystrobin encapsulated in 10 different NLPs as measured in a resazurin-based Fusarium inhibition assay. Assay 1 was performed in duplicate (Screening 1-1 and Screening 1-2) to evaluate NLP580, NLP533, NLP551, NLP487, and NLP608. Assay 2 was performed in duplicate (2-1 and 2-2) to evaluate NLP600, NLP603, NLP530, NLP527, and NLP532. [Figure 35] Figure 1 shows the percentage of deltamethrin recovered from soil in a soil mobility assay after incubating deltamethrin or NLPs containing deltamethrin in soil for 0 hours, 24 hours, and 7 days. The NLP compositions were generated using the HHPH method. Data shown are the average of three replicate columns per treatment. DETAILED DESCRIPTION OF THE INVENTION

[0045] Featured herein are agricultural compositions that include a plurality of naturally occurring lipid particles (NLPs), each of which includes a phospholipid component, a non-phospholipid component, and at least one surface modifier. In some embodiments, the surface modifier enhances mobility of the NLPs through soil compared to NLPs that do not include the surface modifier. The NLPs may optionally include an agent (e.g., a heterofunctional agent (e.g., a heterogeneous agricultural agent (e.g., a pesticide, fertilizer, herbicide, plant modifier, insect attractant, plant growth promoter, biostimulant, or plant immunity inducer) or a heterogeneous therapeutic agent (e.g., an antifungal, antioomycete, antibacterial, virucide, virucide, insecticide, nematicide, antiparasitic, or insect repellent). In some embodiments, the heterofunctional agent is hydrophobic. In some embodiments, the heterofunctional agent is encapsulated. In some embodiments, the encapsulated heterofunctional agent is a pyrethroid, such as deltamethrin. In some embodiments, the encapsulated pyrethroid is more mobile in soil compared to unencapsulated pyrethroid, thus providing a solution to the problem of deltamethrin binding to soil.

[0046] Some embodiments relate to methods and compositions for modifying the movement of heterofunctional agents through soil. In some embodiments, heterofunctional agents with high soil affinity are encapsulated in NLPs comprising a phospholipid component, a non-phospholipid component, and at least one surface modifier, where the surface modifier alters the characteristics of the NLP to promote mobility through soil compared to NLPs that do not contain the surface modifier. In other embodiments, heterofunctional agents with low soil affinity are encapsulated in NLPs comprising a phospholipid component, a non-phospholipid component, and at least one surface modifier, where the surface modifier alters the characteristics of the NLP to increase soil affinity compared to NLPs that do not contain the surface modifier.

[0047] Further featured herein are compositions comprising mixtures of NLPs, each containing a different heterofunctional agent. Further featured herein are mixtures comprising multiple NLPs and an unencapsulated agent. The NLP compositions and methods described herein can be used in a variety of agricultural and therapeutic methods.

[0048] definition As used herein, an "agriculturally acceptable" carrier is one that is suitable for use in agriculture, e.g., for use on plants. In certain embodiments, an agriculturally acceptable carrier does not have undue harmful side effects to plants, the environment, or humans or animals that consume the resulting produce, commensurate with a reasonable benefit / risk ratio.

[0049] As used herein, "delivering" or "contacting" refers to applying an NLP composition described herein directly to or adjacent to a plant, animal (e.g., insect, nematode, etc.), fungus, or bacterium in an area where the composition is effective to alter the fitness of the plant, animal, fungus, or bacterium. In methods where the composition is directly contacted with the plant, animal, fungus, or bacterium, the composition may contact the entire plant, animal, fungus, or bacterium, or only a portion of the plant, animal, fungus, or bacterium. In some embodiments, the NLP composition may be ingested by a plant pest, such as an insect or nematode.

[0050] As used herein, "reducing plant fitness" refers to any disruption of plant (e.g., weed) physiology as a result of administration of a composition described herein (e.g., an NLP composition comprising a heterologous functional agent described herein), including, but not limited to, reducing the population of the plant (e.g., weed) by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more. Reduction in plant fitness can be determined relative to a plant to which the composition has not been administered.

[0051] As used herein, the terms "effective amount," "effective concentration," or "concentration effective for" refer to the amount of a heterologous functional agent provided in the NLP compositions described herein sufficient to affect a recited result or to reach a target level (e.g., a predetermined or threshold level) in or on a target organism.

[0052] As used herein, "increasing plant fitness" refers to an increase in plant production, such as improved yield, improved plant vigor, or improved quality of the harvested product from a plant, as a result of administering a composition described herein (e.g., an NLP composition comprising a heterologous functional agent described herein). Improved plant yield refers to an increase in the yield of a plant's product (e.g., as measured by plant biomass, grain, seed, or fruit yield, protein content, carbohydrate or oil content, or leaf area) by a measurable amount over the yield of the same product of a plant produced under the same conditions without application of the composition or compared to application of a conventional agricultural chemical. For example, the yield can be increased by at least about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, or more than 100%. Yield can be expressed in terms of the amount by weight or volume of the plant or plant product on some basis. The basis can be expressed in terms of time, area under cultivation, weight of plants produced, or amount of raw material used. An increase in plant fitness can also be measured by other means, such as a measurable or significant increase or improvement in vigor rating, an increase in stand number (number of plants per unit area), an increase in plant height, an increase in stem circumference, an increase in plant canopy, an improvement in appearance (such as visually measured green leaf color), an improvement in root rating, an increase in seedling emergence, an increase in protein content, an increase in leaf size, an increase in leaf number, a decrease in basal leaf mortality, an increase in tiller strength, a decrease in nutrient or fertilizer requirements, an increase in seed germination, an increase in tiller productivity, an increase in flowering, an increase in seed or grain maturation or seed maturation, a decrease in plant lodging (lodging), an increase in shoot growth, or any combination of these factors, for the same factor in plants produced under the same conditions but without the administration of the composition or a conventional agricultural chemical.

[0053] As used herein, the term "heterologous" refers to an agent that is exogenous to the plant or plant part that comes into contact with the NLP (e.g., a pesticide derived from a source other than the plant itself, e.g., incorporated into the NLP, may be heterologous). In some embodiments, one or more components of the NLP are heterologous.

[0054]

[0023] As used herein, the term "functional agent" refers to an agent (e.g., an agricultural agent (e.g., a pesticide, insecticide, fungicide, nematicide, fertilizer, herbicide, plant modifier, insect attractant, etc.) or a therapeutic agent (e.g., an antifungal, antioomycete, antibacterial, virucide, antiviral, insecticide, nematicide, antiparasitic, or insect repellent)) that is associated with or can be associated with an NLP composition described herein (e.g., loaded into or onto an NLP (e.g., encapsulated by, embedded in, or conjugated to an NLP)) and that is capable of producing a recited result (e.g., increasing or decreasing the fitness of a plant, plant pest, plant symbiont, animal (e.g., human) pathogen, or animal pathogen vector) in accordance with the present compositions or methods. In some embodiments, the functional agent is a polynucleotide. In some embodiments, the functional agent is a polypeptide. In some embodiments, the functional agent is a small molecule. In some embodiments, the functional agent is a volatile agent and has a high vapor pressure. In some embodiments, the volatile functional agent is a fumigant. In some embodiments, the volatile functional agent is a pheromone. In some embodiments, the volatile functional agent is an essential oil. As used herein, the term "agricultural agent" refers to an agent that can act on plants, plant pests, or plant microorganisms (e.g., plant symbionts), such as a pesticide, pest repellent, fertilizer, plant modifier, plant growth promoter, biostimulant, plant immunity inducer, or plant microorganism regulator.

[0055] As used herein, the term "fertilizer" refers to an agent (e.g., a plant nutrient or a plant growth regulator) that can increase the fitness of a plant. In some embodiments, the fertilizer is a plant symbiont (e.g., a nitrogen-fixing bacterium).

[0056] As used herein, the term "pesticide" refers to an agent, composition, or substance therein that controls or reduces the fitness (e.g., kills or inhibits the growth, proliferation, division, reproduction, or spread) of agricultural, environmental, or household / residential pests, such as insects, mollusks, nematodes, fungi, oomycetes, bacteria, weeds, or viruses. Pesticides are understood to include natural or synthetic insecticides (larvicides or adulticides), insect growth regulators, miticides (acaricides), molluscicides, nematicides, ectoparasiticides, bactericides, fungicides, or herbicides. The term "pesticide" may further encompass antibiotics, antivirals, insecticides, antifungals, antiparasitics, nutrients, and / or other biologically active molecules, such as agents that stop or slow insect movement.

[0057] As used herein, the term "plant modifier" refers to an agent that can alter the genetic (e.g., increase gene expression, decrease gene expression, or otherwise change the nucleotide sequence of DNA or RNA), epigenetic, or biochemical characteristics of a plant so as to result in a change (e.g., increase or decrease) in plant fitness.

[0058] As used herein, the term "therapeutic agent" refers to an agent that can act on an animal, e.g., a mammal (e.g., a human), an animal pathogen, or a pathogen vector, such as an antifungal, antibacterial, virucide, antiviral, insecticide, nematicide, antiparasitic, or insect repellent.

[0059] As used herein, the term "formulated for delivery to plants" refers to an NLP composition that includes an agriculturally acceptable carrier. As used herein, an "agriculturally acceptable" carrier is one that is suitable for use in agriculture without undue harmful side effects to plants, the environment, or humans or animals consuming the resulting produce, commensurate with a reasonable benefit / risk ratio. Non-limiting examples of agriculturally acceptable carriers or excipients are known in the art; see, e.g., the Compendium of Herbicide Adjuvants.

[0060] As defined herein, the terms "nucleic acid" and "polynucleotide" are interchangeable and refer to linear or branched, single- or double-stranded RNA or DNA, or hybrids thereof, regardless of length (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 100, 150, 200, 250, 500, 1000, or more nucleic acids). The term also encompasses RNA / DNA hybrids. While nucleotides are typically linked in nucleic acids by phosphodiester bonds, the term "nucleic acid" also encompasses nucleic acid analogs having other types of linkages or backbones (e.g., phosphoroamide, phosphorothioate, phosphorodithioate, O-methylphosphoramidate, morpholino, locked nucleic acid (LNA), glycerol nucleic acid (GNA), threose nucleic acid (TNA), and peptide nucleic acid (PNA) linkages or backbones, among others. Nucleic acids can be single-stranded, double-stranded, or contain portions of both single-stranded and double-stranded sequence. Nucleic acids can contain any combination of deoxyribonucleotides and ribonucleotides, as well as any combination of modified or non-standard bases, including, for example, adenine, thymine, cytosine, guanine, uracil, and modified or non-standard bases, including hypoxanthine, xanthine, 7-methylguanine, 5,6-dihydrouracil, 5-methylcytosine, and 5-hydroxymethylcytosine.

[0061] As used herein, the term "pest" refers to an organism that causes damage to plants or other organisms, that is present where it is not wanted, or that is otherwise harmful to humans, for example, by adversely affecting human agricultural methods or produce. Pests include, for example, invertebrates (e.g., insects, nematodes, or mollusks), microorganisms (e.g., plant pathogens, endophytes, obligate parasites, facultative parasites, or facultative saprophytes), such as bacteria, fungi, oomycetes, or viruses, or weeds.

[0062] As used herein, the term "pesticidal agent" or "pesticide" refers to an agent, composition, or substance therein that controls or reduces the fitness (e.g., kills or inhibits the growth, reduces reproduction, growth, division, propagation, or spread) of agricultural, environmental, or household / residential pests, such as insects, mollusks, nematodes, fungi, bacteria, weeds, or viruses. Pesticides are understood to include natural or synthetic insecticides (larvicides or adulticides), insect growth regulators, miticides (acaricides), molluscs, nematicides, ectoparasiticides, bactericides, fungicides, or herbicides. The term "pesticide" may further include other biologically active molecules, such as antibiotics, antiviral pesticides, antifungals, antiparasitics, nutrients, and / or agents that stop or slow insect movement.

[0063] As used herein, the term "repellent" refers to an agent, composition, or substance therein that discourages pests from accessing or remaining on a plant. A repellent may, for example, reduce the number of pests on or near a plant, but not necessarily kill or reduce the fitness of the pests.

[0064] As used herein, the terms "peptide," "protein," or "polypeptide" encompass any chain of natural or unnatural amino acids (either D- or L-amino acids), regardless of length (e.g., at least 2, 3, 4, 5, 6, 7, 10, 12, 14, 16, 18, 20, 25, 30, 40, 50, 100 or more amino acids), the presence or absence of post-translational modifications (e.g., glycosylation or phosphorylation), or the presence of, e.g., one or more non-aminoacyl groups (e.g., sugars, lipids, etc.) covalently linked to the peptide, including, e.g., naturally occurring proteins, synthetic or recombinant polypeptides and peptides, hybrid molecules, peptoids, or peptidomimetics.

[0065] As used herein, the "percent identity" between two sequences is determined by the BLAST 2.0 algorithm, which is described in Altschul et al. (1990) J. Mol. Biol. 215:403-410. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information.

[0066] As used herein, the term "plant" refers to whole plants, plant parts, plant organs, plant tissues, seeds, plant cells, seeds, and their progeny. Plant cells include, but are not limited to, seeds, suspension cultures, embryos, meristematic (e.g., meristem) regions, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen, and microspores. Plant parts include differentiated and undifferentiated tissues, including, but not limited to, roots, stems, shoots, leaves, pollen, seeds, fruits, harvested products, tumor tissue, sap (e.g., xylem sap and phloem sap), and various forms of cells and cultures (e.g., single cells, protoplasts, embryos, and callus tissue).

[0067] As used herein, the term "NLP" refers to a composition comprising a plurality of naturally-derived lipid particles, wherein the NLP comprises at least one phospholipid (e.g., phosphatidylcholine), at least one non-polar lipid (e.g., lemon oil), and at least one surface modifier (e.g., a PEGylated compound). In some embodiments, the NLP further comprises a co-solvent (e.g., DCM). In some embodiments, the NLP further comprises an excipient. In some embodiments, the NLP further comprises a heterologous functional agent. In some embodiments, the NLP encapsulates a heterologous functional agent. The NLP may be modified in vitro or in vivo, for example, in a plant. As used herein, the term "soil mobility" refers to the ability of an agent, e.g., an NLP, a heterofunctional agent, or an NLP containing a heterofunctional agent, to move from the site of application in the soil to a location some distance from the site of application in the soil (e.g., 1 mm, 5 mm, 1 cm, 5 cm, 10 cm, 15 cm, 20 cm, 25 cm, 30 cm, 35 cm, 40 cm, 45 cm, 50 cm, 55 cm, 60 cm, 65 cm, 70 cm, 75 cm, 80 cm, 85 cm, 90 cm, 95 cm, 1 meter, 1.5 meters, 2 meters, 2.5 meters, 3 meters, 3.5 meters, 4 meters, 4.5 meters, 5 meters, etc.). When an ingredient is mobile in soil, it means that the ingredient generally moves freely through the soil to reach a location at least distant from the site of application. The location distant from the site of application can be, for example, anywhere from about 1 mm, 1 cm, 10 cm, 50 cm, 1 meter, or 5 meters from the site of application.

[0068] As used herein, "soil affinity" refers to the attraction of a component to soil, such as the attraction between a positively charged heterofunctional agent and a negatively charged component in soil. Typically, affinity is the result of an equilibrium between the binding constant and the dissociation constant, and when there is an equilibrium between the binding constant and the dissociation constant, it can be quantified by an affinity constant. Affinity can arise from, for example, electrostatic interaction, hydrogen bond formation, hydrophobic interaction, and van der Waals interaction, or a combination thereof.

[0069] As used herein, "leachate" refers to the fraction of NLPs that do not bind to soil when an NLP preparation is contacted with soil. For example, without being limited by theory, if an NLP preparation is mixed with a soil suspension and 40% of the preparation does not bind to the soil, the 40% is referred to as "leachate." Similarly, if an NLP preparation is applied to a column containing soil and 40% of the NLP preparation elutes from the column after application of a mobile phase, such as artificial rain, 40% is said to leach from the column. Thus, the eluate is a leachate.

[0070] As used herein, the term "soil" or "mud" refers to a matrix that includes solids, gases, and liquids. In some embodiments, the solids include organic matter (e.g., proteins), inorganic matter (e.g., silicates), microorganisms, and voids that contain gases (e.g., oxygen) and liquids (e.g., water). The composition of soil varies depending on its location on the Earth. Soil is typically negatively charged, which is due to the negative charge of the materials contained in the soil. The negative charge of soil can be quantified as its "cation exchange capacity."

[0071] As used herein, the "cation exchange capacity" of a soil is defined as the amount of positive charge in the soil that can be exchanged per mass of soil. Cation exchange capacity can be determined, for example, by displacing the cations bound to the soil with a concentrated solution of another cation, such as ammonium, and then measuring the displaced cations in the solution.

[0072] As used herein, "encapsulated heterofunctional agent" refers to a heterofunctional agent that is encapsulated or incorporated into another structure, such as an NLP described herein. In some embodiments, there is no contact between the encapsulated heterofunctional agent and that surrounding the NLP, such as water. In some embodiments, the encapsulated functional agent is provided in oil. In other embodiments, the encapsulated functional agent is provided in a hydrophilic liquid. In some embodiments, the encapsulated heterofunctional agent is in contact with the NLP core (e.g., the hydrophobic core). In some embodiments, the encapsulated heterofunctional agent is in contact with the phospholipid membrane. In some embodiments, the encapsulated heterofunctional agent is incorporated into the phospholipid membrane.

[0073] As used herein, the term "encapsulating" refers to the process of incorporating an agent into another entity, for example, the process of incorporating a heterofunctional agent (e.g., deltamethrin) into an NLP. As used herein, an "unencapsulated heterofunctional agent" refers to a heterofunctional agent that is free in solution, e.g., dissolved or dispersed, and can interact directly with its surroundings, e.g., soil.

[0074] As used herein, the term "cellular uptake" refers to the uptake of an NLP or a portion or component thereof (e.g., a heterologous functional agent carried by the NLP) by a cell, such as an animal cell, plant cell, bacterial cell, or fungal cell. For example, uptake can include the movement of the NLP or a portion of its component from the extracellular environment into or across the cell membrane, cell wall, extracellular matrix, or intracellular environment of the cell. Cellular uptake of NLPs can occur via active or passive cellular mechanisms. Cellular uptake includes aspects in which the entire NLP is taken up by the cell, for example, by endocytosis. In embodiments, after endocytosis and endosomal escape, one or more heterologous functional agents (e.g., polynucleotides, polypeptides, small molecule chemicals, etc.) are exposed to the cytoplasm of the target cell. In some embodiments, NLPs (e.g., NLPs comprising charged surface modifiers (e.g., polycarboxylates)) have an increased endosomal escape rate compared to unmodified NLPs. Cellular uptake also includes aspects in which the NLP fuses with the membrane of the target cell. In some embodiments, one or more heterologous functional agents (e.g., polynucleotides, polypeptides, small molecule chemicals, etc.) are exposed to the cytoplasm of the target cell after membrane fusion. In some embodiments, NLPs comprising a surface modifier (e.g., NLPs comprising PEGylated lipids) have an increased rate of fusion with the membrane of the target cell (e.g., are more fusogenic) compared to NLPs without a surface modifier.

[0075] As used herein, the term "cell permeation agent" refers to an agent that alters the properties (e.g., permeability) of the cell wall, extracellular matrix, or cell membrane of a cell (e.g., an animal cell, a plant cell, a bacterial cell, or a fungal cell) in a manner that promotes increased cellular uptake compared to cells not in contact with the cell permeation agent.

[0076] As used herein, the term "NLP" refers to a lipid structure (e.g., a micellar structure, lipid bilayer, monolayer, multilamellar structure, e.g., a vesicular lipid structure) having a diameter of about 5-2000 nm (e.g., at least 5-1000 nm, at least 5-500 nm, at least 400-500 nm, at least 25-250 nm, at least 50-150 nm, or at least 70-120 nm) comprising at least one phospholipid (e.g., phosphatidic acid, PA), at least one nonpolar lipid (e.g., triglyceride), and at least one surface modifier (e.g., Atlox500L). In some embodiments, NLPs are produced by adding energy to a mixture of phospholipid, nonpolar lipid, and surface modifier components provided in an organic phase and an aqueous phase. In some embodiments, the surface modifier component may be added after the NLPs are formed. Non-limiting examples of means for adding energy include one or more of sonicating, vortexing, mixing, or heating a mixture of an organic solution and an aqueous solution to form NLPs. In some embodiments, the NLPs may be further subjected to sonication, freeze / thaw treatment, and / or lipid extrusion, e.g., to reduce the size of the NLPs. In some embodiments, the NLPs may be produced using a microfluidic device (such as the NanoAssemblr® IGNITE™ microfluidic instrument (Precision NanoSystems)). Some embodiments relate to NLP compositions that comprise 10% to 100% of their lipids derived from plant sources; for example, in some embodiments, the NLPs may contain at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% lipids derived from plant sources. Some embodiments may relate to NLPs described herein that comprise all or a portion of the lipid species present in the lipid structure of a plant source.In some embodiments, the NLPs described herein may contain at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% of the lipid species present in the lipid structures from a plant source. In some embodiments, the NLPs described herein may contain none, some, or all of the protein species present in the lipid structures from a plant source. In some embodiments, the NLPs described herein may contain 0%, less than 1%, less than 5%, less than 10%, less than 15%, less than 20%, less than 30%, less than 40%, less than 50%, less than 60%, less than 70%, less than 80%, less than 90%, less than 100%, or 100% of the protein species present in the lipid structures from a plant source. In some embodiments, the NLPs described herein may comprise a lipid bilayer. In some embodiments, the lipid composition of the NLPs may comprise 0%, less than 1%, or at least 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more than 95% exogenous lipids. Non-limiting examples of exogenous lipids include charged lipids (e.g., ionizable and / or cationic lipids). In some embodiments, the exogenous lipid may be a cell-penetrating agent that can increase delivery of a heterologous functional agent (e.g., an agricultural or therapeutic agent) to cells by the NLP and / or may increase the loading (loading efficiency or loading capacity) of a heterologous functional agent (e.g., an agricultural or therapeutic agent) into the NLP. In some embodiments, the NLPs described herein may comprise an exogenous lipid selected from a sterol and a PEGylated lipid.

[0077] NLPs may optionally include additional agents, such as heterofunctional agents, e.g., cell-penetrating agents, pesticides, fertilizers, plant modifiers, therapeutic agents, polynucleotides, polypeptides, small molecules, etc. In some embodiments, NLPs may carry or associate with one or more heterofunctional agents in a variety of ways that enable delivery of the agent to a target plant, for example, by encapsulating the heterofunctional agent, incorporating the heterofunctional agent into the lipid bilayer structure, or associating the heterofunctional agent with the surface of the lipid bilayer structure of the NLP (e.g., by conjugation). Heterofunctional agents may be incorporated into NLPs either in vivo (e.g., in plants) or in vitro (e.g., in tissue culture, cell culture, or synthetically incorporated). In some embodiments, the heterofunctional agent is a pyrethroid. Pyrethroids are hydrophobic agents that have a high affinity for soil and sedimentary particulate matter. The affinity of pyrethroids for soil is thought to be primarily due to their non-polar nature and lack of water solubility. When pyrethroids are dispersed in water, they tend to bind to natural organic matter, such as proteins and clay components found in soil. As a result, pyrethroids have low mobility in soil. General information regarding the persistence and mobility of specific pyrethroid solids can be found in the National Pesticide Telecommunications Network's General and Technical Fact Sheets; the e-pesticide manual, Ver. 5; British Crop Protection Council and Laskowski DA, "Physical and Chemical Properties of Pyrethroids," Rev. Environ. Contam. Toxicol. 2002; 174:49-170 (incorporated herein by reference).

[0078] As used herein, the term "surface modifier" refers to a compound that can alter one or more characteristics of an NLP. In some embodiments, the NLPs described herein may comprise one or more surface modifiers that affect the surface properties of the NLP (e.g., the zeta potential of the NLP). In some embodiments, the surface modifier affects the mobility of the NLP (or a heterofunctional agent contained in the NLP) in soil. In some embodiments, the surface modifier alters the interaction of the NLP with one of the components in the soil compared to an NLP that does not comprise the surface modifier. In some embodiments, the surface modifier increases binding of the NLP to a plant or plant part compared to an NLP that does not comprise the surface modifier. In some embodiments, the surface modifier increases the biodistribution of a heterofunctional agent contained in the NLP upon contact of the NLP with a plant or plant part compared to the biodistribution of a heterofunctional agent contained in the NLP that does not comprise the surface modifier. In some embodiments, the NLPs described herein comprise Atlox 500L as a surface modifier that alters binding of the NLP (or a heterofunctional agent contained in the NLP) to soil. In some embodiments, the surface modifier imparts a charge to the NLP, for example, making the NLP more negatively or more positively charged. In some embodiments, the surface alters the zeta potential of the NLP compared to the NLP without the surface modifier. In some embodiments, the zeta potential predicts the binding properties of the NLP to soil. In some embodiments, the surface modifier is an amphiphilic molecule. In some embodiments, the surface modifier is a zwitterionic agent. In some embodiments, the surface modifier is a cationic agent, for example, a cationic lipid. In some embodiments, the surface modifier is an anionic polymer, for example, a polycarboxylate. In some embodiments, the surface modifier is a lipid compound, for example, a rhamnolipid, a sophorolipid, etc., having attached glycosylation moieties. In some embodiments, the surface modifier is charged (e.g., cationic or anionic) or includes a group (e.g., a head group) that can be ionized under given conditions (e.g., pH) to generate one or more charged species.Non-limiting examples of surface modifiers that affect the soil mobility of the NLPs described herein are rhamnolipids, sophorolipids, PEG2000-C18, PEG5000-C18, Atlox 500L, Atlox 4917, and Atlox CS100B.

[0079] As used herein, a "co-solvent" refers to an agent that aids in dissolving a heterofunctional agent. For example, DCM is a co-solvent that aids in dissolving deltamethrin in the organic (hydrophobic) phase. In some embodiments, the co-solvent may be selected from the group consisting of a fatty acid methyl ester, a non-ionic emulsifier, propylene glycol, ethyl lactate, a non-ionic block copolymer surfactant, or a non-ionic polyalkylene glycol ether. In some embodiments, the co-solvent is dichloromethane (DCM). In some embodiments, the co-solvent is isopropyl myristate (IPM). In some embodiments, the co-solvent is an emulsifier. In some embodiments, the addition of a co-solvent during NLP production increases the solubility of the heterofunctional agent by at least 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold compared to the solubility of the heterofunctional agent in NLPs produced without the addition of a co-solvent.

[0080] As used herein, the term "stable NLP composition" (e.g., a composition comprising loaded or unloaded NLPs) refers to a composition that remains stable over a period of time (e.g., at least 24 hours, at least 48 hours, at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 30 days, at least 60 days, or at least 90 days), optionally within a predetermined temperature range (e.g., at least 24°C (e.g., at least 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, or 30°C), at least 20°C (e.g., at least 20°C, 29°C, or 30°C). at least 5% (e.g., at least 5%, 10%, 15%, 20%, 25%) of the initial number of NLPs (e.g., NLPs per mL of solution) compared to the number of NLPs in the NLP starting material (e.g., at the time of production or formulation) at a temperature of at least 1°C, 22°C, or 23°C), at least 4°C (e.g., at least 5°C, 10°C, or 15°C), at least -20°C (e.g., at least -20°C, -15°C, -10°C, -5°C, or 0°C), or -80°C (e.g., at least -80°C, -70°C, -60°C, -50°C, -40°C, or -30°C), 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or optionally within a predetermined temperature range (e.g., at least 24°C (e.g., at least 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, or 30°C), at least 20°C (e.g., at least 20°C, 21°C, 22°C, or 23°C), at least 4°C (e.g., at least 5°C, 10°C, or 15°C), at least -20°C (e.g., at least -20°C, -15°C, -10°C, -5°C, or refers to an NLP composition that retains at least 5% (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%) of its activity (e.g., cell wall penetrating activity and / or pesticidal and / or repellent activity) compared to the initial activity of the NLP (e.g., at the time of production or formulation) at a temperature of 0°C or -80°C (e.g., at least -80°C, -70°C, -60°C, -50°C, -40°C or -30°C).

[0081] As used herein, the term "untreated" refers to a plant, animal, fungus, or bacterium that has not been contacted with or delivered with an NLP composition described herein, including a separate plant, animal, fungus, or bacterium that has not had the NLP composition delivered to it, the same plant, animal, fungus, or bacterium that has undergone a treatment that has been evaluated at a time prior to delivery of the NLP composition, or the same plant, animal, fungus, or bacterium that has undergone a treatment that has been evaluated on an untreated portion of the plant, animal, fungus, or bacterium.

[0082] As described herein, the term "hydrophobic core" refers to the innermost portion of an NLP that is hydrophobic in nature. In some embodiments, the hydrophobic core comprises one or more non-polar, non-liposome-forming lipids (e.g., triglycerides). In some embodiments, a hydrophobic drug (e.g., deltamethrin) is dissolved in the hydrophobic core. In some embodiments, the hydrophobic core comprises a trace amount of a co-solvent (e.g., DCM) used during NLP production. In some embodiments, the NLP comprises a hydrophobic dye (e.g., Exalite 594) in its hydrophobic core. Some embodiments relate to NLPs having a hydrophobic core encapsulated by an outer phospholipid-containing monolayer, with the fatty acid chains of the phospholipids facing inward and in contact with the hydrophobic core. In some embodiments, the fatty acid chains are part of the hydrophobic core.

[0083] As used herein, the term "hydrophilic core" refers to the innermost portion of an NLP that is polar in nature. In some embodiments, the hydrophilic core is aqueous in nature (e.g., water or a salt solution in water). In some embodiments, the hydrophilic core comprises a water-soluble agent, such as a polynucleotide, a polypeptide, a hydrophilic small molecule, etc. Some embodiments relate to NLPs having a hydrophilic core encapsulated by a phospholipid bilayer, wherein the phosphorylated head groups of the phospholipids that form the inner phospholipid layer of the bilayer are in contact with the hydrophilic core.

[0084] I. NLP Composition and Characteristics A. NLP Composition Some embodiments relate to NLP compositions comprising at least one phospholipid, at least one non-polar lipid, and at least one surface modifier, wherein the NLP has a hydrophobic core. In some embodiments, the NLP composition further comprises a co-solvent. In some embodiments, the NLP composition further comprises one or more excipients. In some embodiments, the NLP composition further comprises one or more heterofunctional agents. In some embodiments, the heterofunctional agent is a hydrophobic agent. In some embodiments, the hydrophobic heterofunctional agent is a pesticide (e.g., deltamethrin). In some embodiments, the NLP composition further comprises a dye. In some embodiments, the NLP composition comprises a lipid layer isolated from a natural source (e.g., natural phospholipids), semi-synthetic, or fully synthetic lipid.

[0085] a) Phospholipids (PL) Some embodiments relate to NLP compositions comprising at least one phospholipid. In some embodiments, the phospholipids in the NLP form one or more phospholipid layers (e.g., a phospholipid monolayer, a phospholipid bilayer, etc.). In some embodiments, the phospholipids form a micellar structure with a hydrophilic core surrounded by a phospholipid bilayer. In some embodiments, the NLP comprises several layers of phospholipid layers similar to the layers of an onion. In some embodiments, the NLP is a sealed structure in the micron and submicron range dispersed in an aqueous solution. In some embodiments, the NLP comprises one or more bilayers (lamellae) that separate the external aqueous solution from an internal phase or "core." In some embodiments, the core is hydrophobic. In other embodiments, the core is hydrophilic. In some embodiments, the one or more phospholipid layers (e.g., a monolayer, a bilayer, etc.) comprise one or more amphiphilic agents. Amphiphilic agents comprise both polar and nonpolar regions. When amphiphilic agents are present in an aqueous phase, they self-aggregate such that their hydrophilic portions face the aqueous phase while their hydrophobic domains are "shielded" from the aqueous phase. In some embodiments, NLPs can comprise a phospholipid bilayer in which the hydrophobic domains face each other. In some embodiments, NLPs can comprise a phospholipid monolayer in which the hydrophobic domains face the hydrophobic core of the NLP. In some embodiments, NLPs are formed by organizing amphiphilic agents, such as phospholipids, into a lamellar phase, where the lamellae form closed structures and organize into vesicles.

[0086] Some embodiments relate to NLP compositions used as carriers to facilitate the movement of heterologous functional agents through soil. In some embodiments, NLP compositions comprising two or more types of liposomes are used to facilitate the movement of heterologous functional agents through soil. In some embodiments, the liposomes can be any one or combination of vesicles selected from the group consisting of small unilamellar vesicles (SUVs), large unilamellar vesicles (LUVs), multilamellar vesicles (MLVs), multivesicular vesicles (MVVs), large multivesicular vesicles (LMVs) (sometimes referred to as giant multivesicular vesicles, ("GMVs")), oligolamellar vesicles (OLVs), and the like.

[0087] Some embodiments relate to NLP compositions comprising at least one phospholipid, at least one of which is a liposome-forming phospholipid. Without being limited by theory, the amount of phospholipid in the NLP can be measured as organophosphates by a modified Bartlett method (Shmeeda H, Even-Chen S, Honen R, Cohen R, Weintraub C, Barenholz 2003. Enzymatic assays for quality control and pharmacokinetics of liposome formulations: comparison with nonenzymatic conventional methodologies. Methods Enzymol 367:272-92).

[0088] In some embodiments, the NLP composition comprises at least one phospholipid selected from glycerophospholipids and sphingomyelins. Glycerophospholipids have a glycerol backbone in which at least one, and preferably two, hydroxyl groups in the head group are replaced by one or two hydrocarbon tails (chains), typically an acyl, alkyl, or alkenyl tail, and the third hydroxyl group is replaced by a phosphate (phosphatidic acid) or phosphoester, such as a phosphocholine group (as exemplified in phosphatidylcholine), which is the polar head group of glycerophospholipids, or any combination of the above and / or derivatives thereof, and may contain a chemically reactive group (such as an amine, acid, ester, aldehyde, or alcohol).Examples of glycerophospholipids include phosphoglycerols (PG) including dimyristoylphosphatidylglycerol (DMPG), egg yolk phosphatidylcholine, soybean PC, sunflower PC, rapeseed PC, krill PC, canola PC, flaxseed lecithin, wheat lecithin, dimyristoylphosphatidylcholine (DMPC, Tm 24°C), 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), hydrogenated soybean phosphatidylcholine (HSPC, Tm 65°C), phosphatidylcholine (PC) including distearoylphosphatidylcholine (DSPC, Tm 55°C), dilauroyl-sn-glycero-2-phosphocholine (DLPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC, Tm 41°C), 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine, 1,2-diarachidoyl-sn-glycero-3-phosphocholine (DBPC), 1,2-dihenarachidoyl-sn-glycero-3-phosphocholine, 1,2-dibehenoyl-sn-glycero-3-phosphocholine, 1,2-ditricosanoyl-sn-glycero-3-phosphocholine, 1,2-dilignoceroyl-sn-glycero-3-phosphocholine, 1-myristoyl-2-stearoyl- Examples of sphingomyelin include, but are not limited to, sn-glycero-3-phosphocholine, 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine (PSPC), 1-stearoyl-2-palmitoyl-sn-glycero-3-phosphocholine (SPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC-1.7°C), phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylserine (PS), and phosphatidylethanolamine (PE). Sphingomyelin consists of a ceramide (N-acylsphingosine) unit with a phosphocholine moiety attached to the 1-position as the polar head group. The terms "sphingomyelin" or "SPM" as used herein refer to any N-acetylsphingosine conjugated to a phosphocholine group, which subsequently forms the polar head group of sphingomyelin (N-acylsphingosylphosphocholine).The acyl chain attached to the primary amino group of sphingosine (to form the ceramide) can be saturated or unsaturated, branched or unbranched.

[0089] In some embodiments, the NLP compositions comprise phospholipids having one or two C14-C24 hydrocarbon tails (e.g., acyl, alkyl, or alkenyl chains) with varying degrees of saturation, from fully saturated to fully, partially, or non-hydrogenated lipids. In some embodiments, naturally occurring phospholipids can be further converted to saturated phospholipids by hydrogenation or can be further treated with enzymes, for example, to remove partial fatty acids (e.g., using phospholipase A2) or to convert polar head groups (e.g., using phospholipase D). Saturated phospholipids are considered naturally occurring phospholipids because the resulting saturated lipids are also naturally occurring (e.g., nature-identical).

[0090] In some embodiments, the NLP composition comprises at least one phospholipid comprising a polar head group. In some embodiments, the polar head group comprises an alcohol moiety. In some embodiments, the polar head group comprises a serine moiety. In some embodiments, the polar head group comprises a choline moiety. In some embodiments, the polar head group comprises an ethanolamine. In some embodiments, the polar head group comprises glycerol.

[0091] In some embodiments, the NLP composition comprises at least one phospholipid comprising a polar inositol head group. In some embodiments, the phospholipid comprising an inositol head group is selected from the group consisting of phosphatidylinositol (PI), PI(4)P, PI(3)P, PI(3,4,5)P3, PI(4,5)P2, PI(3,5)P2, and PI(3,4)P2. In some embodiments, at least one phospholipid has an acidic head group. In some embodiments, the acidic head group comprises a moiety selected from the group consisting of glycerol, hydroxyl, carboxyl, amine, and phosphorus groups.

[0092] In some embodiments, the NLP composition comprises phosphatidylglycerol (PG), such as dilauroylphosphatidylglycerol (DLPG), dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylglycerol (DSPG), dioleoylphosphatidylglycerol (DOPG), egg yolk phosphatidylglycerol (egg yolk PG), hydrogenated egg yolk phosphatidylglycerol, phosphatidylinositol ( and at least one acidic phospholipid, including a natural or synthetic lipid selected from phosphatidylinositol, dimyristoylphosphatidylinositol, dipalmitoylphosphatidylinositol (DPPI), distearoylphosphatidylinositol (DSPI), dioleoylphosphatidylinositol (DOPI), soybean phosphatidylinositol (soybean PI), hydrogenated soybean phosphatidylinositol, phosphoinositides, sphingomyelin, and phosphatidic acid. Each of these acidic phospholipids can be used alone or in combination of two or more in the NLPs of the present disclosure.

[0093] In some embodiments, at least one phospholipid in the NLPs is derived from lecithin, which is described in the United States Pharmacopeia (USP) as a complex mixture of acetone-insoluble phosphatides, composed primarily of PC, PE, phosphatidylserine, and phosphatidylinositol, isolated from crude vegetable oil sources, combined with varying amounts of other substances, such as triglycerides, fatty acids, and carbohydrates.

[0094] In some embodiments, about 5% to 50% (w / w) of the lipids in the NLP composition are phospholipids (e.g., about 10% to 20% of the lipids in the NLP composition are phospholipids, e.g., about 10%, 12.5%, 16%, or 20% of the lipids in the NLP composition are phospholipids). In some embodiments, about 30% to 75% of the lipids in the NLP composition are phospholipids (e.g., about 35% or about 50% phospholipids). In some embodiments, about 35% to 50% (e.g., about 36%, 36.5%, 37%, 37.5%, 38%, 38.5%, 39%, 39.5%, 40%, 40.5%, 41%, 41.5%, 42%, 42.5%, 43%, 43.5%, 44%, 44.5%, 45%, 45.5%, 46%, 46.5%, 47%, 47.5%, 48%, 48.5%, 49%, 49.5%) is phospholipids.

[0095] In some embodiments, the phospholipid is selected from the group consisting of crude lemon lipids, purified lemon phospholipids, phosphatidylethanolamine (PE), LIPOID H PS 70 (phosphatidylserine), 14C-PA (phosphatidic acid), LIPOID H90 (phosphatidylcholine), sunflower lecithin, soybean lecithin, and deoiled soybean lecithin.

[0096] b) Non-polar lipids (NP) Some embodiments relate to NLP compositions comprising at least one non-polar lipid. Non-polar lipids are understood to be non-amphiphilic and non-liposome-forming. Non-liposome-forming lipids refer to lipids that do not spontaneously form into vesicles when introduced into an aqueous medium. In some embodiments, the non-polar lipids are derived from natural sources. In some embodiments, the non-polar lipids are derived from plant sources. In some embodiments, the NLP compositions comprise one or more natural / plant-derived non-polar lipids obtained from vegetable sources such as seed oils (soybean, rapeseed (canola), wheat germ, sunflower, flax, cotton, corn, coconut, peanut, sesame), pulp oils (palm, olive, avocado pulp), desert shrub, tobacco, kidney bean, and carrot. In some embodiments, the non-polar lipids comprise triglycerides each comprising at least one fatty acid typically selected from the group consisting of C6:0, C8:0, C10:0, C12:0, C14:0, C15:0, C16:0, C17:0, C18:0, C20:0, C22:0, and C24:0, and the saturated fatty acids are C16:1(n-7), C16:1(n-9), C17:1(n-7), C18:1(n-9), C19:1(n-9), C20:1(n-9), C22:1(n-9), C24 ... : 1(n-7), C20: 1(n-7), C20: 1(n-9), C22: (n-9) and C24: 1(n-9) and monounsaturated fatty acids C18: 2(n-6), C18: 3(n-3), C18: 3(n-6), C18: 4(n-3), C20: 2(n-6), C20: 3(n-6), C20: 4(n-6), C20: 5(n-3), C22: 2(n-6) and C22: 4(n-6). The fatty acid profile types of 80 vegetable oils are described by Dubois et al., Eur. J. Lipid Sci. Technol. 109 (2007) 710-732 (incorporated herein by reference).

[0097] In some embodiments, the NLP composition comprises a non-polar lipid comprising 40% of at least one fatty acid chain selected from the group consisting of polyunsaturated fatty acids, monounsaturated fatty acids, and saturated fatty acids. In some embodiments, the NLP composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, or more than 60% (w / w) oil (e.g., soybean oil), for example, 1%-10%, 10%-20%, 20%-30%, 30%-40%, 40%-50%, or 50%-60% (w / w) soybean oil. In some embodiments, the NLP composition comprises about 35%-50% oil (e.g., soybean oil), for example, a molar ratio of about 36%, 38.5%, 42.5%, or 46.5% oil. In some embodiments, the NLP composition comprises about 20% to 60% oil.

[0098] In some embodiments, the NLP composition comprises one or more lipids that do not naturally form vesicles but can be incorporated into vesicles. Non-limiting examples of non-vesiculating lipids include sterols, sphingolipids (e.g., sphingomyelin), and lipoproteins. In some embodiments, the NLP composition comprises one or more sterols selected from the group consisting of β-sitosterol, β-sitostanol, stigmasterol, stigmastanol, campesterol, campestanol, ergosterol, avenasterol, brassicasterol, fucosterol, cholesterol (CHOL), cholesteryl hemisuccinate, and cholesteryl sulfate, or any combination of two or more of these sterols. In some embodiments, the sterol is a plant-derived sterol (e.g., a phytosterol). The NLP composition comprises one or more plant sterols selected from the group consisting of β-sitosterol, β-sitostanol, stigmasterol, stigmastanol, campesterol, campestanol, ergosterol, avenasterol, brassicasterol, and any combination of two or more of these sterols. In some embodiments, the NLP composition comprises one or more plant sterols selected from the group consisting of β-sitosterol, stigmasterol, and ergosterol.

[0099] In some embodiments, the NLP composition comprises one or more lipid membranes comprising a molar ratio between phospholipids and non-polar lipids of 10%:90% to 90%:10%, optionally a molar ratio of 20%:80% to 80%:20%, optionally a molar ratio of 30%:70% to 70%:30%, optionally a molar ratio of 20%:80% to 50%:50%, and optionally a molar ratio of 20:80 to 40%:60%.

[0100] In some embodiments, the non-polar lipid is selected from the group consisting of sunflower oil, canola oil, soybean oil, olive oil, palm oil, and refined lemon lipids.

[0101] c) Surface modifier Some embodiments relate to NLP compositions comprising at least one surface modifier, which alters the mobility of the NLP composition through soil compared to an NLP composition without the surface modifier. In some embodiments, the surface modifier stabilizes the NLP composition. The NLP compositions described herein may comprise one or more surface modifiers (e.g., may be loaded, encapsulated, conjugated with one or more surface modifiers) or may be formulated with one or more surface modifiers (e.g., may be suspended or resuspended in a solution containing one or more surface modifiers). In some embodiments, the surface modifier affects the binding of any of the components of the NLP composition to any components present in the soil. In some embodiments, the one or more surface modifiers are incorporated into one or more of the phospholipid layers of the NLP. In some embodiments, the NLP composition comprises at least one surface modifier selected from Table 1. In some embodiments, the NLP composition comprises at least two, three, four, five, or more surface modifiers selected from Table 1.

[0102] In some embodiments, the NLP composition comprises at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more than 90% of one or more surface modifiers. In some embodiments, the NLP composition comprises at least 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than 90% weight / weight of a synthetic chemical surface modifier (e.g., pegylated compounds, polycarboxylates, etc.). In some embodiments, the NLP composition comprises at least 1%-10%, 10%-20%, 20%-30%, 30%-40%, 40%-50%, 50%-60%, 60%-70%, 70%-80%, or 80%-90% weight / weight of one or more surface modifiers (e.g., polycarboxylates). In some embodiments, the NLP composition comprises at least about 30%-75% weight / weight of polycarboxylate surface modifiers. In some embodiments, the NLP contains up to 5 mol% of a surface modifier. In some embodiments, the NLP composition comprises about 0.1 mol%-5 mol%, about 0.5 mol%-4 mol%, or about 1 mol%-3 mol% of one or more surface modifiers. In some embodiments, the NLP composition comprises 25% Atlox 500L. In some embodiments, the NLP composition comprises 35% Atlox 500L. In some embodiments, the NLP composition comprises a molar ratio of 50% Atlox 500L.

[0103] Some embodiments relate to NLP compositions comprising one or more surface modifiers that affect the mobility of the NLP composition in soil. In some embodiments, the surface modifier is a synthetic compound that affects the mobility of the NLP composition in soil, and the NLP composition optionally includes a heterofunctional agent. In some embodiments, the surface modifier alters one or more surface properties of the NLP composition. In some embodiments, the surface modifier structurally alters the NLP, for example, by adding chemical groups to the outer surface of the NLP. In some embodiments, the NLP composition comprises a glycolipid moiety exposed at the outer surface of the NLP composition. In some embodiments, the NLP composition comprises at least one glycoprotein embedded in the outer surface of the NLP composition. In some embodiments, at least a portion of the surface modifier is incorporated into a lipid membrane of the NLP, for example, a lipid domain embedded in a phospholipid membrane. In some embodiments, at least a portion of the surface modifier is exposed to the exterior of the NLP, for example, facing air, soil, or a solution in which the NLP is dispersed. In some embodiments, the surface modifier is an emulsifier. In some embodiments, the surface modifier is amphiphilic in nature, e.g., comprising a hydrophobic portion and a hydrophilic portion chemically linked in one molecule. In some embodiments, the surface modifier is a surfactant. In some embodiments, the surface modifier affects the surface charge of the NLPs, e.g., by making the surface charge of the NLPs more or less negative. In some embodiments, the surface charge of the NLPs is expressed as the zeta potential of the NLPs. In some embodiments, the surface charge of the NLPs affects the affinity of the NLPs for a charged matrix. In some embodiments, the charged matrix is ​​soil. In some embodiments, the surface charge affects the affinity of the NLPs for one or more components present in the soil (e.g., silicates, clays, biological components, etc.). In some embodiments, the surface charge of the NLPs affects the mobility of the NLPs in soil. In some embodiments, the zeta potential predicts the retention of the NLPs in soil. In some embodiments, the zeta potential predicts the mobility of the NLPs in soil.In some embodiments, the NLP composition comprises at least one surface modifier selected from Table 1 that affects the mobility of the NLP composition in soil. In some embodiments, the NLP composition comprises at least two, three, four, five, or more surface modifiers selected from Table 1 that affect the mobility of the NLP composition in soil.

[0104] Some embodiments relate to NLP compositions comprising one or more surface modifiers that increase uptake of the NLP composition by a plant or plant part (e.g., roots, leaves, plant cells, etc.). In some embodiments, the one or more surface modifiers increase uptake of the NLP composition as a whole. In some embodiments, the one or more surface modifiers increase uptake of a portion or component of the NLP composition, such as the uptake of a heterologous functional agent (e.g., a heterologous agricultural agent (e.g., a pesticide, fertilizer, herbicide, plant modifier, plant growth promoter, biostimulant, or plant immunity inducer) carried by the NLP. The extent to which uptake is increased can vary depending on the plant or plant part to which the NLP composition is delivered. In some embodiments, the one or more surface modifiers increase uptake of the NLP composition by a plant or plant part by at least 1%, 2%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70% or more compared to an NLP composition lacking the one or more surface modifiers. , 80%, 90% or 100%. In some embodiments, the one or more surface modifiers may increase uptake of the NLP composition by the plant or plant part by at least 2-fold, 4-fold, 5-fold, 10-fold, 100-fold or 1000-fold compared to an NLP composition lacking the one or more surface modifiers. In some embodiments, the NLP composition comprises at least one surface modifier selected from Table 1 that increases uptake of the NLP composition in the plant or plant part. In some embodiments, the NLP composition comprises at least two, three, four, five or more surface modifiers selected from Table 1 that increase uptake of the NLP composition in the plant or plant part.

[0105] Some embodiments relate to NLP compositions comprising one or more surface modifiers that increase uptake of the NLP composition by cells, such as plant cells. In some embodiments, the one or more surface modifiers increase uptake of the NLP composition as a whole. In some embodiments, the one or more surface modifiers increase uptake of a portion or component of the NLP composition, such as the uptake of a heterologous functional agent (e.g., a bactericide) carried by the NLP composition. The extent to which uptake is increased may vary depending on the bacterial cells to which the NLP composition is delivered. In some embodiments, the one or more surface modifiers may increase uptake of the NLP composition by bacterial cells by at least 1%, 2%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to an NLP composition lacking the one or more surface modifiers. In some embodiments, the NLP composition comprises at least one surface modifier selected from Table 1 that increases uptake of the NLP composition in bacterial cells. In some embodiments, the NLP composition comprises at least two, three, four, five, or more surface modifiers selected from Table 1 that increase uptake of the NLP composition in bacterial cells.

[0106] Some embodiments relate to NLP compositions comprising one or more surface modifiers that increase uptake of the NLP composition by fungal cells. In some embodiments, the one or more surface modifiers increase uptake of the NLP composition as a whole. In some embodiments, the one or more surface modifiers increase uptake of a portion or component of the NLP composition, such as the uptake of a heterologous functional agent (e.g., a fungicide) carried by the NLP composition. The extent to which uptake is increased may vary depending on the fungal cells to which the NLP composition is delivered. In some embodiments, the one or more surface modifiers may increase uptake of the NLP composition by fungal cells by at least 1%, 2%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to an NLP composition lacking the one or more surface modifiers. In some embodiments, the NLP composition comprises at least one surface modifier selected from Table 1 that increases uptake of the NLP composition in fungal cells. In some embodiments, the NLP composition comprises at least two, three, four, five, or more surface modifiers selected from Table 1 that increase uptake of the NLP composition in fungal cells.

[0107] In some embodiments, the surface modifier can be an anionic agent, a cationic agent, or a zwitterionic agent. In some embodiments, the surface modifier can be a pegylated compound, a glycolipid, an anionic polymer, a polycarboxylate, a polysaccharide, or any combination thereof. In some embodiments, the surface modifier can be a polysaccharide having a lipid chain. In some embodiments, the surface modifier is a pegylated surface modifier selected from the group consisting of a pegylated block copolymer (e.g., a poloxamer) or a cocamide derivative. In some embodiments, the surface modifier is a pegylated compound selected from the group consisting of PEG2000-C18 and PEG5000-C18. In some embodiments, the surface modifier is a rhamnolipid. In some embodiments, the surface modifier is a sophorolipid. In some embodiments, the surface modifier is an anionic polymer. In some embodiments, the surface modifier is Atlox 500L. In some embodiments, the surface modifier is an anionic polymer. In some embodiments, the surface modifier is a styrene-acrylic copolymer. In some embodiments, the surface modifier is Atlox 4917. In some embodiments, the surface modifier is a polycarboxylate. In some embodiments, the surface modifier is Atlox CS100B. In some embodiments, the surface modifier is a polysaccharide, such as a C8-C10 alkyl polysaccharide. In some embodiments, the surface modifier is Atlox AL2575. In some embodiments, the surface modifier is an emulsifier. In some embodiments, the surface modifier is selected from the examples of surface modifiers suitable for NLP production provided in Table 1.

[0108] [Table 1]

[0109] In some embodiments, the surface modifier may be a lipopolymer. As used herein, the term lipopolymer refers to a lipid material modified by incorporating a hydrophilic polymer into its polar head group. In some embodiments, the polymer head group of the lipopolymer is water-soluble. In some embodiments, the hydrophilic polymer has a molecular weight of 750 Da or greater. There are many polymers that can be attached to lipids to form lipopolymers, non-limiting examples of which include polyethylene glycol (PEG), polysialic acid, polylactic acid (also known as polylactide), polyglycolic acid (also known as polyglycolide), polylactic-polyglycolic acid, polyvinyl alcohol, polyvinylpyrrolidone, polymethoxazoline, polyethyloxazoline, polyhydroxyethyloxazoline, polyhydroxypropyloxazoline, polyaspartamide, polyhydroxypropylmethacrylamide, polymethacrylamide, polydimethylacrylamide, polyvinylmethylether, polyhydroxyethylacrylate, and derivatized celluloses (e.g., hydroxymethylcellulose, hydroxyethylcellulose, etc.). The polymers may be used as homopolymers or block or random copolymers. The lipids derivatized to the lipopolymer can be neutral, negatively charged, and positively charged.

[0110] In some embodiments, the surface modifier is a PEGylated lipid. The length of the polyethylene glycol (PEG) can vary from 1 kDa to 10 kDa. In some embodiments, the NLP composition comprises one or more PEGylated lipids having a PEG length of 2 kDa. In some embodiments, the NLP composition comprises one or more PEGylated lipids independently selected from C14-PEG2k, C18-PEG2k, and DMPE-PEG2k. In some embodiments, the PEGylated lipid is a PEG5K PEGylated lipid (e.g., C14-PEG5k, C18-PEG5k, or DMPE-PEG5K). In some embodiments, the NLP composition comprises at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.5%, 4%, 4.5%, 5%, 10%, 20%, 30%, 40%, 50% or greater than 50% molar ratio of one or more PEGylated lipids (e.g., C14-PEG2k, C18-PEG2k, C18-PEG5K, DMPE-PEG2k, etc.). In some embodiments, the NLP composition comprises one or more PEGylated lipids in a molar ratio of at least 0.1%-0.5%, 0.5%-1%, 1%-1.5%, 1.5%-2.5%, 2.5%-3.5%, 3.5%-5%, 5%-10%, 10%-20%, 20%-30%, 30%-40%, or 30%-50%. In some embodiments, the NLP composition comprises about 0.1% to about 10% (w / w) PEGylated lipid (e.g., C14-PEG2k, C18-PEG2k, DMPE-PEG2k, etc.). In some embodiments, the NLP composition comprises about 1%-3% of one or more PEGylated lipids. In some embodiments, the NLP composition comprises about 1.5% of one or more PEGylated lipids. In some embodiments, the NLP composition comprises about 2.5% of one or more PEGylated lipids. In some embodiments, NLP compositions comprising one or more PEGylated lipids have altered mobility in soil compared to NLP compositions that do not contain one or more PEGylated lipids.In some embodiments, an NLP composition comprising one or more PEGylated lipids has an altered affinity for soil compared to an NLP composition that does not contain one or more PEGylated lipids.

[0111] In some embodiments, the NLP composition comprises one or more phospholipids. In some embodiments, about 5% to 50% (w / w) of the lipids in the NLP composition are phospholipids (e.g., about 10% to 20% of the lipids in the NLP composition are phospholipids, e.g., about 10%, 12.5%, 16%, or 20% of the lipids in the NLP composition are phospholipids). In some embodiments, about 30% to 75% of the lipids in the NLP composition are phospholipids (e.g., about 35% or about 50% phospholipids). In some embodiments, about 35% to 50% (e.g., about 36%, 36.5%, 37%, 37.5%, 38%, 38.5%, 39%, 39.5%, 40%, 40.5%, 41%, 41.5%, 42%, 42.5%, 43%, 43.5%, 44%, 44.5%, 45%, 45.5%, 46%, 46.5%, 47%, 47.5%, 48%, 48.5%, 49%, 49.5%) is phospholipids.

[0112] In some embodiments, the NLP composition comprises one or more PEGylated lipids. In some embodiments, about 5% to 50% (w / w) of the lipids in the NLP composition are PEGylated lipids (e.g., about 10%-20% of the lipids in the NLP composition are PEGylated lipids, e.g., about 10%, 12.5%, 16%, or 20% of the lipids in the NLP composition are PEGylated lipids). In some embodiments, about 30%-75% of the lipids in the NLP composition are PEGylated lipids (e.g., about 35% or about 50% PEGylated lipids). In some embodiments, about 35%-50% (e.g., about 36%, 36.5%, 37%, 37.5%, 38%, 38.5%, 39%, 39.5%, 40%, 40.5%, 41%, 41.5%, 42%, 42.5%, 43%, 43.5%, 44%, 44.5%, 45%, 45.5%, 46%, 46.5%, 47%, 47.5%, 48%, 48.5%, 49%, 49.5%) of the lipids in the NLP composition are phospholipids.

[0113] In some embodiments, NLP compositions comprising one or more PEGylated lipids have enhanced uptake compared to NLP compositions without one or more PEGylated lipids. In some embodiments, NLP compositions comprising one or more PEGylated lipids have altered stability (e.g., improved stability, decreased stability, etc.) compared to NLP compositions without one or more PEGylated lipids. In some embodiments, NLP compositions comprising one or more PEGylated lipids have altered particle size compared to NLP compositions without one or more PEGylated lipids. In some embodiments, NLP compositions comprising one or more PEGylated lipids are less likely to be phagocytosed by cells than NLP compositions without one or more PEGylated lipids. In some embodiments, NLP compositions comprise one or more surface modifiers comprising one or more PEG moieties having a head group molecular weight of about 750 Da to about 20,000 Da. In some embodiments, NLP compositions comprise one or more surface modifiers comprising one or more PEG moieties having a head group molecular weight of about 750 Da to about 12,000 Da. In some embodiments, NLP compositions comprise one or more surface modifiers comprising one or more PEG moieties having a head group molecular weight of about 1,000 Da to about 5,000 Da. In some embodiments, NLP compositions comprise one or more neutral (uncharged) lipopolymers. In some embodiments, NLP compositions comprise one or more positively charged lipopolymers. In some embodiments, NLP compositions comprise one or more negatively charged lipopolymers. In some embodiments, NLP compositions comprise one or more neutral distearoylglycerols and negatively charged distearoylphosphatidylethanolamines, both covalently attached to methoxypoly(ethylene glycol) (mPEG or PEG) of Mw 750, 2,000, 5,000, or 12,000.

[0114] In some embodiments, the surface modifier is a glycolipid. In some embodiments, one or more glycolipids are rhamnolipids. In some embodiments, one or more glycolipids are sophorolipids. In some embodiments, about 5%-50% (w / w) of the lipids in the NLP composition are glycolipids (e.g., about 10%-20% of the lipids in the NLP composition are glycolipids, e.g., about 10%, 12.5%, 16%, or 20% of the lipids in the NLP composition are glycolipids). In some embodiments, about 30%-75% of the lipids in the NLP composition are glycolipids (e.g., about 35% or about 50% glycolipids). In some embodiments, about 35%-50% (e.g., about 36%, 36.5%, 37%, 37.5%, 38%, 38.5%, 39%, 39.5%, 40%, 40.5%, 41%, 41.5%, 42%, 42.5%, 43%, 43.5%, 44%, 44.5%, 45%, 45.5%, 46%, 46.5%, 47%, 47.5%, 48%, 48.5%, 49%, 49.5%) of the lipids in the NLP composition are glycolipids.

[0115] In some embodiments, the NLPs comprise any of the modifiers (SM) listed in Table 1. In some embodiments, the NLPs comprise any combination of non-polar lipids (NPs), polar lipids (PLs), and surface modifiers (SMs) listed in Table 2. In some embodiments, the NPs, PLs, and SMs are mixed in any of the ratios shown in Table 2 and subjected to any of the fabrication methods disclosed herein (e.g., the DCM method, the HHPH method, or the NanoAssemblr® IGNITE™ method) to form lipid nanoparticles, as outlined in the Examples. Those skilled in the art will recognize additional methods for forming lipid nanoparticles. Exemplary NLPs that can be produced by any of the methods are listed in Table 2.

[0116] [Table 2]

[0117] [Table 3]

[0118] Table 4

[0119] Table 5

[0120] Table 6

[0121] Table 7

[0122] Table 8

[0123] Table 9

[0124] d)Co-solvent Some embodiments relate to NLP compositions comprising one or more cosolvents. In some embodiments, one or more cosolvents are included in the NLP composition to improve the encapsulation efficiency of a cargo (e.g., a heterofunctional agent) compared to an NLP without the one or more cosolvents. In some embodiments, an NLP composition comprising one or more cosolvents has a cargo encapsulation efficiency that is at least 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or greater than 99% greater than the cargo encapsulation efficiency of an NLP composition without the one or more cosolvents. In some embodiments, the cargo is a heterofunctional agent. In some embodiments, the cargo is a pyrethroid. Without wishing to be bound by theory, the inclusion of one or more cosolvents increases the solubility of the cargo (e.g., a heterofunctional agent (e.g., a pyrethroid)). Without wishing to be bound by any particular theory, the inclusion of one or more co-solvents prevents precipitation of the cargo (e.g., a heterofunctional agent (e.g., a pyrethroid). In some embodiments, the one or more co-solvents are water-immiscible fluids. Without wishing to be bound by any particular theory, the inclusion of one or more water-immiscible co-solvents prevents precipitation of a hydrophobic cargo (e.g., a hydrophobic heterofunctional agent (e.g., a pyrethroid)). In some embodiments, any of the NLP compositions in Table 2 includes one or more water-immiscible co-solvents selected from Table 3. In some embodiments, the co-solvent is dichloromethane (DCM). In some embodiments, the co-solvent is isopropyl myristate (IPM). In some embodiments, trace amounts of the co-solvent remain after evaporation during production of the NLP composition. In some embodiments, the co-solvent is water-miscible. In some embodiments, the co-solvent is water-immiscible. In some embodiments, the non-polar lipid phase of the NLP is represented solely by the co-solvent (e.g., Genagen, as in NLP683). 4296).

[0125] [Table 10]

[0126] e) excipients In some embodiments, NLP compositions may further comprise one or more stabilizing molecules that increase the stability of the NLPs compared to compositions lacking the one or more stabilizing molecules (e.g., for at least one day at room temperature, at least one week at 4°C, etc.). In some embodiments, NLP compositions comprising one or more excipients are more stable at room temperature than NLP compositions without excipients. In some embodiments, NLP compositions comprising one or more excipients are more stable in soil than NLP compositions without excipients. In some embodiments, NLP compositions comprise one or more excipients that prevent aggregation of the NLPs. In some embodiments, one or more excipients are added during NLP production to increase the stability of the resulting NLPs. In some embodiments, one or more excipients are added after NLP production to increase the stability of the resulting NLPs. In some embodiments, one or more excipients are encapsulated within the liposomal inner core (e.g., hydrophilic core, hydrophobic core) of the NLPs. In some embodiments, one or more excipients are embedded in the lipid membrane of the NLPs. In some embodiments, one or more excipients are provided in a solution in which the one or more NLPs are suspended. Non-limiting examples of excipients that can be included in the NLP compositions are the non-ionic block copolymer surfactants and non-ionic polyalkylene glycol ether surfactants listed in Table 3.

[0127] f) Heterofunctional drugs 1. Functional drugs In some embodiments, the NLP composition comprises one or more heterofunctional agents. In some embodiments, the one or more heterofunctional agents are encapsulated within the inner liposomal core (e.g., hydrophilic core, hydrophobic core, etc.) of the NLP. In some embodiments, the one or more heterofunctional agents are embedded in the lipid membrane of the NLP.

[0128] In some embodiments, the one or more heterofunctional agents included in the NLP composition can be any of the pesticides disclosed herein. In some embodiments, the pesticide can be a natural or synthetic insecticide (e.g., a larvicide, an adulticide). In some embodiments, the pesticide can be a natural or synthetic insect growth regulator. In some embodiments, the pesticide can be a natural or synthetic acaricide (miticide). In some embodiments, the pesticide can be a natural or synthetic molluscicide, nematicide, ectoparasiticide, bactericide, fungicide, or herbicide. The term "pesticide" can further include antibiotics, antiviral pesticides, antifungals, antiparasitics, agents that stop or slow nutrients and / or insect movement, reproduction, etc. In some embodiments, the heterologous functional agent can be a therapeutic agent (e.g., a cell-penetrating agent, an antifungal agent, an antibacterial agent, a virucide, an antiviral agent, an insecticide, a nematicide, an antiparasitic agent, an insect repellent, etc.). In some embodiments, the NLP compositions described herein can include one or more heterologous functional agents listed in Tables 4-6.

[0129] In some embodiments, one or more heterofunctional agents included in the NLP compositions described herein is a pyrethroid (e.g., deltamethrin). In some embodiments, one or more heterofunctional agents included in the NLP compositions described herein is emamectin. In some embodiments, one or more heterofunctional agents are provided in the hydrophobic core of the NLP. In some embodiments, one or more heterofunctional agents are provided in the hydrophilic core of the NLP. In some embodiments, one or more heterofunctional agents are provided in the lipid membrane of the NLP. In some embodiments, deltamethrin is included in the hydrophobic core of the NLP. In some embodiments, emamectin is included in the hydrophobic core of the NLP.

[0130] 2. Volatile functional agents. Volatile functional agents are agents with high vapor pressure. Examples of volatile agents used in agriculture are herbicides, which are typically applied as foliar sprays, fumigants applied to the soil to kill insects, pheromones to disrupt mating, and essential oils to repel insects. High vapor pressure means high environmental exposure and potential environmental hazards for agricultural workers and off-target plants (e.g., grapevines) and off-target insects (e.g., honeybees). Spray drift is a common concern for off-target damage. It occurs when droplets containing bioactive substances are transferred to off-target vegetation during the process of treating the target site. Furthermore, undesirable diffusion of herbicides, for example, can occur when the spray solution settles on-site and then transforms into a gas phase and is carried away from the site by wind. NLP encapsulation of volatile bioactive substances provides a means to reduce their risk and unwanted effects on off-targets.

[0131] In some embodiments, encapsulation of volatile bioactive substances in NLPs alters the environmental exposure of the volatile bioactive substances. By varying the chemical nature of the NLP's components, NLPs of various sizes, stabilities, and degrees of permeability can be formed. These factors determine the rate at which the encapsulated volatile bioactive substance is released, which in turn affects the residual properties, rate of action, and environmental exposure of the bioactive substance. In some embodiments, compositions include a mixture of NLPs containing one or more volatile bioactive substances, facilitating the controlled release of one or more bioactive substances over time.

[0132] In some embodiments, NLP encapsulation of any of the volatile functional agents described herein (e.g., volatile insecticides, volatile herbicides, volatile fumigants, or volatile essential oils) facilitates their use in spray form. In some embodiments, the NLP composition comprising a volatile functional agent is a sprayable formulation.

[0133] Insecticides In some embodiments, the volatile functional agent is an insecticide. In some embodiments, the volatile functional agent is an insecticide described herein that has a high vapor pressure. In some embodiments, NLP encapsulation of any of the volatile insecticides described herein facilitates their use in spray form. In some embodiments, an NLP composition comprising a volatile insecticide is a sprayable insecticide formulation. In some embodiments, the volatile insecticide is a pyrethroid. In some embodiments, the pyrethroid is tefluthrin.

[0134] 2.2. Herbicides Exemplary volatile herbicides used in agriculture that can be encapsulated in any of the NLPs by any of the methods described herein include, but are not limited to, dicamba and 2.4-D. In some embodiments, the dicamba formulation is XtendiMax, Engenia, and Tavium, or FeXapan. In some embodiments, the volatility of the herbicide is reduced by encapsulating the herbicide in any of the NLP compositions disclosed herein. In some embodiments, NLP encapsulation of any of the volatile herbicides described herein facilitates their use in spray form. In some embodiments, the NLP composition comprising a volatile herbicide is a sprayable herbicide formulation. In some embodiments, the volatile herbicide is dicamba.

[0135] 2.3. Fumigants Exemplary fumigants that may be encapsulated in any of the NLPs by any of the methods described in this application include, but are not limited to, 1,2-dibromo-3-chloropropane, 1,3-dichloropropene, aluminum phosphide, atrazine, azinphosmethyl, benomyl, carbaryl (Sevin®), carbofuran, carbon disulfide, chlordane, chloropicrin, chlorpyrifos, dazomet (Basamid®), DD, 1,3-dichloropropene, diazinon, dichlorvos (DDVP), dicrotophos, dieldrin, diquat (inhalable), endosulfan, endrin, epichlorohydrin, ethyl p-nitrophenyl phenol, Examples of suitable fumigant compositions include phenylphosphorothioate (EPN), ethion, ethylene dibromide (EDB), fenamiphos, fenthion, fonofos (diphonate), formaldehyde, heptachlor, iodoform, hydrogen cyanide, hydrogen disulfide, malathion, metam sodium, methoxychlor, methyl bromide, methyl iodide, methyl isocyanate, methyl isothiocyanate, methyl parathion, mevinphos (phosdrin), naled, paraquat, parathion, phosphine, picloram, ronnel, rotenone, sodium tetrathiocarbonate, sulfotep (TEDP), sulfuryl fluoride, temephos, thiram, trichlorophenoxyacetic acid, warfarin, or tefluthrin. In some embodiments, the volatility of the fumigant is reduced by encapsulation of the fumigant in any of the NLP compositions disclosed herein. In some embodiments, NLP encapsulation of any of the fumigant compositions described herein facilitates their use in spray form.

[0136] 2.4. Pheromones. Exemplary insect pheromones that may be encapsulated into any of the NLPs by any of the methods described in this application include, but are not limited to, (E)-2-decen-1-ol, (E,E)-10,12-tetradecadien-1-ol, (E)-2-decenyl acetate, (E,E)-10,12-tetradecadienyl acetate, (E)-2-decenal, (E,E)-10,12-tetradecadienal, (Z)-2-decen-1-ol, (E,Z)-10,12-tetradecadienyl acetate, (Z)-2-decenyl acetate, (Z,E)-1 0,12-Tetradecadienyl acetate, (Z)-2-decenal, (Z,Z)-10,12-tetradecadien-1-ol, (E)-3-decen-1-ol, (Z,Z)-10,12-tetradecadienyl acetate, (Z)-3-decenyl acetate, (E,Z,Z)-3,8,11-tetradecatrienyl acetate, (Z)-3-decen-1-ol, (E)-8-pentadecen-1-ol, (Z)-4-decen-1-ol (E)-8-pentadecenyl acetate, (E)-4-decenyl acetate, (Z)-8-pentadecen- 1-ol, (Z)-4-decenyl acetate, (Z)-8-pentadecenyl acetate, (Z)-4-decenal, (Z)-9-pentadecenyl acetate, (E)-5-decen-1-ol, (E)-9-pentadecenyl acetate, (E)-5-decenyl acetate, (Z)-10-pentadecenyl acetate, (Z)-5-decen-1-ol, (Z)-10-pentadecenal, (Z)-5-decenyl acetate, (E)-12-pentadecenyl acetate, (Z)-5-decenal, (Z)-12-pentadecenyl acetate, (E)- 7-decenyl acetate, (Z,Z)-6,9-pentadecadien-1-ol, (Z)-7-decenyl acetate, (Z,Z)-6,9-pentadecadienyl acetate, (E)-8-decen-1-ol, (Z,Z)-6,9-pentadecadienal, (E,E)-2,4-decadienal, (E,E)-8,10-pentadecadienyl acetate, (E,Z)-2,4-decadienal, (E,Z)-8,10-pentadecadien-1-ol, (Z,Z)-2,4-decadienal, (E,Z)-8,10-pentadecadienyl acetate,(E,E)-3,5-decadienyl acetate, (Z,E)-8,10-pentadecadienyl acetate, (Z,E)-3,5-decadienyl acetate, (Z,Z)-8,10-pentadecadienyl acetate, (Z,Z)-4,7-decadien-1-ol, (E,Z)-9,11-pentadecadienal, (Z,Z)-4,7-decadienyl acetate, (Z,Z)-9,11-pentadecadienal, (E)-2-undecenyl acetate, (Z)-3-hexadecenyl acetate, (E)-2-undecenal, (E)-5-hexadecene 1-ol, (Z)-5-undecenyl acetate, (E)-5-hexadecenyl acetate, (Z)-7-undecenyl acetate, (Z)-5-hexadecen-1-ol (Z)-8-undecenyl acetate, (Z)-5-hexadecenyl acetate, (Z)-9-undecenyl acetate, (E)-6-hexadecenyl acetate, (E)-2-dodecenal, (E)-7-hexadecen-1-ol, (Z)-3-dodecen-1-ol, (E)-7-hexadecenyl acetate, (E)-3-dodecenyl acetate, (E)-7-hexadecen nal, (Z)-3-dodecenyl acetate, (Z)-7-hexadecen-1-ol, (E)-4-dodecenyl acetate, (Z)-7-hexadecenyl acetate, (E)-5-dodecen-1-ol, (Z)-7-hexadecenal, (E)-5-dodecenyl acetate, (E)-8-hexadecenyl acetate, (Z)-5-dodecen-1-ol, (E)-9-hexadecen-1-ol, (Z)-5-dodecenyl acetate, (E)-9-hexadecenyl acetate, (Z)-5-dodecenal, (E)-9-hexadecenal, (E)- 6-dodecen-1-ol, (Z)-9-hexadecen-1-ol, (Z)-6-dodecenyl acetate, (Z)-9-hexadecenyl acetate, (E)-6-dodecenal, (Z)-9-hexadecenal, (E)-7-dodecen-1-ol, (E)-10-hexadecen-1-ol, (E)-7-dodecenyl acetate, (E)-10-hexadecenal, (E)-7-dodecenal, (Z)-10-hexadecenyl acetate, (Z)-7-dodecen-1-ol, (Z)-10-hexadecenal, (Z)-7-dodecenyl acetate,(E)-11-hexadecen-1-ol, (Z)-7-dodecenal, (E)-11-hexadecenyl acetate, (E)-8-dodecen-1-ol, (E)-11-hexadecenal, (E)-8-dodecenyl acetate, (Z)-11-hexadecen-1-ol, (E)-8-dodecenal, (Z)-11-hexadecenyl acetate, (Z)-8-dodecen-1-ol, (Z)-11-hexadecenal, (Z)-8-dodecenyl acetate, (Z)-12-hexadecenyl acetate, (E)-9-dodecen-1-ol, (Z)- 12-Hexadecenal, (E)-9-Dodecenyl acetate, (E)-14-Hexadecenal, (E)-9-Dodecenal, (Z)-14-Hexadecenyl acetate, (Z)-9-Dodecen-1-ol, (E,E)-1,3-Hexadecadien-1-ol, (Z)-9-Dodecenyl acetate, (E,Z)-4,6-Hexadecadien-1-ol, (Z)-9-Dodecenal (E,Z)-4,6-Hexadecadienyl acetate, (E)-10-Dodecen-1-ol, (E,Z)-4,6-Hexadecadienal, (E)-10-Dodecen Hexadecadienyl acetate, (E,Z)-6,11-hexadecadienyl acetate, (E)-10-dodecenal, (E,Z)-6,11-hexadecadienal, (Z)-10-dodecen-1-ol, (Z,Z)-7,10-hexadecadien-1-ol, (Z)-10-dodecenyl acetate, (Z,Z)-7,10-hexadecadienyl acetate, (E,Z)-3,5-dodecadienyl acetate, (Z,E)-7,11-hexadecadien-1-ol, (Z,E)-3,5-dodecadienyl acetate, (Z,E)-7,11-hexadecadien-1-ol dienyl acetate, (Z,Z)-3,6-dodecadien-1-ol, (Z,E)-7,11-hexadecadienal, (E,E)-4,10-dodecadienyl acetate, (Z,Z)-7,11-hexadecadien-1-ol, (E,E)-5,7-dodecadien-1-ol, (Z,Z)-7,11-hexadecadienyl acetate, (E,E)-5,7-dodecadienyl acetate, (Z,Z)-7,11-hexadecadienal, (E,Z)-5,7-dodecadien-1-ol, (Z,Z)-8,10-hexadecadienyl acetate,(E,Z)-5,7-dodecadienyl acetate, (E,Z)-8,11-hexadecadienal, (E,Z)-5,7-dodecadienal, (E,E)-9,11-hexadecadienal, (Z,E)-5,7-dodecadien-1-ol, (E,Z)-9,11-hexadecadienyl acetate, (Z,E)-5,7-dodecadienyl acetate, (E,Z)-9,11-hexadecadienal, (Z,E)-5,7-dodecadienal, (Z,E)-9,11-hexadecadienal, (Z,Z)-5,7-dodecadienyl acetate, ( (Z,Z)-9,11-hexadecadienal, (Z,Z)-5,7-dodecadienal, (E,E)-10,12-hexadecadien-1-ol, (E,E)-7,9-dodecadienyl acetate, (E,E)-10,12-hexadecadienyl acetate, (E,Z)-7,9-dodecadien-1-ol, (E,E)-10,12-hexadecadienal, (E,Z)-7,9-dodecadienyl acetate, (E,Z)-10,12-hexadecadien-1-ol, (E,Z)-7,9-dodecadienal, (E,Z)-10,12-hexadecadienal Hexadecadienyl acetate, (Z,E)-7,9-dodecadien-1-ol, (E,Z)-10,12-hexadecadienal, (Z,E)-7,9-dodecadienyl acetate, (Z,E)-10,12-hexadecadienyl acetate, (Z,Z)-7,9-dodecadien-1-ol, (Z,E)-10,12-hexadecadienal, (Z,Z)-7,9-dodecadienyl acetate, (Z,Z)-10,12-hexadecadienal, (E,E)-8,10-dodecadien-1-ol, (E,E)-11,13-hexadecadien -1-ol, (E,E)-8,10-dodecadienyl acetate, (E,E)-11,13-hexadecadienyl acetate, (E,E)-8,10-dodecadienal, (E,E)-11,13-hexadecadienal, (E,Z)-8,10-dodecadien-1-ol, (E,Z)-11,13-hexadecadien-1-ol, (E,Z)-8,10-dodecadienyl acetate, (E,Z)-11,13-hexadecadienyl acetate, (E,Z)-8,10-dodecadienal, (E,Z)-11,13-hexadecadienal,(Z,E)-8,10-dodecadien-1-ol, (Z,E)-11,13-hexadecadien-1-ol, (Z,E)-8,10-dodecadienyl acetate, (Z,E)-11,13-hexadecadienyl acetate, (Z,E)-8,10-dodecadienal, (Z,E)-11,13-hexadecadienal, (Z,Z)-8,10-dodecadien-1-ol, (Z,Z)-11,13-hexadecadienal Sadecadien-1-ol, (Z,Z)-8,10-dodecadienyl acetate, (Z,Z)-11,13-hexadecadienyl acetate, (Z,E,E)-3,6,8-dodecatrien-1-ol, (Z,Z)-11,13-hexadecadienal, (Z,Z,E)-3,6,8-dodecatrien-1-ol, (E,E)-10,14-hexadecadienal, (E)-2-tridecenyl acetate, (Z,E)-11,14-hexadecadienyl acetate, (Z)-2-tridecenyl acetate, (E,E,Z)-4,6,10-hexadecatrien-1-ol, (E)-3-tridecenyl acetate, (E,E,Z)-4,6,10-hexadecatrien-1-ol, (E)-4-tridecenyl acetate, (E,Z,Z)-4,6,10-hexadecatrien-1-ol, (Z ...al (E,E,Z)-4 ,6,11-Hexadecatrienyl acetate, (E)-6-tridecenyl acetate (E,E,Z)-4,6,11-hexadecatrienal (Z)-7-tridecenyl acetate (Z,Z,E)-7,11,13-hexadecatrienal (E)-8-tridecenyl acetate, (E,E,E)-10,12,14-hexadecatrienyl acetate, (Z)-8-tridecenyl acetate, (E,E,E)-10,12,14-hexadecatrienal, (E)-9-tridecenyl acetate, (E,E,Z)-10,12,14-hexadecatrienal trienyl acetate, (Z)-9-tridecenyl acetate, (E,E,Z)-10,12,14-hexadecatrienal, (Z)-10-tridecenyl acetate, (E,E,Z,Z)-4,6,11,13-hexadecatetraenal, (E)-11-tridecenyl acetate, (E)-2-heptadecenal, (Z)-11-tridecenyl acetate, (Z)-2-heptadecenal, (E,Z)-4,7-tridecadienyl acetate, (E)-8-heptadecen-1-ol, (Z,Z)-4,7-tridecadien-1-ol, (E)-8-heptadecenyl acetate, (Z,Z)-4,7-tridecadienyl acetate, (Z)-8-heptadecen-1-ol, (E,Z)-5,9-tridecadienyl acetate, (Z)-9-heptadecenal, (Z,E)-5,9-tridecadienyl acetate, (E)-10-heptadecenyl acetate, (Z,Z)-5,9-tridecadienyl acetate, (Z)-11-heptadecen-1-ol, (Z,Z)-7,11-tridecadienyl acetate, (Z)-11-heptadecenyl acetate, (E,Z,Z)-4,7,10-Tridecatrienyl acetate, (E,E)-4,8-heptadecadienyl acetate, (E)-3-tetradecen-1-ol, (Z,Z)-8,10-heptadecadien-1-ol, (E)-3-tetradecenyl acetate, (Z,Z)-8,11-heptadecadienyl acetate, (Z)-3-tetradecen-1-ol, (E)-2-octadecenyl acetate, (Z)-3-tetradecenyl acetate, (E)-2-octadecenal, (E)-5-tetradecen-1-ol, (Z)-2-octadecenyl acetate, (E)-5- Tetradecenyl acetate, (Z)-2-octadecenal, (E)-5-tetradecenal, (E)-9-octadecen-1-ol, (Z)-5-tetradecen-1-ol, (E)-9-octadecenyl acetate, (Z)-5-tetradecenyl acetate, (E)-9-octadecenal, (Z)-5-tetradecenal, (Z)-9-octadecen-1-ol, (E)-6-tetradecenyl acetate, (Z)-9-octadecenyl acetate, (Z)-6-tetradecenyl acetate, (Z)-9-octadecenal, (E)-7-tetradecenyl acetate octadecen-1-ol, (E)-11-octadecen-1-ol, (E)-7-tetradecenyl acetate, (E)-11-octadecenal, (Z)-7-tetradecen-1-ol, (Z)-11-octadecen-1-ol, (Z)-7-tetradecenyl acetate, (Z)-11-octadecenyl acetate, (Z)-7-tetradecenal, (Z)-11-octadecenal, (E)-8-tetradecenyl acetate, (E)-13-octadecenyl acetate, (Z)-8-tetradecen-1-ol, (E)-13-octadecenal, (Z )-8-tetradecenyl acetate, (Z)-13-octadecen-1-ol, (Z)-8-tetradecenal, (Z)-13-octadecenyl acetate, (E)-9-tetradecen-1-ol, (Z)-13-octadecenal, (E)-9-tetradecenyl acetate, (E)-14-octadecenal, (Z)-9-tetradecen-1-ol, (E,Z)-2,13-octadecadien-1-ol, (Z)-9-tetradecenyl acetate, (E,Z)-2,13-octadecadienyl acetate, (Z)-9-tetradecenal, (E,Z)-2,13-octadecadienal, (E)-10-tetradecenyl acetate, (Z,E)-2,13-octadecadienyl acetate, (Z)-10-tetradecenyl acetate, (Z,Z)-2,13-octadecadien-1-ol, (E)-11-tetradecen-1-ol, (Z,Z)-2,13-octadecadienyl acetate, (E)-11-tetradecenyl acetate, (E,E)-3,13-octadecadienyl acetate, (E)-11-tetradecenal, (E,Z)-3,13-octadecadienyl acetate, (Z)-1 1-Tetradecen-1-ol, (E,Z)-3,13-octadecadienal, (Z)-11-tetradecenyl acetate, (Z,E)-3,13-octadecadienyl acetate, (Z)-11-tetradecenal, (Z,Z)-3,13-octadecadienyl acetate, (E)-12-tetradecenyl acetate, (Z,Z)-3,13-octadecadienal, (Z)-12-tetradecenyl acetate, (E,E)-5,9-octadecadien-1-ol, (E,E)-2,4-tetradecadienal, (E,E)-5,9-octadecadien nyl acetate, (E,E)-3,5-tetradecadienyl acetate, (E,E)-9,12-octadecadien-1-ol, (E,Z)-3,5-tetradecadienyl acetate, (Z,Z)-9,12-octadecadienyl acetate, (Z,E)-3,5-tetradecadienyl acetate, (Z,Z)-9,12-octadecadienal, (E,Z)-3,7-tetradecadienyl acetate, (Z,Z)-11,13-octadecadienal, (E,Z)-3,8-tetradecadienyl acetate, (E,E)-11,14-octadecadienal ol, (E,Z)-4,9-tetradecadienyl acetate, (Z,Z)-13,15-octadecadienal, (E,Z)-4,9-tetradecadienal, (Z,Z,Z)-3,6,9-octadecatrienyl acetate, (E,Z)-4,10-tetradecadienyl acetate, (E,E,E)-9,12,15-octadecatrien-1-ol, (E,E)-5,8-tetradecadienal, (Z,Z,Z)-9,12,15-octadecatrienyl acetate, (Z,Z)-5,8-tetradecadien-1-ol, (Z,Z,Z)-9,12,15-Octadecatrienal, (Z,Z)-5,8-Tetradecadienyl acetate, (Z,Z)-5,8-Tetradecadienal, (E,E)-8,10-Tetradecadien-1-ol, (E,E)-8,10-Tetradecadienyl acetate, (E,E)-8,10-Tetradecadienal, (E,Z)-8,10-Tetradecadienyl ol acetate, (E,Z)-8,10-tetradecadienal, (Z,E)-8,10-tetradecadien-1-ol, (Z,E)-8,10-tetradecadienyl acetate, (Z,Z)-8,10-tetradecadienal, (E,E)-9,11-tetradecadienyl acetate, (E,Z)-9,11-tetradecadienyl acetate , (Z,E)-9,11-tetradecadien-1-ol, (Z,E)-9,11-tetradecadienyl acetate, (Z,E)-9,11-tetradecadienal, (Z,Z)-9,11-tetradecadien-1-ol, (Z,Z)-9,11-tetradecadienyl acetate, (Z,Z)-9,11-tetradecadienal, (E,E)-9,12-tetradecadienyl acetate, (Z,E)-9,12-tetradecadien-1-ol, (Z,E)-9,12-tetradecadienyl acetate, (Z,E)-9,12-tetradecadienal, (Z,Z)-9,12-tetradecadien-1-ol and (Z,Z)-9,12-tetradecadienyl acetate. In some embodiments, the volatile material is an insect repellent. In some embodiments, the volatility of the pheromone is reduced by encapsulation of the pheromone in any of the NLP compositions disclosed herein. In some embodiments, NLP encapsulation of any of the pheromones listed herein extends their effectiveness through sustained release of the pheromone.

[0137] 2.4.Essential oil Exemplary essential oils that may be encapsulated in any of the NLPs by any of the methods described herein include, but are not limited to, cinnamon, cedar, castor, clove, geranium, lemongrass, mint, thyme, turmeric, wintergreen, rosemary, anise, cardamom, chamomile, coriander, cumin, dill, mint, parsley, lavender, basil, camphor, citronella, eucalyptus, fennel, ginger, grapefruit, lemon, mandarin, orange, pine needle, pepper, rose, sweet orange, tangerine, tea tree, tea seed, caraway, garlic, peppermint, onion, and spearmint oil. In some embodiments, the essential oil is a volatile oil. In some embodiments, the volatility of the essential oil is reduced by encapsulating the essential oil in any of the NLP compositions disclosed herein. In some embodiments, NLP encapsulation of any of the essential oils described herein extends their effectiveness through sustained release of the essential oil.

[0138] 2.5. Combinations of Volatile Bioactive Substances In some embodiments, the NLP compositions provided herein comprise a mixture of NLP compositions each comprising a different heterofunctional agent (e.g., two different bioactive substances). In some embodiments, the stability of an NLP composition comprising a first functional agent (e.g., deltamethrin) differs from the stability of an NLP composition comprising a second functional agent (e.g., a herbicide). In some embodiments, one or more of the NLP compositions comprises a volatile functional agent (e.g., tefluthrin). In some embodiments, an NLP comprises two or more bioactive substances. In some embodiments, at least one of the bioactive substances is a volatile bioactive substance.

[0139] g) Signs To aid in analysis and characterization, the NLP compositions may include a detectable label to monitor the mobility of the NLP compositions in the soil, to assess the affinity of the NLP compositions for any components in the soil, cellular uptake, etc. In some embodiments, the label is a fluorescent protein (e.g., green fluorescent protein). In some embodiments, the label is a protein or polynucleic acid conjugated to a fluorophore. In some embodiments, the NLP compositions may include a dye (e.g., a fluorescent dye). In some embodiments, the dye may be added to the organic or aqueous phase during production of the NLP compositions, depending on the chemical properties of the dye. In some embodiments, the NLP compositions may include a fluorescent dye such as 3,3'-dihexyloxacarbocyanine iodide (DIOC6), a fluorescent lipophilic dye, PKH67 (Sigma Aldrich), Alexa Fluor® 488 (Thermo Fisher Scientific), DyLight™ 800 (Thermo Fisher), Exalite 594, Nile Red, Exalite 428, Coumarin 481, Coumarin 486, DiD' solid, DiIC18(5) solid (1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate), DiO™ solid, DiOΔ9,12-C18(3), ClO4 (3,3'-dilinoleyloxacarbocyanine perchlorate), DiI™ oil, DiIΔ9,12-C18(3), ClO4 (1,1'-dilinoleyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), DiI™ solid, DiIΔ9,12-C 18(3), CBS (1,1'-dilinoleyl-3,3,3',3'-tetramethylindocarbocyanine, 4-chlorobenzenesulfonate), DiIC12(3) (1,1'-didodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), DiIC16(3) (1,1'-dihexadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), and DiR', DiIC18(7) (1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbocyanine iodide).In some embodiments, Exalyte 594 is added to the organic phase during production of the NLP composition. Some embodiments relate to the use of a label to quantitate total membrane content, which can be used to indirectly measure the concentration of NLPs. Some embodiments relate to the use of a label (e.g., a fluorescent marker) to detect cellular uptake of the NLP composition.

[0140] Furthermore, the production methods described herein can be supplemented with any quantitative or qualitative method known in the art to characterize or identify NLPs at any step in the production process. NLPs can be characterized by various analytical methods to estimate NLP yield, NLP concentration, NLP purity, NLP composition, or NLP size. NLPs can be evaluated by several methods known in the art that allow visualization, quantitative, or qualitative characterization (e.g., composition identification) of NLPs, such as microscopy (e.g., transmission electron microscopy), dynamic light scattering, nanoparticle tracking, spectroscopy (e.g., Fourier transform infrared analysis), or mass spectrometry (protein and lipid analysis). In some examples, methods (e.g., mass spectrometry) can be used to identify plant EV markers present on the NLPs, such as the plant EV markers disclosed in WO2021041301A1. To aid in the analysis and characterization of NLP fractions, NLPs can be further labeled or stained. For example, NLPs can be stained with 3,3'-dihexyloxacarbocyanine iodide (DIOC6), a fluorescent lipophilic dye, PKH67 (Sigma Aldrich), Alexa Fluor® 488 (Thermo Fisher Scientific), or DyLight™ 800 (Thermo Fisher). In the absence of advanced forms of nanoparticle tracking, this relatively simple technique can be used to quantify total membrane content and indirectly measure the concentration of NLPs (Rutter and Innes, Plant Physiol. 173(1):728-741, 2017; Rutter et al., Bio. Protoc. 7(17):e2533, 2017). For more precise measurements and to assess the size distribution of NLPs, nanoparticle tracking can be used.

[0141] B.NLP size In some embodiments, the NLPs described herein have a diameter of about 5-50 nm, about 50-100 nm, about 100-150 nm, about 150-200 nm, about 200-250 nm, about 250-300 nm, about 300-350 nm, about 350-400 nm, about 400-450 nm, about 450-500 nm, about 500-550 nm, about 550-600 nm. m, about 600 to 650 nm, about 650 to 700 nm, about 700 to 750 nm, about 750 to 800 nm, about 800 to 850 nm, about 850 to 900 nm, about 900 to 950 nm, about 950 to 1000 nm, about 1000 to 1250 nm, about 1250 to 1500 nm, about 1500 to 1750 nm, or about 1750 to 2000 nm. In some embodiments, the NLPs described herein have an average diameter of about 5-950 nm, about 5-900 nm, about 5-850 nm, about 5-800 nm, about 5-750 nm, about 5-700 nm, about 5-650 nm, about 5-600 nm, about 5-550 nm, about 5-500 nm, about 5-450 nm, about 5-400 nm, about 5-350 nm, about 5-300 nm, about 5-250 nm, about 5-200 nm, about 5-150 nm, about 5-100 nm, about 5-50 nm, or about 5-25 nm. In some embodiments, the NLPs described herein have an average diameter of about 50-200 nm. In some embodiments, the NLPs described herein have an average diameter of about 50-300 nm. In some embodiments, the NLPs described herein have an average diameter of about 200-500 nm. In some embodiments, the NLPs described herein have an average diameter of about 30-150 nm, hi some embodiments, the NLPs described herein have an average diameter of at least 5 nm, at least 50 nm, at least 100 nm, at least 150 nm, at least 200 nm, at least 250 nm, at least 300 nm, at least 350 nm, at least 400 nm, at least 450 nm, at least 500 nm, at least 550 nm, at least 600 nm, at least 650 nm, at least 700 nm, at least 750 nm, at least 800 nm, at least 850 nm, at least 900 nm, at least 950 nm, or at least 1000 nm.In some embodiments, the NLPs described herein have an average diameter of less than 1000 nm, less than 950 nm, less than 900 nm, less than 850 nm, less than 800 nm, less than 750 nm, less than 700 nm, less than 650 nm, less than 600 nm, less than 550 nm, less than 500 nm, less than 450 nm, less than 400 nm, less than 350 nm, less than 300 nm, less than 250 nm, less than 200 nm, less than 150 nm, less than 100 nm, or less than 50 nm. Various methods standard in the art (e.g., dynamic light scattering) can be used to measure the particle size of NLPs.

[0142] In some embodiments, the NLP is 77 nm 2 ~3.2×10 6 nm 2 (e.g., 77-100 nm 2 , 100~1000nm 2 , 1000 to 1 × 10 4 nm 2 , 1×10 4 ~1×10 5 nm 2 , 1×10 5 ~1×10 6 nm 2 or 1 x 10 6 ~3.2×10 6 nm 2 In some embodiments, the NLPs have an average surface area of ​​65 nm 3 ~5.3×10 8 nm 3 (e.g., 65 to 100 nm 3 , 100~1000nm 3 , 1000 to 1 × 10 4 nm 3 , 1×10 4 ~1×10 5 nm 3 , 1×10 5 ~1×10 6 nm 3 , 1×10 6 ~1×10 7 nm 3 , 1×10 7 ~1×10 8 nm 3 , 1×10 8 ~5.3×108 nm 3 In some embodiments, the NLPs have an average volume of at least 77 nm 2 , (e.g., at least 77 nm 2 , at least 100 nm 2 , at least 1000 nm 2 , at least 1 x 10 4 nm 2 , at least 1 x 10 5 nm 2 , at least 1 x 10 6 nm 2 or at least 2 x 10 6 nm 2 In some embodiments, the NLPs have an average surface area of ​​at least 65 nm 3 (e.g., at least 65 nm 3 , at least 100 nm 3 , at least 1000 nm 3 , at least 1 x 10 4 nm 3 , at least 1 x 10 5 nm 3 , at least 1 x 10 6 nm 3 , at least 1 x 10 7 nm 3 , at least 1 x 10 8 nm 3 , at least 2 × 10 8 nm 3 , at least 3 × 10 8 nm 3 , at least 4 × 10 8 nm 3 or at least 5 × 108 nm 3 The plant EVs or segments, parts or extracts thereof may comprise plant EVs having an average volume of

[0143] In some embodiments, the size of the NLPs can be determined after loading with one or more heterofunctional agents or other modifications to the NLPs. In some embodiments, NLPs containing one or more heterofunctional agents can be sized to within 77 nm. 2 ~1.3×10 7 nm 2 (e.g., 77-100 nm2 , 100~1000nm 2 , 1000 to 1 × 10 4 nm 2 , 1×10 4 ~1×10 5 nm 2 , 1×10 5 ~1×10 6 nm 2 or 1 x 10 6 ~1.3×10 7 nm 2 ) average surface area.

[0144] In some embodiments, the NLPs comprising heterologous functional agents are 65 nm 3 ~4.2×10 9 nm 3 (e.g., 65 to 100 nm 3 , 100~1000nm 3 , 1000 to 1 × 10 4 nm 3 , 1×10 4 ~1×10 5 nm 3 , 1×10 5 ~1×10 6 nm 3 , 1×10 6 ~1×10 7 nm 3 , 1×10 7 ~1×10 8 nm 3 , 1×10 8 ~1×10 9 nm 3 or 1 x 10 9 ~4.2×10 9 nm 3 In some embodiments, the NLPs may have an average volume of at least 77 nm 2 , (e.g., at least 77 nm 2 , at least 100 nm 2 , at least 1000 nm 2 , at least 1 x 10 4 nm 2 , at least 1 x 10 5 nm 2 , at least 1 x 10 6 nm 2or at least 1 × 10 7 nm 2 In some embodiments, the NLPs have an average surface area of ​​at least 65 nm 3 (e.g., at least 65 nm 3 , at least 100 nm 3 , at least 1000 nm 3 , at least 1 x 10 4 nm 3 , at least 1 x 10 5 nm 3 , at least 1 x 10 6 nm 3 , at least 1 x 10 7 nm 3 , at least 1 x 10 8 nm 3 , at least 1 x 10 9 nm 3 , at least 2 × 10 9 nm 3 , at least 3 × 10 9 nm 3 Or at least 4 x 10 9 nm 3 ) has an average volume of

[0145] C. Generation method In some embodiments, the NLP compositions described herein can be produced by one of the four methods described in Examples 1 and 2. Additional methods of preparing NLPs are available to those of skill in the art. In some embodiments, the NLP compositions described herein can be prepared by mixing an organic phase containing at least one nonpolar lipid and at least one phospholipid with an aqueous phase. In some embodiments, one or more surface modifiers are added to either the organic or aqueous phase, depending on the chemical nature of the surface modifier. In some embodiments, a hydrophobic agent, rhamnolipid, is added to the organic phase. In some embodiments, a polycarboxylate, Atlox 500L, is added to the aqueous phase. In some embodiments, a co-solvent (e.g., DCM, IPM, etc.) is added to the organic phase. In some embodiments, a co-solvent is added during production of the NLP composition to increase the solubility of the organic compound (e.g., deltamethrin). In some embodiments, the co-solvent is evaporated in the final step of NLP production. In some embodiments, the NLP composition contains trace amounts of a co-solvent. In some embodiments, excipients are added during production of the NLP composition to increase the stability of the NLPs, hi some embodiments, one or more excipients are added to either the organic phase or the aqueous phase during production of the NLP composition, depending on the chemical properties of the excipient.

[0146] D. Medication filling Some embodiments relate to NLP compositions comprising one or more heterofunctional agents (e.g., a cell-penetrating agent, an agricultural agent (e.g., a pesticide, a fertilizer, a herbicide, a plant modifier, a plant growth promoter, a biostimulant, or a plant immunity inducer), a therapeutic agent (e.g., an antifungal agent, an antioomycete, an antibacterial agent, a virucide, an antiviral agent, an insecticide, a nematicide, an antiparasitic agent, an insect repellent), etc.). The NLPs described herein may possess or be associated with one or more heterofunctional agents by a variety of means, such as encapsulating the heterofunctional agent, incorporating the heterofunctional agent into a lipid layer (e.g., a lipid bilayer), or associating (e.g., by conjugation) the one or more heterofunctional agents with the surface of the lipid layer of the NLP. In some embodiments, one or more heterofunctional agents (e.g., a cell-penetrating agent, a pesticide, etc.) are included in the NLP composition as described in Section IB herein. In some embodiments, the one or more heterologous functional agents are stably associated with the NLP composition before and after delivery, e.g., to the soil, plant roots, pests, etc. In some embodiments, the one or more heterologous functional agents dissociate (e.g., are released) from the NLP composition after delivery, e.g., to the soil, plant roots, pests, etc.

[0147] In some embodiments, one or more heterofunctional agents are incorporated into the NLPs during their formation using a microfluidic device. Some embodiments involve incorporating one or more heterofunctional agents into the NLPs by providing one or more phospholipids and oils in an organic phase and one or more surface modifiers and heterofunctional agents in an aqueous phase, where the organic and aqueous phases are combined (e.g., in a microfluidic device) to produce an NLP composition comprising the heterofunctional agents. Some embodiments involve incorporating one or more heterofunctional agents into the NLPs by providing one or more phospholipids, oils, and surface modifiers in an organic phase and one or more heterofunctional agents in an aqueous phase, where the organic and aqueous phases are combined (e.g., in a microfluidic device) to produce an NLP composition comprising the heterofunctional agents. Some embodiments relate to incorporating one or more heterofunctional agents into NLPs by providing one or more phospholipids, oils, and heterofunctional agents in an organic phase and one or more surface modifiers in an aqueous phase, where the organic and aqueous phases are combined (e.g., in a microfluidic device) to produce NLPs containing the heterofunctional agents.

[0148] Some embodiments relate to incorporating one or more heterologous functional agents into NLPs by loading the one or more heterologous functional agents into preformed NLPs by any method known in the art that allows for direct or indirect association between the NLP and the one or more heterologous functional agents. Heterologous functional agents may be loaded onto or into the NLP (e.g., encapsulated by the NLP) using, but not limited to, physical, chemical, and / or biological methods. In some embodiments, one or more heterologous functional agents may be introduced into the NLP by one or more of electroporation, sonication, passive diffusion, agitation, lipid extraction, and extrusion. However, it should be understood by those skilled in the art that loading of a substance of interest into an NLP is not limited to the methods set forth above. Loaded NLPs can be evaluated to confirm the presence or level of loaded agents using a variety of methods, such as HPLC (e.g., to evaluate small molecules), immunoblotting (e.g., to evaluate proteins), and / or quantitative PCR (e.g., to evaluate nucleotides).

[0149] Some embodiments relate to NLP compositions comprising one or more heterofunctional agents conjugated to an NLP. In some embodiments, the one or more heterofunctional agents are indirectly linked or bound to the NLP. In some embodiments, the one or more heterofunctional agents are directly linked or bound to the NLP. In some embodiments, the one or more heterofunctional agents are chemically bound to the NLP. In some embodiments, the one or more heterofunctional agents are directly bound (e.g., by covalent or ionic bonds) to the lipid structure (e.g., lipid bilayer) of the NLP.

[0150] Some embodiments relate to methods of conjugating one or more heterofunctional agents to an NLP, the method comprising incubating the one or more heterofunctional agents with a suitable cross-linking agent (e.g., N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC). Without being bound by theory, EDC may be used as a carboxyl activating agent for amide bonds with primary amines and reacts with phosphate groups in a suitable solvent for a period of time sufficient to couple the one or more heterofunctional agents to the cross-linking agent and incubate the one or more heterofunctional agents coupled to the cross-linking agent with the NLP composition). In some embodiments, a mixture of the one or more heterofunctional agents coupled to the cross-linking agent and the NLP composition is provided on a sucrose gradient (e.g., an 8, 30, 45, or 60% sucrose gradient) and centrifuged to separate the one or more free heterofunctional agents, the free NLP composition, and the heterofunctional agents conjugated to the NLP composition. In some embodiments, the NLP composition conjugated to a heterologous functional agent is collected, washed, and dissolved in a solution suitable for use as described herein.

[0151] In some embodiments, compositions comprising NLPs are formulated or one or more NLP compositions are loaded to provide compositions comprising NLPs with varying concentrations of one or more heterofunctional agents depending on the particular drug or application. In some embodiments, compositions comprising NLPs are formulated or one or more NLP compositions are loaded such that compositions comprising NLPs disclosed herein contain about 0.001, 0.01, 0.1, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 95 (or any range between about 0.001 and 95) or more weight percent of one or more heterofunctional agents. In some embodiments, the NLP composition is loaded or formulated such that the NLP composition comprises about 95, 90, 80, 70, 60, 50, 40, 30, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1.0, 0.1, 0.01, 0.001 (or any range between about 95 and 0.001) or less of one or more heterofunctional agents. In some embodiments, the NLP composition may comprise about 0.001 to about 0.01 wt%, about 0.01 to about 0.1 wt%, about 0.1 to about 1 wt%, about 1 to about 5 wt%, or about 5 to about 10 wt%, or about 10 to about 20 wt% of one or more heterofunctional agents. In some embodiments, the NLP compositions may be loaded or formulated with about 1, 5, 10, 50, 100, 200, 500, 1,000, 2,000 (or any range from about 1 to 2,000) or more pg / ml of one or more heterologous functional agents. In some embodiments, the NLP compositions may be loaded or formulated with about 2,000, 1,000, 500, 200, 100, 50, 10, 5, 1 (or any range from about 2,000 to 1) or more pg / ml of one or more heterologous functional agents.

[0152] In some embodiments, the NLP composition is formulated or loaded such that the NLP composition comprises at least 0.001%, at least 0.01%, at least 0.1%, at least 1.0%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% by weight of one or more heterologous functional agents. In some embodiments, the NLP composition may be loaded or formulated with at least 1 pg / ml, at least 5 pg / ml, at least 10 pg / ml, at least 50 pg / ml, at least 100 pg / ml, at least 200 pg / ml, at least 500 pg / ml, at least 1,000 pg / ml, or at least 2,000 pg / ml of one or more heterologous functional agents.

[0153] In some embodiments, the NLP compositions are formulated with one or more heterofunctional agents by suspending (e.g., by vigorously mixing) the NLP compositions in a solution comprising or consisting essentially of the one or more heterofunctional agents. In some embodiments, the one or more heterofunctional agents (e.g., antifungals, antioomycetes, antibacterial agents, insecticides, molluscicides, nematicides, herbicides, virucides, peptides, polypeptides, nucleic acids, polynucleotides, etc.) may comprise less than 1% or at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the solution in which the one or more NLP compositions are suspended.

[0154] E. Generation of NLP Compositions Using Microfluidics In some embodiments, the NLP compositions are produced by a process involving microfluidics. In some embodiments, the NLP compositions are produced by mixing a lipid solution and an aqueous phase in any suitable ratio in a microfluidic device. In some embodiments, the NLP compositions are produced by mixing a lipid solution and an aqueous phase in a 1:3 volumetric ratio in a microfluidic device. In some embodiments, the NLP compositions are produced by mixing a lipid solution and an aqueous phase in a 1:1, 1:2, 1:3, 1:4, or 1:5 volumetric ratio in a microfluidic device.

[0155] In some embodiments, one or more lipids included in the NLP composition are extracted from multiple lipid sources (e.g., lipids are extracted using the Bligh-Dyer method (Bligh and Dyer, J Biolchem ​​Physiol, 37:911-917, 1959)). In some embodiments, one or more lipids included in the NLP composition are extracted from a plant source (e.g., soybean, citrus fruits (e.g., lemon, orange, grapefruit, etc.), avocado, tomato, corn, etc.). In some embodiments, one or more extracted lipids may be provided as a stock solution (e.g., a solution in chloroform-methanol). In some embodiments, one or more extracted lipids are processed to produce a lipid film. In some embodiments, the lipid film is produced by evaporation of a solvent using a stream of inert gas (e.g., nitrogen). In some embodiments, the lipid phase used during NLP production may comprise one or more phospholipids. In some embodiments, the lipid phase used during NLP production may comprise one or more non-polar lipids. In some embodiments, the lipid phase used during NLP production may comprise one or more phospholipids and one or more non-polar lipids. In some embodiments, the lipid phase used during NLP production may comprise one or more hydrophobic heterofunctional agents. In some embodiments, the lipid phase used during NLP production may be selected from the group consisting of antifungals, antibacterials, insecticides, molluscicides, nematicides, herbicides, virucides, peptides, polypeptides, nucleic acids, and polynucleotides, or any combination thereof. In some embodiments, the lipid phase used during NLP production may comprise one or more proteins. In some embodiments, the lipid phase used during NLP production may comprise one or more ribonucleoproteins. In some embodiments, the lipid phase used during NLP production may comprise one or more surface modifiers. In some embodiments, the lipid phase used during NLP production may comprise one or more co-solvents. In some embodiments, the lipid phase used during NLP production may comprise one or more excipients.

[0156] In some embodiments, the aqueous phase used during NLP production may be a citrate buffer (e.g., a citrate buffer having a pH of about 3.2). In some embodiments, the aqueous phase used during NLP production may be deionized water. In some embodiments, the aqueous phase used during NLP production may be phosphate buffered saline (PBS). In some embodiments, the aqueous phase used during NLP production may comprise one or more hydrophilic heterofunctional agents. In some embodiments, the aqueous phase used during NLP production may comprise one or more hydrophilic heterofunctional agents selected from the group consisting of antifungals, antibacterial agents, insecticides, molluscicides, nematicides, herbicides, virucides, peptides, polypeptides, nucleic acids, and polynucleotides, or any combination thereof. In some embodiments, the aqueous phase used during NLP production may comprise one or more proteins. In some embodiments, the aqueous phase used during NLP production may comprise one or more ribonucleoproteins. In some embodiments, the aqueous phase used during NLP production may comprise one or more nucleic acids. In some embodiments, the aqueous phase used during NLP production may comprise one or more cationic molecules. In some embodiments, the aqueous phase used during NLP production may include one or more surface modifiers. In some embodiments, the aqueous phase used during NLP production may include one or more co-solvents. In some embodiments, the aqueous phase used during NLP production may include one or more excipients.

[0157] In some embodiments, the NLP composition may comprise at least one phospholipid. In some embodiments, the NLP composition may comprise one or more phospholipids selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid, phosphatidylserine, and 1,2-dimyristoyl-sn-glycero-3-phosphate, or any combination thereof. In some embodiments, the NLP composition may comprise at least one phospholipid selected from the group consisting of soybean lecithin and sunflower lecithin. In some embodiments, the NLP composition may comprise at least one nonpolar lipid. In some embodiments, the NLP composition may comprise at least one nonpolar lipid comprising 40% of at least one fatty acid chain selected from the group consisting of polyunsaturated fatty acids, monounsaturated fatty acids, and saturated fatty acids, or any combination thereof. In some embodiments, the NLP composition may comprise one or more phospholipids and one or more nonpolar lipids. In some embodiments, the NLP composition may comprise at least 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more than 90% phospholipids (w / w) of the total lipids in the NLP composition. In some embodiments, the NLP composition may comprise at least 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more than 90% non-polar lipids (w / w) of the total lipids in the NLP composition. In some embodiments, an NLP composition may comprise at least 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than 90% phospholipids and non-polar lipids (w / w) of the total lipids in the NLP composition. In some embodiments, an NLP composition may comprise one or more non-polar lipids in an amount of 25%-40% (w / w) of the total lipids in the preparation.

[0158] In some embodiments, the NLP composition comprises one or more surface modifiers. In some embodiments, the NLP composition comprises one or more surface modifiers selected from the group consisting of PEGylated moieties, PEGylated block copolymers (e.g., as poloxamers), cocamide derivatives, glycolipids, polysaccharides and polysaccharides with lipid chains, or any combination thereof. In some embodiments, the one or more surface modifiers may account for about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more than 90% (w / w) of the total lipids and sterols in the NLP composition.

[0159] In some embodiments, the NLP composition comprises one or more co-solvents. In some embodiments, the co-solvent is an organic solvent. In some embodiments, the co-solvent is DCM. In some embodiments, the co-solvent is IPM. In some embodiments, trace amounts of the co-solvent remain in the NLP composition after evaporation of the co-solvent. In some embodiments, the co-solvent comprises 1-10% (w / w) of the total weight of the NLP. In some examples, the one or more co-solvents provided in the NLP composition are dimethylformamide:methanol (DMF:MeOH). In some embodiments, the NLP composition comprises one or more co-solvents selected from the group consisting of acetonitrile, acetone, ethanol, methanol, dimethylformamide, tetrahydrofuran, 1-butanol, dimethyl sulfoxide, acetonitrile:ethanol, acetonitrile:methanol, acetone:methanol, methyl tert-butylethenepropanol, tetrahydrofuranmethanol, dimethyl sulfoxide:methanol, and dimethylformamide:methanol, or any combination thereof.

[0160] In some embodiments, the NLP composition may include one or more excipients in the organic phase. In some embodiments, one or more excipients may be included in the NLP composition to stabilize the NLP composition.

[0161] F. Zeta potential An NLP composition comprising a phospholipid, a non-polar lipid, and a surface modifier can have a zeta potential of, for example, less than 0 mV, less than -5 mV, less than -10 mV, less than -20 mV, less than -30 mV, less than -40 mV, less than -50 mV, less than -60 mV, less than -70 mV, less than -80 mV, less than -90 mV, or less than -100 mV in the absence of cargo. In some embodiments, an NLP composition comprising a phospholipid, a non-polar lipid, and a surface modifier can have a zeta potential of, for example, less than 0 mV, less than -5 mV, less than -10 mV, less than -20 mV, less than -30 mV, less than -40 mV, less than -50 mV, less than -60 mV, less than -70 mV, less than -80 mV, less than -90 mV, or less than -100 mV in the presence of cargo. In some embodiments, the zeta potential of an NLP comprising a phospholipid, a non-polar lipid, a surface modifier, and a cargo (e.g., a heterofunctional agent) is in the range of -10 mV to -60 mV, -20 mV to -50 mV, or -30 mV to -50 mV.

[0162] The zeta potential of an NLP composition can be measured using any method known in the art. Zeta potential is generally measured indirectly and calculated using a theoretical model from data obtained using methods and techniques known in the art, such as electrophoretic mobility or dynamic electrophoretic mobility. Electrophoretic mobility is typically measured using microelectrophoresis, electrophoretic light scattering, or tunable resistive pulse sensing. Electrophoretic light scattering is based on dynamic light scattering. Typically, zeta potential can be accessed from photon correlation spectroscopy or dynamic light scattering (DLS), also known as quasi-elastic light scattering.

[0163] G. Preparation i. Agricultural formulations In some embodiments, the NLP compositions described herein may be formulated with other materials to allow for ease of application, handling, transportation, storage, effective activity, etc. In some embodiments, the NLP compositions may be formulated in, for example, baits, concentrated emulsions, dusts, emulsifiable concentrates, fumigants, gels, granules, microencapsulated formulations, seed treatments, suspension concentrates, suspoemulsions, tablets, water-soluble liquids, water-dispersible granules or dry flowables, wettable powders, and concentrated low-volume solutions. In some embodiments, the NLP compositions described herein may be formulated as the formulation types described in "Catalogue of Pesticide Formulation Types and International Coding System," Technical Monograph no. 2, 5th Edition by CropLife International (2002), which is incorporated herein in its entirety.

[0164] In some embodiments, the NLP compositions described herein may be formulated as aqueous suspensions or emulsions. In some embodiments, the NLP compositions described herein may be formulated as aqueous suspensions or emulsions prepared from concentrated formulations. In some embodiments, the concentrated NLP formulations may be water-soluble, water-suspendable, or emulsifiable. In some embodiments, the concentrated NLP formulations may be solids, such as wettable powders or water-dispersible granules, or liquids, such as emulsifiable concentrates or aqueous suspensions. In some embodiments, the concentrated NLP formulations may be wettable powders, which may be compressed to form hydratable granules comprising a homogeneous mixture of one or more NLP compositions, one or more carriers, and, optionally, one or more surfactants. In some embodiments, the NLP compositions described herein may be formulated with one or more carriers selected from the group consisting of attapulgite clay, montmorillonite clay, diatomaceous earth, and purified silicates, or any combination thereof. In some embodiments, the NLP compositions described herein may be formulated with one or more surfactants, including about 0.5% to about 10% of a formulation (e.g., wettable powder) comprising one or more of sulfonated lignin, condensed naphthalene sulfonate, naphthalene sulfonate, alkyl benzene sulfonate, alkyl sulfate, and a nonionic surfactant (e.g., ethylene oxide adduct of an alkyl phenol).

[0165] In some embodiments, the NLP compositions described herein can be formulated as emulsifiable concentrates. In some embodiments, emulsifiable concentrates comprise one or more NLP compositions described herein at a concentration of about 50 to about 500 grams per liter of liquid dissolved in a carrier (e.g., an organic solvent, a water-miscible solvent, a mixture of a water-immiscible organic solvent and an emulsifier, etc.). In some embodiments, emulsifiable concentrates comprise one or more NLP compositions dissolved in an organic solvent. In some embodiments, emulsifiable concentrates comprise one or more NLP compositions dissolved in an organic solvent selected from aromatic solvents (e.g., xylene, petroleum fractions (e.g., the high-boiling naphthalene and olefin portions of petroleum, such as heavy aromatic naphtha), terpene solvents (e.g., rosin derivatives), aliphatic ketones such as cyclohexanone, and complex alcohols such as 2-ethoxyethanol. In some embodiments, emulsifiable concentrates comprise one or more NLP compositions and one or more suitable emulsifiers, such as anionic and nonionic surfactants.

[0166] In some embodiments, the NLP compositions described herein may be formulated as an aqueous suspension. In some embodiments, the aqueous suspension comprises one or more water-insoluble NLP compositions dispersed in an aqueous carrier at a concentration ranging from about 5% to about 50% by weight. In some embodiments, the aqueous suspension is prepared by pulverizing a dry formulation of one or more NLP compositions and vigorously mixing it with an aqueous carrier (e.g., water) and, optionally, one or more surfactants. In some embodiments, the one or more NLP compositions may be formulated in an aqueous carrier comprising one or more inorganic salts, synthetic gums, natural gums, and the like, which may be added to increase the density and viscosity of the aqueous carrier.

[0167] In some embodiments, the NLP compositions described herein may be formulated as granular compositions. In some embodiments, the granular composition comprises about 0.5% to about 10% by weight of one or more NLP compositions dispersed in a carrier such as clay, starch, silicate, etc. In some embodiments, the granular composition is prepared by dispersing one or more NLP compositions in a suitable solvent and applying it to a granular carrier preformed to an appropriate particle size in the range of about 0.5 to about 3 mm. In some embodiments, the granular composition is prepared by making a dough or paste of the carrier and one or more NLP compositions, crushing and drying to obtain the desired granular particle size.

[0168] In some embodiments, the NLP compositions described herein can be formulated as a powder. In some embodiments, the powder is formulated by mixing one or more NLP compositions described herein provided in powder form with a suitable dusty agricultural carrier, such as kaolin clay, crushed volcanic rock, etc. In some embodiments, a powder formulation of one or more NLP compositions described herein comprises a suitable dusty agricultural carrier at a concentration of about 1% to about 10%. In some embodiments, a powder formulation of one or more NLP compositions can be applied as a seed coating or as a foliar spray using a dust blower machine.

[0169] In some embodiments, the NLP compositions described herein may be formulated in an organic solvent (e.g., petroleum oil, such as spray oils commonly used in agricultural chemistry).

[0170] In some embodiments, the NLP compositions described herein can be formulated to be applied in the form of an aerosol composition. In some embodiments, one or more NLP compositions are dissolved or dispersed in a carrier and packaged in a container that contains a pressure-generating propellant mixture. In some embodiments, NLP compositions formulated as aerosol compositions are packaged in a container from which the mixture is dispensed through an atomizing valve.

[0171] In some embodiments, the NLP compositions described herein can be formulated as an oil-in-water emulsion. In some embodiments, the NLP compositions described herein can be formulated as an oil-in-water emulsion comprising oil droplets each comprising a lamellar liquid crystal coating dispersed in an aqueous phase, each oil droplet comprising at least one heterogeneous active agent, and each oil droplet individually coated with a monolayer or oligolayer comprising (1) at least one nonionic lipophilic surfactant, (2) at least one nonionic hydrophilic surfactant, and (3) at least one ionic surfactant, the droplets having an average particle size of less than 800 nanometers. Further information about this embodiment is disclosed in U.S. Patent Application Publication No. 20070027034, published February 1, 2007. For ease of use, this embodiment is referred to as an "OIWE."

[0172] In some embodiments, the NLP compositions described herein may be formulated with one or more of a wetting agent, a spreading agent, an adhesive, a penetrating agent, a buffering agent, a sequestering agent, a drift reducing agent, a compatibilizer, an anti-foaming agent, a detergent, and an emulsifier.

[0173] As used herein, the term "wetting agent" or "wetter" refers to a substance that, when added to a liquid, increases the spreading or penetration power of the liquid by reducing the interfacial tension between the liquid and the surface it is spreading on. Without wishing to be bound by any particular theory, wetting agents can be used in formulations (e.g., agricultural formulations) for two primary functions: to increase the wetting rate of powders in water during processing and manufacturing to produce concentrates or suspension concentrates for soluble liquids, and to decrease the wetting time of wettable powders during mixing of the product with water in the spray tank, improving the penetration of water into water-dispersible granules. Non-limiting examples of wetting agents include sodium lauryl sulfate, sodium dioctyl sulfosuccinate, alkylphenol ethoxylates, and fatty alcohol ethoxylates.

[0174] As used herein, the term "dispersant" refers to a substance that adsorbs onto the surface of particles, helps keep them dispersed, and prevents them from reagglomerating. Without wishing to be bound by any particular theory, dispersants have the ability to strongly adsorb onto particle surfaces and provide charge or steric hindrance to particle reagglomeration. In some embodiments, dispersants may be used in formulations (e.g., agricultural formulations) of the NLP compositions described herein to facilitate dispersion and suspension during manufacturing and / or ensure that particles redisperse in water in a spray tank. In some embodiments, dispersants may be used in wettable powders, suspension concentrates, and water-dispersible granules. In some embodiments, one or more surfactants may be used as dispersants. In some embodiments, anionic surfactants, nonionic surfactants, or a mixture of anionic and nonionic surfactants may be used as dispersants. In some embodiments, one or more dispersants selected from the group consisting of sodium lignosulfonate, polyelectrolytes (e.g., sodium naphthalenesulfonate formaldehyde condensates), tristyrylphenol ethoxylate phosphate esters, alkylarylethylene oxide condensates, fatty alcohol ethoxylates, alkyl ethoxylates, EO-PO (ethylene oxide-propylene oxide) block and graft copolymers, or any combination thereof, optionally in combination with an anionic surfactant, may be used in formulations (e.g., agricultural compositions) comprising one or more NLP compositions described herein. In some embodiments, one or more high molecular weight polymeric surfactants may be used as dispersants. In some embodiments, the high molecular weight polymeric surfactant has a long hydrophobic "backbone" and multiple ethylene oxide chains that form the "teeth" of a "comb" surfactant. Without wishing to be bound by any particular theory, it is believed that high molecular weight polymers may provide long-term stability to suspension concentrates because the hydrophobic backbone has many anchoring points on the particle surface.

[0175] As used herein, the term "emulsifying agent" or "emulsifier" refers to a substance that stabilizes the suspension of droplets of one liquid phase in another liquid phase. In some embodiments, an emulsifier may be used in formulations (e.g., agricultural formulations) of the NLP compositions described herein to prevent separation of two liquids into two immiscible liquid phases. In some embodiments, the emulsifier is one or more emulsifiers selected from the group consisting of alkylphenols, fatty alcohols (e.g., fatty alcohols having 12 or more ethylene oxide units), and oil-soluble calcium salts of dodecylbenzenesulfonic acid, or any combination thereof. In some embodiments, a small amount of an EO-PO block copolymer surfactant is provided along with an emulsifier in formulations (e.g., agricultural formulations) of the NLP compositions described herein to improve emulsion stability. Some embodiments relate to formulations (e.g., agricultural formulations) of the NLP compositions described herein having a hydrophilic-lipophilic balance ("HLB") value in the range of 8 to 18.

[0176] In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein include a surfactant as a solubilizing agent that will form micelles in water at concentrations above the critical micelle concentration. Without wishing to be bound by any particular theory, the micelles in the formulations (e.g., agricultural formulations) of the NLP compositions can dissolve or solubilize water-insoluble substances within the hydrophobic compartment of the NLP composition. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein include one or more surfactants selected from the group consisting of nonionic surfactants, sorbitan monooleate, sorbitan monooleate ethoxylate, and methyl oleate, or any combination thereof.

[0177] In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein comprise a surfactant, alone or in combination with other excipients such as mineral oil or vegetable oil, that enhances the biological activity of the NLP composition against its target. The type of surfactant used to enhance the biological activity of an NLP composition generally depends on the nature and mode of action of the NLP composition. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein comprise one or more surfactants selected from the group consisting of non-ionic surfactants (e.g., alkyl ethoxylates), linear fatty alcohol ethoxylates, and fatty amine ethoxylates, or any combination thereof.

[0178] In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein include a carrier or diluent in an amount necessary to adjust the concentration, strength, and / or biological activity. In some embodiments, one or more substances with high absorption capacity are provided as a carrier in formulations (e.g., agricultural formulations) of the NLP compositions described herein. In some embodiments, one or more substances with low absorption capacity are provided as a diluent in formulations (e.g., agricultural formulations) of the NLP compositions described herein. In some embodiments, one or more carriers and / or diluents are provided in formulations of dust, wettable powder, granule, and water-dispersible granule formulations of the NLP compositions described herein.

[0179]

[0013] Some embodiments relate to the use of one or more organic solvents in the formulation of one or more NLP compositions as emulsifiable concentrates, oil-in-water emulsions, suspoemulsions, concentrated low-volume NLP formulations, and granular NLP formulations. In some embodiments, formulations of NLP compositions described herein (e.g., agricultural formulations) comprise a mixture of organic solvents. In some embodiments, formulations of NLP compositions described herein (e.g., agricultural formulations) comprise an aliphatic paraffinic oil such as kerosene or refined paraffin. In some embodiments, formulations of NLP compositions described herein (e.g., agricultural formulations) comprise an aromatic solvent such as xylene and higher molecular weight fractions of C9 and C10 aromatic solvents. In some embodiments, formulations of NLP compositions described herein (e.g., agricultural formulations) comprise a mixture of aliphatic paraffinic oil and an aromatic solvent. In some embodiments, formulations of NLP compositions described herein (e.g., agricultural formulations) comprise one or more co-solvents. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein comprise one or more chlorinated hydrocarbons as co-solvents to prevent crystallization of the NLP composition when formulated as an emulsion in water. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein comprise one or more alcohols as co-solvents. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein comprise one or more solvents selected from the group consisting of vegetable oils, seed oils, esters of vegetable oils and esters of seed oils, or any combination thereof.

[0180] Some embodiments relate to the use of one or more thickening agents, gelling agents, and / or anti-settling agents in the formulation of one or more NLP compositions as suspension concentrates, emulsions, or suspoemulsions to modify the rheology or flow properties of the liquid formulation and prevent separation and settling of the dispersed NLP compositions. In some embodiments, formulations (e.g., agricultural formulations) of NLP compositions described herein comprise one or more water-insoluble particles (e.g., clay, silica, montmorillonite, bentonite, magnesium aluminum silicate, attapulgite, etc.). In some embodiments, formulations (e.g., agricultural formulations) of NLP compositions described herein comprise one or more water-soluble polymers (e.g., polysaccharides) as thickening agents, gelling agents, and / or anti-settling agents. In some embodiments, formulations (e.g., agricultural formulations) of NLP compositions described herein comprise one or more polysaccharides extracted from seeds or seaweed. In some embodiments, formulations (e.g., agricultural formulations) of NLP compositions described herein comprise one or more cellulose derivatives. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein comprise one or more cellulose derivatives selected from the group consisting of xanthan gum, guar gum, locust bean gum, carrageenan, alginate, methylcellulose, sodium carboxymethylcellulose (SCMC), and hydroxyethylcellulose (HEC). In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein comprise one or more anti-settling agents selected from the group consisting of modified starch, polyacrylates, polyvinyl alcohol, and polyethylene oxide.

[0181] Microorganisms can cause spoilage or degradation of the NLP compositions described herein. In some embodiments, one or more preservatives are used to eliminate or reduce the effect of microorganisms on the NLP compositions. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein include one or more antimicrobial agents. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein include one or more of propionic acid, sodium salt of propionic acid, sorbic acid, sodium salt of sorbic acid, potassium salt of sorbic acid, benzoic acid, sodium salt of benzoic acid, sodium salt of p-hydroxybenzoic acid, methyl p-hydroxybenzoate, and 1,2-benzisothiazolin-3-one (BIT).

[0182] The presence of surfactants in aqueous formulations of NLP compositions can cause foaming during the mixing process during manufacturing and application via spray tanks. To reduce the tendency to foam, antifoaming agents can be added either during the manufacturing stage or before filling into bottles. Generally, there are two types of antifoaming agents: silicone and non-silicone. Silicones are usually aqueous emulsions of dimethylpolysiloxane, while non-silicone antifoaming agents are water-insoluble oils such as octanol and nonanol or silica. In either case, the function of the antifoaming agent is to remove the surfactant from the air-water interface.

[0183] Some embodiments relate to the use of one or more "green" agents (e.g., adjuvants, surfactants, solvents) that reduce the overall environmental footprint of crop protection formulations. Green agents are biodegradable and generally derived from natural and / or sustainable sources, such as plant and animal sources. In some embodiments, formulations (e.g., agricultural formulations) of the NLP compositions described herein comprise one or more green agents selected from the group consisting of vegetable oils, vegetable oil esters, seed oils, seed oil esters, and alkoxylated alkyl polyglucosides.

[0184] In some embodiments, the NLP compositions described herein may be freeze-dried or lyophilized. See, e.g., U.S. Pat. No. 4,311,712, incorporated herein by reference. In some embodiments, the freeze-dried or lyophilized NLP compositions may be reconstituted with water or another liquid. In some embodiments, the freeze-dried or lyophilized NLP compositions may be reconstituted with a solution comprising one or more heterologous functional agents, agriculturally acceptable carriers, solvents, co-solvents, dispersants, emulsifiers, and / or other ingredients.

[0185] In some embodiments, the NLP compositions described herein may be formulated with a carrier or delivery vehicle that protects the NLP composition from UV and / or acidic conditions. In some embodiments, the NLP compositions described herein may be formulated with a delivery vehicle containing a pH buffer. In some embodiments, the NLP compositions described herein may be formulated to have a pH in the range of about 4.5 to 9.0, including, for example, any one of the approximate pH ranges of about 5.0 to about 8.0, about 6.5 to about 7.5, or about 6.5 to about 7.0. In some embodiments, the NLP compositions described herein may be formulated as described in "Chemistry and Technology of Agrochemical Formulations," edited by D.A. Knowles, copyright 1998 by Kluwer Academic Publishers, incorporated herein by reference. In some embodiments, the NLP compositions described herein may be formulated as described in "Insecticides in Agriculture and Environment - Retrospects and Prospects" by A S Perry, I. Yamamoto, I. Ishaaya, and R. Perry, copyright 1998 by Springer-Verlag, which is incorporated herein by reference.

[0186] ii. Pharmaceutical preparations Some embodiments relate to pharmaceutical formulations of one or more NLP compositions described herein. The NLP compositions described herein may be formulated according to conventional pharmaceutical practice. In some embodiments, the NLP compositions described herein may be formulated into pharmaceutical compositions suitable for administration to animals (e.g., humans). In some embodiments, pharmaceutical formulations of one or more NLP compositions described herein may be administered to animals (e.g., humans) with pharmaceutically acceptable diluents, carriers, and / or excipients. Depending on the mode of administration and dosage, the NLP compositions described herein may be formulated into pharmaceutical compositions suitable for easy delivery. In some embodiments, an effective amount of one or more NLP compositions described herein may be formulated as a single dose, optionally in a unit dosage form. The concentration of the NLP in the pharmaceutical formulation may vary depending on several factors, including the dosage of the heterologous active agent contained in the NLP to be administered and the route of administration.

[0187] In some embodiments, the NLP compositions described herein may be formulated with one or more excipients and / or carriers. In some embodiments, the NLP compositions may be formulated with a pharmaceutical carrier suitable for oral, intravenous (e.g., injection or infusion), or subcutaneous administration to an animal. In some embodiments, the NLP compositions may be formulated with a pharmaceutical carrier described in Remington: The Science and Practice of Pharmacy, 22nd ed., (2012) or ASHP Handbook on Injectable Drugs, 18th ed., (2014), both of which are incorporated by reference. Pharmaceutically acceptable carriers and excipients suitable for use in pharmaceutical formulations of NLP compositions are non-toxic to recipients at the dosages and concentrations employed. In some embodiments, the NLP compositions may be formulated with one or more of a pharmaceutically acceptable buffer (e.g., phosphate buffer, citrate buffer, HEPES, TAE, etc.), antioxidant (e.g., ascorbic acid, methionine, etc.), preservative (e.g., hexamethonium chloride, octadecyldimethylbenzylammonium chloride, resorcinol, benzalkonium chloride, etc.), protein (e.g., serum albumin, gelatin, dextran, immunoglobulin, etc.), hydrophilic polymer (e.g., polyvinylpyrrolidone, etc.), amino acid (e.g., glycine, glutamine, histidine, lysine, etc.), and carbohydrate (e.g., glucose, mannose, sucrose, sorbitol, etc.).

[0188] In some embodiments, the NLP compositions described herein may contain any of a variety of additives, including but not limited to inert diluents, fillers (e.g., sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, starches (e.g., potato starch), calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, sodium phosphate, etc.), granulating agents, disintegrating agents (e.g., cellulose derivatives (e.g., microcrystalline cellulose), starches (e.g., potato starch), croscarmellose sodium, alginates, alginic acid), binders (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, etc.), and the like. , sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, polyethylene glycol, etc.), lubricants, glidants, anti-adherents (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oils or talc), colorants, flavorings, plasticizers, humectants and buffering agents, or any combination thereof.

[0189] For oral administration to animals, the NLP compositions may be prepared in the form of oral formulations. Formulations for oral use may include tablets, caplets, capsules, syrups, or oral liquid dosage forms containing one or more NLP compositions in a mixture with non-toxic pharmaceutically acceptable excipients. In some embodiments, the NLP compositions described herein may be formulated for oral delivery (e.g., chewable tablets, non-chewable tablets, caplets, capsules (e.g., as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, or as soft gelatin capsules in which the NLP composition is mixed with water or an oil vehicle). In some embodiments, the NLP compositions described herein may be formulated as immediate-release, sustained-release, or delayed-release formulations.

[0190] For parenteral administration to animals, NLP compositions can be formulated in the form of a liquid solution or suspension and administered by a parenteral route (e.g., subcutaneous, intravenous, or intramuscular). In some embodiments, the NLP compositions described herein can be formulated for injection or infusion. Pharmaceutical formulations of NLP compositions for parenteral administration can be formulated using a sterile solution or any pharmaceutically acceptable liquid as a vehicle. In some embodiments, pharmaceutical formulations of NLP compositions suitable for parenteral administration comprise one or more pharmaceutically acceptable vehicles selected from the group consisting of sterile water, saline, and cell culture media (e.g., Dulbecco's Modified Eagle's Medium (DMEM), Modified Eagle's Medium (α-MEM), F-12 medium). Formulation methods are known in the art; see, for example, Gibson (ed.) Pharmaceutical Preformulation and Formulation (2nd ed.) Taylor & Francis Group, CRC Press (2009).

[0191] II. Heterofunctional Drugs Some embodiments relate to NLP compositions comprising one or more heterologous functional agents, such as heterologous agricultural agents (e.g., pesticides, fertilizers, herbicides, plant modifiers, etc.) or heterologous therapeutic agents (e.g., antifungals, antioomycetes, antibacterial agents, virucides, antivirucides, insecticides, nematicides, antiparasitics, insect repellents, etc.). In some embodiments of the NLP compositions described herein, the NLP may encapsulate the heterologous functional agent. In some embodiments of the NLP compositions described herein, the heterologous functional agent may be embedded on or conjugated to the surface of the NLP. In some embodiments, the NLP composition may comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different heterologous functional agents. The heterologous functional agents may be added at any step during the manufacturing process effective to introduce the agent into the NLP composition.

[0192] In some embodiments, a heterologous functional agent (e.g., a heterologous agricultural agent (e.g., a pesticide, a fertilizer, a herbicide, a plant modifier, a heterologous nucleic acid, a heterologous polypeptide, a heterologous small molecule, etc.) or a heterologous therapeutic agent (e.g., an antifungal agent, an antioomycete, an antibacterial agent, a virucide, an antiviral agent, a nematicide, an antiparasitic agent, an insect repellent, etc.) can be modified. For example, the modification can be a chemical modification, such as conjugation to a marker, such as a fluorescent marker or a radioactive marker. In some embodiments, the modification can include conjugation or operably linking the heterologous functional agent to a moiety that enhances the stability, delivery, targeting, bioavailability, or half-life of the agent, such as a lipid, a glycan, a polymer (e.g., PEG), or a cationic moiety.

[0193] Examples of heterofunctional agents that can be loaded into the NLP compositions described herein are outlined below.

[0194] A. Heterogeneous agricultural chemicals The NLP compositions described herein may include one or more heterologous agricultural agents. Non-limiting examples of heterologous agricultural agents include antifungals, antibacterials, insecticides, molluscicides, nematicides, herbicides, virucides, peptides, polypeptides, nucleic acids, polynucleotides, and ribonucleoproteins.

[0195] For example, in some instances, the NLP composition may include one or more pesticides. Non-limiting examples of pesticides include antifungals, antibacterials, insecticides, molluscicides, nematicides, and virucides. In some embodiments, the pesticide may be a chemical agent (e.g., deltamethrin), such as those known in the art. In some embodiments, the pesticide may be a peptide, polypeptide, nucleic acid, polynucleotide, or small molecule. In some embodiments, the pesticide may be an agent capable of reducing the fitness of various plant pests or an agent that targets one or more specific target plant pests (e.g., a specific species or genus of plant pest).

[0196] In some examples, the NLP composition may include one or more heterologous fertilizing agents. Non-limiting examples of heterologous fertilizing agents include plant nutrients and plant growth regulators, such as those known in the art. In some embodiments, the fertilizing agent may be a mineral, peptide, polypeptide, nucleic acid, or polynucleotide capable of increasing the fitness of a plant or plant microorganism (e.g., a plant symbiont). In some embodiments, the fertilizing agent may be an agent capable of increasing the fitness of various plants or plant microorganisms, or may be an agent that targets one or more specific target plants or plant microorganisms (e.g., a specific species or genus of plant or plant microorganism).

[0197] In some embodiments, the NLP compositions may include one or more heterologous plant modifiers. In some examples, the plant modifier may include a peptide or a nucleic acid.

[0198] B. Heterogeneous Therapeutics The NLP compositions described herein can include one or more heterologous therapeutic agents (e.g., an animal (e.g., a mammal, e.g., a human), an animal pathogen, or a pathogen vector thereof). Non-limiting examples of heterologous therapeutic agents include therapeutic peptides, therapeutic nucleic acids (e.g., therapeutic RNA), therapeutic small molecules, and pathogen control agents (e.g., antifungals, antibacterials, virucides, antivirals, insecticides, nematicides, antiparasitics, insect repellents, etc.). In some embodiments, NLP compositions loaded with heterologous therapeutic agents can be formulated with a pharmaceutically acceptable carrier for delivery to an animal, animal pathogen, or a pathogen vector thereof.

[0199] C. Antibacterial agents In some embodiments, the NLP compositions described herein may comprise an antimicrobial agent. In some embodiments, the NLP compositions comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different antimicrobial agents. In some embodiments, the NLP compositions described herein may comprise an antimicrobial agent that reduces the fitness (e.g., reduces the growth or kills) of a bacterial pathogen (e.g., a bacterial plant pathogen, a bacterial animal pathogen). In some embodiments, a target bacterium or a plant or animal infected with a target bacterium may be contacted with an NLP composition comprising an antibiotic described herein in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of antibiotic concentration in or on the target bacterium, and (b) reduce the fitness of the target bacterium. The antimicrobial agent may be loaded into the NLP composition according to any of the methods described herein, and in some instances may be associated with the surface of the NLP.

[0200] As used herein, the term "antimicrobial agent" refers to any material that kills or inhibits the growth, proliferation, division, reproduction, or spread of bacteria, such as plant pathogenic bacteria, and includes bactericides (e.g., disinfecting compounds, antiseptic compounds, antibiotics, etc.) and bacteriostatic agents (e.g., growth or reproduction inhibiting compounds or antibiotics). Bactericides kill bacteria, while bacteriostatic agents only slow their growth or reproduction. Bactericides can include disinfectants, antiseptics, or antibiotics. Non-limiting examples of disinfectants include active chlorine (e.g., hypochlorites (e.g., sodium hypochlorite), chloramines, dichloroisocyanurate and trichloroisocyanurate, wet chlorine, chlorine dioxide, active oxygen, peroxides (e.g., peracetic acid, potassium persulfate, sodium perborate, sodium percarbonate, urea perhydrate, etc.), iodine, iodopovidone, povidone-iodine, betadine, Lugol's solution, tincture of iodine, iodinated nonionic surfactants, concentrated alcohols (e.g., ethanol, 1-propanol (also called n-propanol), 2-propanol (also called isopropanol), 2-phenoxyethanol, 1-phenoxypropanol, 2-phenoxypropanol, etc.), phenolics (e.g., phenols (also called carboxylic acids)), cresols (combined with liquid potassium soap), These include chlorinated and brominated phenols, such as hexachlorophene, triclosan, trichlorophenol, tribromophenol, pentachlorophenol, dibromol, and their salts; cationic surfactants, such as some quaternary ammonium cations (e.g., benzalkonium chloride, cetyltrimethylammonium bromide or chloride, didecyldimethylammonium chloride, cetylpyridinium chloride, benzethonium chloride); non-quaternary compounds, such as chlorhexidine, glucoprotamine, and octenidine dihydrochloride; strong oxidizing agents, such as ozone and permanganate solutions; heavy metals and their salts, such as colloidal silver, silver nitrate, mercury chloride, phenylmercuric salts, copper sulfate, copper oxide-chloride, copper hydroxide, copper octoate, copper oxychloride sulfate, and copper sulfate pentahydrate.Concentrated strong acids (eg, phosphoric acid, nitric acid, sulfuric acid, amidosulfonic acid, toluenesulfonic acid, etc.) and alkalis (eg, sodium hydroxide, potassium hydroxide, calcium hydroxide, etc.) can also be used as disinfectants.

[0201] As used herein, the term "antiseptic" refers to an antimicrobial or disinfectant that can be applied to an animal (e.g., skin, mucous membranes, wounds, etc.) under appropriate conditions (e.g., concentration, pH, temperature, etc.). Non-limiting examples of preservatives include dilute chlorine preparations (e.g., Dakin's solution, 0.5% sodium or potassium hypochlorite solution adjusted to pH 7-8, 0.5-1% solution of benzenesulfochloramide (chloramine B), etc.), iodine preparations (e.g., iodopovidone in various galenicals (ointments, solutions, wound plasters)), Lugol's solution, peroxide, urea perhydrate solution, pH-buffered 0.1-0.25% peracetic acid solution, alcohol with or without preservative additives, weak organic acids (e.g., sorbic acid, benzoic acid, lactic acid, salicylic acid), phenolic compounds (e.g., hexachlorophene, triclosan, dibromide, etc.), cationically active compounds (e.g., 0.05-0.5% benzalkonium, 0.5-4% chlorhexidine, 0.1-2% octenidine solution).

[0202] Some embodiments relate to NLP compositions described herein that include one or more antibiotics. Antibiotics are generally classified based on their mechanism of action, chemical structure, or spectrum of activity. Antibiotics can target any bacterial function or growth process and can be either bacteriostatic or bacteriocidal. Antibiotics also vary in their level of target specificity (e.g., narrow or broad spectrum). In some instances, an antibiotic is a narrow-spectrum antibiotic, thus targeting a specific type of bacteria, such as gram-negative or gram-positive bacteria. Alternatively, an antibiotic can be a broad-spectrum antibiotic, targeting a wide range of bacteria.

[0203] Any antibiotic known in the art may be included in and / or formulated with the NLP compositions described herein. In some embodiments, the NLP compositions described herein may comprise one or more bactericidal antibiotics. Non-limiting examples of bactericidal antibiotics include antibiotics that target bacterial cell walls (e.g., penicillins, cephalosporins), antibiotics that target cell membranes (e.g., polymyxins), antibiotics that inhibit essential bacterial enzymes (e.g., rifamycins, lipiamycins, quinolones, sulfonamides), and aminoglycosides (e.g., kasugamycin). In some embodiments, the NLP compositions described herein may comprise one or more bacteriostatic antibiotics. Non-limiting examples of bacteriostatic antibiotics include antibiotics that target protein synthesis (e.g., macrolides, lincosamides, tetracyclines). Additional classes of antibiotics that may be included in NLP compositions include, but are not limited to, cyclic lipopeptides (such as daptomycin), glycylcyclines (such as tigecycline), oxazolidinones (such as linezolid), and lipiamycins (such as fidaxomicin). In some embodiments, the NLP compositions described herein may include one or more antibiotics selected from the group consisting of rifampicin, ciprofloxacin, doxycycline, ampicillin, and polymyxin B. In some embodiments, the NLP compositions described herein may include one or more antibiotics listed in Table 4. One of skill in the art will understand that the appropriate concentration of antibiotics in an NLP composition or formulation comprising one or more NLP compositions will depend on factors such as the effectiveness of the antibiotics, stability, number of individual antibiotics, formulation, and method of application of the composition.

[0204] [Table 11]

[0205] Some embodiments relate to NLP compositions comprising antibiotics that can be administered to an animal (e.g., a human) in an amount and for a time sufficient to achieve a target level (e.g., a predetermined or threshold level) of antibiotic concentration in or on the animal and / or to treat or prevent a bacterial infection in the animal. Non-limiting examples of antibacterial agents suitable for treating animals that can be included in and / or formulated with the NLP compositions described herein include penicillin (amoxicillin, ampicillin, bacampicillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, nafcillin, oxacillin, penicillin g, crysticillin 300). AS, Pentids, Permapen, Pfizerpen, Pfizerpen-AS, Wycillin, Penicillin V, Piperacillin, Pivampicillin, Pivmecillinam, Ticarcillin, Cephalosporins (Cefacetrile (Cephacetrile)), Cefadroxil (Cefadroxil (Cefadroxil)), Cephalexin (Cephalexin)), Cefaloglycin (Cephaloglycin)), Cephalonium (Cephalonium), Cephaloridine (Cefalorazine), Cephalothin (Ceph) alotin (cephalothin), cephapirin (cefapirin (cephapirin)), cefatrizine, cefazaflur, cefazedone, cefazolin (cefazolin (cephazolin)), cefradine (cefradine (cephradine)), cefroxadine, ceftezole, cefaclor, cefamandole, cefmetazole, cefonicid, cefotetan, cefoxitin, cefprozil (cefproxil), cefuroxime, cefuzonam, cefcapene, cefdaloxime (cefdaloxime), cefdinir, cefditoren, cefetamet, cefixime, cefmenoxime, cefodizime, cefotaxime, cefpimizole, cefpodoxime, cefteram, ceftibuten, ceftiofur,Ceftiolene, ceftizoxime, ceftriaxone, cefoperazone, ceftazidime, cefclidine, cefepime, cefluprenam, cefoselis, cefozopran, cefpirome, cefquinome, ceftobiprole, ceftaroline, cefaclomezine, cefaloram, cefaparole, cefcanel, cefedrolol, cefempidone, Cefetrizole, cefivitril, cefmatilen, cefmepidium, cefovecin, cefoxazol, cefrotil, cefsumide, cefracetime, ceftioxide, combinations i.e. ceftazidime / avibactam, ceftolozane / tazobactam, monobactam (aztreonam), carbapenems (imipenem, imipenem / cilastatin), Benzene, doripenem, ertapenem, meropenem, meropenem / vaborbactam), macrolides (azithromycin, erythromycin, clarithromycin, dirithromycin, roxithromycin, telithromycin), lincosamides (clindamycin, lincomycin), streptogramins (pristinamycin, quinupristin / dalfopristin), aminoglycosides (amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, streptomycin, tobramycin), quinolones (flumequine, nari Dixic acid, oxolinic acid, piromidic acid, pipemidic acid, losoxacin, second generation i.e. ciprofloxacin, enoxacin, lomefloxacin, nadifloxacin, norfloxacin, ofloxacin, pefloxacin, rufloxacin, balofloxacin, gatifloxacin, grepafloxacin, levofloxacin, moxifloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, besifloxacin, delafloxacin, clinafloxacin, gemifloxacin, prulifloxacin, sitafloxacin,trovafloxacin), sulfonamides (sulfamethizole, sulfamethoxazole, sulfisoxazole, trimethoprim-sulfamethoxazole), tetracyclines (demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline, tigecycline), other (lipopeptides, fluoroquinolones, lipidated glycopeptides, cephalosporins, macrocyclics, chloramphenicol, metronidazole, tinidazole, nitrofurantoin, glycopeptides, vancomycin, teicoplanin, glycolipid peptides, telavancin, oxazolidinone, linezolid, cycloserine 2, rifamycin, rifampin, rifabutin, rifapentine, rifalazil, polypeptides, bacitracin, polymyxin B, tuberactinomycin, viomycin, capreomycin). Those skilled in the art will understand that the appropriate concentration of each antibiotic in the composition will depend on factors such as the efficacy, stability of the antibiotics, the number of individual antibiotics, the formulation and method of application of the composition, etc.

[0206] D. Antifungal agents Some embodiments relate to NLP compositions described herein comprising one or more antifungal agents. In some examples, the NLP compositions may comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different antifungal agents. Some embodiments relate to methods of reducing the fitness (e.g., reducing the growth or killing) of a fungus (e.g., a fungal plant pathogen) by contacting a fungus or a fungal-infested plant or animal with an NLP composition comprising an antifungal agent in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of antifungal agent concentration in or on the target fungus, and (b) reduce the fitness of the target fungus. The antifungal agent may be loaded / incorporated into the NLP compositions described herein and / or formulated with the NLP compositions by any of the methods described herein.

[0207] As used herein, the terms "fungicide" or "antifungal agent" refer to a substance that kills or inhibits the growth, proliferation, division, reproduction or spread of fungi, such as plant pathogenic fungi or animal pathogenic fungi. Non-limiting examples of antifungal agents that may be included in and / or formulated with the NLP compositions described herein include azoxystrobin, mancozeb, prothioconazole, folpet, tebuconazole, difenoconazole, captan, bupirimate or fosetyl-AI, strobilurins, azoxystrobin, dimoxystrobin, enestrobin, fluoxastrobin, kresoxim-methyl, metominostrobin, picoxystrobin, pyraclostrobin, trifloxystrobin, thiazolinone ... Strobin, oryzastrobin, carboxamide, carboxanilide, benalaxyl, benalaxyl-M, benodanil, carboxin, mebenil, mepronil, fenfuram, fenhexamid, flutolanil, furalaxyl, flucarbanil, furametpyr, metalaxyl, metalaxyl-M (mefenoxam), metofuroxam, metsulfovax, ofrace, oxadixyl, oxycarboxin, penthiopyrad, pyracarbollide, salicylanilide, tecloftalam, thifluzamide, thiazinil Benzamide, N-biphenylamide, bixafen, boscalid, carboxylic acid morpholide, dimethomorph, flumorph, benzamide, flumetober, fluopicolide (picobenzamide), zoxamide, carboxamide, carpropamid, diclocymet, mandipropamide, silthiofam, azoles, triazoles, bitertanol, bromuconazole, cyproconazole, diniconazole, enilconazole, epoxiconazole, fenbuconazole, flusilazole, fluquinconazole, flutriazole Aphor, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propiconazole, prothioconazole, simeconazole, tetraconazole, triadimenol, triadimefon, triticonazole, imidazole, cyazofamid, imazalil, pefurazoate, prochloraz, triflumizole, benzimidazole, benomyl, carbendazim, fuberidazole, thiabendazole, ethaboxam, etridiazole, hymexazole,Nitrogen-containing heterocyclyl compounds, pyridine, fadinam, pyrifenox, pyrimidine, cyprodinil, ferimzone, fenarimol, mepanipyrim, nuarimol, pyrimethanil, piperazine, triforine, pyrrole, fludioxonil, fenpiclonil, morpholine, aldimorph, dodemorph, fenpropimorph, tridemorph, dicarboximide, iprodione, procymidone, vinclozolin, acibenzolar-S-methyl, anilazine, captan, captafol, dazomet, diclomedine, fenoxa Nil, folpet, fenpropizin, famoxadone, fenamidone, octhilinone, probenazole, proquinazide, pyroquilon, quinoxyfen, tricyclazole, carbamate, dithiocarbamate, ferbam, mancozeb, maneb, metiram, metam, propineb, thiram, zineb, ziram, diethofencarb, flubenthiavalicarb, iprovalicarb, propamocarb, guanidine, dodine, iminoctadine, guazatine, kasugamycin, polyoxin, streptomycin, validamycin A , organometallic compounds, fentin salts, sulfur-containing heterocyclyl compounds, isoprothiolane, dithianon, organophosphorus compounds, edifenphos, fosetyl, fosetyl-aluminum, iprobenfos, pyrazophos, tolclofos-methyl, organochlorine compounds, thiophanate-methyl, chlorothalonil, dichlofluanid, tolylfluanid, flusulfamide, phthalide, hexachlorobenzene, pencycuron, quintozene, nitrophenyl derivatives, binapacryl, dinocap, dinobuton, spiroxamine, cyflufenamid , cymoxanil, metrafenone, N-2-cyanophenyl-3,4-dichloroisothiazole-5-carboxamide (isothianil), N-(3',4',5'-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methylpyrazole-4-carboxamide, 3-[5-(4-chlorophenyl)-2,3-dimethylisoxazolidin-3-yl]-pyridine, N-(3',4'-dichloro-4-fluorobiphenyl-2-yl)-3-difluoromethyl-1-methylpyrazole-4-carboxamide,5-chloro-7-(4-methylpiperidin-1-yl)-6-(2,4,6-trifluorophenyl)-[1,2,4]triazolo[1,5-a]pyrimidine, 2-butoxy-6-iodo-3-propylchromen-4-one, N,N-dimethyl-3-(3-bromo-6-fluoro-2-methylindole-1-sulfonyl)-[1,2,4]triazole-1-sulfonamide, methyl-(2-chloro-5-[1-(3-methylbenzyloxyimino)-ethyl]benzyl)carbamate, methyl methyl-(2-chloro-5-[1-(6-methylpyridin-2-ylmethoxy-imino)ethyl]benzyl)carbamate, methyl 3-(4-chlorophenyl)-3-(2-isopropoxycarbonylamino-3-methylbutyryl-amino)propionate, 4-fluorophenyl N-(1-(1-(4-cyanophenyl)ethanesulfonyl)but-2-yl)carbamate, N-(2-(4-[3-(4-chlorophenyl)prop-2-ynyloxy]-3-methoxyphenyl)ethyl)-2-meth ethanesulfonylamino-3-methylbutyramide, N-(2-(4-[3-(4-chlorophenyl)prop-2-ynyloxy]-3-methoxyphenyl)ethyl)-2-ethanesulfonylamino-3-methylbutyramide, N-(4'-bromobiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, N-(4'-trifluoromethylbiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, N-(4'-chloro- Antifungal agents that may be used include (3'-fluorobiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, methyl 2-(ortho-((2,5-dimethylphenyloxy-methylene)phenyl)-3-methoxyacrylate, or oxathiapiproline. One of skill in the art will understand that the suitable concentration of each antifungal agent in an NLP composition or a formulation comprising one or more NLP compositions will depend on factors such as efficacy, stability of the antifungal agents, the number of individual antifungal agents, and the formulation and method of application of the composition.

[0208] Some embodiments relate to NLP compositions comprising an antifungal agent that can be administered to an animal (e.g., a human) in an amount and for a time sufficient to achieve a target level (e.g., a predetermined or threshold level) of antifungal concentration in or on the animal and / or to treat or prevent a fungal infection in the animal. Non-limiting examples of antifungal agents suitable for treating animals that can be included in and / or formulated with the NLP compositions described herein include allylamines (amorolfine, butenafine, naftifine, terbinafine), imidazoles (bifonazole, butoconazole, clotrimazole, econazole, fenticonazole, ketoconazole, isoconazole, luliconazole, miconazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, terconazole), triazoles (albaconazole, efinaconazole, fluconazole, isavuconazole), Antifungal agents include benzoyl peroxidase (BPO), ...

[0209] E. Insecticides Some embodiments relate to NLP compositions described herein that comprise one or more insecticides. In some examples, the NLP composition comprises two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different insecticides. Some embodiments relate to methods of reducing the fitness of a target insect (e.g., reducing growth or killing) by contacting the target insect or a plant or animal infested or infested by the target insect with an NLP composition that comprises an insecticide in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of insecticide concentration in or on the target insect, and (b) reduce the fitness of the target insect. The insecticide may be loaded / incorporated into the NLP compositions described herein and / or formulated with the NLP composition by any of the methods described herein.

[0210] As used herein, the term "insecticide" or "insecticidal agent" refers to an agent that reduces the fitness (e.g., kills, inhibits growth, inhibits proliferation, inhibits reproduction, inhibits spread, inhibits feeding, reduces reproduction, etc.) of insects, such as agricultural insect pests (e.g., corn rootworms, stink bugs, canola flea beetles, thrips, armyworms, etc.), insect vectors of animal pathogens, or insect parasites. Non-limiting examples of insecticides that may be included in and / or formulated with the NLP compositions described herein are listed in Tables 5A and 5B. Further non-limiting examples of insecticides are flupyradifurone, spirotetramat, spiromesifen, fluopyram, imidacloprid, flubendiamide, permethrin, thiacloprid, fluopyram, sulfoxaflor, spinetoram, chlorpyrifos, spinosad, cyantraniliprole, indoxacarb, chlorantraniliprole, cypermethrin, emamectin benzoate, pymetrozine, thiamethoxam, lambda cyhalothrin, and cyclaniliprole.

[0211] Further non-limiting examples of suitable insecticides that may be included in and / or formulated with the NLP compositions described herein include biological insecticides, such as: inhibitory RNAs that target essential insect genes; hormones, such as ecdysteroids and juvenile hormones; pheromones, such as codrumone; insecticidal proteins (e.g., Bacillus thuringiensis (Bt) crystal proteins (Cry) (e.g., Cry1Ab, Cry1Fa, Cry2Ab, etc.); vegetative insecticidal proteins (Vip) (e.g., Vip1, Vip2, Vip3 (e.g., Vip3Aa), Vip4, etc.); IPD083Aa, IPD083Cb, etc.); insecticidal plant extracts such as neem oil and azadirachtin; insecticidal bacteria, such as Bacillus species (e.g., Bacillus thuringiensis); thuringiensis), Beauveria spp., Metarrhizium spp., Saccharopolyspora spp., Paecilomyces spp., and Verticillium spp. In some embodiments, the NLP compositions described herein may include or be formulated with one or more insecticidal active compounds with an unknown or unspecified mode of action, such as fumigants (e.g., aluminum phosphide, methyl bromide, sulfuryl fluoride, etc.) and selective antifeedants (e.g., cryolite, flonicamid, pymetrozine, etc.). One of skill in the art will understand that the appropriate concentration of each insecticide in the NLP composition will depend on factors such as efficacy, stability of the insecticide, number of individual insecticides, formulation, and method of application of the NLP composition.

[0212] [Table 12]

[0213] [Table 13]

[0214] [Table 14]

[0215] [Table 15]

[0216] F. Nematicides Some embodiments relate to NLP compositions described herein comprising one or more nematicides. In some examples, the NLP compositions may comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different nematicides. Some embodiments relate to methods of reducing the fitness (e.g., reducing growth or killing) of targeted nematodes by contacting the targeted nematodes or a plant or animal infested or parasitized by the targeted nematodes with an NLP composition comprising one or more nematicides in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of nematicide concentration in or on the targeted nematode, and (b) reduce the fitness of the targeted nematode. The nematicides may be loaded / incorporated into the NLP compositions described herein and / or formulated with the NLP compositions by any of the methods described herein.

[0217] As used herein, the term "nematicide" or "nematocidal agent" refers to a substance that reduces the fitness (e.g., kills, inhibits growth, inhibits proliferation, inhibits reproduction, inhibits spread, inhibits feeding, etc.) of nematodes, such as agricultural nematode pests or parasitic nematodes. Non-limiting examples of nematicides that may be included in and / or formulated with the NLP compositions described herein are listed in Table 6. One of skill in the art will understand that the appropriate concentration of each nematicide in the NLP composition will depend on factors such as nematicide efficacy, stability, number of individual nematicides, formulation, and method of application of the NLP composition.

[0218] [Table 16]

[0219] G. Antiparasitic Agents Some embodiments relate to NLP compositions described herein that include one or more antiparasitic agents. In some examples, the NLP composition includes two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different antiparasitic agents. In some embodiments, an animal is contacted with an NLP composition that includes one or more antiparasitic agents in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of antiparasitic agent concentration in or on a parasite (e.g., a parasitic nematode, a parasitic insect, a protozoan) or an animal infected therewith, and (b) reduce the fitness of the parasite. This may be useful for treating or preventing parasites in the animal. The antiparasitic agent may be formulated with the NLP composition by any of the methods described herein, and in certain examples, may be associated with or encapsulated by the NLP.

[0220] As used herein, the term "antiparasitic" or "antiparasitic agent" refers to a substance that kills or inhibits the growth, proliferation, reproduction or spread of parasites, such as parasitic protozoa, parasitic nematodes or parasitic insects. Non-limiting examples of antiparasitic agents that may be included in and / or formulated with the NLP compositions described herein include antihelminthic drugs (bephenium, diethylcarbamazine, ivermectin, niclosamide, piperazine, praziquantel, pyrantel, pyrivinium, benzimidazole, albendazole, flubendazole, mebendazole, thiabendazole, levamisole, nitazoxanide, monopantel, emodepside, spiroindole), scabicides, Examples of antiparasitic agents include drugs (benzyl benzoate, benzyl benzoate / disulfiram, lindane, malathion, permethrin), pediculicides (piperonyl butoxide / pyrethrin, spinosad, moxidectin), scabicides (crotamiton), cestodes (niclosamide, plungequantel, albendazole), amoebicides (rifampin, amphotericin B), or antiprotozoals (melarsoprol, eflornithine, metronidazole, tinidazole, miltefosine, artemisinin). In certain examples, NLP compositions comprising antiparasitic agents may be used to treat or prevent infections in livestock animals. In some embodiments, the NLP compositions described herein may include or be formulated with one or more of levamisole, fenbendazole, oxifendazole, albendazole, moxidectin, eprinomectin, doramectin, ivermectin, and clorsulon. One of skill in the art will understand that the suitable concentration of each antiparasitic agent in the NLP composition will depend on factors such as the efficacy of the antiparasitic agent, stability, the number of individual antiparasitic agents, and the formulation and method of application of the NLP composition.

[0221] H. Molluscicides Some embodiments relate to NLP compositions described herein comprising one or more molluscicides. In some examples, the NLP composition comprises two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) different molluscicides. Some embodiments relate to methods of reducing the fitness of a target mollusc (e.g., killing or reducing the growth) by contacting the target mollusc or a plant infested by the target mollusc with an NLP composition comprising a molluscicide in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of molluscicide concentration in or on the target mollusc, and (b) reduce the fitness of the target mollusc. The molluscicide may be loaded / incorporated into the NLP compositions described herein and / or formulated with the NLP compositions by any of the methods described herein.

[0222] As used herein, the term "molluscicide" or "molluscicidal agent" refers to a substance that reduces the fitness of (e.g., kills, inhibits growth, inhibits proliferation, inhibits reproduction, inhibits spreading, inhibits feeding, etc.) molluscs, such as agricultural mollusc pests. Non-limiting examples of molluscs that may be included in and / or formulated with the NLP compositions described herein include metal salts such as iron(III) phosphate, aluminum sulfate and ferric sodium, EDTA, metaldehyde, methiocarb, and acetylcholinesterase inhibitors. One of skill in the art will understand that the appropriate concentration of each molluscicide in an NLP composition will depend on factors such as efficacy, stability of the molluscicide, number of individual molluscicides, formulation, and method of application of the NLP composition.

[0223] I. Antiviral Agents Some embodiments relate to NLP compositions described herein that further comprise one or more antiviral agents. In some examples, the NLP composition comprises two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different antiviral agents. Some embodiments relate to methods of treating (e.g., reducing the growth or killing) a viral infection by contacting a virus or a virus-infected plant or animal with an NLP composition comprising an antiviral agent in an amount and for a time sufficient to (a) reach a target level (e.g., a predetermined or threshold level) of antiviral agent concentration, and (b) reduce or eliminate the target virus. In some embodiments, an NLP composition comprising an antiviral agent can be administered to an animal in an amount and for a time sufficient to reach a target level (e.g., a predetermined or threshold level) of antiviral concentration in or on the animal and / or treat or prevent a viral infection in the animal. The antiviral agent can be loaded / incorporated into the NLP compositions described herein and / or formulated with the NLP composition by any of the methods described herein.

[0224] As used herein, the terms "virucide" or "antiviral" or "antiviral agent" refer to any substance that inactivates a virus, destroys a virus, or interferes with or inhibits any stage of the viral life cycle (e.g., prevents infection (e.g., attachment to a host cell), viral uncoating, integration, transcription, translation, replication, assembly, or release). In some embodiments, the vector is a viral plant pathogen. Several agents can be used as virucidal agents, including chemical and biological agents (e.g., biomimetics, nucleic acids (e.g., dsRNA, morpholinos, etc.)).Non-limiting examples of antiviral agents that may be included in and / or formulated with the NLP compositions described herein include abacavir, acyclovir (acyclovir), adefovir, amantadine, amprenavir (agenerase), ampligen, arbidol, atazanavir, atripla, baravir, cidofovir, combivir, dolutegravir, darunavir, delavirdine, didanosine, docosanol, edoxudine, efavirenz, emtricitabine, enfuvirtide, entecavir, ecoliever, famciclovir, fomivirsen, fosamprenavir, foscarnet, phosphonet, fusion inhibitors, ganciclovir, ibacitabine, immunovir, idoxuridine, imiquimod, indinavir, inosine, integrase inhibitors, interferon type III, interferon Interferon type II, interferon type I, interferon, lamivudine, lopinavir, loviride, maraviroc, moroxydine, methisazone, nelfinavir, nevirapine, nexavir, nitazoxanide, nucleoside analogues, norvir, oseltamivir (Tamiflu), peginterferon alfa-2a, penciclovir, peramivir, pleconaril, podophyllotoxin, raltegravir, ribavirin, rimantadine, ribavirin These include tetanus, pyramidine, saquinavir, sofosbuvir, stavudine, synergistic enhancer (antiretroviral), telaprevir, tenofovir, tenofovir disoproxil, tipranavir, trifluridine, trizivir, tromantadine, Truvada, valacyclovir (Valtrex), valganciclovir, vicriviroc, vidarabine, viramidine, zalcitabine, zanamivir (Relenza), and zidovudine. One of skill in the art will understand that the appropriate concentration of each antiviral agent in the NLP composition will depend on factors such as efficacy, stability of the virucidal agents, number of individual virucidal agents, formulation, and method of application of the NLP composition.

[0225] J. Herbicides Some embodiments relate to NLP compositions described herein that comprise one or more herbicides. In some examples, the NLP composition comprises two or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different herbicides. Some embodiments relate to methods of reducing the fitness of a plant (e.g., a weed) by contacting the plant with an NLP composition that comprises a herbicide in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of herbicide concentration on the plant, and (b) reduce the fitness of the plant. The herbicide may be loaded or incorporated into the NLP compositions described herein and / or may be formulated with the NLP composition by any of the methods described herein.

[0226] As used herein, the term "herbicide" or "herbicidal agent" refers to a substance that reduces the fitness of plants (e.g., kills, inhibits growth, inhibits proliferation, inhibits reproduction, inhibits spread, etc.). Non-limiting examples of herbicides that may be included in and / or formulated with the NLP compositions described herein include benzoic acid herbicides such as glufosinate, propaquizafop, metamitron, metazachlor, pendimethalin, flufenacet, diflufenican, clomazone, nicosulfuron, mesotrione, pinoxaden, sulcotrione, prosulfocarb, sulfentrazone, bifenox, quinmerac, triallate, terbuthylazine, atrazine, oxyfluorfen, diuron, trifluralin, chlorotoluron, and dicamba esters; phenoxyalkanoic acid herbicides such as 2,4-D, MCPA, and 2,4-DB esters; clodinafop, cyhalofop, fenoxaprop, fluazifop, and the like. herbicides such as aryloxyphenoxypropionic acid herbicides, such as haloxyfop and quizalofop esters; pyridine carboxylic acid herbicides, such as aminopyralid, picloram, and clopyralid esters; pyrimidine carboxylic acid herbicides, such as aminocyclopyrachlor esters; pyridyloxyalkanoic acid herbicides, such as fluoroxypyr and triclopyr esters; hydroxybenzonitrile herbicides, such as bromoxynil and ioxynil esters; esters of arylpyridine carboxylic acids; and arylpyrimidine carboxylic acids of the general structure disclosed in U.S. Pat. Nos. 7,314,849, 7,300,907, and 7,642,220, the entireties of which are incorporated herein by reference.In certain embodiments, the NLP compositions described herein may be selected from the group consisting of 2,4-D, 2,4-DB, acetochlor, acifluorfen, alachlor, ametryn, amitrole, asulam, atrazine, azafenidin, benefin, bensulfuron, bensulide, bentazon, bromacil, butyrate, carfentrazone, chloramben, chlorimuron, chlorpropham, chlorsulfuron, clethodim, clopyralid, chloransulam, cyanazine, cycloate, DCPA, desmedipham, diclofenac, cyclohexyl benzoate ... Benil, diclofop, diclosulam, diethathyl, difenzoquat, diflufenzopyr, dimethenamid-p, diquat, DSMA, endosole, EPTC, ethalfluralin, ethametsulfuron, ethofumesate, fluazifop-P, flucarbazone, flumetsulam, flumiclorac, flumioxazin, fluometuron, fluroxypyr, fluthiacet, fomesafen, foramsulfuron, glufosinate, glyphosate, halosulfuron, haloxyfop, hexazinone, imazametha Benz, imazamox, imazapic, imazaquin, imazethapyr, isoxaben, isoxaflutole, lactofen, linuron, MCPA, MCPB, mesotrione, methazole, metolachlor-s, metribuzin, metsulfuron, molinate, MSMA, napropamide, naptalam, norflurazon, oryzalin, oxadiazon, oxasulfuron, oxyfluorfen, paraquat, pebulate, pelargonic acid, pendimethalin, phenmedipham, primisulfuron, prodiamine, The herbicide may include one or more herbicides selected from the group consisting of prometryn, pronamide, propachlor, propanil, prosulfuron, pyrazone, pyridate, pyrithiobac, quinclorac, quizalofop, rimsulfuron, sethoxydim, siduron, simazine, sulfometuron, sulfosulfuron, tebuthiuron, terbacil, thiazopyr, thifensulfuron, thiobencarb, tralkoxydim, triallate, triasulfuron, tribenuron, triclopyr, triflusulfuron, and vernolate.In certain embodiments, the NLP compositions described herein comprise one or more nucleic acids that induce silencing or reduce expression of essential plant genes (e.g., siRNA, miRNA, dsRNA, RNA / DNA hybrids, etc.). One of skill in the art will understand that the appropriate concentration of each herbicide in the NLP composition will depend on factors such as herbicide efficacy, stability, the number of individual herbicides, the formulation, and method of application of the NLP composition.

[0227] When a herbicide is included in the NLP or composition thereof, the method can further be used to reduce the fitness of weeds or kill weeds. In such cases, the method can be effective to reduce the fitness of weeds by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more compared to the fitness of untreated weeds (weeds to which the NLP composition has not been administered). For example, the method can be effective to kill weeds, thereby reducing the weed population by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more compared to untreated weeds. In some examples, the method substantially eliminates weeds. Examples of weeds that may be treated with NLPs according to the present compositions and methods are described in WO2021041301A1, which is incorporated herein by reference in its entirety.

[0228] K. Repellents Some embodiments relate to NLP compositions described herein that comprise one or more repellents. In some examples, the NLP compositions comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different repellents. Some embodiments relate to methods of repelling a target pest by contacting the target pest, an environment occupied by the target pest, a plant, or an animal with an NLP composition or formulation that includes a repellent in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of repellent concentration, and (b) reduce the level of the pest on the plant, animal, or environment compared to an untreated plant, animal, or environment. Repellents can be loaded / incorporated into the NLP compositions described herein and / or formulated with the NLP compositions by any of the methods described herein.

[0229] As used herein, the term "repellent" refers to any substance that acts to repel or prevent the infestation of pests (e.g., insects, nematodes, mollusks, endophytes, fungi, weeds, etc.). In some examples, the repellent is an insect repellent. Non-limiting examples of repellents that may be included in and / or formulated with the NLP compositions described herein include benzyl, benzyl benzoate, 2,3,4,5-bis(butyl-2-ene)tetrahydrofurfural (MGK Repellent 11), butoxypolypropylene glycol, N-butylacetanily, normal-butyl-6,6-dimethyl-5,6-dihydro-1,4-pyrone-2-carboxylate (Indalone), dibutyl adipate, dibutyl phthalate, di-normal-butyl succinate (Tabatrex), N,N-diethyl-meta-toluamide (DEET), dimethylcarbamate (endo,endo)-dimethylbicyclo[2.2.1]hept-5-ene-2,3-dicarboxylate), dimethyl phthalate, 2-ethyl-2-butyl-1,3-propanediol, 2-ethyl-1,3-hexanediol (Rutgers 612), di-n-propyl isocinchomeronate (MGK Repellent 326), 2-phenylcyclohexanol, p-methane-3,8-diol and n-propyl N,N-diethylsuccinamate, citronella oil, dimethyl phthalate, n-butyl mesityl oxide oxalate and 2-ethylhexanediol-1,3 (see Kirk-Othmer Encyclopedia of Chemical Technology, 2nd Ed., Vol. 11:724-728 and The Condensed Chemical Dictionary, 8th Ed., p 756).

[0230] Insect repellents that may be included in and / or formulated with the NLP compositions described herein may be synthetic or non-synthetic insect repellents. Examples of synthetic insect repellents include methyl anthranilate and other anthranilate insect repellents, benzaldehyde, DEET (N,N-diethyl-m-toluamide), dimethylcarbamate, dimethyl phthalate, icaridin (e.g., picaridin, Bayrepel, and KBR 3023), indalone (e.g., "6-2-2" mixture (60% dimethyl phthalate, 20% indalone, 20% ethyl hexanediol), IR3535 (3-[N-butyl-N-acetyl]-aminopropionic acid, ethyl ester), metofluthrin, permethrin, SS220, or tricyclodecenyl allyl ether. Examples of natural insect repellents include bark of red bark (Callicarpa) leaves, birch bark, and bay willow (Myrica Gale), catnip oil (e.g., nepetalactone), citronella oil, lemon eucalyptus (Corymbia citriodora) essential oil, p-menthane-3,8-diol (PMD), neem oil, lemongrass, tea tree oil from tea tree (Melaleuca alternifolia) leaves, tobacco or extracts thereof.

[0231] L. Fertilizer Some embodiments relate to NLP compositions described herein that include one or more heterologous fertilizing agents. In some instances, the heterologous fertilizing agent is associated with a formula that includes the NLP. In some embodiments, the NLP may encapsulate the heterologous fertilizing agent. In some embodiments, the heterologous fertilizing agent may be embedded in or conjugated to the surface of the NLP.

[0232] As used herein, the term "heterologous fertilizer" refers to a substance that can increase the fitness of plants or plant microorganisms (e.g., plant symbionts). Heterologous fertilizers include any material of natural or synthetic origin that is applied to soil or plant tissue to increase the fitness of plants and plant microorganisms. Heterologous fertilizers can stimulate the growth and activity of soil microbial populations. Increasing soil microbial populations (e.g., plant symbionts) can have a significant beneficial effect on the physical and chemical properties of the soil and can increase disease resistance and pest resistance. In some examples, heterologous fertilizers can be modified. For example, the modification can be a chemical modification, such as conjugation to a marker, such as a fluorescent marker or a radioactive marker. In other examples, the modification can include conjugation or operably linking to a moiety that enhances the stability, delivery, targeting, bioavailability, or half-life of the drug, such as a lipid, glycan, polymer (e.g., PEG), or cationic moiety.

[0233] Non-limiting examples of heterologous fertilizing agents that may be included in and / or formulated with the NLP compositions described herein include plant nutrients and plant growth regulators. In some embodiments, the heterologous fertilizing agent may be a peptide, polypeptide, nucleic acid, or polynucleotide capable of increasing the fitness of a plant or plant microorganism (e.g., plant symbiont). In some embodiments, the heterologous fertilizing agent may be a plant nutrient selected from macronutrients, micronutrients, or combinations thereof. Non-limiting examples of macronutrients include nitrogen, phosphorus, potassium, calcium, magnesium, and sulfur. Non-limiting examples of micronutrients include copper, iron, manganese, molybdenum, zinc, boron, silicon, cobalt, and vanadium. In some embodiments, the heterologous fertilizing agent may be a nitrogen fertilizer, including, but not limited to, urea, ammonium nitrate, ammonium sulfate, non-pressurized nitrogen solution, aqueous ammonia, anhydrous ammonia, ammonium thiosulfate, sulfur-coated urea, urea-formaldehyde, IBDU, polymer-coated urea, calcium nitrate, ureaform, and methylene urea. In some embodiments, the heterologous fertilizer may be a phosphorus fertilizer, such as diammonium phosphate, monoammonium phosphate, ammonium polyphosphate, concentrated superphosphate, and triple superphosphate. In some embodiments, the heterologous fertilizer may be a potassium fertilizer, such as potassium chloride, potassium sulfate, potassium-magnesium sulfate, and potassium nitrate. Such heterologous fertilizers may be present in the NLP composition or in the formulation of the NLP composition as free salts or ions.

[0234] In some embodiments, heterogeneous fertilizer agents can be classified as organic fertilizers or inorganic fertilizers. In some embodiments, inorganic fertilizers are derived from or produced from non-living materials. Non-limiting examples of heterogeneous inorganic fertilizer agents include ammonium nitrate, ammonium sulfate, urea, potassium chloride, potash, ammonium phosphate, anhydrous ammonia, and other phosphate salts. Organic fertilizers include fertilizers with molecular skeletons having a carbon backbone. In some embodiments, organic fertilizers are derived from or produced from living organisms. Non-limiting examples of heterogeneous organic fertilizer agents include animal manure, compost, bone meal, feather meal, and blood meal. One skilled in the art will understand that the exact amount of a given element in a fertilizer agent can be calculated and administered to plants or soil.

[0235] Some embodiments relate to methods and compositions for modifying plant nutrient mobility through soil by providing the plant nutrients in an NLP composition described herein. In some embodiments, a plant is contacted with an NLP composition comprising one or more plant nutrients in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of plant nutrient concentration in or on the plant, and (b) increase the fitness of the plant compared to an untreated plant. In some embodiments, a plant microorganism (e.g., a bacterial or fungal endosymbiont) is contacted with an NLP composition comprising one or more plant nutrients in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of plant nutrient concentration in or on the plant, and (b) increase the fitness of the plant microorganism compared to an untreated plant microorganism. In some embodiments, an NLP composition comprising one or more plant nutrients is applied to soil and migrates through the soil to contact the plant or plant microorganism.

[0236] Non-limiting examples of plant growth regulators that may be included in and / or formulated with the NLP compositions described herein include auxins, cytokinins, gibberellins (e.g., gibberellic acid), abscisic acid, amidochlor, ancymidol, 6-benzylaminopurine, brassinolide, butralin, chlormequat (e.g., chlormequat chloride), choline chloride, cyclanilide, daminozide, dikegulac, dimethipin, 2,6-dimethylpridine, ethephon, flumetralin, flumethicone ... Examples of plant growth regulators that can be incorporated into NLP compositions include luprimidol, fluthiacet, forchlorfenuron, inabenfide, indole-3-acetic acid, maleic hydrazide, mefluidide, mepiquat (mepiquat chloride), naphthaleneacetic acid, N-6-benzyladenine, paclobutrazol, prohexadione (prohexadione-calcium), prohydrojasmone, thidiazuron, triapentenol, tributyl phosphorotrithioate, 2,3,5-tri-iodobenzoic acid, trinexapac-ethyl, and uniconazole. Other plant growth regulators that can be incorporated into NLP compositions are described in U.S. Patent Application Publication No. 2012 / 0108431, which is incorporated by reference in its entirety.

[0237] M. Plant Modifier Some embodiments relate to NLP compositions described herein that comprise one or more heterologous plant modifiers. In some examples, the NLP composition comprises two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different plant modifiers. In some examples, the heterologous plant modifier is associated with a formula that includes the NLP. In some embodiments, the NLP may encapsulate the heterologous plant modifier. In some embodiments, the heterologous plant modifier may be embedded in or conjugated to the surface of the NLP.

[0238] As used herein, the term "plant modifier" refers to a substance that alters the phenotype of a plant to which it is provided, compared to a plant that has not been administered the plant modifier. In some examples, the plant modifier is a peptide. In some examples, the plant modifier is a nucleic acid. In some examples, the plant modifier may alter the phenotype of a variety of plants or target a phenotype to one or more specific plants (e.g., a specific species or genus of plant). In some examples, a heterologous plant modifier (e.g., an agent comprising a nucleic acid molecule or peptide) may be modified. For example, the modification may be a chemical modification, such as conjugation to a marker, such as a fluorescent marker or a radioactive marker. In some embodiments, the modification may include conjugation or operably linkage to a moiety that enhances the stability, delivery, targeting, bioavailability, or half-life of the plant modifier, such as a lipid, glycan, polymer (e.g., PEG), cationic moiety, etc. Non-limiting examples of heterologous plant modifiers (e.g., peptides, nucleic acids) that can be used are in the NLP compositions and methods disclosed herein.

[0239] N. Polypeptide Some embodiments relate to NLP compositions described herein that comprise one or more heterologous polypeptides. In some examples, the NLP composition comprises two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) heterologous polypeptides. In some examples, the NLP compositions described herein comprise a polypeptide or a functional fragment or derivative thereof that modifies a plant characteristic (e.g., increases plant fitness, confers insect resistance, enhances herbicide tolerance, etc.). In some examples, the heterologous polypeptide (or functional fragment or derivative thereof) is associated with a formula that comprises the NLP. In some embodiments, the NLP may encapsulate this heterologous polypeptide (or functional fragment or derivative). In some embodiments, this heterologous polypeptide (or functional fragment or derivative) may be embedded or conjugated to the surface of the NLP. Some embodiments relate to methods of modifying a characteristic of a plant by contacting the plant with an NLP composition comprising a heterologous polypeptide in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of polypeptide concentration, and (b) modify the plant (e.g., increase the fitness of the plant). One of skill in the art will understand that the suitable concentration of each heterologous polypeptide (or functional fragment or derivative thereof) in the NLP composition will depend on factors such as efficacy, stability of the polypeptide, number of individual polypeptides, formulation, and method of application of the NLP composition. In some examples, each polypeptide in a liquid formulation of the NLP composition is from about 0.1 ng / ml to about 100 mg / ml. In some examples, each polypeptide in a solid formulation of the NLP composition is from about 0.1 ng / g to about 100 mg / g.

[0240] Non-limiting examples of heterologous polypeptides (or functional fragments or derivatives thereof) that may be included in and / or formulated with the NLP compositions described herein include enzymes (e.g., metabolic recombinases, helicases, integrases, RNAses, DNAses, or ubiquitinating proteins), pore-forming proteins, signaling ligands, cell-penetrating peptides, transcription factors, receptors, antibodies, nanobodies, gene-editing proteins (e.g., CRISPR-associated proteins (Cas), TALENs, zinc fingers, meganucleases, etc.), riboproteins, protein aptamers, insecticidal proteins, and chaperones. Polypeptides that may be included in and / or formulated with the NLP compositions described herein may include naturally occurring or recombinantly produced polypeptides, functional fragments, and variants thereof. In some examples, the polypeptide may be a functional fragment or variant of a naturally occurring polypeptide (e.g., an enzymatically active fragment or variant thereof).

[0241] In some embodiments, the NLP composition comprises one or more antibodies and / or antigen-binding fragments thereof. In some embodiments, the heterologous functional agent included in the NLP compositions described herein may be an antibody that blocks or enhances a plant activity and / or function. In some embodiments, the antibody or functional fragment thereof included in the NLP composition may act as an antagonist or agonist of a polypeptide (e.g., an enzyme or cellular receptor) in the plant.

[0242] O. Nucleic acid Some embodiments relate to NLP compositions described herein that comprise one or more heterologous nucleic acids. In some examples, the NLP compositions comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) different heterologous nucleic acids. Non-limiting examples of nucleic acids that may be loaded / incorporated into the NLP compositions described herein and / or formulated with the NLP compositions by any of the methods described herein include deoxyribose nucleic acids (DNA), cDNA, such as plasmids, expression cassettes, protein-coding sequences, expression elements, editing templates, etc., ribose nucleic acids (RNA), such as mRNA, guide RNA (gRNA), inhibitory RNA molecules (e.g., siRNA, shRNA, microRNA (miRNA), etc.), DNA / RNA hybrid molecules, such as hybrid DNA / RNA guide molecules, peptide nucleic acids (PNAs), and precursors or derivatives of any of the foregoing (e.g., phosphorothioate-based molecules, such as deoxyribonucleic acid guanidine (DNG) and ribonucleic acid guanidine (RNG)). Nucleic acids that may be loaded / incorporated into the NLP compositions described herein and / or formulated with the NLP compositions by any of the methods described herein may be single-stranded, double-stranded, partially double-stranded, partially single-stranded, circular, linear, and / or comprise one or more non-standard, modified, or synthetic nucleotides. In some embodiments, nucleic acids included in and / or formulated with the NLP compositions described herein may be chemically modified (e.g., 2'-fluoro, 2'-o-methyl, 2'-deoxy, unlocked nucleic acid, 2'-hydroxy, phosphorothioate, 2'-thiouridine, 4'-thiouridine, 2'-deoxyuridine, etc.). In some embodiments, nucleic acids included in and / or formulated with the NLP compositions described herein comprise physiologically labile linkers that undergo chemical transformation (e.g., cleavage) when present in certain physiological conditions (e.g., disulfide bonds that are cleaved in the reducing environment of the cytoplasm of a cell). In some embodiments, the nucleic acids included in and / or formulated with the NLP compositions described herein are linked to the polymer via a physiologically labile bond or linker.

[0243] The NLP compositions described herein may include and / or be formulated with any number or type of heterologous nucleic acid. In some embodiments, the NLP compositions described herein may comprise and / or be formulated with one or more nucleic acids selected from the group consisting of a plasmid, a DNA molecule encoding an RNA, a DNA molecule encoding a polypeptide, an expression element, an expression vector, mRNA, siRNA, tRNA, a Dicer substrate small interfering RNA (dsiRNA), antisense RNA, small interfering RNA (siRNA), siRNA precursors (e.g., one or more strands of RNA that hybridize intermolecularly or intramolecularly to form at least partially double-stranded RNA), short hairpin RNA (shRNA), microRNA (miRNA), miRNA precursor, asymmetric interfering RNA (aiRNA), peptide nucleic acid (PNA), morpholino, locked nucleic acid (LNA), piwi-interacting RNA (piRNA), ribozyme, deoxyribozyme (DNAzyme), DNA aptamer, RNA aptamer, DNA / RNA hybrid aptamer, circular RNA (circRNA), guide RNA (gRNA), tracrRNA, CRISPR RNA (crRNA), single guide RNA (sgRNA), and precursors of any of the foregoing. Those skilled in the art will understand that the appropriate concentration of each nucleic acid in an NLP composition will depend on factors such as the efficacy and stability of the nucleic acids, the number of nucleic acids, the type of nucleic acid, the formulation, and the method of application of the NLP composition. Some embodiments relate to methods of providing an NLP composition comprising one or more nucleic acids to a plant, animal, plant cell, or animal cell by contacting the plant, animal, plant cell, or animal cell with the NLP composition comprising one or more nucleic acids in an amount and for a time sufficient to (a) achieve a target level (e.g., a predetermined or threshold level) of nucleic acid concentration, and (b) modify a characteristic of the plant, animal, plant cell, or animal cell (e.g., increase fitness).

[0244] Some embodiments relate to methods of reducing target gene expression levels and / or reducing protein levels in plants by providing to the plant surface (e.g., leaves, roots, seeds, etc.) an effective amount of an NLP composition comprising an inhibitory RNA (RNAi) molecule. Examples of RNAi molecules include, but are not limited to, small interfering RNA (siRNA), double-stranded RNA (dsRNA), short hairpin RNA (shRNA), partial duplex (meroduplex), Dicer substrate, and microRNA.

[0245] Some embodiments relate to methods for increasing the level of a protein in a plant by providing to the surface of the plant (e.g., leaves, roots, seeds, etc.) an effective amount of an NLP composition comprising a nucleic acid molecule encoding the protein. In some embodiments, an NLP composition comprising an enzyme (e.g., a metabolic recombinase, a helicase, an integrase, an RNAse, a DNAse, or a ubiquitinating protein), a pore-forming protein, a signaling ligand, a cell-penetrating peptide, a transcription factor, a receptor, an antibody, a nanobody, a gene-editing protein (e.g., a CRISPR-associated protein (e.g., Cas9, Cas12a, C1C2, etc.), a TALEN, a zinc finger, etc.), a riboprotein, a protein aptamer, or a chaperone is provided to the plant.

[0246] P. Gene Editing Some embodiments relate to NLP compositions described herein that include one or more components of a gene editing system. In some embodiments, the NLP compositions described herein include one or more zinc finger nucleases (ZFNs). In some embodiments, the NLP compositions described herein include one or more transcription activator-like effector-based nucleases (TALENs). In some embodiments, the NLP compositions described herein include one or more components of a clustered regularly interspaced short palindromic repeats (CRISPR) system. In some embodiments, the NLP compositions described herein comprise one or more components of a CRISPR gene editing system selected from the group consisting of a CRISPR-associated (Cas) protein (e.g., Cas9, Cas12a (also known as Cpfl), C2C1, C2C3, MAD7, etc.), a CRISPR RNA (crRNA), a trans-activating crRNA (tracrRNA), a chimeric single guide RNA (sgRNA), a prime editing guide RNA (pegRNA), and a donor template (e.g., a single-stranded or double-stranded DNA template comprising a desired nucleotide sequence to be inserted or knocked in at the double-stranded break). Non-limiting examples of CRISPR-associated proteins that may be included in and / or formulated with the NLP compositions described herein include Cas9 (e.g., wild-type Cas9, nickase Cas9 (e.g., Cas9 D10A), dead (catalytically inactive) Cas9 (dCas9), eSpCas9, etc.), Cas12a (e.g., AsCas12a (from Acidaminococcus sp.), LbCas12a (from Lachnospiraceae), etc.), Cas12b (e.g., AsCas12a (from Acidaminococcus sp.), LbCas12a (from Lachnospiraceae), etc.), Cas12c (e.g., Cas12b (from Lachnospiraceae), etc.), Cas12d (e.g., Cas12c (from Lachnospiraceae), etc.), Cas12e (e.g., Cas12a (from Lachnospiraceae), etc.), Cas12f (e.g., Cas12a (from Lachnospiraceae), etc.), Cas12g ... Examples of such dCas9 include Cas9 derived from Caspase sp.), CasX (Cas12e), Cas13a, Cas14, C2C1, C2C3, CasΦ, dCas9 conjugated with an effector that suppresses the expression of a target gene (CRISPRi) (e.g., a KRAB domain, SID4X, etc.), dCas9 conjugated with an effector that activates the expression of a target gene (CRISPRa) (e.g., VP64, p65 activation domain (p65D), etc.), and nucleic acids encoding any of the above.

[0247] III.How to use The NLP compositions described herein are useful in a variety of agricultural or therapeutic methods. Examples of methods of using the NLP compositions are further described below.

[0248] A. Delivery to Plants Some embodiments relate to methods of delivering NLP compositions and / or formulations comprising one or more NLP compositions to a plant, for example, by contacting the plant, parts thereof, or the environment in which the plant is located (e.g., soil) with the NLP compositions and / or formulations comprising the NLP compositions. In some embodiments, the NLP compositions described herein comprise one or more heterofunctional agents selected from the group consisting of pesticides, antibacterial agents, antifungal agents, nematicides, molluscicides, virucides, herbicides, pest control agents (e.g., repellents), fertilizers, and plant modifiers.

[0249] Some embodiments relate to methods of increasing the fitness of a plant, the methods comprising delivering to a plant an effective amount of one or more NLP compositions described herein to increase the fitness of the plant compared to an untreated plant (e.g., a plant to which the NLP composition has not been delivered). The increase in plant fitness as a result of delivery of an NLP composition can, in some methods, be manifested as, for example, improved yield (e.g., biomass, grain yield, seed yield, fruit yield, protein content, carbohydrate content, oil content, leaf area, etc.), increased plant vigor (e.g., increased tolerance of abiotic or biotic stress, increased resistance to pests, increased germination rate, etc.), or an increase in the quality of the product harvested from the plant by a measurable amount over the fitness of the plant compared to not applying an NLP composition or applying a conventional agricultural chemical. In some embodiments, delivery of an effective amount of one or more NLP compositions described herein to a plant can increase yield by at least about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, or more than 100%. Yield may be expressed in terms of the amount by weight or volume of the plant or product of the plant on any basis. Increased plant fitness as a result of delivery of an NLP composition can also be measured by other methods such as vigor assessment, stand size (number of plants per unit area), plant height, stem circumference, stem length, leaf number, leaf size, plant canopy, appearance (such as green leaf color), root assessment, emergence, protein content, increased tillers, larger leaves, more leaves, more dead basal leaves, stronger tillers, reduced fertilizer requirements, reduced seed requirements, more productive tillers, earlier flowering, earlier grain or seed maturity, reduced plant lodging (lodging), increased shoot growth, earlier germination, or any combination of these factors, in a measurable or significant amount for the same factor in plants produced under the same conditions but without administration of the composition or application of a conventional agricultural chemical.In some embodiments, delivering an effective amount of one or more NLP compositions described herein to a plant introduces or increases (e.g., about 1%, 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more than 100%) a beneficial trait in the plant compared to an untreated plant. In some examples, the increase in plant fitness is an increase (e.g., about 1%, 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more) in disease resistance, drought tolerance, heat tolerance, cold tolerance, salt tolerance, metal tolerance, herbicide tolerance, chemical tolerance, water use efficiency, nitrogen utilization, tolerance to nitrogen stress, nitrogen fixation, pest resistance, herbivore resistance, pathogen resistance, yield, yield under water-limited conditions, vigor, growth, photosynthetic capacity, nutrition, protein content, carbohydrate content, oil content, biomass, shoot length, root length, root structure, seed weight or amount of harvestable produce.

[0250] Some embodiments relate to methods of increasing the fitness of a plant, comprising contacting one or more of a seed, protoplast, embryo, leaf, root, stem, tissue (e.g., meristematic or meristem) of the plant with an effective amount of an NLP composition disclosed herein, wherein the fitness of the plant is increased (e.g., by about 1%, 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more than 100%) compared to an untreated plant.

[0251] Some embodiments relate to a method of reducing the fitness of a plant (e.g., a weed) or plant part (e.g., reproductive tissue), comprising contacting one or more of a plant's seeds, protoplasts, embryos, leaves, roots, stems, or tissues (e.g., meristems, reproductive tissues) with an effective amount of an NLP composition comprising one or more herbicides. When a herbicide is included in the NLP composition, the method may further be used to reduce weed fitness or kill weeds. When a herbicide is included in the NLP composition provided to a plant's reproductive tissue, the method may further be used to prevent pollen production. In some embodiments, the method may be effective to reduce weed fitness by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more compared to untreated weeds (e.g., weeds to which the NLP composition has not been administered). For example, the methods may be effective in killing weeds, thereby reducing weed populations by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more compared to untreated weeds. In some examples, the methods substantially eliminate weeds. Examples of weeds that can be treated according to the methods are further described herein. When the NLP composition provided to the reproductive tissue of a plant includes a herbicide, the methods may further be used to prevent pollen production or germination. In some embodiments, the methods may be effective in reducing pollen production by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more compared to an untreated plant (e.g., a plant to which the NLP composition has not been administered).

[0252] The uptake of NLPs into plant cells can be measured by various methods known in the art. For example, the NLP or a component thereof can be labeled with a marker (e.g., a fluorescent marker) that can be detected in isolated cells to confirm uptake. For example, cellular uptake can be detected based on intracellular fluorescence intensity, which can be determined, for example, by microscopy, e.g., using confocal microscopy. Uptake can also be determined as a measure of fitness, e.g., the fitness of a plant containing treated cells. For example, the efficacy of the present compositions and methods can be determined by comparing the change in fitness of plants treated with an NLP containing a heterologous functional agent, e.g., a herbicide, to treatment of plants treated with an NLP without a herbicide.

[0253] i.Plants Various plants can be contacted with one or more NLP compositions described herein. The NLP compositions described herein can be provided to plants according to any method known in the art. In some embodiments, the NLP compositions described herein can be provided to whole plants, including but not limited to vegetative organs / structures of shoots (e.g., leaves, stems, and tubers), roots, flowers and floral organs / structures (e.g., bracts, sepals, petals, stamens, carpels, anthers, and ovules), seeds (including embryos, endosperm, cotyledons, and seed coats) and fruits (mature ovaries), plant tissues (e.g., meristems, vascular tissue, ground tissue, etc.), and cells (e.g., guard cells, egg cells, etc.) and their progeny. Classes of plants that can be treated with the methods or NLP compositions disclosed herein include angiosperms (monocotyledons and dicotyledons), gymnosperms, ferns, horsetails, ancient spiraea, microphytes, bryophytes, and algae (e.g., multicellular or unicellular algae).Plants that may be treated according to the present method further include any vascular plant, such as a monocotyledonous or dicotyledonous or gymnosperm, including, but not limited to, alfalfa, apple, Arabidopsis, banana, barley, canola, castor, chrysanthemum, clover, cocoa, coffee, cotton, cottonseed, corn, crambe, cranberry, Brassicaceae, cucumber, Dendrobium, yam, Eucalyptus, fescue, flax, gladiolus, liliacea, linseed, millet, muskmelon, mustard, oat, oil palm, canola or rapeseed, papaya, peanut, pineapple, ornamentals, Phaseolus, potato, rapeseed, rice, rye, ryegrass, safflower, sesame, sorghum, soybean, sugar Included are: sugarcane, sunflower, strawberry, tobacco, tomato, turfgrass, wheat and vegetable crops such as lettuce, celery, broccoli, cauliflower, cucurbits, fruit and nut trees such as apple, pear, peach, orange, grapefruit, lemon, lime, almond, pecan, walnut, hazel, vines such as grapes (e.g., vineyard), kiwi, hops, cannabis, fruit bushes and brambles, such as raspberry, blackberry, gooseberry, forest trees such as ash, pine, fir, maple, oak, chestnut, poplar, as well as alfalfa, canola, castor, corn, cotton, crambe, flax, linseed, mustard, oil palm, rapeseed, peanut, potato, rice, safflower, sesame, soybean, sugar beet, sunflower, tobacco, tomato and wheat.

[0254] Some embodiments relate to methods of providing one or more NLP compositions described herein to crop plants. Crop plants include, for example, plants grown for fodder, plants grown for oil (e.g., oilseeds), plants grown for grain (e.g., wheat, millet, barley, rye, etc.), plants grown for fruit, vegetables, plants grown for fiber, spice crops, plants grown for nuts, plants grown for timber, etc. In particular examples, the crop plant treated with the method is a soybean plant. In other particular examples, the crop plant is wheat. In particular examples, the crop plant is corn. In particular examples, the crop plant is cotton. In particular examples, the crop plant is alfalfa. In particular examples, the crop plant is sugar beet. In particular examples, the crop plant is rice. In particular examples, the crop plant is potato. In particular examples, the crop plant is tomato.

[0255] Examples of such crop plants include, but are not limited to, monocotyledonous and dicotyledonous plants, including, but not limited to, Acer, Allium, Amaranthus, Ananas comosus, celery (Apium graveolens), Arachis, Asparagus (Asparagus officinalis), beet (Beta vulgaris), Brassica (e.g., Brassica napus, Brassica rapa (canola, rapeseed, turnip rape)), Camellia sinensis, Canna indica, Cannabis sativa, Capsicum, Castanea, endive, Cichorium endivia, Citrullus lanatus, Citrus genus, Cocos genus, Coffea genus, Coriander genus, Corylus genus, Crataegus genus, Cucurbita genus, Cucumis genus, Daucus carota genus, Fagus genus, Ficus carica genus, Fragaria genus, Ginkgo biloba genus, Glycine genus (e.g., Glycine max, Soja hispida, or Soja max), Gossypium hirsutum genus, hirsutum, Helianthus (e.g., Helianthus annuus), Hibiscus, Hordeum (e.g., Hordeum vulgare), Ipomoea batatas, Juglans, lettuce (Lactuca sativa), flax (Linumusitatissimum, Litchi chinensis, Lotus, Luffa acutangula, Lupinus, Lycopersicon (e.g., Lycopersicon esculenturn, Lycopersicon lycopersicum, Lycopersicon pyriforme), Malus, Alfalfa (Medicago sativa), Mentha, Miscanthus sinensis, Morus nigra, Musa, Nicotiana, Olea, Oryza (e.g., Oryza sativa), sativa, Oryza latifolia, Panicum miliaceum, Panicum virgatum, Passiflora edulis, Petroselinum crispum, Phaseolus, Pinus, Pistacia vera, Pisum, Poa, Populus, Prunus, Pyrus communis, Quercus, Raphanus sativus, Rheum rhabarbarum, Ribes, Ricinus communis, Rubus, Saccharum, Salix, Sambucus, Secale cereale, Sesamum, Sinapis, Solanum (e.g., potato, Solanum tuberosum, Solanum integrifolium),integrifolium or tomato (Solanum lycopersicum), sorghum (Sorghum bicolor), Sorghum halepense, spinach (Spinacia), Tamarindus indica, cacao (Theobroma cacao), Trifolium, Triticosecale rimpaui, Triticum (e.g. Triticum aestivum, Triticum durum, Triticum turgidum, Triticum hybernum, Triticum macha, Triticum sativum or Triticum vulgare), The present invention also includes a forage or fodder legume, ornamental plant, food crop, tree, or shrub selected from the group consisting of Vaccinium vulgare, Vicia, Vigna, Viola odorata, Vitis, and Zea mays. In certain embodiments, the crop plant is rice, oilseed rape, canola, soybean, corn (maize), cotton, sugarcane, alfalfa, sorghum, or wheat. In certain examples, the compositions and methods may be used to treat post-harvest plants or plant parts, food, or feed products.

[0256] In some examples, the food or feed product is a non-plant food or feed product (eg, a product that can be consumed by humans, animals, or livestock (eg, a mushroom)).

[0257] Some embodiments relate to methods and compositions for reducing weed fitness or killing weeds. In some embodiments, NLP compositions comprising one or more herbicides are used to reduce weed fitness or kill weeds. In some embodiments, NLP compositions comprising one or more herbicides reduce weed fitness by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more. In some embodiments, NLP compositions comprising one or more herbicides are used to reduce weed populations in a treated area compared to an untreated area by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. In some embodiments, the NLP compositions comprising one or more herbicides are used to combat monocotyledonous plants (e.g., Agrostis, Alopecurus, Avena, Bromus, Cyperus, Digitaria, Echinochloa, Lolium, Monochoria, Rottboellia, Sagittaria, Scirpus, Setaria, Sicirpus ... da) or Sorghum) or dicotyledonous plants (Abutilon, Amaranthus, Chenopodium, Chrysanthemum, Conyza, Galium, Ipomoea, Nasturtium, Sinapis, Solanum, Stellaria, Veronica, Viola or Xanthium).In some embodiments, an NLP composition comprising one or more herbicides is applied to control Lolium rigidum, Amaranthus palmeri, Abutilon theopratsi, Sorghum halepense, Conyza canadensis, Setaria verticillata, Capsella pastoris, and Cyperus rotundas.

[0258] Plants or plant parts that can be contacted with the NLP compositions described herein include plants at any stage of plant development. In certain instances, delivery can occur during germination, seedling growth, vegetative growth, and reproductive growth. In certain instances, delivery to a plant occurs during the vegetative and reproductive growth stages. Alternatively, delivery can occur to a seed. Vegetative and reproductive growth stages are also referred to herein as "adult" or "mature" plants.

[0259] B. Delivery to Plant Pests Some embodiments relate to methods and compositions for controlling plant pests. In some embodiments, the plant pest is contacted with an NLP composition comprising a heterofunctional agent, such as a pesticide (e.g., an antibacterial agent, antifungal agent, nematicide, molluscicide, virucide, herbicide, etc.) or a pest control agent (e.g., a repellent). Some embodiments relate to methods and compositions for reducing the fitness of the pest, for example, for preventing or treating pest infestation as a result of delivery of the NLP composition.

[0260] Some embodiments relate to methods and compositions for reducing a fungal infection in a plant having a fungal infection. In some embodiments, the plant is contacted with an NLP composition comprising one or more antifungal agents. In some examples, the antifungal agent is a nucleic acid that inhibits expression of genes (e.g., dell and dcl2 (e.g., dcH / 2)) in the fungus causing the fungal infection. In some examples, the fungal infection is caused by a fungus belonging to the genus Sclerotinia spp. (e.g., Sclerotinia sclerotiorum), Botrytis spp. (e.g., Botrytis cinerea), Aspergillus spp., Fusarium spp., or Penicillium spp.In some embodiments, the NLP compositions comprising one or more antifungal agents are effective against white rust (Albugo candida), downy mildew, powdery mildew, clubroot (Plasmodiophora brassicae), Pythium species, Sclerotinia rot (e.g., S. sclerotiorum, S. minor), Sclerotium rolfsii, S. cepivorum), Fusarium wilts and rot (various Fusarium species, including F. solani and F. oxysporum), gray mold (e.g., Botrytis cinerea), and The method is applied to prevent or treat a fungal disease selected from the group consisting of Pythium, Rhizoctonia, Phytophthora, Fusarium, Aphanomyces, Pythium sulcatum, Alternaria solani, leaf rust, Fusarium head blight, wheat leaf spot, stripe rust, spot blotch, and yellow spot. In some examples, the method reduces or substantially eliminates the fungal infection.

[0261] Some embodiments relate to methods and compositions for reducing bacterial infection in plants having a bacterial infection. In some embodiments, the plant is contacted with an NLP composition comprising one or more antimicrobial agents. In some examples, the antimicrobial agent is streptomycin. In some examples, the bacterial infection is caused by a bacterium belonging to the genus Pseudomonas (e.g., Pseudomonas syringae). In some examples, the antimicrobial agent is oxytetracycline (OTC). In some examples, the bacterial infection causes citrus greening (huanglongbing). In some examples, the antimicrobial agent is selected from the group consisting of amoxicillin, enrofloxacin, chloramphenicol, penicillin, oxytetracycline, quinolones, sulfonamides, sulfamethazine, sulfadimidine, sulfamethoxazole, and tetracycline. In some embodiments, the NLP compositions comprising one or more antimicrobial agents are selected from the group consisting of Actinobacteria, Agrobacterium tumefaciens, Burkholderiaceae, Clavibacter (e.g., Clavibacter michiganensis, Clavibacter sepedonicus), Corynebacterium, Dickeya (e.g., Dickeya dadantii, Dickeya solani), Erwinia (e.g., Erwinia amylovora, E. carotovora), and the like.carotovora), Enterobacteriaceae, Microbacteriaceae, Pectobacterium (e.g., Pectobacterium carotovorum, Pectobacterium atrosepticum), Pseudomonas (e.g., Pseudomonas syringae, Pseudomonas savastanoi), Proteobacteria, Rhizobiaceae, Ralstonia The method is applied to prevent or treat bacterial diseases caused by bacterial pathogens selected from the group consisting of Bacillus solanacearum, Streptomyces, Xanthomonas (e.g., Xanthomonas axonopodis, Xanthomonas campestris, Xanthomonas oryzae), and Xylella (e.g., Xylella fastidiosa). In some examples, the method reduces or substantially eliminates bacterial infection.

[0262] Some embodiments relate to methods and compositions for reducing the fitness of insect plant pests. In some embodiments, the insect plant pest is contacted with an NLP composition comprising one or more insecticides. In some embodiments, the plant is contacted with the NLP, which then delivers the insecticide to the insect pest, for example, upon ingestion of the plant by the insect pest. In some examples, the insecticide is a peptide nucleic acid. In some examples, the insecticide is an insecticidal peptide. In some examples, the insect plant pest is an aphid. In some examples, the insect plant pest is a lepidopteran (e.g., Spodoptera frugiperda). In some examples, the insect plant pest is an arachnid, such as a mite. In some embodiments, an NLP composition comprising one or more insecticides is provided to an insect pest selected from the group consisting of Asian citrus psyllid, Asian longhorned beetle, emerald borer, grasshopper, false codling moth, armyworm, brown marmorated stink bug, cotton bollworm, silverleaf whitefly, diamondback moth, red flour beetle, green peach aphid, cotton aphid, taro caterpillar, thrips, flea beetle, Colorado potato beetle, corn rootworm, tomato hornworm, weevil, and brown planthopper. In some examples, the method reduces the fitness of the insect plant pest compared to an untreated insect plant pest.

[0263] Some embodiments relate to methods and compositions for reducing the fitness of nematode plant pests. In some embodiments, the nematode plant pest is contacted with an NLP composition comprising a nematicide. In some embodiments, a plant at risk of or having a nematode infestation is contacted with an NLP composition comprising a nematicide. In some examples, the nematicide is a neuropeptide (e.g., Mi-NLP-15b). In some examples, the nematode plant pest is a root-knot nematode. In some embodiments, the NLP compositions comprising one or more nematicides are effective against root-knot nematodes (e.g., Meloidogyne spp.), cyst nematodes (e.g., Heterodera and Globodera spp.), root-lesion nematodes (e.g., Pratylenchus spp.), banana root-miner nematode (Radopholus similis), stem nematode (Ditylenchus dipsaci), pine wood nematode (e.g., Bursaphelenchus xylophilus), false root-knot nematode (e.g., Rotylenchulus reniformis), grape spur nematode (Xiphinema index), false root-knot nematode (Nacobbus In some instances, the method reduces the fitness of the nematode plant pest compared to an untreated nematode plant pest.

[0264] Some embodiments relate to methods and compositions for reducing the fitness of weeds. In some embodiments, weeds are contacted with an NLP composition comprising a herbicide (e.g., glufosinate). In some examples, the weed is goosegrass. In some examples, the NLP composition comprises an agent (e.g., an RNAi agent targeting 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase) that increases the susceptibility of the weed to the herbicide compared to untreated weeds. In some embodiments, an NLP composition comprising one or more herbicides (and / or agents that increase the susceptibility of a weed to a herbicide) is provided to weeds selected from the group consisting of dandelion, tiger nut, horsetail, horsenettle, kochia, thistle, artemisia, morning glory, prairie cup grass, pokeweed, powell amaranth, quackgrass, ragweed, sedge, wild buckwheat, dayflower species, pigweed (Palmer amaranth), water chestnut, vetch, and native plants.

[0265] Some embodiments relate to methods and compositions for reducing the fitness of mollusks. In some embodiments, a mollusk is contacted with an NLP composition comprising an active agent that reduces the fitness of the mollusk. In some embodiments, the NLP compositions described herein are delivered to a mollusk by contacting the mollusk with the NLP composition. In some embodiments, the NLP compositions described herein are delivered to a plant that is at risk of or has a mollusk infestation. In some embodiments, an NLP composition comprising one or more active agents that reduce the fitness of a mollusk is provided to a mollusk selected from the group consisting of Achatinidae, Agriolimacidae, Ampullariidae, Arionidae, Bradybaenidae, Helicidae, Hydromiidae, Lymnaeidae, Milacidae, Urocyclidae, or Veronicellidae.

[0266] Some embodiments relate to methods and compositions that are effective in reducing the ability of a pest to carry or transmit a plant pathogen (e.g., a plant virus (e.g., TYLCV) or a plant bacterium (e.g., Agrobacterium) compared to a pest that has not been administered an NLP composition. The methods and compositions provided herein can be effective in reducing the ability of a pest to carry or transmit a plant pathogen (e.g., a plant virus (e.g., TYLCV) or a plant bacterium (e.g., Agrobacterium)) by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% compared to a baseline level (e.g., the level found in a pest that has not been administered an NLP composition).

[0267] C. Delivery to Plant Microbes Some embodiments relate to methods and compositions for increasing the fitness of plant microorganisms, such as symbionts that are beneficial to plant fitness (e.g., bacterial endosymbionts, fungal endosymbionts, pollinating insects, etc.). In some embodiments, a plant or plant symbiont is contacted with an NLP composition comprising a heterofunctional agent (e.g., a fertilizer, a nutritional supplement, etc.) that increases the fitness of the microorganism compared to an untreated microorganism (e.g., a microbial symbiont that has not received the NLP composition). In one aspect, an endosymbiont is contacted with an NLP composition described herein comprising a heterofunctional agent that increases the fitness of the symbiont compared to an untreated endosymbiont. In some examples, a method or composition provided herein may be effective in increasing the resistance of a symbiont to a parasite or pathogen (e.g., a fungal, bacterial, or viral pathogen, or a parasitic mite (e.g., Varroa destructor mite in honeybees)) compared to a symbiont that has not received the NLP composition. In some examples, a method or composition provided herein may be effective to increase the resistance of a symbiont to a pathogen or parasite (e.g., a fungal, bacterial, or viral pathogen or a parasitic mite (e.g., Varroa destructor dite in honeybees)) by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% compared to a reference level (e.g., the level found in a symbiont that does not receive an NLP composition). In some embodiments, the plant symbiont treated with the NLP compositions described herein is an endosymbiotic fungus, such as an Aspergillaceae, Ceratobasidiaceae, Coniochaetaceae, Cordycipitaceae, Corticiaceae, Cystofilobasidiaceae, Davidiellaceae, Debaryomycetaceae, Dothioraceae, Erysiphaceae, Filobasidiaceae,The fungus may be of the genus Glomerellaceae, Hydnaceae, Hypocreaceae, Leptosphaeriaceae, Montagnulaceae, Mortierellaceae, Mycosphaerellaceae, Nectriaceae, Orbiliaceae, Phaeosphaeriaceae, Pleosporaceae, Pseudeurotiaceae, Rhizopodaceae, Sclerotiniaceae, Stereaceae or Trichocomacea. In some embodiments, the plant symbionts treated with the NLP compositions described herein are endobiotic bacteria, such as Acetobacteraceae, Acidobacteriaceae, Acidothermaceae, Aerococcaceae, Alcaligenaceae, Alicyclobacillaceae, Alteromonadaceae, Anaerolineaceae, Aurantimonadaceae, Bacillus subtilis ... Bacillaceae, Bacteriovoracaceae, Bdellovibrionaceae, Bradyrhizobiaceae, Brevibacteriaceae, Brucellaceae, Burkholderiaceae, Carboxydocellaceae, Caulobacteraceae, Cellulomonadaceae, Chitinophagaceae,Chromatiaceae, Chthoniobacteraceae, Chthonomonadaceae, Clostridiaceae, Comamonadaceae, Corynebacteriaceae, Coxiellaceae, Cryomorphaceae, Cyclobacteriaceae, Cytophagaceae, Deinococcaceae, Dermabacteraceae, Der macoccaceae, Enterobacteriaceae, Enterococcaceae, Erythrobacteraceae, Fibrobacteraceae, Flammeovirgaceae, Flavobacteriaceae, Frankiaceae, Fusobacteriaceae, Gaiellaceae, Gemmatimonadaceae, Geodermatophilaceae, Glycorniellace ... corny cetaceae, Haliangiaceae, Halomonadaceae, Holosporaceae, Hyphomicrobiaceae, Iamiaceae, Intrasporangiaceae, Kineosporiaceae, Koribacteraceae, Lachnospiraceae, Lactobacillaceae, Legionellaceae, Leptospiraceae, Leuconostocaceae,Methylobacteriaceae, Methylocystaceae, Methylophilaceae, Microbacteriaceae, Micrococcaceae, Micromonosporaceae, Moraxellaceae, Mycobacteriaceae, Mycoplasmataceae ae), Myxococcaceae, Nakamurellaceae, Neisseriaceae, Nitrosomonadaceae, Nocardiaceae, Nocardioidaceae, Oceanospirillaceae, Opitutaceae, Oxalobacteraceae, Paenibacillaceae acillaceae), Parachlamydiaceae, Pasteurellaceae, Patulibacteraceae, Peptostreptococcaceae, Phyllobacteriaceae, Piscirickettsiaceae, Planctomycetaceae, Planococcaceae, Polyangiaceae, Porphyromonadaceae, Prevotellaceae, Promicromonosporaceae, Pseudomonadaceae, Pseudonocardiaceae, Rhizobiaceae, Rhodobacteraceae, Rhodospirillaceae,Roseiflexaceae, Rubrobacteriaceae, Sandaracinaceae, Sanguibacteraceae, Saprospiraceae, Segniliparaceae, Shewanellaceae, Sinobacteraceae, Solibacteraceae, Solimonadaceae, Solirubrobacteraceae, Sphingobacteriaceae, Sphingomonadaceae, The bacterium may be from the family Spiroplasmataceae, Sporichthyaceae, Sporolactobacillaceae, Staphylococcaceae, Streptococcaceae, Streptomycetaceae, Syntrophobacteraceae, Veillonellaceae, Verrucomicrobiaceae, Weeksellaceae, Xanthobacteraceae or Xanthomonadaceae.

[0268] Some embodiments relate to methods and compositions for increasing the fitness of insects beneficial to plants (e.g., insect symbionts of plants). In some embodiments, the beneficial insects are contacted with an NLP composition comprising a heterologous functional agent that increases the fitness of the beneficial insect compared to an untreated insect. For example, the host can include insects used in agricultural applications, including insects that aid in crop pollination, seed dispersal, or pest control. In some examples, the insect is a plant pollinating insect. For example, the insect can be of the genus Hymenoptera or Diptera. In some examples, the insect of the genus Hymenoptera is a bee. In other examples, the insect of the genus Diptera is a fly.

[0269] D. Delivery to Animal Pathogens Some embodiments relate to methods and compositions for controlling animal (e.g., human) pathogens. In some embodiments, the animal pathogen is contacted with an NLP composition comprising a heterofunctional agent (e.g., an antibacterial, antifungal, insecticide, nematicide, antiparasitic, antiviral, etc.). As used herein, the term "pathogen" refers to an animal, such as a microorganism or invertebrate, that causes disease or disease symptoms in an animal host by, for example, (i) directly infecting the animal, (ii) producing a substance that causes disease or disease symptoms in the animal (e.g., bacteria that produce pathogenic toxins, etc.), and / or (iii) eliciting an immune (e.g., inflammatory response) in the animal (e.g., biting insects, e.g., bed bugs). Non-limiting examples of pathogens that can be controlled by application of NLP compositions comprising heterologous functional agents include bacteria (e.g., Streptococcus spp., Pneumococcus spp., Pseudomonas spp., Salmonella spp., Campylobacter spp. or Escherichia spp. or Candida spp.), parasitic insects (e.g., Cimex spp.), parasitic nematodes (e.g., Heligmosomoides spp.), parasitic protozoans (e.g., Trichomoniasis spp.), protozoans, fungi, nematodes, insects, viroids, and viruses. In some embodiments, NLP compositions comprising heterologous functional agents may be useful for reducing the fitness of animal pathogens, for example, preventing or treating pathogen infection or controlling the spread of the pathogen as a result of delivery of the NLP composition. In some embodiments, the method comprises delivering the NLP composition to at least one habitat where the pathogen grows, survives, reproduces, feeds, or infests. In some examples, the NLP composition is delivered as a pathogen-ingestible composition for ingestion by the pathogen. Pathogen fitness may be assessed using any standard method in the art. In some examples, pest fitness may be assessed by assessing individual pathogens.Alternatively, pest fitness can be assessed by assessing pathogen populations. For example, a decrease in pathogen fitness can be manifested as a decrease in successful competition with other pathogens, thereby resulting in a decrease in pathogen population size. The NLP compositions and related methods described herein are useful for reducing the fitness of animal pathogens, thereby treating or preventing infectious diseases in animals.

[0270] Some embodiments relate to methods and compositions for preventing or treating fungal infections in animals. In some embodiments, an NLP composition comprising one or more antifungal agents is provided to a fungus or an animal at risk of having a fungal infection. The NLP compositions and related methods described herein are effective against fungi from the phylum Ascomycota (Fusarium oxysporum, Pneumocystis jirovecii, Aspergillus spp., Coccidioides immitis / posadasii, Candida albicans), Basidiomycota (Filobasidiella neoformans, Trichosporon), Chytridiomycota, Microsporidia (Encephalitozoon cuniculi), and the phylum Pseudomonas (Pseudomonas aeruginosa, P ... The present invention is suitable for the treatment or prevention of fungal infections in animals, including infections caused by fungi belonging to the ...

Claims

1. 1. An agricultural composition comprising: at least one phospholipid, at least one non-polar lipid, and At least one surface modifier 1. An agricultural composition comprising a plurality of naturally occurring lipid particles (NLPs) each comprising:

2. 2. The agricultural composition according to claim 1, wherein the at least one phospholipid is selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid, phosphatidylserine and 1,2-dimyristoyl-sn-glycero-3-phosphate.

3. 2. The agricultural composition of claim 1, wherein the at least one phospholipid is derived from lecithin.

4. 2. The agricultural composition of claim 1, wherein the at least one non-polar lipid comprises at least one fatty acid chain selected from the group consisting of polyunsaturated fatty acid chains, monounsaturated fatty acid chains, and saturated fatty acid chains.

5. 2. The agricultural composition of claim 1, wherein the NLP comprises at least one phospholipid layer.

6. 6. The agricultural composition of claim 5, wherein the NLP comprises at least one phospholipid bilayer.

7. 2. The agricultural composition of claim 1, wherein the NLP has a micellar structure.

8. 2. The agricultural composition of claim 1, wherein the hydrophobic core comprises at least one non-polar lipid.

9. 10. The agricultural composition of claim 1, wherein the hydrophobic core is a solid.

10. 10. The agricultural composition of claim 1, wherein the surface modifier is incorporated into a phospholipid layer.

11. 2. The agricultural composition of claim 1, wherein the surface modifier is selected from the group consisting of glycolipids, polysaccharides, fatty acid ethoxylates, linear alcohol ethoxylates, cetyltrimethyl, linear isopropylamine dodecylbenzenesulfonate, tristyrylphenol ethoxylate phosphate esters, modified styrene acrylic copolymers, hydrophobically modified polycarboxylate polymers, anionic polymers, nonionic acrylic copolymers, nonionic combination polymers, tristyrylphenol polyalkylene oxide block copolymers or headgroup-modified PEG lipids.

12. 12. The agricultural composition of claim 11, wherein the head group-modified PEG lipid is PEG2000-C18 or PEG5000-C18.

13. 12. The agricultural composition according to claim 11, wherein the glycolipid is a rhamnolipid or a sophorolipid.

14. 12. The agricultural composition of claim 11, wherein the anionic polymer is Atlox 500L, Atlox 4917 or Atlox CS100B.

15. 12. The agricultural composition according to claim 11, wherein the polysaccharide is a C8 to C10 alkyl polysaccharide.

16. 10. The agricultural composition of claim 1, wherein the surface modifier stabilizes the integrity of the NLP.

17. 10. The agricultural composition of claim 1, wherein the surface modifier affects the binding of the NLP to one or more components present in soil.

18. 10. The agricultural composition of claim 1, wherein the surface modifier affects the affinity of the NLP for one or more components present in soil.

19. 10. The agricultural composition of claim 1, wherein the surface modifier affects the surface charge of the NLP.

20. 10. The agricultural composition of claim 1, wherein the NLP exhibits a negative surface charge as evidenced by a negative zeta potential.

21. 21. The agricultural composition according to claim 20, wherein the negative zeta potential is in the range of −10 to −100 mV.

22. 22. The agricultural composition of claim 21, wherein the negative zeta potential increases the mobility of the NLPs through the soil.

23. 10. The agricultural composition of claim 1, further comprising a co-solvent.

24. 24. The agricultural composition of claim 23, wherein the co-solvent is selected from the group consisting of fatty acid methyl esters, non-ionic emulsifiers, propylene glycol, ethyl lactate, non-ionic block copolymer surfactants or non-ionic polyalkylene glycol ethers, dichloromethane and isopropyl myristate.

25. 10. The agricultural composition of claim 1, further comprising one or more excipients.

26. 26. The agricultural composition of claim 25, wherein the one or more excipients are selected from the group consisting of ethyl lactate, Atlas G5002L and polyethylene glycol.

27. 27. The agricultural composition according to any one of claims 1 to 26, further comprising at least one heterofunctional agent.

28. 28. The agricultural composition of claim 27, wherein the heterofunctional agent is selected from the group consisting of pesticides, fertilizers, herbicides, plant modifiers, insect attractants, plant growth promoters, biostimulants and plant immunity inducers.

29. 29. The agricultural composition of claim 28, wherein the pesticide is selected from the group consisting of antifungals, antioomycetes, antibacterials, insecticides, molluscicides, nematicides, herbicides and virucides.

30. (a) The antifungal agent is selected from the group consisting of azoxystrobin, mancozeb, prothioconazole, folpet, tebuconazole, difenoconazole, captan, bupirimate, fosetyl-AI, strobilurin, dimoxystrobin, enestrobrin, fluoxastrobin, kresoxim-methyl, metominostrobin, picoxystrobin, pyraclostrobin, trifloxystrobin, oryzastrobin, carboxamide, carboxanilide, benalaxyl, benalaxyl-M, benodanil, carboxin, mebenil, mepronil, fenfuram, and furan. Enhexamide, flutolanil, furalaxyl, flucarbanil, furametpyr, metalaxyl, metalaxyl-M, metofuroxam, metsulfovax, ofrace, oxadixyl, oxycarboxin, penthiopyrad, pyracarbollide, salicylanilide, tecloftalam, thifluzamide, tiadinil, N-biphenylamide, bixafen, boscalid, carboxylic acid morpholide, dimethomorph, flumorph, benzamide, flumetobir, fluopicolide, zoxamide, carpropamid, diclocymet, mandipropamide, silthiofa azoles, triazoles, bitertanol, bromuconazole, cyproconazole, diniconazole, enilconazole, epoxiconazole, fenbuconazole, flusilazole, fluquinconazole, flutriafol, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propiconazole, prothioconazole, simeconazole, tetraconazole, triadimenol, triadimefon, triticonazole, imidazole, cyazofamid, imazalil, pefurazoate, promethazine Rochloraz, triflumizole, benzimidazole, benomyl, carbendazim, fuberidazole, thiabendazole, ethaboxam, etridiazole, hymexazole, pyridine, fadinam, pyrifenox, pyrimidine, cyprodinil, ferimzone, fenarimol, mepanipyrim, nuarimol, pyrimethanil, piperazine, triforine, pyrrole, fludioxonil, fenpiclonil, morpholine, aldimorph, dodemorph, fenpropimorph, tridemorph, dicarboximide, iprodione, procymidone, vinclozolin,Acibenzolar-S-methyl, Anilazine, Captafol, Dazomet, Diclomezine, Fenoxanil, Folpet, Fenpropizine, Famoxadone, Fenamidone, Octilinone, Probenazole, Proquinazide, Pyroquilon, Quinoxyfen, Tricyclazole, Carbamate, Dithiocarbamate, Felbam, Maneb, Methiram, Metam, Propineb, Thiram, Zineb, Ziram, Diethofencarb, Flubenthiavalicarb, Iprovalicarb, Propamocarb, Guanidine, Dodine, Iminoctadine, Guazatine, Kasugamycin, Poly Oxine, streptomycin, validamycin A, fentin salts, sulfur-containing heterocyclyl compounds, isoprothiolane, dithianon, organophosphorus compounds, edifenphos, fosetyl, fosetyl-aluminum, iprobenfos, pyrazophos, tolclofos-methyl, organochlorine compounds, thiophanate-methyl, chlorothalonil, dichlofluanid, tolylfluanid, flusulfamide, phthalide, hexachlorobenzene, pencycuron, quintozene, nitrophenyl derivatives, binapacryl, dinocap, dinobuton, spiroxamine, cyflufenamid, Cymoxanil, metrafenone, N-2-cyanophenyl-3,4-dichloroisothiazole-5-carboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methylpyrazole-4-carboxamide, 3-[5-(4-chlorophenyl)-2,3-dimethylisoxazolidin-3-yl]-pyridine, N-(3',4'-dichloro-4-fluorobiphenyl-2-yl)-3-difluoromethyl-1-methylpyrazole-4-carboxamide, 5-chloro-7-(4-methylpiperidin-1-yl)- 6-(2,4,6-trifluorophenyl)-[1,2,4]triazolo[1,5-a]pyrimidine, 2-butoxy-6-iodo-3-propylchromen-4-one, N,N-dimethyl-3-(3-bromo-6-fluoro-2-methylindole-1-sulfonyl)-[1,2,4]triazole-1-sulfonamide, methyl-(2-chloro-5-[1-(3-methylbenzyloxyimino)-ethyl]benzyl)carbamate, methyl-(2-chloro-5-[1-(6-methylpyridin-2-ylmethoxy-imino)ethyl]benzyl)carbamate,Methyl 3-(4-chlorophenyl)-3-(2-isopropoxycarbonylamino-3-methylbutyryl-amino)propionate, 4-fluorophenyl N-(1-(1-(4-cyanophenyl)ethanesulfonyl)but-2-yl)carbamate, N-(2-(4-[3-(4-chlorophenyl)prop-2-ynyloxy]-3-methoxyphenyl)ethyl)-2-methanesulfonylamino-3-methylbutyramide, N-(2-(4-[3-(4-chlorophenyl)prop-2-ynyloxy]-3-methoxyphenyl)ethyl)-2-ethanesulfonylamino-3-methylbutyl amide, N-(4'-bromobiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, N-(4'-trifluoromethylbiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, N-(4'-chloro-3'-fluorobiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, methyl 2-(ortho-((2,5-dimethylphenyloxy-methylene)phenyl)-3-methoxyacrylate, oxathiapiproline, and esters and salts thereof; (b) The antibacterial agent is selected from the group consisting of hypochlorite, sodium hypochlorite, chloramine, dichloroisocyanurate, trichloroisocyanurate, wet chlorine, chlorine dioxide, peroxide, peracetic acid, potassium persulfate, sodium perborate, sodium percarbonate, urea hydrogen peroxide, iodine, iodopovidone, ethanol, 1-propanol, 2-propanol, 2-phenoxyethanol, phenol, cresol, halogenated phenols, hexachlorophene, triclosan, trichlorophenol, tribromophenol, pentachlorophenol, cationic surfactants, benzalkonium chloride, cetyltrimethylammonium bromide, cetyltrimethylammonium chloride, didecyldimethylammonium chloride, cetylpyridinium chloride, benzethonium chloride, chlorhexidine, glucoprotamine, octenidine dihydrochloride, ozone solution, colloidal silver, silver nitrate, mercury chloride, phenyl mercury salts, copper sulfate, copper oxide-chloride, copper hydroxide, copper octanoate, copper oxychloride sulfate, copper sulfate pentahydrate, phosphoric acid, nitric acid, sulfuric acid, amidosulfonic acid, toluenesulfonic acid, sodium hydroxide, potassium hydroxide, calcium hydroxide, sorbic acid, benzoic acid, lactic acid, salicylic acid, penicillin, cephalosporin, vancomycin, polymyxin, rifamycin, lipiamycin, quinolone, sulfonamide, aminoglycoside, kasugamycin, macrolide, lincosamide, tetracycline, cyclic lipopeptide, daptomycin, glycylcycline, tigecycline, oxazolidinone, linezolid, fidaxomicin, rifampicin, ciprofloxacin, doxycycline, ampicillin, polymyxin B, gramicidin, isoniazid, pyrazinamide, ethambutol, myanbutol, streptomycin, and esters and salts thereof; (c) The insecticide is a chloronicotinyl, a neonicotinoid, an acetamiprid, a clothianidin, a dinotefuran, an imidacloprid, a nitenpyram, a nithiazine, a thiacloprid, a thiamethoxam, an imidaclothiz, an (2E)-1-[(2-chloro-1,3-thiazol-5-yl)methyl]-3,5-dimethyl-N-nitro-1,3,5-triazinan-2-imine, an acetylcholinesterase (AChE) inhibitor, a carbamate, an alanycarb, an aldicarb, an aldoxycarb, an alixycarb, an aminocarb, a bendiocarb, or a benfuracarb , bufencarb, butacarb, butocarboxim, butoxycarboxim, carbaryl, carbofuran, carbosulfan, cloethocarb, dimethylane, ethiofencarb, fenobucarb, fenothiocarb, formetanate, furathiocarb, isoprocarb, metam-sodium, methiocarb, methomyl, metolcarb, oxamyl, phosphocarb, pirimicarb, promecarb, propoxur, thiodicarb, thiofanox, triazamate, trimethacarb, XMC, xylylcarb, organophosphate esters, acephate, azamethic Phos, Azinphos (-methyl, -ethyl), Bromophos-ethyl, Bromfenvinphos (-methyl), Butathiophos, Cadusafos, Carbophenothion, Chlorethoxyphos, Chlorfenvinphos, Chlormephos, Chlorpyrifos (-methyl / -ethyl), Coumaphos, Cyanofenphos, Cyanophos, Demeton-S-methyl, Demeton-S-methylsulfone, Dialifos, Diazinon, Diclofenthion, Dichlorvos / DDVP, Dicrotophos, Dimethoate, Dimethylvinphos, Dioxabenzophos, Disulfoton, EPN, Ethion, Ethopropyl phos, etrimphos, fanfur, fenamiphos, fenitrothion, fensulfothion, fenthion, flupyrazophos, fonofos, formothion, fosmetilan, fosthiazate, heptenophos, iodofenphos, iprobenfos, isazophos, isofenphos, isopropyl O-salicylate, isoxathion, malathion, mecarbam, methacrifos, methamidophos, methidathion, mevinphos, monocrotophos, naled, omethoate, oxydemeton-methyl, parathion (-methyl / -ethyl), phenthoate, phorate, phosalone,Phosmet, phosphamidon, phosphocarb, phoxim, pirimiphos (-methyl / -ethyl), profenofos, propafos, propetamphos, prothiofos, prothoate, pyraclofos, pyridaphenthion, pyridathion, quinalphos, cebufos, sulfotep, sulprofos, tebupirimfos, temephos, terbufos, tetrachlorvinphos, thiometon, triazophos, trichlorfon, vamidothion, pyrethroids, acrinathrin, allethrin (d-cis-trans, d-trans), cypermethrin (α-, β-, θ-, ζ-), permethrin (cis-, trans-), β-cyfluthrin, bifenthrin, bioallethrin, bioallethrin-S-cyclopentyl-isomer, bioethanomethrin, biopermethrin methacrylate, bioresmethrin, clovaportrin, cis-cypermethrin, cis-resmethrin, cis-permethrin, clocithrin, cycloprothrin, cyfluthrin, cyhalothrin, cyphenothrin, DDT, deltamethrin, empenthrin (1R-isomer), esfenvalerate, etofenprox, fenfluthrin, fenpropathrin, fenpyrithrin, fenvalerate, flubrocythrinate, flucythrinate, flufenprox, flumethrin, fluvalinate, fubufenprox, γ-cyhalothrin, imiprothrin, kadethrin, λ, metofluthrin, fenothrin (1R-trans isomer), prallethrin, profluthrin, protrifenbut, pyresmethrin, resmethrin, RU 15525, silafluofen, tau-fluvalinate, tefluthrin, telalethrin, tetramethrin (1R-isomer), tralocitrin, tralomethrin, transfluthrin, ZXI 8901, pyrethrins, pyrethrum, oxadiazine, indoxacarb, acetylcholine receptor modulators, spinosyn, spinosad, cyclodiene, camphechlor, chlordane, endosulfan, gamma-HCH, HCH, heptachlor, organochlorines, lindane, methoxychlor, fiprole, acetoprole, ethiprole, vaniliprole, fipronil, mectin, abamectin, avermectin, emamectin, emamectin benzoate, fenoxycarb, hydroprene, kinoprene, methoprene, ivermectin, lepimectin,Epofenonane, pyriproxyfen, milbemectin, milbemycin, triplen, diacylhydrazine, chromafenozide, halofenozide, methoxyfenozide, tebufenozide, benzoylurea, bistrifluron, chlorfluazuron, diflubenzuron, fluazuron, flucycloxuron, flufenoxuron, hexaflumuron, lufenuron, novaluron, noviflumuron, penfluoron, teflubenzuron, triflurane Lumulon, organotins, azocyclotin, cyhexatin, fenbutatin oxide, pyrrole, chlorfenapyr, dinitrophenol, pinapacryl, dinobuton, dinocap, DNOC, METI, fenazaquin, fenpyroximate, pyrimidifen, pyridaben, tebufenpyrad, tolfenpyrad, rotenone, acequinocyl, fluacrypyrim, microbial disruptors of insect gut membranes, Bacillus thuringiensis thuringiensis strains, lipid synthesis inhibitors, tetronic acid, tetramic acid, spirodiclofen, spiromesifen, spirotetramat, cis-3-(2,5-dimethylphenyl)-8-methoxy-2-oxo-1-azaspiro[4.5]dec-3-en-4-ylethyl carbonate, carboxamide, flonicamide, octopamine agonists, amitraz, inhibitors of magnesium-stimulated ATPase, propargite, ryanodine receptor agonists, phthalamide, linaxapyr, N2-[1,1-dimethyl-2-(methylsulfonyl)ethyl]-3-iodo-N1-[2-methyl-4-[1,2,2,2-tetrafluoro-1-(trifluoromethyl) ethyl]phenyl]-1,2-benzenedi-carboxamide, spidoxamato, nicofluprole, tetraniliprole, thioxazafen, flupyradifurone, fluopyram, flubendiamide, deltamethrin, permethrin, ginpropylidaz, brofuranilide, afidopiropen, fluopyram, fluazaindolizine, triflumezopyrim, sulfoxaflor, spinetoram, chlorpyrifos, spinosad, cyantraniliprole, chlorantraniliprole, cypermethrin, prinazoline, cyclobutrifluram, spiropydione, fluensulfone, pymetrozine, thiamethoxam, lambda cyhalothrin, oxazosulfil, benzpyrimoxane,at least one of dichloromezothiaz, flupentiofenox, fluhexafon, fluxamethamide, flupirimine, cyhalodiamide, acinonapyr, cyclaniliprole, cetopyrafen, ciprofuranilide, tetrachlorantraniliprole, isocycloceram, brofuranilide, spiropidione, and esters and salts thereof; (d) the molluscicide comprises at least one of a metal salt, iron phosphate, aluminum sulfate, sodium ferric EDTA, metaldehyde, methiocarb, and an acetylcholinesterase inhibitor; (e) the nematicide comprises at least one of a fumigant, D-D, 1,3-dichloropropene, ethylene dibromide, 1,2-dibromo-3-chloropropane, methyl bromide, chloropicrin, metam sodium, dazomet, methyl isothiocyanate (MITC), sodium tetrathiocarbonate, carbamate, aldicarb, aldoxycarb, carbofuran, oxamyl, creotocarb, organophosphate ester, ethoprophos, fenamiphos, cadusafos, fosthiazate, fensulfothion, thionazine, isazophos, and a biochemical; and (f) The herbicide is selected from the group consisting of glufosinate, propaquizafop, metamitron, metazachlor, pendimethalin, flufenacet, diflufenican, clomazone, nicosulfuron, mesotrione, pinoxaden, sulcotrione, prosulfocarb, sulfentrazone, bifenox, quinmerac, triallate, terbuthylazine, atrazine, oxyfluorfen, diuron, trifluralin, chlorotoluron, benzoic acid herbicides, dicamba, phenoxyalkanoic acid herbicides, 2,4-D, MCPA, 2,4-DB esters, aryloxypheno Xypropionic acid herbicides, clodinafop, cyhalofop, fenoxaprop, fluazifop, haloxyfop, quizalofop esters, pyridinecarboxylic acid herbicides, aminopyralid, picloram, clopyralid esters, pyrimidinecarboxylic acid herbicides, aminocyclopyrachlor esters, pyridyloxyalkanoic acid herbicides, fluoroxypyr, triclopyr, hydroxybenzonitrile herbicides, bromoxynil, ioxynil, arylpyridinecarboxylic acids, arylpyrimidinecarboxylic acids, acetochlor, acifluorfen, alachlor, Ametryn, Amitrole, Ashlam, Azafenidin, Benefin, Bensulfuron, Bensulide, Bentazon, Bromacil, Butyrate, Carfentrazone, Chloramben, Chlorimuron, Chlorpropham, Chlorsulfuron, Clethodim, Clopyralid, Cloransulam, Cyanazine, Cycloate, DCPA, Desmedipham, Dichlobenil, Diclofop, Diclosulam, Diethathyl, Difenzoquat, Diflufenzopyr, Dimethenamid-p, Diquat, DSMA, Endosole, EPTC, Ethalfluralin, Ethamethosulfuron , ethofumesate, fluazifop-P, flucarbazone, flumetsulam, flumiclorac, flumioxazin, fluometuron, fluroxypyr, fluthiaset, fomesafen, foramsulfuron, glyphosate, halosulfuron, haloxyfop, hexazinone, imazamethabenz, imazamox, imazapic, imazaquin, imazethapyr, isoxaben, isoxaflutole, lactofen, linuron, MCPB, methazole, metolachlor-s, metribuzin, metsulfuron, molinate, MSMA, napropamide, naptalam,30. The agricultural composition of claim 29, comprising at least one of norflurazon, oryzalin, oxadiazon, oxasulfuron, oxyfluorfen, paraquat, pebulate, pelargonic acid, pendimethalin, phenmedipham, primisulfuron, prodiamine, prometryn, pronamide, propachlor, propanil, prosulfuron, pyrazone, pyridate, pyrithiobac, quinclorac, quizalofop, rimsulfuron, sethoxydim, siduron, simazine, sulfometuron, sulfosulfuron, tebuthiuron, terbacil, thiazopyr, thifensulfuron, thiobencarb, tralkoxydim, triallate, triasulfuron, tribenuron, triflusulfuron, vernolate, and esters and salts thereof.

31. 31. The agricultural composition of claim 30, wherein the pyrethroid is deltamethrin.

32. 28. The agricultural composition of claim 27, wherein the heterofunctional agent comprises a plant modifier.

33. 30. The agricultural composition of claim 27, wherein the heterofunctional agent comprises an insect modifier.

34. 34. The agricultural composition according to any one of claims 27 to 33, wherein the heterologous functional agent is encapsulated in the NLP.

35. 10. The agricultural composition of claim 1, formulated for application to soil.

36. 10. The agricultural composition of claim 1, wherein the at least one surface modifier modifies the mobility of the agricultural composition through soil compared to a composition that does not contain the surface modifier.

37. 37. The agricultural composition of claim 36, wherein the mobility of the heterofunctional agent in the soil is increased.

38. 37. The agricultural composition of claim 36, wherein the mobility of the heterofunctional agent in the soil is reduced.

39. 10. The agricultural composition of claim 1, formulated for delivery to a plant, plant part, or plant pest.

40. 40. The agricultural composition of claim 39, wherein the plant part is a plant seed.

41. 40. The agricultural composition of claim 39, wherein the NLP is detected in germinating seeds.

42. 10. The agricultural composition of claim 1, wherein the heterofunctional agent is a volatile agent.

43. 43. The method of any one of claims 1 to 42, wherein the volatile heterofunctional agent is a pesticide, a fertilizer, a herbicide, a plant modifier, an insect attractant, a plant growth promoter, a biostimulant, or a plant immunity inducer.

44. 43. The method of claim 42, wherein the volatile pesticide is selected from the group consisting of antifungals, antioomycetes, antibacterials, insecticides, molluscicides, nematicides, herbicides and virucides.

45. 44. The method of claim 43, wherein the herbicide is dicamba.

46. 45. The method of claim 44, wherein the insecticide is tefluthrin.

47. 47. The agricultural composition of any one of claims 1 to 46, wherein the at least one surface modifier enhances uptake of the agricultural composition by plants or plant parts compared to a composition not comprising the surface modifier.

48. 47. The agricultural composition of any one of claims 1 to 46, wherein the at least one surface modifier enhances biodistribution of the agricultural composition by plants or plant parts compared to a composition not comprising the surface modifier.

49. 47. The agricultural composition of any one of claims 1 to 46, wherein the NLPs are targeted to the meristematic region.

50. 47. The agricultural composition of any one of claims 1 to 46, wherein the encapsulated heterofunctional agent is protected from ultraviolet light.

51. 1. An agricultural composition comprising: a) a first plurality of NLPs, at least one phospholipid, at least one non-polar lipid, and at least one surface modifier; and First heterofunctional agent a first plurality of NLPs including: b) a second plurality of NLPs, at least one phospholipid, at least one non-polar lipid, and at least one surface modifier; and Second heterofunctional agent a second plurality of NLPs including wherein said first plurality of NLPs comprises a hydrophobic core.

52. 52. The agricultural composition of claim 51, wherein the first plurality of NLPs and the second plurality of NLPs differ in stability.

53. 1. An agricultural composition comprising: a) a plurality of NLPs, at least one phospholipid, at least one non-polar lipid, and at least one surface modifier; and First heterofunctional agent and several NLPs, including b) a second, heterologous functional agent that is not encapsulated; and wherein the plurality of NLPs comprises a hydrophobic core.

54. An agricultural composition comprising a plurality of NLPs each containing a different functional agent, wherein the NLPs are at least one phospholipid, at least one non-polar lipid, at least one surface modifier; Heterofunctional drugs, and aqueous solution wherein the plurality of NLPs comprises a hydrophobic core.

55. 1. A method of making an agricultural composition comprising a plurality of NLPs, each comprising a heterologous functional agent, the method comprising: at least one phospholipid, at least one non-polar lipid, at least one surface modifier; Heterofunctional drugs, and aqueous solution wherein the NLP comprises a hydrophobic core.

56. 1. A method for modifying the binding of a heterofunctional agent to at least one component in soil, comprising: Applying an NLP composition containing a heterologous functional agent encapsulated in an NLP to soil The NLP comprises: at least one phospholipid, at least one non-polar lipid, and At least one surface modifier wherein the binding of the encapsulated heterofunctional agent to the soil is different from the binding of an unencapsulated heterofunctional agent to the soil.

57. 1. A method for modifying the mobility of a heterofunctional agent in soil, comprising: Applying an NLP composition containing a heterologous functional agent encapsulated in an NLP to soil The NLP comprises: at least one phospholipid, at least one non-polar lipid, and At least one surface modifier wherein the mobility of the encapsulated heterofunctional agent in soil is different from the mobility of the unencapsulated heterofunctional agent in soil.

58. A method for reducing rootworm survival rate, comprising: For soil infested with rootworms, at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs applying an NLP composition comprising wherein the heterologous functional agent contacts the rootworms, thereby reducing survival rate of the rootworms.

59. 59. The method of claim 58, wherein the NLP composition is applied to the soil as a soil drench.

60. 59. The method of claim 58, wherein the NLP composition is applied to the soil in a planting furrow.

61. 59. The method of claim 58, wherein the rootworm is a member of the genus Diabrotica.

62. 59. The method of claim 58, wherein the rootworm is the western corn rootworm (Diabrotica virgifera virgifera).

63. 1. A method for reducing fungal viability, comprising: For soil containing fungi, at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs applying an NLP composition comprising wherein the heterofunctional agent contacts the fungus, thereby reducing viability of the fungus in the soil.

64. 64. The method of claim 63, wherein the fungus belongs to a family selected from the group consisting of Sciaridae, Diadocidiidae, Ditomyiidae, Keroplatidae, Bolitophilidae, and Mycetophilidae, and Sclerotiniaceae.

65. 65. The method of claim 64, wherein the fungus is Botrytis cinerea.

66. 1. A method for preventing plants from developing diseases caused by plant pests, comprising: at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs applying to the soil an NLP composition comprising Including, NLPs contain a hydrophobic core, The heterofunctional agent contacts the plant pest, thereby killing the pest and thereby preventing the plant from developing the disease.

67. 67. The method of claim 66, wherein the NLP composition is applied to the soil as a soil drench.

68. 67. The method of claim 66, wherein the NLP composition is applied to the soil in a planting furrow.

69. 67. The method of claim 66, wherein the plant pest is a member of the order Coleoptera or Hemiptera.

70. 67. The method of claim 66, wherein the plant pest is a member of the genus Diabrotica.

71. 71. The method of claim 70, wherein the plant pest is the western corn rootworm (Diabrotica virgifera virgifera).

72. 67. The method of claim 66, wherein the plant pest is a fungus.

73. 69. The method of claim 68, wherein the fungus belongs to a family selected from the group consisting of Sciaridae, Diadocidiidae, Ditomyiidae, Keroplatidae, Bolitophilidae, and Mycetophilidae, and Sclerotiniaceae.

74. 73. The method of claim 72, wherein the fungus is Botrytis cinerea.

75. 1. A method for increasing uptake of a heterologous functional agent by a plant or plant part, comprising: contacting a plant or plant part with a composition comprising a heterologous functional agent encapsulated in an NLP; The NLP comprises: at least one phospholipid, at least one non-polar lipid, and At least one surface modifier Including, The NLP comprises a hydrophobic core, A method wherein uptake of the encapsulated heterologous functional agent by the plant or plant part is higher than uptake of an unencapsulated heterologous functional agent by the plant or plant part.

76. 1. A method for delivering a heterologous functional agent to a plant or plant part, comprising: The plant or plant part at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs wherein the NLP comprises a hydrophobic core, thereby delivering the heterologous functional agent to the plant. A method comprising:

77. 1. A method for delivering a heterologous functional agent to a meristem, comprising: contacting a plant or plant part with a composition comprising a heterologous functional agent encapsulated in an NLP; The NLP comprises: at least one phospholipid, at least one non-polar lipid, and At least one surface modifier A method comprising:

78. 78. The method of any one of claims 75 to 77, wherein the plant part is a plant seed.

79. 1. A method for distributing a heterofunctional agent in soil, comprising: Plant seeds, at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs wherein the NLP comprises a hydrophobic core; and incubating said plant seeds in soil, thereby distributing said heterologous functional agent in said soil. A method comprising:

80. 1. A method for distributing a heterologous functional agent in a plant, comprising: Plant seeds, at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs wherein the NLP comprises a hydrophobic core; and incubating said plant seeds under conditions that cause germination, thereby distributing said heterologous functional agent throughout said plants. A method comprising:

81. 81. The method of claim 80, wherein said contacting is by injecting said composition into said plant or plant part.

82. 82. The method of claim 81, wherein the composition is injected or infiltrated into one or more leaves.

83. 81. The method of claim 80, wherein the composition is injected into the tree.

84. 82. The method of claim 81, wherein the composition is injected into the tree at several locations.

85. 1. A method for treating a plant disease, comprising: The plant or plant part at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs wherein the NLP comprises a hydrophobic core, thereby treating said disease in the plant. A method comprising:

86. 86. The method of claim 85, wherein the disease is caused by Candidatus Liberibacter asiaticus (CLas).

87. 86. The method of claim 85, wherein the disease is citrus greening.

88. 86. The method of claim 85, wherein the disease is caused by Xylella.

89. A method for preventing a plant from developing a disease, comprising: The plant or plant part at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs wherein the NLP comprises a hydrophobic core, thereby preventing plant disease. A method comprising:

90. 10. A method for reducing the volatility of a heterofunctional agent, comprising encapsulating the volatile heterofunctional agent in the composition of claim 1.

91. 10. A method for sequestering a volatile heterofunctional agent, comprising encapsulating the heterofunctional agent in the composition of claim 1.

92. A method for the controlled release of a volatile heterofunctional agent into an environment, comprising encapsulating the heterofunctional agent in the composition of claim 1, wherein the release of the agent is inversely proportional to the stability of the NLP.

93. 49. A method for controlled release of at least one volatile heterofunctional agent into an environment, comprising encapsulating at least one volatile heterofunctional agent in the composition of claim 48, wherein the release of the agent is inversely proportional to the stability of the at least one plurality of NLPs.

94. 94. The method of any one of claims 90 to 93, wherein the volatile heterofunctional agent is a pesticide, a fertilizer, a herbicide, a plant modifier, an insect attractant, a plant growth promoter, a biostimulant, or a plant immunity inducer.

95. 95. The method of claim 94, wherein the volatile pesticide is selected from the group consisting of antifungals, antioomycetes, antibacterials, insecticides, molluscicides, nematicides, herbicides, and virucides.

96. 95. The method of claim 94, wherein the herbicide is dicamba.

97. 96. The method of claim 95, wherein the insecticide is tefluthrin.

98. 1. A kit comprising an agricultural composition, the composition comprising: at least one phospholipid, at least one non-polar lipid, at least one surface modifier, and Heterofunctional drugs wherein the NLP comprises a hydrophobic core.

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