Pesticidal compounds and their uses

Abstract

Provided herein are compositions comprising yeast particles containing a pesticide and yeast cell wall components, and methods for controlling plant pests using the compositions. Also provided herein are compositions comprising a pesticide and one or more polysaccharides selected from beta-glucan, mannan oligosaccharide, and / or laminarin; and methods for controlling plant pests using the compositions.

Description

[Technical Field]

[0001] Related Applications This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. USSN 63 / 443,179, filed February 3, 2023, and U.S. Provisional Patent Application No. USSN 63 / 450,333, filed March 6, 2023, each of which is incorporated herein by reference.

[0002] FIELD OF THE INVENTION This disclosure relates to agriculture and plant protection. More particularly, this disclosure relates to compositions and delivery systems for pesticides.

[0003] Electronic Sequence Listing Reference The contents of the electronic sequence listing (G083070046WO00-SEQ-MSB.xml; size: 17,390 bytes; and creation date: February 1, 2024) are incorporated herein by reference in their entirety. [Background technology]

[0004] background Plant crops are targets of pathogen (e.g., fungal, bacterial, and insect) attack worldwide. According to the Food and Agriculture Organization of the United Nations, farmers globally lose 30–40% of their crops to pests and diseases. Crop maintenance and crop health are essential for agricultural yield and quality and ultimately require long-term strategies to minimize pest and disease outbreaks. The annual cost of controlling crop-pathogenic pests (e.g., Lepidoptera, Diptera, Coleoptera, Hemiptera, etc.) is estimated to be in the tens of millions of dollars, and if left uncontrolled, the estimated annual cost of crop loss reaches billions of dollars.

[0005] Although chemical pesticides are one approach to eradicating pest infestations, alternative and more environmentally safe solutions are needed. Chemical pesticides are harmful to the environment, lack specificity and selectivity, and can ultimately lead to off-target effects. Furthermore, given the slow metabolism and potential accumulation of chemical pesticides, resistance may develop. Thus, there has been a long-standing need for more environmentally friendly compositions and methods (including those requiring a reduced amount of chemical pesticide for efficacy) for controlling or eradicating pest infestations and diseases (e.g., more selective, environmentally safe, biodegradable, and / or capable of delaying the development of resistance by pests). Summary of the Invention

[0006] overview The inventors of the present disclosure have determined that compositions comprising a pesticide and a biological adjuvant (e.g., yeast particles containing cell wall components, or one or more polysaccharides) function in some embodiments to enhance (increase) the efficacy of the pesticide (e.g., by increasing the ability of the pesticide to control plant pests such as insects). The efficacy of a pesticide is enhanced by a biological adjuvant if a lower amount or concentration of pesticide is required to control a plant pest (e.g., an insect) in the presence of the biological adjuvant (compared to the amount or concentration of pesticide required to control the plant pest in the absence of the biological adjuvant), or if the same amount of pesticide provides greater control (e.g., increased mortality) when applied in conjunction with the biological adjuvant. In other words, the presence (or addition) of a biological adjuvant to a composition containing a pesticide allows, for example, a lower effective dose of the pesticide required to provide beneficial protection of plants infested by plant pests, or allows greater protection of the plant when a biological adjuvant is applied. By allowing users to reduce the application rate of a given biological or chemical pesticide sprayed on crops while maintaining high efficacy of the pesticide against plant pests, these inventions result in more environmentally friendly compositions at a lower cost. The use of yeast particles according to the claimed methods may further enable improved efficacy of biopesticides expressed by genetically modified crops against both susceptible and resistant insect populations, extending the effectiveness of these transformed crops in the marketplace. The inventors have discovered that such effective compositions may include a biological adjuvant, which may be yeast particles containing yeast cell wall components or one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharides, and laminarin.

[0007] In some aspects, the present disclosure provides compositions comprising yeast particles containing a pesticide and yeast cell wall components, wherein the pesticide is a biopesticide or a pesticidal polynucleotide. In other aspects, the present disclosure provides methods of applying yeast particles containing yeast cell wall components to a transformed crop that has been genetically modified to express one or more pesticidal proteins, resulting in improved efficacy of the pesticidal proteins against plant pests.

[0008] Another aspect of the present disclosure is a composition comprising a yeast particle comprising a pesticide and a yeast cell wall component, wherein the pesticide is a chemical pesticide and the pesticide is selected from the group consisting of ryanodine receptor modulators (IRAC class 28), chitin biosynthesis inhibitors acting on CHS1 (IRAC class 15), nicotinic acetylcholine receptor (nACHR) allosteric modulators-site 1 (IRAC class 5), nACHR competitive modulators (IRAC class 4), sodium channel modulators (IRAC class 3), acetylcholinesterase inhibitors (IRAC class 4), and acetylcholinesterase inhibitors (IRAC class 5). Enzyme (ACHE) inhibitors (IRAC class 1), GABA-gated chloride channel blockers (IRAC class 2), glutamate-gated chloride channel (GLUCL) allosteric modulators (IRAC class 6), juvenile hormone receptor modulators (IRAC class 7), other nonspecific (multi-site) inhibitors (IRAC class 8), chordotonal organ TRPV channel modulators (IRAC class 9), mite growth inhibitors acting on CHS1 (IRAC class 10), mitochondrial ATP biosynthesis enzyme inhibitors (IRAC class 11), class 12), oxidative phosphorylation uncouplers that disrupt the proton gradient (IRAC class 13), nicotinic acetylcholine receptor (nACHR) channel blockers (IRAC class 14), chitin biosynthesis inhibitors, type 1 (IRAC class 16), molting disruptors Diptera (IRAC class 17), ecdysone receptor agonists (IRAC class 18), and octopamine receptor agonists (IRAC class 19), mitochondrial complex III electron transport inhibitors QO site (IRAC class 20), mitochondrial complex I electron transport inhibitors (IRAC class 21). Class 21), voltage-gated sodium channel blockers (IRAC class 22), acetyl-CoA carboxylase inhibitors (IRAC class 23), mitochondrial complex IV electron transport inhibitors (IRAC class 24), mitochondrial transport inhibitors (IRAC class 25), chordotonal organ nicotinamidase inhibitors (IRAC class 29), GABA-gated chloride channel allosteric modulators (IRAC class 30), nicotinic acetylcholine receptor (NACHR) allosteric modulators - site II (IRAC class 32),The composition is a calcium-activated potassium channel (KCa2) modulator (IRAC class 33), a mitochondrial complex III electron transport inhibitor QI site (IRAC class 34), or a chordotonal organ modulator - target site undefined (IRAC class 36). In some embodiments, the chemical pesticide is selected from the group consisting of chlorantraniliprole, novaluron, spinosad, acetamiprid, cypermethrin, diflubenzuron, and bifenthrin.

[0009] In some embodiments, the yeast particle is selected from the group consisting of yeast cell wall particle (YCWP), yeast cell particle (YCP), yeast glucan particle (GP or YGP), yeast glucan mannan particle (GMP or YGMP), yeast glucan chitin particle (GCP or YGCP), yeast glucan chitin mannan particle (GCMP or YGCMP), yeast glucan lipid particle (GLP or YGLP), whole glucan particle (WGP), and combinations thereof.

[0010] In some embodiments, the yeast particles enhance the efficacy of the pesticide, and optionally, the yeast particles enhance the ability of the pesticide to control plant pests (e.g., insects) by at least 5%, 10%, 20%, 30%, 40%, or 50% compared to a control composition that does not include the yeast particles.

[0011] In some embodiments, the yeast particles are commercially available yeast particles. In some embodiments, the yeast particles comprise β-glucan and / or mannan oligosaccharides. In some embodiments, the yeast particles comprise β-1,3-glucan, β-1,6-glucan, and / or β-1,3 / 1,6-glucan, and optionally, the β-1,3-glucan, β-1,6-glucan, and / or β-1,3 / 1,6-glucan are branched. In some embodiments, the yeast particles comprise intact or fragmented yeast cell walls. In some embodiments, the yeast particles comprise β-1,6-glucan, β-1,3-glucan, mannan oligosaccharide, mannoprotein, chitin, and / or lipid. In some embodiments, the lipid is phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), and / or cytidine diphosphate diacylglycerol (CDP-DAG).

[0012] In some embodiments, the yeast particles do not include yeast extract. In some embodiments, the yeast particles do not consist of yeast extract.

[0013] In some embodiments, the pesticide is not encapsulated in the yeast particle. In some embodiments, the pesticide is encapsulated in the yeast particle.

[0014] Some aspects of the present disclosure provide compositions comprising a pesticide and one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharides, and laminarin, wherein the pesticide is a biopesticide or a pesticidal polynucleotide.

[0015] Some aspects of the disclosure relate to a composition comprising a pesticide and one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharides, and laminarin, wherein the pesticide is a chemical pesticide and the pesticide is selected from the group consisting of ryanodine receptor modulators (IRAC class 28), chitin biosynthesis inhibitors acting on CHS1 (IRAC class 15), nicotinic acetylcholine receptor (nACHR) allosteric modulators-site 1 (IRAC class 5), nACHR competitive modulators (IRAC class 4), and the like. , sodium channel modulators (IRAC class 3), acetylcholinesterase (ACHE) inhibitors (IRAC class 1), GABA-gated chloride channel blockers (IRAC class 2), glutamate-gated chloride channel (GLUCL) allosteric modulators (IRAC class 6), juvenile hormone receptor modulators (IRAC class 7), other nonspecific (multi-site) inhibitors (IRAC class 8), chordotonal organ TRPV channel modulators (IRAC class 9), and DAs acting on CHS1. Class II growth inhibitors (IRAC class 10), mitochondrial ATP biosynthesis enzyme inhibitors (IRAC class 12), oxidative phosphorylation uncouplers that disrupt the proton gradient (IRAC class 13), nicotinic acetylcholine receptor (nACHR) channel blockers (IRAC class 14), chitin biosynthesis inhibitors, type 1 (IRAC class 16), molting disruptors Diptera (IRAC class 17), ecdysone receptor agonists (IRAC class 18), and octopamine receptor agonists (IRAC class 19), mitochondrial complex III Electron transport inhibitors QO site (IRAC class 20), mitochondrial complex I electron transport inhibitors (IRAC class 21), voltage-gated sodium channel blockers (IRAC class 22), acetyl-CoA carboxylase inhibitors (IRAC class 23), mitochondrial complex IV electron transport inhibitors (IRAC class 24), mitochondrial transport inhibitors (IRAC class 25), chordotonal organ nicotinamidase inhibitors (IRAC class 29), GABA-gated chloride channel allosteric modulators (IRAC class 30),The composition is a nicotinic acetylcholine receptor (NACHR) allosteric modulator - site II (IRAC class 32), a calcium-activated potassium channel (KCa2) modulator (IRAC class 33), a mitochondrial complex III electron transport inhibitor QI site (IRAC class 34), or a chordotonal organ modulator - target site undefined (IRAC class 36). In some embodiments, the chemical pesticide is selected from the group consisting of chlorantraniliprole, novaluron, spinosad, acetamiprid, cypermethrin, diflubenzuron, and bifenthrin.

[0016] In some embodiments, the one or more polysaccharides enhance the efficacy of the pesticide, and optionally, the one or more polysaccharides enhance the ability of the pesticide to control plant pests by at least 5%, 10%, 20%, 30%, 40%, or 50% compared to a control composition that does not include the one or more polysaccharides.

[0017] In some embodiments, the one or more polysaccharides comprise β-1,6-glucan, β-1,3-glucan, β-1,3 / 1,6-glucan, and / or mannan oligosaccharides, and optionally, the β-1,3-glucan, β-1,6-glucan, and / or β-1,3 / 1,6-glucan is branched. In some embodiments, the one or more polysaccharides are derived from yeast cell walls. In some embodiments, the one or more polysaccharides comprise β-glucan derived from yeast cell walls, optionally β-glucan derived from Saccharomyces cerevisiae. In some embodiments, the one or more polysaccharides are derived from a source other than yeast, and optionally, the non-yeast source is a bacterium, fungus, algae, lichen, or plant. In some embodiments, the one or more polysaccharides comprise β-1,3-glucan derived from algae, optionally β-1,3-glucan derived from Euglena gracilis.

[0018] In some embodiments, the pesticide is a biopesticide. In some embodiments, the biopesticide is a microorganism. In some embodiments, the microorganism is a naturally occurring microorganism or an engineered microorganism.

[0019] In some embodiments, the microorganism produces one or more proteins that control a plant pest, and optionally, the plant pest is a fungus or an insect. In some embodiments, the plant pest is an insect. In some embodiments, the one or more proteins produced by the microorganism are activated in the insect gut. In some embodiments, the one or more proteins produced by the naturally occurring microorganism are selected from the group consisting of crystal proteins, vegetative insecticidal proteins, and toxin complex proteins. In some embodiments, the one or more proteins produced by the naturally occurring microorganism are Bacillus thuringiensis (Bt) crystal proteins. In some embodiments, the microorganism is a Bacillus thuringiensis microorganism. In some embodiments, the biopesticide comprises a commercially available biopesticide, wherein the biopesticide controls a plant pest, and optionally, the plant pest is a fungus or an insect.

[0020] In some embodiments, the biopesticide comprises isolated cellular components derived from a microorganism, optionally the microorganism is a naturally occurring or engineered microorganism. In some embodiments, the isolated cellular components comprise one or more proteins or polynucleotides that control a plant pest, optionally the plant pest is a fungus or an insect. In some embodiments, the one or more proteins or polynucleotides are activated in the insect gut. In some embodiments, the one or more proteins are selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth. In some embodiments, the biopesticide comprises a Bacillus thuringiensis (Bt) protein, optionally the Bt protein is a Bt crystal protein, and further optionally the Bt protein is derived from a microorganism.

[0021] In some embodiments, the composition further comprises a pesticidal polynucleotide. In some embodiments, the composition comprises a biopesticide and a pesticidal polynucleotide. In some embodiments, the composition comprises a chemical pesticide and a pesticidal polynucleotide.

[0022] In some embodiments, the composition comprises a pesticidal polynucleotide.

[0023] In some embodiments, the pesticidal polynucleotide inhibits expression of a target gene in a plant pest, and optionally, the plant pest is an insect. In some embodiments, the pesticidal polynucleotide is single-stranded RNA (ssRNA) or double-stranded RNA (dsRNA). In some embodiments, the composition comprises a yeast particle, and the pesticidal polynucleotide is encapsulated within the yeast particle.

[0024] In some embodiments, the composition further comprises a cationic polymer and / or a nuclease inhibitor, and the pesticidal polynucleotide is encapsulated within the yeast particle. In some embodiments, the encapsulation efficiency is greater than about 80% or greater than 90%.

[0025] In some embodiments, the composition further comprises a biopesticide, optionally the biopesticide comprises a Bacillus thuringiensis (Bt) protein, optionally the Bt protein is a Bt crystal protein.

[0026] In some embodiments, the pesticidal polynucleotide comprises a first strand that is identical to or complementary to a region of a messenger RNA (mRNA) encoded by one or more target genes, hi some embodiments, the pesticidal polynucleotide is a dsRNA comprising a second strand that is complementary to the first strand. In some embodiments, the target gene comprises one or more genes selected from the group consisting of proteasome beta 5 (PSMB5) gene, vacuolar ATPase E (vATPase E) gene, calmodulin gene, inhibitor of apoptosis protein (IAP) gene, soluble NSF attachment protein (a-SNAP) gene, Ras opposite (ROP) gene, proteasome alpha type 2 (PTSA2) gene, secretion associated Ras related GTPase 1 (SAR1) gene, PBAN, ATPase, wings up A (wupA), CP4S3_DROME, and C12C1_DROME. In some embodiments, the target gene is PSMB5 or IAP.

[0027] In some embodiments, the target gene has the sequence of SEQ ID NO: 1. In some embodiments, the pesticidal polynucleotide comprises at least 20, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 contiguous nucleotides that are complementary to or comprise at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity to a segment of the target gene. In some embodiments, the pesticidal polynucleotide comprises at least 20, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 contiguous nucleotides that are complementary to or comprise at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity to a segment of a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0028] In some embodiments, the pesticidal polynucleotide comprises at least 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 nucleotides.

[0029] In some embodiments, the pesticide comprises at least 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, or 1000 amino acids.

[0030] In some embodiments, the ryanodine receptor modulator (IRAC class 28) comprises a diamide selected from the group consisting of chlorantraniliprole, tetraniliprole, cyclaniliprole, brofuranilide, cyantraniliprole, imidacloprid, or flubendiamide.

[0031] In some embodiments, chitin biosynthesis inhibitors (IRAC class 15) acting on CHS1 comprise a benzoyl urea selected from the group consisting of bistrifluron, chlorfluazuron, diflubenzuron, flucycloxuron, flufenoxuron, hexaflumuron, lufenuron, novaluron, noviflumuron, teflubenzuron, and triflumuron.

[0032] In some embodiments, the nACHR allosteric modulator-site 1 (IRAC class 5) is a spinosyn, and optionally, the spinosyn is spinetoram or spinosad.

[0033] In some embodiments, the nACHR competitive modulator (IRAC class 4) is a neonicotinoid, and optionally the neonicotinoid is selected from the group consisting of acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, thiacloprid, and thiamethoxam.

[0034] In some embodiments, the sodium channel modulator (IRAC class 3) is a pyrethroid or pyrethrin, and optionally the pyrethroid or pyrethrin is acrinathrin, allethrin, d-cis-trans-allethrin, d-trans-allethrin, bifenthrin, bioallethrin, bioallethrin S-cyclopentenyl, bioresmethrin, cycloprothrin, cyfluthrin, beta-cyfluthrin, cyhalothrin, lambda-cyhalothrin, gamma-cyhalothrin, cypermethrin, alpha-cypermethrin, beta-cypermethrin, theta-cypermethrin, zeta-cypermethrin. phosphate, cyphenothrin [(1R)-trans-isomer], deltamethrin, empenthrin [(EZ)-(1R)-isomer], esfenvalerate, etofenprox, fenpropathrin, fenvalerate, flucythrinate, flumethrin, tau-fluvalinate, kadethrin, pyrethrins (pyrethram), halfenprox, fenothrin [(1R)-trans-isomer], prallethrin, resmethrin, silafluofen, tefluthrin, tetramethrin, tetramethrin [(1R)-isomer], tralomethrin, transfluthrin, and permethrin.

[0035] In some embodiments, the composition controls or is intended to control a plant pest, and optionally the plant pest is a fungus or an insect. In some embodiments, the plant pest is an insect. In some embodiments, the plant pest is pesticide-resistant.

[0036] In some embodiments, the plant pest is an insect belonging to the order Lepidoptera, Coleoptera, Hemiptera, Diptera, or Acari. In some embodiments, the plant pest is resistant to Bacillus thuringiensis (Bt) or Bt proteins.

[0037] In some embodiments, plant pests belonging to the order Lepidoptera include Nymphalidae (nymphalians), Danaidae (monarch butterflies), Pieridae (pierid and ceratopsian butterflies), Papilionidae (swallowtail butterflies), Lycaenidae (lycaenid butterflies), Hesperiidae (skippers), Tineidae (moths), and Sesiidae (clematid moths). The species are of a family selected from the group consisting of: Esiidae (cleanse moths), Pyralidae (pyralid moths), Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawk moths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (looper moths, noctuids, and underwing moths), and Plutellidae (diamond moths).

[0038] In some embodiments, the lepidopteran insect species is a Plutella species, and optionally the Plutella species insect is Plutella xylostella.

[0039] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera frugiperda.

[0040] In some embodiments, the lepidopteran insect species is a Chrysodeixis species, and optionally, the Chrysodeixis species insect is Chyrysodeixis includens (the soybean looper).

[0041] In some embodiments, the lepidopteran insect species is a Helicoverpa species, and optionally, the Helicoverpa species insect is Helicoverpa armigera (cotton boll worm).

[0042] In some embodiments, the lepidopteran insect species is a Helicoverpa species, and optionally, the Helicoverpa species insect is Helicoverpa armigera (cotton boll worm).

[0043] In some embodiments, the lepidopteran insect species is a Plutella species, and optionally the Plutella species insect is Plutella xylostella.

[0044] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera frugiperda.

[0045] In some embodiments, the lepidopteran insect species is a Chrysodeixis species, and optionally, the Chrysodeixis species insect is Chrysodeixis includens (the soybean looper).

[0046] In some embodiments, the lepidopteran insect species is a Helicoverpa species, and optionally the Helicoverpa species insect is Helicoverpa zea (tomato fruitworm).

[0047] In some embodiments, the lepidopteran insect species is Cydia spp., and optionally, the Cydia spp. insect is Cydia pomonella (the fruit moth).

[0048] In some embodiments, the lepidopteran insect species is a Trichoplusia species, and optionally the Trichoplusia species insect is Trichoplusia ni (cabbage looper).

[0049] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera exigua (beet armyworm).

[0050] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera litura (tobacco rootworm).

[0051] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera litoralis (the tomato moth).

[0052] In some embodiments, the lepidopteran insect species is a Pieris species, and optionally, the Pieris species insect is Pieris rapae (imported caterpillar).

[0053] In some embodiments, the lepidopteran insect species is a Tuta species, and optionally, the Tuta species insect is Tuta absoluta (tomato tea moth).

[0054] In some embodiments, the lepidopteran insect species is a Helicoverpa species, and optionally, the Helicoverpa species insect is Helicoverpa armigera (cotton boll worm).

[0055] In some embodiments, the lepidopteran insect species is a Chrysodeixis species, and optionally, the Chrysodeixis species insect is Chyrysodeixis acuta (tomato semilooper).

[0056] In some embodiments, the lepidopteran insect species is Paramyelois species, and optionally, the Paramyelois species insect is Paramyelois transitella (navel orangeworm).

[0057] In some embodiments, the lepidopteran insect species is Lobesia spp., and optionally the Lobesia spp. insect is Lobesia botrana (European grapevine moth).

[0058] In some embodiments, the insect belongs to the order Coleoptera, and the Coleoptera insect is a species selected from the group consisting of Leptinotarsa ​​spp., Phyllotreta spp., Cerotoma spp., Diabrotica spp., Tribolium spp., Anthonomus spp., and Alticini spp.

[0059] In some embodiments, the Colepotera insect species is a Phyllotreta species, and optionally the Phyllotreta species insect is Phyllotreta cruciferae (canola flea beetle).

[0060] In some embodiments, the Colepotera insect species is a Phyllotreta species, and optionally the Phyllotreta species insect is Phyllotreta striolata (flea beetle).

[0061] In some embodiments, the Colepotera insect species is a Psylliodes species, and optionally the Psylliodes species insect is Psylliodes chrysocephala (cabbage stem flea beetle).

[0062] In some embodiments, the Coleoptera insect species is a Leptinotarsa ​​species, and optionally, the Leptinotarsa ​​species insect is a Colorado potato beetle.

[0063] In some embodiments, the composition further comprises one or more compounds selected from the group consisting of cationic lipids, cationic polymers, non-cationic polymers, organic carriers, nuclease inhibitors, surfactants, antifoaming agents, and biocides.

[0064] In some embodiments, the composition further comprises a cationic polymer and a nuclease inhibitor.

[0065] In some embodiments, the cationic polymer is polyethyleneimine (PEI), poly-L-lysine (PLL), cationic gelatin, cationic chitosan, cationic cellulose, cationic dextran, poly(2-N,N-dimethylaminoethyl methacrylate), or poly(amidoamine).

[0066] In some embodiments, the nuclease inhibitor is ethylenediaminetetraacetic acid (EDTA), polyvinylsulfonic acid (PVSA), sodium hexametaphosphate (SHMP), sodium tripolyphosphate (TPP), diethylpyrocarbonate, aurintricarboxylic acid (ATA), formamide, macaloid, proteinase K, heparin, hydroxylamine-oxygen-copper(II) ion, bentonite, ammonium sulfate, dithiothreitol (DTT), beta-mercaptoethanol, cysteine, dithioerythritol, or tris(2-carboxyethyl)phosphene hydrochloride.

[0067] In some embodiments, the composition further comprises one or more of an acidifying agent, a buffering agent, an antifoaming agent, an anti-transpirant, a biocide preservative, a dye and brightener, a compatibilizer, a crop oil concentrate, a surfactant, a deposition agent, a drift reducing agent, a feeding stimulant, a spreading agent, a weighting agent, an adhesive, a suspending agent, a gelling agent, a synergist, a wetting agent, an emulsifier, a dispersing agent, a penetrating agent, a neutralizing agent, a water absorbing agent, and / or a water softener.

[0068] In some embodiments, the composition is formulated as a spray, a solution, an emulsifiable concentrate, a solid, a suspension, a colloid, a micelle, or an emulsion, a soluble liquid concentrate, a wettable powder, a water dispersible granule, an emulsion, an aerosol, a homogeneous mixture, or a heterogeneous mixture.

[0069] Some aspects of the present disclosure include: A yeast particle comprising (i) one or more Bacillus thuringiensis (Bt) proteins, and (ii) a yeast cell wall component. A composition comprising:

[0070] In some embodiments, the composition causes at least about 10% greater mortality in plant-infesting insect pests compared to one or more Bt proteins alone.

[0071] In some embodiments, the concentration of the yeast particles or one or more polysaccharides is about 2 g / L to about 15 g / L, about 5 g / L to about 12 g / L, or about 10 g / L; or about 2% w:w to about 15% w:w, about 5% w:w, about 12% w:w, or about 10% w:w.

[0072] In some embodiments, less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, or less than 5% of the pesticide is encapsulated within the yeast particle or one or more polysaccharides, hi some embodiments, 1-25% or 1-10% of the pesticide is encapsulated within the yeast particle or one or more polysaccharides.

[0073] Some aspects of the present disclosure provide kits comprising any one of the compositions described herein, optionally, the kit further comprising instructions for use.

[0074] Some aspects of the present disclosure provide methods for controlling plant pests, the method comprising delivering any one of the compositions described herein to a plant, soil, a plant pest, or the food of a plant pest.

[0075] Some aspects of the present disclosure provide methods for controlling insect infestation of a plant, the method comprising delivering any one of the compositions described herein to the plant, the soil, the insect's food, or the insect.

[0076] Some aspects of the present disclosure provide methods for controlling infestation of plants by lepidopteran insects, the method comprising delivering any one of the compositions described herein to the plant, the soil, the insect's food, or the insect. Some aspects of the present disclosure provide methods for controlling insect infestation of a plant, the method comprising delivering a composition to the plant, soil, insect food, or insect, wherein the composition comprises yeast particles comprising a pesticidal polynucleotide and a yeast cell wall component, and optionally the insect is a lepidopteran insect.

[0077] In some embodiments, the pesticidal polynucleotide is encapsulated in a yeast cell wall particle.

[0078] In some embodiments, the plant is a transgenic plant engineered to express a biopesticide or pesticidal polynucleotide, hi some embodiments, the transgenic plant is engineered to express a biopesticide, wherein the biopesticide comprises one or more Bt proteins.

[0079] Some aspects of the present disclosure provide methods for controlling plant pests, the method comprising delivering yeast particles comprising yeast cell wall components to a transgenic plant engineered to express a biopesticide or pesticidal polynucleotide.

[0080] In some embodiments, the yeast particle is selected from the group consisting of yeast cell wall particle (YCWP), yeast cell particle (YCP), yeast glucan particle (GP or YGP), yeast glucan mannan particle (GMP or YGMP), yeast glucan chitin particle (GCP or YGCP), yeast glucan chitin mannan particle (GCMP or YGCMP), yeast glucan lipid particle (GLP or YGLP), whole glucan particle (WGP), and combinations thereof.

[0081] In some embodiments, the yeast particles enhance the efficacy of the pesticide, and optionally, the yeast particles enhance the ability of the pesticide to control plant pests by at least 5%, 10%, 20%, 30%, 40%, or 50% compared to a control composition that does not include the yeast particles.

[0082] In some embodiments, the yeast particles are commercially available yeast particles. In some embodiments, the yeast particles comprise β-glucan and / or mannan oligosaccharides. In some embodiments, the yeast particles comprise β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan, and optionally, the β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan is branched. In some embodiments, the yeast particles comprise intact or fragmented yeast cell walls. In some embodiments, the yeast particle comprises β-1,6-glucan, β-1,3-glucan, mannan oligosaccharide, mannoprotein, chitin, and / or lipid, optionally, the lipid being phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), and / or cytidine diphosphate diacylglycerol (CDP-DAG).

[0083] In some embodiments, the yeast particles do not include yeast extract. In some embodiments, the yeast particles do not consist of yeast extract.

[0084] In some embodiments, the yeast particles are in a composition further comprising one or more of an acidifying agent, a buffering agent, an antifoaming agent, an anti-transpirant, a biocide preservative, a dye and brightener, a compatibilizer, a crop oil concentrate, a surfactant, a deposition agent, a drift reducing agent, a feeding stimulant, a spreading agent, a bulking agent, an adhesive, a suspending agent, a gelling agent, a synergist, a wetting agent, an emulsifier, a dispersing agent, a penetrating agent, a neutralizing agent, a water absorbing agent, and / or a water softener. In some embodiments, the yeast particles are in a composition further comprising one or more of a surfactant, an antifoaming agent, and a biocide preservative. In some embodiments, the yeast particles are in a composition formulated as a spray, a liquid (including homogeneous mixtures such as soluble liquid concentrates and heterogeneous mixtures such as suspensions, colloids, micelles, and emulsions), a wettable powder, a water dispersible granule, a suspension, an emulsion, an aerosol, an emulsifiable concentrate, or a solid.

[0085] In some embodiments, the yeast particles are delivered in a composition that does not contain a pesticide. In some embodiments, the yeast particles are delivered in a composition that does not contain a Bt pesticide. In some embodiments, the yeast particles are delivered in a composition that does not contain a biopesticide. In some embodiments, the yeast particles are delivered in a composition that does not contain a chemical pesticide.

[0086] In some embodiments, the yeast particles are applied at a dosage of about 50 to about 300 g yeast particles / ha; or about 200 to about 300 g yeast particles / ha; or about 100 to about 500 g yeast particles / ha; or about 100 to about 1,000 g yeast particles / ha; or about 200 g yeast particles / ha; or about 250 g yeast particles / ha; or about 300 g yeast particles / ha; or about 350 g yeast particles / ha; or about 400 g yeast particles / ha; or about 450 g yeast particles / ha; or about 500 g yeast particles / ha; or about 550 g yeast particles / ha.

[0087] Some aspects of the present disclosure provide methods for controlling insect infestation of a plant, the method comprising delivering a composition to the plant, to the soil, to the insect's food, or to the insect, the composition comprising a pesticidal polynucleotide and one or more polysaccharides, and optionally, the insect is a lepidopteran insect.

[0088] Some aspects of the present disclosure provide methods for controlling plant pests, the methods comprising delivering one or more polysaccharides to a transgenic plant engineered to express a biopesticide or pesticidal polynucleotide.

[0089] In some embodiments, the one or more polysaccharides comprise a β-1,3-glucan, a β-1,6-glucan, and / or a β-1,3-1,6-glucan, and optionally, the β-1,3-glucan, the β-1,6-glucan, and / or the β-1,3-1,6-glucan is branched.

[0090] In some embodiments, the one or more polysaccharides comprise a β-glucan derived from yeast, optionally a β-glucan derived from Saccharomyces cerevisiae.

[0091] In some embodiments, the one or more polysaccharides are derived from a source other than yeast, and optionally the non-yeast source is a bacterium, a fungus, an algae, a lichen, or a plant.

[0092] In some embodiments, the one or more polysaccharides comprise a β-1,3-glucan derived from algae, optionally a β-1,3-glucan derived from Euglena gracilis.

[0093] In some embodiments, the transgenic plant expresses one or more proteins that control a plant pest, and optionally the plant pest is a fungus or an insect. In some embodiments, the plant pest is an insect.

[0094] In some embodiments, one or more proteins expressed by the transgenic plant are activated in the insect gut. In some embodiments, one or more proteins expressed by the transgenic plant are selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth. In some embodiments, one or more proteins expressed by the transgenic plant are Bacillus thuringiensis (Bt) crystal proteins. In some embodiments, one or more proteins expressed by the transgenic plant are selected from the group consisting of VIP3a, Cry1F, Cry1AB, Cry1Ac, Cry1.A105, Cry2AB, Cry1B.868, Cry1C, Cry1Da7, Cry2Ae, and Cry1Fa. In some embodiments, the one or more proteins expressed by the transgenic plant are selected from the group consisting of Cry1F, Cry1AB, Cry1Ac, Cry1.A105, Cry2AB, Cry1B.868, Cry1C, Cry1Da7, Cry2Ae, and Cry1Fa. In some embodiments, the one or more proteins expressed by the transgenic plant are selected from the group consisting of Cry1F, Cry1Fa, and Cry1Ab.

[0095] In some embodiments, the transgenic plant is selected from the group consisting of a Solanaceae plant, a Brassicaceae plant, a Poaceae plant, a Cucurbitaceae plant, a Fabaceae plant, a Fagaceae plant, an Asteraceae plant, an Amaryllidaceae plant, a Umbelliferae plant, an Apiaceae plant, an Amranthaceae plant, and a Malvaceae plant, optionally, the plant is selected from the group consisting of corn, soybean, and cotton.

[0096] In some embodiments, the transgenic plant is in the family Malvaceae. In some embodiments, the plant in the family Malvaceae is a cotton seed.

[0097] In some embodiments, the transgenic plant is in the Poaceae family. In some embodiments, the plant in the Poaceae family is a maize species. In some embodiments, the transgenic plant in the Poaceae family is a rice species.

[0098] In some embodiments, the transgenic plant is a plant in the Fabaceae family. In some embodiments, the Fabaceae plant is soybean.

[0099] In some embodiments, the pesticidal polynucleotide inhibits the expression of a target gene in a plant pest, and optionally, the plant pest is an insect. In some embodiments, the pesticidal polynucleotide is a single-stranded RNA (ssRNA) or a double-stranded RNA (dsRNA). In some embodiments, the pesticidal polynucleotide comprises a first strand that is identical to or complementary to a region of a messenger RNA (mRNA) encoded by one or more target genes. In some embodiments, the pesticidal polynucleotide is a dsRNA comprising a second strand that is complementary to the first strand.

[0100] In some embodiments, the target gene comprises one or more genes selected from the group consisting of proteasome beta 5 (PSMB5) gene, vacuolar ATPase E (vATPase E) gene, calmodulin gene, inhibitor of apoptosis protein (IAP) gene, soluble NSF attachment protein (a-SNAP) gene, Ras opposite (ROP) gene, proteasome alpha type 2 (PTSA2) gene, secretion associated Ras related GTPase 1 (SAR1) gene, PBAN, ATPase, wings up A (wupA), CP4S3_DROME, and C12C1_DROME. In some embodiments, the target gene is an IAP.

[0101] In some embodiments, the target gene has the sequence of SEQ ID NO:1.

[0102] In some embodiments, the pesticidal polynucleotide comprises at least 20, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 contiguous nucleotides that are complementary to or comprise at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity to a segment of the target gene. In some embodiments, the pesticidal polynucleotide comprises at least 20, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 contiguous nucleotides that are complementary to or comprise at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity to a segment of a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0103] In some embodiments, the composition is delivered to the leaves, stems, branches, seeds, fruits, flowers, roots, or soil of the plant. In some embodiments, delivering the composition comprises topically applying the composition to the plant or soil. In some embodiments, delivering the composition comprises spraying the composition onto the leaves, stems, branches, seeds, fruits, flowers, roots, or soil of the plant, and optionally, delivering comprises spraying the composition onto the leaves of the plant. In some embodiments, delivering the composition to the plant comprises seed treatment.

[0104] In some embodiments, the plant is selected from the group consisting of Solanaceae plants, Brassicaceae plants, Poaceae plants, Cucurbitaceae plants, Fabaceae plants, Fagaceae plants, Asteraceae plants, Amaryllidaceae plants, Umbelliferae plants, Apiaceae plants, Amranthaceae plants, and Malvaceae plants, and optionally, the plant is selected from the group consisting of corn, soybean, and cotton.

[0105] In some embodiments, the plant pest is an insect. In some embodiments, the plant pest is an insect pest belonging to the order Lepidoptera, Coleoptera, Hemiptera, Diptera, or Acari.

[0106] In some embodiments, the plant pest is an insect that is resistant to one or more biopesticides, optionally, the one or more biopesticides comprise one or more Bt proteins, and optionally, the one or more Bt proteins comprise one or more Bt crystal proteins.

[0107] In some embodiments, plant pests belonging to the order Lepidoptera include Nymphalidae (nymphalians), Danaidae (monarch butterflies), Pieridae (pierid and ceratopsian butterflies), Papilionidae (swallowtail butterflies), Lycaenidae (lycaenid butterflies), Hesperiidae (skippers), Tineidae (moths), Sesiidae (cleaners). The present invention also includes species of families selected from the group consisting of: Pyralidae (pyralid moths), Pyralidae (pyralid moths), Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawk moths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (inchworms, noctuids, and underwing moths), Tortricidae (tortricid moths), and Plutellidae (diamond moths).

[0108] In some embodiments, the lepidopteran insect species is one or more of a Plutella species, a Spodoptera species, a Helicoverpa species, or a Chyrysodeixis species.

[0109] In some embodiments, the lepidopteran insect species is a Plutella species, and optionally the Plutella species insect is Plutella xylostella.

[0110] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera frugiperda.

[0111] In some embodiments, the lepidopteran insect species is a Chrysodeixis species, and optionally, the Chrysodeixis species insect is Chrysodeixis includens (the soybean looper).

[0112] In some embodiments, the lepidopteran insect species is a Helicoverpa species, and optionally the Helicoverpa species insect is Helicoverpa zea (tomato fruitworm).

[0113] In some embodiments, the lepidopteran insect species is Cydia spp., and optionally, the Cydia spp. insect is Cydia pomonella (the fruit moth).

[0114] In some embodiments, the lepidopteran insect species is a Trichoplusia species, and optionally the Trichoplusia species insect is Trichoplusia ni (cabbage looper).

[0115] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera exigua (beet armyworm).

[0116] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera litura (tobacco rootworm).

[0117] In some embodiments, the lepidopteran insect species is a Spodoptera species, and optionally, the Spodoptera species insect is Spodoptera litoralis (the tomato moth).

[0118] In some embodiments, the lepidopteran insect species is a Pieris species, and optionally, the Pieris species insect is Pieris rapae (imported caterpillar).

[0119] In some embodiments, the lepidopteran insect species is a Tuta species, and optionally, the Tuta species insect is Tuta absoluta (tomato tea moth).

[0120] In some embodiments, the lepidopteran insect species is a Helicoverpa species, and optionally, the Helicoverpa species insect is Helicoverpa armigera (cotton boll worm).

[0121] In some embodiments, the lepidopteran insect species is a Chrysodeixis species, and optionally, the Chrysodeixis species insect is Chyrysodeixis acuta (tomato semilooper).

[0122] In some embodiments, the lepidopteran insect species is Paramyelois species, and optionally, the Paramyelois species insect is Paramyelois transitella (navel orangeworm).

[0123] In some embodiments, the lepidopteran insect species is Lobesia spp., and optionally the Lobesia spp. insect is Lobesia botrana (European grapevine moth).

[0124] In some embodiments, the insect belongs to the order Coleoptera, and the Coleopteran insect is of a species selected from the group consisting of Leptinotarsa ​​spp., Phyllotreta spp., Cerotoma spp., Diabrotica spp., Tribolium spp., Anthonomus spp., and Alticini spp.

[0125] In some embodiments, the Coleoptera insect species is a Phyllotreta species, and optionally the Phyllotreta species insect is Phyllotreta cruciferae (canola flea beetle).

[0126] In some embodiments, the Coleoptera insect species is a Phyllotreta species, and optionally the Phyllotreta species insect is Phyllotreta striolata (flea beetle).

[0127] In some embodiments, the Coleopotera insect species is a Psylliodes species, and optionally the Psylliodes species insect is Psylliodes chrysocephala (cabbage stem flea beetle).

[0128] In some embodiments, the percent mortality of the plant pest is increased by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the yeast particles, and optionally, the mortality of the plant pest is increased by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0129] In some embodiments, the percent plant part consumption by the plant pest is reduced by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the yeast particles, and optionally, the percent plant part consumption by the plant pest is reduced by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0130] In some embodiments, the percent plant part surface area affected by the plant pest is reduced by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the yeast particles, and optionally, the percent plant part surface area affected by the plant pest is reduced by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0131] In some embodiments, the effective amount of pesticide required to produce a desired outcome is reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 60% compared to a control method, optionally wherein the control method does not include yeast particles, beta-glucan, mannan oligosaccharides, and / or laminarin.

[0132] In some embodiments, the desired outcome is (a) an increase in mortality of plant pests by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 75% after delivery; (b) a decrease in plant part consumption by plant pests by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 75% after delivery; and / or (c) a decrease in plant part surface area affected by plant pests by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 75% after delivery.

[0133] In some embodiments, the method includes delivering an effective amount of a pesticide. In some embodiments, the effective amount of the pesticide required to control plant pests in a composition comprising yeast particles or one or more polysaccharides is lower than the effective amount of the pesticide in a composition that does not comprise yeast particles or one or more polysaccharides.

[0134] In some embodiments, the method further comprises delivering a pesticide having a different mode of action than the composition to the plant, the ground, the plant pest, or the food of the plant pest.

[0135] The foregoing and other features and advantages of the present disclosure will be more fully understood from the following detailed description of illustrative embodiments taken in conjunction with the accompanying drawings. [Brief explanation of the drawings]

[0136] Brief description of the drawings [Figure 1] Figure 1 graphically depicts mortality of newly hatched first-instar larvae of the diamondback moth (DBM) (Plutella xylostolla) after 5, 7, and 10 days of infestation on Chinese cabbage leaf discs treated with naked dsRNA or complex core A (CCA)-encapsulated dsRNA. GS134: control, non-insecticidal dsRNA. GS329: insecticidal dsRNA. Naked dsRNA was used at a concentration of 2.5 g / L, and CCA-encapsulated dsRNA was used at a concentration of 0.5 g / L.

[0137] [Figure 2] Figure 2 graphically depicts mortality of newly hatched first-instar diamondback moth (DBM) larvae after 5, 7, and 10 days of infestation on Chinese cabbage leaf disks treated with a combination of a commercial Bacillus thuringiensis (Bt) product and 0.5 g / L of either naked dsRNA or CCA-encapsulated dsRNA. GS134: control, non-insecticidal dsRNA. GS329: insecticidal dsRNA. Bt product alone (compound only) was used as a control.

[0138] [Figure 3] FIG. 3 graphically depicts mortality of newly hatched first instar larvae of the diamondback moth (DBM) after 5, 7, and 10 days of infestation on Chinese cabbage leaf discs treated with a combination of a commercial Bacillus thuringiensis (Bt) product and empty yeast particles (YP) at various concentrations.

[0139] [Figure 4] FIG. 4 graphically depicts mortality of newly hatched first-instar larvae of Spodoptera frugiperda (Fallen Armyworm, FAW) after 5, 7, and 10 days of infestation on soybean leaf discs treated with a combination of a commercial Bacillus thuringiensis (Bt) product and empty yeast particles (YP) at various concentrations.

[0140] [Figure 5] FIG. 5 graphically depicts dose-response curves for Bt (control) and Bt+YP indicating predicted mortality of Plutella xlylostella after 5, 7, and 11 days.

[0141] [Figure 6] Figure 6 graphically depicts the rating scale scores (a measure of insect mortality) of first instar larvae of the fall armyworm (FAW) susceptible to the Bacillus thuringiensis (Bt) Cry1Ab protein after 12 days of infestation on transgenic corn plants expressing the Bacillus thuringiensis (Bt) Cry1Ab protein (transformed plants) and standard corn plants (non-transformed plants) treated with yeast particles.

[0142] [Figure 7]7 is a graphical depiction of the number of insects per plant (a measure of insect mortality) of first-instar larvae of the fall armyworm (FAW) resistant to the Bacillus thuringiensis (Bt) Cry1Ab protein after 14 days of infestation on transgenic corn plants expressing the Bt Cry1Ab protein (transformed plants) and standard corn plants (non-transformed plants) that were treated with yeast particles. The corn plants were sprayed with yeast particles once or twice.

[0143] [Figure 8] 8 graphically depicts the number of insects per plant (a measure of insect mortality) of first instar larvae of the fall armyworm (FAW) resistant to the Bacillus thuringiensis (Bt) Cry1Ab protein after 14 days of infestation on transgenic corn plants expressing the Bt Cry1Ab protein (transformed plants) and standard corn plants (non-transformed plants) treated with yeast particles with or without Coragen® (chlorantraniliprole). The yeast particles were sprayed on the corn plants once or twice.

[0144] [Figure 9] 9 graphically depicts infestation of transgenic corn plants expressing the Bacillus thuringiensis (Bt) Cry1Ab protein by first-instar larvae of the fall armyworm (FAW) resistant to the Bacillus thuringiensis (Bt) Cry1Ab protein and defoliation scores after treatment with Coragen® (chlorantraniliprole). The addition of yeast particles in the delivered composition provided superior results at low concentrations of Coragen® (chlorantraniliprole).

[0145] [Figure 10]10 graphically depicts the average percent defoliation (left) and average number of insects per cage (a measure of insect mortality; right) following infestation of soybeans with soybean looper insects and treatment with Coragen® (chlorantraniliprole). The addition of yeast particles in the delivered composition provided superior results compared to the same concentration of Coragen® without the yeast particles.

[0146] [Figure 11] 11 graphically depicts the average percent defoliation (left) and average number of insects per cage (a measure of insect mortality; right) following infestation of soybeans with soybean looper insects and treatment with Rimon (Novaluron). The addition of yeast particles in the delivered composition provided superior results compared to the same concentration of Rimon without the yeast particles.

[0147] [Figure 12] 12 graphically depicts the average percent defoliation (left) and average number of insects per cage (a measure of insect mortality; right) following infestation of soybeans with soybean looper insects and treatment with Entrust (spinosad). The addition of yeast particles in the delivered composition provided superior results compared to the same concentration of Entrust without the yeast particles.

[0148] [Figures 13A-13B] 13A-13B are graphical depictions of infestation of cabbage plants by diamondback moth insects and percent plant damage after treatment with diflubenzuron. The addition of yeast particles in the delivered composition provided superior results compared to the same concentration of diflubenzuron without the yeast particles.

[0149] [Figure 14] 14 is a graphical depiction of infestation of cauliflower plants by diamondback moth insects and percent plant damage after treatment with cypermethrin. The addition of yeast particles in the delivered composition provided superior results compared to the same concentration of cypermethrin without the yeast particles.

[0150] [Figure 15] 15 is a graphical depiction of infestation of cauliflower plants by diamondback moth insects and percent plant damage following treatment with Bacillus thuringiensis (Bt) microorganisms. The addition of yeast particles in the delivered composition provided superior results compared to the same concentration of Bt microorganisms without the yeast particles.

[0151] [Figure 16] 16 is a graphical depiction of infestation of soybean plants by diamondback moth insects and percent plant damage after treatment with acetamiprid. The addition of yeast particles in the delivered composition provided superior results compared to the same concentration of acetamiprid without the yeast particles.

[0152] [Figure 17] FIG. 17 is a graphical depiction of infestation of cabbage plants by diamondback moth insects and percent mortality following treatment with a composition comprising Bacillus thuringiensis (Bt) microorganisms to assess the ability of polysaccharide and yeast particles to enhance the efficacy of a biopesticide.

[0153] [Figure 18] FIG. 18 is a graphical depiction of the ability of compositions comprising yeast particles encapsulating a pesticidal polynucleotide and a biopesticide to control plant pests.

[0154] [Figure 19] FIG. 19 is a graphical depiction of the ability of compositions comprising yeast particles encapsulating pesticidal polynucleotides to control plant pests. DETAILED DESCRIPTION OF THE INVENTION

[0155] Detailed Description The present disclosure provides compositions, methods, and systems for controlling plant infestation by plant pests. Compositions of the present disclosure containing a mixture of a pesticide (e.g., a biological pesticide, a pesticidal polynucleotide, or a chemical pesticide) and a biological adjuvant (e.g., yeast particles containing yeast cell wall components, β-glucan, mannan oligosaccharides, and / or laminarin) have demonstrated increased efficacy against plant pests (e.g., insect pests, fungal pests) compared to the pesticide alone.

[0156] definition Unless otherwise defined, all technical features and all technical and scientific terms used have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs.When a term is provided in the singular, the inventors also contemplate aspects of this disclosure described by the plural of that term.If there is a discrepancy in the terms and definitions used in references incorporated by reference, the terms used in this application shall have the definitions given herein.Other technical terms used have their ordinary meaning in the technical field in which they are used, as exemplified by various technology-specific dictionaries, such as The American Heritage® Science Dictionary (Editors of the American Heritage Dictionaries, 2011, Houghton Mifflin Harcourt, Boston and New York), the McGraw-Hill Dictionary of Scientific and Technical Terms (6th edition, 2002, McGraw-Hill, New York), or the Oxford Dictionary of Biology (6th edition, 2008, Oxford University Press, Oxford and New York). We do not intend to be limited by the mechanism or mode of action, and references thereto are provided for illustrative purposes only.

[0157] In order that the present disclosure may be more readily understood, selected terms are defined below.

[0158] As used herein, the term "pesticide" (which may be used interchangeably with the term "pesticidal compound") generally describes a compound capable of controlling pests (e.g., plant pests such as insect pests, fungal pests, bacterial pests, parasites, etc.). In some embodiments, a pesticide is a compound capable of inducing physiological or behavioral changes in pests (e.g., plant pests) through a specific biological effect, such as, but not limited to, increased mortality, growth inhibition, reduced or fertility, reduced or cessation of feeding or movement, or reduced or cessation of development of metamorphic stages. In some embodiments, a pesticide is an insecticide (a compound capable of controlling insects) or a fungicide (a compound capable of controlling fungal species). In some embodiments, pesticides can be classified as belonging to a mode of action as defined by the Insecticide Resistance Task Force (IRAC) (e.g., as defined at https: / / irac-online.org / ). The pesticide may be a biological pesticide, a pesticidal polynucleotide, or a chemical pesticide.

[0159] In the context of pesticide compositions and methods, the term "control" or "controlling" in relation to controlling plant pests means inducing a physiological or behavioral change, such as, but not limited to, causing increased mortality, stunted growth, reduced reproductive or fecundity, reduced or cessation of feeding or movement, or reduced or cessation of development of metamorphic stages.

[0160] As used herein, the term "biopesticide" generally refers to a pesticide or pesticidal compound that is a protein or polynucleotide derived from or isolated from a whole microorganism (e.g., whole bacteria), or an animal, plant, microorganism (e.g., bacteria, cyanobacteria, algae, etc.), fungal species, or certain minerals. In some embodiments, the biopesticide is a protein isolated from an animal, plant, microorganism (e.g., bacteria, cyanobacteria, algae, etc.), or fungal species. The biopesticide may comprise a whole organism (e.g., a microorganism, e.g., a Bacillus thuringiensis microorganism) or a cellular component of an organism (e.g., a microorganism, e.g., a Bacillus thuringiensis microorganism). In some embodiments, the cellular component comprises one or more proteins that control plant pests (e.g., a crystal protein from a Bacillus thuringiensis microorganism or an insecticidal protein produced during vegetative growth). In some embodiments, the biopesticide comprises a toxin complex (Tc) protein expressed by Photorhabdus and Xenorhabdus bacteria. In some embodiments, the biopesticide is a protein or nucleic acid obtained from an animal, plant, microorganism (e.g., bacteria, cyanobacteria, algae, etc.), or fungal species. In some embodiments, the biopesticide is a recombinantly or synthetically produced protein. In some embodiments, the biopesticide is a protein expressed from a transgenic organism (e.g., a transgenic plant). In some embodiments, the biopesticide is a polynucleotide isolated from an animal, plant, microorganism (e.g., bacteria, cyanobacteria, algae, etc.), or fungal species. In some embodiments, the biopesticide is a recombinantly or synthetically produced polynucleotide. In some embodiments, the biopesticide is a polynucleotide expressed from a transgenic organism (e.g., a transgenic plant).

[0161] As used herein, the term "pesticidal polynucleotide" generally refers to a pesticide or pesticidal compound that comprises or consists of a polynucleotide molecule. A polynucleotide molecule is a polymer of nucleotide monomers covalently linked in a linear chain. In some embodiments, the pesticidal polynucleotide is a polynucleotide that regulates the expression of a target gene in a plant pest (e.g., to effect polynucleotide-mediated control of the plant pest). In some embodiments, the pesticidal polynucleotide is a polynucleotide that inhibits the expression of a target gene (e.g., inhibits DNA transcription or mRNA translation) in a plant pest (e.g., to effect polynucleotide-mediated control of the plant pest). In some embodiments, the pesticidal polynucleotide is an RNA interference molecule. The pesticidal polynucleotide can function to regulate the expression of a target gene by binding (e.g., transiently binding) to messenger RNA encoded by the target gene (e.g., resulting in inhibition of mRNA translation due to mRNA degradation). In some embodiments, the pesticidal polynucleotide is a double-stranded RNA (dsRNA) molecule. In some embodiments, if there is an epigenetic change near the gene of interest, the pesticidal polynucleotide may inhibit the expression of the mRNA encoded by the target gene. In some embodiments, the pesticidal polynucleotide is a double-stranded RNA (dsRNA) that inhibits the expression of the coding region of the target gene. In other embodiments, the pesticidal polynucleotide is a DNA sequence that encodes an RNA molecule (e.g., a dsRNA molecule). In some embodiments, the pesticidal polynucleotide is an antisense RNA. It should be understood that the sequences disclosed herein as DNA sequences can be converted from DNA sequences to RNA sequences by substituting some or each of the thymidines with uracil.

[0162] As used herein, the term "chemical pesticide" generally refers to a pesticide or pesticidal compound that is a small molecule pesticide. A chemical pesticide can be a natural product small molecule or a synthetic small molecule (e.g., a synthetically produced small molecule). A small molecule can be any organic compound with a low molecular weight (e.g., less than 1000 daltons).

[0163] As used herein, the term "biological adjuvant" refers to a substance derived from an organism, including but not limited to an animal, plant, bacterium, fungus, algae, or lichen, that, when administered to a host in association with one or more pesticides (e.g., biopesticides, pesticidal polynucleotides, chemical pesticides), enhances the efficacy of the one or more biopesticides or pesticidal polynucleotides. In some embodiments, the biological adjuvant is a yeast particle (e.g., a yeast particle comprising yeast cell wall components). In some embodiments, the biological adjuvant is one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharide, and laminarin.

[0164] As used herein, the term "plant" includes plant bodies, plant organs (e.g., leaves, petals, stems, roots, rhizomes, and seeds), plant tissues (e.g., epidermis, phloem, parenchyma, xylem, and vascular bundles), and plant cells. In addition, the term "plant cells" includes seed suspension cultures, embryos, meristematic regions, callus tissue, cells derived from leaves and roots, and gametophytes (embryos and pollen), and their precursors.

[0165] As used herein, the term "plant pest" refers to an organism (e.g., an insect pest or a fungal pest) that damages a plant or a population of plants by living, growing, and / or feeding on the plant or a population of plants. In some embodiments, a plant pest damages a plant or a population of plants by shortening the plant's lifespan, reducing the plant's size (e.g., reducing the size of its leaves, roots, stems), reducing the plant's reproductive capacity, and / or reducing the yield of fruits, vegetables, flowers, seeds, foliage, or other commodities resulting from the population of plants. Plant pests can include viruses, bacteria, fungi, rodents, parasites, and insects.

[0166] As used herein, "Bt products" refers to products that contain compounds derived from Bacillus thuringiensis or Bt.

[0167] As used herein, the term "Bt crystal protein" refers to parasporal crystalline inclusions produced by Bacillus thuringiensis (Bt), many of which are pore-forming toxins that specifically target invertebrates but are harmless to mammals. In some embodiments, the Bt crystal proteins are Cry1F, Cry1AB, Cry1Ac, Cry1.A105, Cry2AB, Cry1B.868, Cry1C, Cry1Da7, Cry2Ae, and Cry1Fa.

[0168] As used herein, the term "recombinant microorganism" refers to a genetically modified microorganism or a microorganism that maintains and replicates recombinant DNA.

[0169] As used herein, the term "transgenic plant" (also called "transformed plant") is a plant that has been genetically modified (or engineered) using recombinant DNA technology. In some embodiments, the transgenic plant has been modified to express a gene that is not native to the plant or to modify an endogenous gene. In some embodiments, the protein encoded by the gene provides a particular trait or characteristic to the plant. In some embodiments, the transgenic plant has been engineered to express a biopesticide or pesticide polynucleotide.

[0170] As used herein, the term "yeast particle" or "YP" refers to a yeast cell or a yeast cell wall component derived from a yeast cell. The yeast particle may contain yeast cell wall components (e.g., β-1,6-glucan, β-1,3-glucan, β-1,3 / 1,6-glucan (also called β-1,3-1,6-glucan), mannan oligosaccharides, mannoproteins, chitin, and / or lipids). In some embodiments, the yeast cell wall component comprises a lipid selected from the group consisting of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), and / or cytidine diphosphate diacylglycerol (CDP-DAG). In some embodiments, the yeast particle is not yeast extract. In some embodiments, the yeast particle consists of yeast extract. Examples of yeast particles (e.g., yeast particles containing yeast cell wall components) include, but are not limited to, heat-killed whole yeast cells, yeast cell walls, and yeast cell wall components remaining after an extraction process. Yeast particles may be, for example, in the form of hollow shells and / or yeast cell wall fragments. Yeast particles include, but are not limited to, commercially available yeast particles (e.g., ACTIVEMOS® and SAFMANNAN®), extracted yeast cell wall particles (YCWP), yeast cell particles (YCP), yeast glucan particles (GP or YGP), yeast glucan mannan particles (GMP or YGMP), yeast glucan chitin particles (GCP or YGCP), yeast glucan chitin mannan particles (GCMP or YGCMP), yeast glucan lipid particles (GLP or YGLP), whole glucan particles (WGP), etc. In some embodiments, a yeast particle (e.g., a yeast particle comprising a yeast cell wall component) comprises β-glucan (e.g., β-1,3-branched glucan, β-1,6-branched glucan, and / or β-1,3-1,6-branched glucan) and / or mannan oligosaccharides.

[0171] Yeast particles containing yeast cell wall components may be prepared from yeast cells by extraction and purification of the insoluble particle fraction from the soluble components of the yeast cells. Fungal cell walls can be produced from the insoluble by-products of yeast extract production. Furthermore, yeast cells can be treated with aqueous hydroxide to digest the proteins and intracellular parts of the cells without destroying the yeast cell walls, and the yeast cell wall components remain free of significant protein contamination and have substantially unchanged cell wall structure of β(1-6) and β(1-3) linked glucans. In some embodiments, the process for preparing yeast particles of the present disclosure is as described in U.S. Patent No. 4,810,646; U.S. Patent No. 6,242,594; U.S. Patent No. 5,401,727; or U.S. Patent No. 5,607,677, the entire contents of which are incorporated herein by reference. In some embodiments, the process for preparing yeast particles of the present disclosure involves preparing yeast glucan particles by alkaline extraction, acid extraction, and then extraction with an organic solvent, and finally drying.

[0172] As used herein, the term "polysaccharide" refers to a long-chain polymeric carbohydrate composed of monosaccharide units linked by glycosidic bonds. In some embodiments, the polysaccharide is a disaccharide or trisaccharide. In some embodiments, the polysaccharide contains at least 3, 5, 7, 9, or 10 monosaccharide units linked by glycosidic bonds. In some embodiments, the polysaccharide is a long-chain polymeric carbohydrate composed of glucan units linked by glycosidic bonds.

[0173] In some embodiments, the polysaccharide is a long-chain polymeric carbohydrate composed of glucan units linked by β-1,3 and / or β-1,6 bonds. In some embodiments, the β-glucan comprises β-1,3-branched glucan, β-1,6-branched glucan, and / or β-1,3-1,6-branched glucan. In some embodiments, the beta-glucan comprises a linear chain of glucose units without branches. In some embodiments, the beta-glucan comprises a branched chain of glucose units. In some embodiments, the molecular weight of the beta-glucan is at least 300,000 daltons, at least 400,000 daltons, or at least 500,000 daltons. In some embodiments, the molecular weight of the beta-glucan is 35,600-650,000 daltons; 200,000-600,000 daltons; 300,000-600,000 daltons; or 400,000-600,000 daltons. In some embodiments, the molecular weight of the beta-glucan is about 500,000.

[0174] In some embodiments, the polysaccharide is a mannan oligosaccharide. In some embodiments, the mannan oligosaccharide is a long-chain polymeric carbohydrate composed of mannose units. In some embodiments, the mannan oligosaccharide is a long-chain polymeric carbohydrate composed of mannose units and galactose units. In some embodiments, the mannan oligosaccharide is a long-chain polymeric carbohydrate composed of mannose units and glucose units. In some embodiments, the mannan oligosaccharide is a long-chain polymeric carbohydrate composed of mannose units, galactose units, and glucose units. In some embodiments, the mannan oligosaccharide contains α-1,2 linkages, α-1,3 linkages, and / or α-1,6 linkages. In some embodiments, the mannan oligosaccharide contains β-1,4 linkages and / or α-1,6 linkages.

[0175] In some embodiments, the polysaccharide is laminarin. In some embodiments, laminarin is a long-chain polymeric carbohydrate composed of glucose units. In some embodiments, laminarin is a linear polysaccharide. In some embodiments, laminarin contains β-1,3 and / or β-1,6 linkages. In some embodiments, laminarin contains β-1,3 and β-1,6 linkages in a 3:1 ratio. Laminarin can be prepared as described in Kadam et al. Extraction, structure and biofunctional activities of laminarin from brown algae. International Journal of Food Science and Technology (2015), the contents of which are incorporated herein by reference. In some embodiments, laminarin is derived from Laminaria digitata.

[0176] As used herein, the term "Coleoptera insects" refers to any insect of the order Coleoptera. Examples of insects of the order Coleoptera include, but are not limited to, the families Chrysomelidae (leaf beetles, broad-shouldered beetles, alligatorweed flea beetles), Curculionidae (weevils), Meloidae (blister beetles), Tenebrionidae (meal beetles), Scarabaeidae (scarab beetles), and Cerambycidae (long-horned beetles). Cerambycidae (pine beetles), Curculionidae (Chinese white pine beetles), Nitidulidae (small hive beetles), Cerambycidae (mulberry longhorn beetles), Phyllotreta (flea beetles), Diabrotica (corn rootworms), Chrysomela ( Cottonwood Leaf Beetle, Hypothenemus (Coffee Berry Borer), Sitophilus (Maize Weevil), Epitrix (Tobacco Flea Beetle), E. cucumeris (Potato Beetle), P. pusilla (Western Black Flea Beetle); Anthonomus (Pepper Weevil), Hemicrep idus (wireworm), Melanotus (wireworm), Ceutorhychus (cabbage pod weevil), Aeolus (wireworm), Horistonotus (sandworm), Sphenophorus (corn billbug), S. zea (timothy billbug), S. parvulus (bluegrass billbug), S.callosus (southern cornbill bug); Phyllophaga (white grub), Chaetocnema (corn flea beetle), Popillia (bean beetle), Epilachna (green lady beetle), Cerotoma (bean leaf beetle), Epicauta (blister beetle), and any combination thereof. Within the Chrysomelidae family, examples may include any species of the genus Leptinotarsa. Leptinotarsa ​​species include, but are not limited to, Leptinotarsa ​​decemlineata (Colorado potato beetle), Leptinotarsa ​​juncta (false potato beetle), Leptinotarsa ​​behrensi, Leptinotarsa ​​collinsi, Leptinotarsa ​​defecta, Leptinotarsa ​​haldemani (Holdman's green potato beetle), Leptinotarsa ​​heydeni, Leptinotarsa ​​juncta (false potato beetle), Leptinotarsa ​​ineolate (burrobrush leaf beetle), Leptinotarsa ​​peninsularis, Leptinotarsa ​​rubiginosa, Leptinotarsa ​​texana, Leptinotarsa ​​tlascalana, Leptinotarsa ​​tumamoca, and Leptinotarsa ​​typographica.

[0177] As used herein, the term "lepidopteran insects" refers to any insect belonging to the order Lepidoptera. Examples of insects within the order Lepidoptera include the families Nymphalidae (nymphalians), Danaidae (monarchs), Pieridae (pierids and ceratopsians), Papilionidae (swallowtails), Lycaenidae (lycaenids), Hesperiidae (skippers), Tineidae (moths), Sesiidae (clematids), and the families The Noctuidae include any species of the genus Spodoptera, including, for example, S. frugiperda (snake moths), S. siidae (clearwing moths), Pyralidae (pyralid moths), Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawkmoths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (looper moths), and Plutellidae (diamond moths). The Noctuidae include any species of the genus Spodoptera, including, for example, S. frugiperda (snake armyworm). In the family Plutellidae, examples include Plutella species such as P. xylostolla (diamond moth).

[0178] As used herein, the term "effective amount" generally refers to the amount or concentration of pesticide(s) required to achieve an observable effect on physiological or behavioral changes in a pest (e.g., a plant pest), such as, but not limited to, increased mortality, growth inhibition, reduced reproductive or fecundity, reduced or cessation of feeding or movement, or reduced or cessation of development of metamorphic stages. In some embodiments, the presence of a biological adjuvant, such as yeast particles containing yeast cell wall components, lowers the amount of pesticide required to achieve the desired effect on physiological or behavioral changes in a pest (e.g., a plant pest).

[0179] As used herein, the term "Complex Core A" (abbreviated as CCA) generally refers to a formulation comprising (i) yeast particles (e.g., yeast particles comprising yeast cell wall components) or polysaccharides (e.g., beta-glucan, mannan oligosaccharides, and / or laminarin) that form spherical particles and (ii) a nuclease inhibitor. In some embodiments, the complex further comprises a cationic polymer (e.g., polyethyleneimine (PEI) or poly-L-lysine (PLL)). In some embodiments, the Complex Core A formulation is 2-10, 2-5, 2-6, 3-10, 3-5, or 3-7 microns in diameter. In some embodiments, the nuclease inhibitor is ethylenediaminetetraacetic acid (EDTA), polyvinylsulfonic acid (PVSA), sodium hexametaphosphate (SHMP), sodium tripolyphosphate (TPP), diethylpyrocarbonate, aurintricarboxylic acid (ATA), formamide, macaloid, proteinase K, heparin, hydroxylamine-oxygen-copper(II) ion, bentonite, ammonium sulfate, dithiothreitol (DTT), beta-mercaptoethanol, cysteine, dithioerythritol, or tris(2-carboxyethyl)phosphene hydrochloride.

[0180] As used herein, the term "complementary" refers to the relationship between two nucleic acid sequences. A first nucleic acid sequence is complementary to a second nucleic acid sequence if it can bind to the second nucleic acid sequence and form a duplex with the second nucleic acid sequence. In some embodiments, a first nucleic acid sequence is considered complementary to a second nucleic acid sequence if it base pairs with at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the second nucleic acid sequence over the length of the first and / or second nucleic acid sequence.

[0181] As used herein, the term "substantially" refers to a variation of plus or minus 10 percent or less (e.g., plus or minus 9%, 8%, 7%, 6%, 5%, 4%, or 3% or less) relative to one or more listed items.

[0182] As used herein, the term "about" refers to numerical values, including, for example, integers, fractions, and percentages, whether or not explicitly indicated. The term "about" generally refers to a range of numerical values ​​(e.g., + / - 5 to 10% of the recited value) that one of ordinary skill in the art would consider equivalent to the recited value (e.g., having the same function or result). In some instances, the term "about" may include numerical values ​​that are rounded to the nearest significant figure.

[0183] composition In some embodiments, the present disclosure provides compositions comprising a pesticide and a biological adjuvant (e.g., yeast particles comprising cell wall components) that function to enhance (increase) the efficacy of the pesticide (e.g., by increasing the ability of the pesticide to control plant pests). In some embodiments, the biological adjuvant enhances the efficacy of the pesticide by at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70% or more. For example, in some embodiments, the biological adjuvant enhances the efficacy of the pesticide by causing the death of at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, or at least 70% of plant pests (e.g., insects). The efficacy of a pesticide is enhanced by a biological adjuvant if the efficacy is enhanced in the presence of the biological adjuvant compared to a similar concentration of the pesticide without the biological adjuvant, or if a lower amount or concentration of the pesticide is required to control the plant pest in the presence of the biological adjuvant (compared to the amount or concentration of the pesticide required to control the plant pest in the absence of the biological adjuvant). In other words, the presence (or addition) of a biological adjuvant to a composition containing a pesticide provides greater efficacy or allows for a lower effective amount of the pesticide to provide beneficial protection of plants infested by plant pests, for example. In some embodiments, the composition enhances the control of insects that have developed resistance to the pesticide.

[0184] In some embodiments, the present disclosure provides a composition comprising a yeast particle that comprises a pesticide and yeast cell wall components, wherein the pesticide is a biopesticide or a pesticide polynucleotide.In still other embodiments, the present disclosure provides a composition comprising a yeast particle that comprises a pesticide and yeast cell wall components, wherein the pesticide is a chemical pesticide, and the pesticide is a ryanodine receptor modulator (IRAC class 28), a chitin biosynthesis inhibitor that acts on CHS1 (IRAC class 15), a nicotinic acetylcholine receptor (nACHR) allosteric modulator-site 1 (IRAC class 5), a nACHR competitive modulator (IRAC class 4), or a sodium channel modulator (IRAC class 3).

[0185] In some embodiments, the present disclosure provides a composition comprising a pesticide and one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharides, and laminarin, wherein the pesticide is a biopesticide or a pesticidal polynucleotide. In yet other embodiments, the present disclosure provides a composition comprising a pesticide and one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharides, and laminarin, wherein the pesticide is a chemical pesticide and the pesticide is a ryanodine receptor modulator (IRAC class 28), a chitin biosynthesis inhibitor acting on CHS1 (IRAC class 15), a nicotinic acetylcholine receptor (nACHR) allosteric modulator-site 1 (IRAC class 5), a nACHR competitive modulator (IRAC class 4), or a sodium channel modulator (IRAC class 3).

[0186] In some embodiments, the present disclosure provides a composition comprising a pesticide and a yeast particle comprising a yeast cell wall component. In some embodiments, the present disclosure provides a composition comprising a pesticide and one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharide, and laminarin.

[0187] In one aspect, the present disclosure provides a composition comprising a biopesticide and a biological adjuvant, wherein the biological adjuvant comprises YP, and wherein the biological adjuvant enhances the efficacy of the biopesticide.

[0188] In one aspect, the present disclosure provides a composition comprising a biopesticide and a biological adjuvant, wherein the biological adjuvant comprises a polysaccharide, and wherein the biological adjuvant enhances the efficacy of the biopesticide.

[0189] Pesticides for use in compositions In some embodiments, biopesticides are preferred because they are typically non-toxic to humans, target specific pests, and biodegrade over time so they do not persist in the environment, which may require multiple applications to achieve sufficient control of the pest.

[0190] Some biopesticides function as bioinsecticides. Some biopesticides inhibit digestion and must be eaten by insects to be effective, while others impair the respiratory function of insects and act upon contact with insects. Many factors other than the ability to cause mortality determine the value of a material as a biopesticide. In the control of plant insect pests, it is necessary to employ chemicals that are non-toxic to humans and non-harmful to vegetation. In some embodiments, biopesticides function as fungicides (e.g., to control fungal species).

[0191] In some embodiments of the present disclosure, the biopesticide can be a naturally occurring microorganism, such as Bacillus thuringiensis. Bacillus thuringiensis (Bt) is a naturally occurring, spore-forming, Gram-positive soil bacterium. Bt strains produce several different types of toxin / insecticidal proteins. Bt produces insecticidal proteins (Vips) that are produced during vegetative growth during normal cell growth. During sporulation, Bt produces insecticidal crystal proteins (Cry proteins). Bt insecticidal proteins are generally selective for insects within a given order and often even more selective for specific insects. Cry proteins are activated by proteases in the high-pH gut of insect larvae. Once activated, the proteins destroy the gut lining of susceptible insects, leading to feeding cessation and eventual death by starvation within 1 to 5 days. Bt toxins and spores are destroyed by the low, acidic pH of the mammalian gut and therefore pose no known risk to humans or other mammals. Target insect specificity is a hallmark of Bt and is the basis for sprayable all-Bt strain products such as DIPEL for insect control in vegetables or major row crops (eg, corn, cotton, soybean).

[0192] Commercial Bt products are available, for example, as liquid suspensions or powders containing a mixture of dried spores and toxin crystals. They are applied to leaves or other environments where insect larvae feed. The Bt products used in this disclosure, DIPEL® and XENTARI®, are produced by Valent Biosciences. Other commercial Bt products include BACTERIOSPEINE ES®, BIOBIT HP®, FLORBAC®, COSTAR WG®, JAVELIN WP®, THURICIDE®, TEKAR®, BACTIMOS®, VECTOLEX®, NOVODOR®, TRIDENT®, and LEPROTEC®.

[0193] For example, isolated Cry proteins or Bt proteins containing vegetatively produced insecticidal proteins (Vip) may also be used as natural organic insecticides to control crop pests, mosquitoes, and blackflies. Cry proteins and / or vegetatively produced insecticidal proteins (Vip) are non-toxic to vertebrates and are EPA-approved for expression in transgenic food crops (e.g., corn, soybean, potato, etc.) and other transgenic plants (e.g., cotton). They are stable and inexpensive to mass-produce. Several transgenic crops genetically modified to express Cry proteins or vegetatively produced insecticidal proteins (Vip) are currently in use. Non-limiting examples include corn, soybean, and cotton.

[0194] Although Bt has several advantages as an environmentally safe, highly specific biological control agent often used in integrated pest management, it has several limitations to its more widespread use. First, it is relatively slow-acting compared to synthetic chemicals. Therefore, improving the rate at which Bt insecticides act is desirable to protect against feeding damage. Second, the efficacy of Bt insecticides varies depending on the target insect. To reduce dosage and application costs, improving efficacy is desirable. Furthermore, shortening application times is desirable because long-term use of Bt can lead to the development of resistance. Importantly, various populations of Bt target insects have developed resistance to Bt insecticides in the field, including sprayable Bt insecticides and Bt insecticides expressed in transgenic plants. Field resistance has been a long-standing problem, identified at least as early as 1991 (Tabashnik et al., Managing Resistance to Bacillus thuringiensis: Lessons from the Diamondback Moth (Lepidoptera: Plutellidae) J. Econ. Entomol. 84(1):49-55 (1991)). Resistance to such Bt toxins produced by various transgenic crops genetically engineered to express Bt toxins has also developed. Tabashnik et al., Global Patterns of Insect Resistance to Transgenic Bt Crops: The First 25 Years, Journal of Econ. Entomol., 2023, 1-13. By 2013, at least 11 pest species, including nine lepidopteran and two coleopteran species, had been documented to have developed or are developing resistance to Bt toxins expressed by transgenic crops.

[0195] In some embodiments, the biopesticide comprises a toxin complex (Tc) protein expressed by Paenibacillus, Photorhabdus, or Xenorhabdus bacteria. Four different toxin complexes (TCs) - Tca, Tcb, Tcc, and Tcd - have been identified in Photorhabdus species. Any protein from one of these toxin complexes can be used as a biopesticide.

[0196] In some embodiments, the biopesticide is a Paenibacillus, Photorhabdus, or Xenorhabdus bacterium (eg, whole microorganism).

[0197] In some embodiments, the biopesticide is a Btvar. israelensis, Btvar. aizawai, Btvar. kurstaki, Btvar. tenebrionensis, or Bacillus sphaericus microorganism (e.g., the whole microorganism). In some embodiments, the biopesticide is an isolated cellular component (e.g., one or more proteins or polynucleotides) derived from a Btvar. israelensis, Btvar. aizawai, Btvar. kurstaki, Btvar. tenebrionensis, or Bacillus sphaericus microorganism.

[0198] Another technology that has shown some effectiveness in controlling certain insects and fungi species is RNA interference (RNAi).The RNAi-based technology for controlling plant pests, which is a type of pesticidal polynucleotide, can reduce the damage caused by pests by delivering ribonucleic acid interference molecules that target (for example, bind to) and interfere with the messenger RNA (mRNA) of pest genes.Laboratory and field studies have confirmed that oral delivery of RNA molecules (for example, double-stranded RNA (dsRNA)), whose mode of action is through the RNAi process, is effective against many insect species, and therefore, topical dsRNA is considered to be a suitable delivery form. (See, e.g., PCT / US2019 / 053129 (published April 2, 2020, and entitled "Control of Coleopteran Insects"), PCT / US2019 / 060389 (published May 14, 2020, and entitled "Control of Insect Infestations"), and PCT / US2021 / 032334 (published November 18, 2021, and entitled "RNA-Based Control of Lepidopteran Pests"), all of which are incorporated by reference herein.) Additionally, several studies have shown that it is possible to silence essential genes to control pathogens via RNA interference without adversely affecting non-target species, allowing growers to target pests precisely and environmentally more efficiently than traditional pesticides (Cagliari et al., 2019). RNAi-based technology has also shown effectiveness in controlling fungal pathogens (see, e.g., International PCT Application PCT / US2022 / 019320, entitled "RNA-Based Control of Powdery Mildew" and published September 15, 2022, which is incorporated herein by reference). RNAi molecules targeting insect or fungal genes may be produced by any method known in the art, including, for example, the cell-free production methods described in U.S. Pat. No. 10,954,541 (entitled "Cell-Free Production of Ribonucleic Acids") and U.S. Pat. No. 10,858,385 (entitled "Methods and Compositions for Nucleoside Triphosphate and Ribonucleic Acid Production"), each of which is incorporated herein by reference.

[0199] For example, challenges exist with oral delivery of insecticidal RNA to certain lepidopteran insect species, including diamondback moth, fall armyworm, cotton bollworm, and soybean looper (Terenius et al. 2010. RNA interference in Lepidoptera: An overview of successful and unsuccessful studies and implications for experimental design doi:10.1016 / j.jinsphys.2010.11.006). Although RNA, such as dsRNA, can be effective in controlling insects when delivered to insect gut cells after gene knockdown and feeding, the insect's natural defenses in the gut environment (e.g., the presence of nucleases, highly acidic environments, highly basic environments) can degrade the RNA and limit its effective delivery. In some embodiments of the present disclosure, dsRNA is encapsulated in YPs to reduce degradation in the gut and allow more dsRNA to be delivered to cells for RNAi. In some embodiments, dsRNA is encapsulated in YPs to reduce degradation in the highly basic environment of the gut of Lepidoptera, e.g., Plutella spp., Plutella xylostella, Spodoptera spp., or S. frugiperda.

[0200] In some embodiments of the present disclosure, the biopesticide is a commercial Bt product such as DIPEL® or XENTARI® or BACTOSPEINE® ES, or BIOBIT® HP or FLORBAC® or COSTAR® WG or JAVELIN® WP or THURICIDE or TEKAR or BACTIMOS or VECTOLEX or NOVODOR or TRIDENT or LEPROTEC, or any combination thereof.

[0201] Some embodiments of the present disclosure provide uses of Bt Cry proteins. In some embodiments, the biopesticides are recombinant microorganisms engineered to produce one or more recombinant proteins that target plant pests.

[0202] In some embodiments, the recombinant microorganism is B. thuringiensis that produces one or more recombinant Cry proteins that are activated in the insect gut. In some embodiments, the one or more Cry proteins include Cry1F, Cry1Aa, Cry1Ab, and Cry1Ac.

[0203] In some embodiments, the recombinant microorganism is B. thuringiensis that produces one or more vegetatively produced insecticidal proteins (Vips). In some embodiments, the one or more Vips include Vip1, Vip2, and / or Vip3.

[0204] In certain embodiments, the biopesticide comprises isolated cellular components derived from naturally occurring or recombinant microorganisms. In certain embodiments, the cellular components are proteins that target plant pests. In certain embodiments, the proteins are activated in the insect gut. In one embodiment, the protein is a crystal protein. In some embodiments, the crystal protein belongs to the Cry protein class. In some embodiments, the protein is an insecticidal protein (Vip) produced during vegetative growth.

[0205] In some embodiments, the pesticidal polynucleotide is RNA. In certain embodiments, the RNA is dsRNA or small interfering RNA (siRNA).

[0206] In another aspect, the present disclosure relates to a composition comprising a pesticidal polynucleotide and a biological adjuvant. In some embodiments, the composition further comprises a biopesticide. In some embodiments, the biological adjuvant is a yeast particle, and the biopesticide is a Bt protein. In some embodiments, the polynucleotide is encapsulated in the yeast particle, for example, in the hollow core of the yeast particle. In some embodiments, the polynucleotide is not encapsulated in the yeast particle.

[0207] In one embodiment, the polynucleotide inhibits the expression of one or more genes essential for the growth and / or development of the plant pest.

[0208] Expression of a gene in a plant pest (e.g., an insect cell) is considered to be inhibited or reduced through contact with a polynucleotide, for example, if the level of mRNA and / or protein encoded by the gene is reduced by at least 10% in the cell compared to a control cell not contacted with the polynucleotide. For example, delivering a pesticidal polynucleotide (e.g., dsRNA) that targets a target gene to a plant pest (e.g., contacting the plant pest with it) may result in a reduction (e.g., at least 10%) in the amount of RNA transcript and / or protein (e.g., encoded by the target gene) compared to a cell not contacted with the polynucleotide.

[0209] In certain embodiments, the target gene (e.g., a gene essential for the growth and / or development of a plant pest) is selected from the group consisting of proteasome beta 5 (PSMB5) gene, vacuolar ATPase E (vATPase E) gene, calmodulin gene, inhibitor of apoptosis protein (IAP) gene, soluble NSF attachment protein (a-SNAP) gene, Ras opposite (ROP) gene, proteasome alpha type 2 (PTSA2) gene, Ras-associated GTPase 1 associated with secretion (SAR1) gene, PBAN, ATPase, wingsup A (wupA) (wupA encodes troponin I), and any of the genes disclosed in CP4S3_DROME, C12C1_DROME, WO20212317912 (incorporated herein by reference), or any combination thereof.

[0210] In some embodiments, the target gene is IAP. In some embodiments, the target gene has the sequence of SEQ ID NO: 1. In some embodiments, the pesticidal polynucleotide comprises or consists of the nucleic acid sequence of any one of SEQ ID NOs: 1-13. [Table A] TIFF2026505896000003.tif247158TIFF2026505896000004.tif249158TIFF2026505896000005.tif130158

[0211] The expression of genes essential for the growth and development of plant pests can also be inhibited by post-transcriptional gene silencing, which can be achieved, for example, using antisense molecules or molecules that mediate RNA interference (RNAi).

[0212] In exemplary embodiments, the RNA that mediates RNAi is double-stranded RNA (dsRNA) that comprises a first strand complementary to a portion of messenger RNA (mRNA) encoded by one or more genes essential for the growth and / or development of the plant pest, and a second strand complementary to the first strand.

[0213] The dsRNA may comprise RNA strands of the same length or different lengths. In some embodiments, the dsRNA comprises a first strand (e.g., an antisense strand) that is the same length as a second strand (e.g., a sense strand). In some embodiments, the dsRNA comprises a first strand (e.g., an antisense strand) that is a different length from the second strand (e.g., a sense strand). The first strand may be about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, or more than 20% longer than the second strand. The first strand may be 1 to 5, 2 to 5, 2 to 10, 5 to 10, 5 to 15, 10 to 20, 15 to 20, or more than 20 nucleotides longer than the second strand.

[0214] dsRNA molecules can also be assembled from a single oligonucleotide of stem-loop structure (wherein the self-complementary sense region and antisense region of the RNA molecule are connected by nucleic acid-based or non-nucleic acid-based linker(s)), and circular single-stranded RNA (wherein the circular RNA can be processed in vivo or in vitro to generate an active RNAi molecule that can mediate RNAi) with a stem that has two or more loop structures and self-complementary sense and antisense strands.RNAi molecules can include a 3' overhang at one end of the molecule, and the other end can be blunt or have an overhang (5' or 3').When RNAi molecules include overhangs at both ends of the molecule, the length of the overhangs can be the same or different.

[0215] Antisense polynucleotides are designed to specifically bind to RNA, resulting in the formation of an RNA-DNA or RNA-RNA hybrid, along with the termination of reverse transcription or messenger RNA translation. Antisense polynucleotides are typically produced intracellularly by expression from antisense constructs containing the antisense strand as the transcribed strand. Antisense polynucleotides will bind to and / or interfere with the translation of corresponding mRNAs. Antisense RNA or antisense oligodeoxynucleotides (antisense ODNs) can both be used and may also be prepared synthetically in vitro or by recombinant DNA technology. ODNs and antisense RNAs may be chemically modified to avoid digestion by DNases. Trans-cleavage catalytic RNAs (ribozymes) are RNA molecules that possess endoribonuclease activity. Ribozymes are specifically designed for specific targets, and the target message must contain a specific nucleotide sequence. They are engineered to site-specifically cleave either RNA species in the background of cellular RNA. The cleavage event destabilizes the mRNA and prevents protein expression.

[0216] The pesticidal polynucleotide (e.g., dsRNA) as provided herein can vary in length. In some embodiments, long RNA (e.g., dsRNA or ssRNA) molecules are applied (e.g., to plants) as pesticides, but it should be understood that after entering cells, this dsRNA is cut by Dicer enzyme into shorter double-stranded RNA fragments, for example, having a length of 15-25 nucleotides. Thus, the pesticidal polynucleotide (e.g., dsRNA) of the present disclosure can be delivered as, for example, 15-25 nucleotide fragments, or they can be delivered as longer double-stranded nucleic acids (e.g., at least 100 nucleotides).

[0217] Thus, in some embodiments, the pesticidal polynucleotide (eg, dsRNA) comprises between 15 and 2000 nucleotides (ssRNA) or nucleotide base pairs (dsRNA). For example, the pesticidal polynucleotides of the present disclosure may be any of the following: 15 to 1000, 15 to 950, 15 to 900, 15 to 850, 15 to 800, 15 to 750, 15 to 700, 15 to 650, 15 to 600, 15 to 500, 15 to 450, 15 to 400, 15 to 350, 15 to 300, 15 to 250, 15 to 200, 15 to 150, 15 to 100, 15 to 50, 19 to 1000, 18 to 950, 18 to 900, 18 to 850, 18 to 800, 18 to 750, 18 to 700, 18 to 650, 18 to 600, 18 to 500, 18 to 1000, 18 to 15 ... 8~450, 18~400, 18~350, 18~300, 18~250, 18~200, 18~180, 18~100, 18~50, 19~1000, 19~950, 19~900, 19~850, 19~800, 19~750, 19~700, 19~650, 19~600, 19~500, 19~450, 19~400, 19~350, 19~300, 19~250, 19~200, 19~190, 19~100, 19~50, 20~1000, 20~950, 20~900, 20~850, 20~800 , 20~750, 20~700, 20~650, 20~600, 20~500, 20~450, 20~400, 20~350, 20~300, 20~250, 20~200, 20~200, 20~100, 20~50, 15211000, 21~950, 21~900, 21~850, 21~800, 21~750, 21~700, 21~650, 21~600, 21~500, 21~450, 21~400, 21~350, 21~300, 21~250, 21~210, 21~210, 21~100, 21 ~50, 22~1000, 22~950, 22~900, 22~850, 22~800, 22~750, 22~700, 22~650, 22~600, 22~500, 22~450, 22~400, 22~350, 22~300, 22~250, 22~220, 22~220, 22~100, 22~50, 23~1000, 23~950, 23~900, 23~850, 23~800, 23~750, 23~700, 23~650, 23~600, 23~500, 23~450, 23~400, 23~350,23~300, 23~250, 23~230, 23~230, 23~100, 23~50, 24~1000, 24~950, 24~900, 24~850, 24~800, 24~750, 24~700, 24~650, 24~600, 24~500, 24~450, 24~400, 24~350, 24~300, 24~250, 24~240, 24~240, 24~100, It may contain 24 to 50, 25 to 1000, 25 to 950, 25 to 900, 25 to 850, 25 to 800, 25 to 750, 25 to 700, 25 to 650, 25 to 600, 25 to 500, 25 to 450, 25 to 400, 25 to 350, 25 to 300, 25 to 250, 25 to 250, 25 to 250, 25 to 100, or 25 to 50 nucleotides or nucleotide base pairs. In some embodiments, the pesticidal polynucleotide comprises or consists of at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 50, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, or at least 1000 nucleotides or nucleotide base pairs.

[0218] In some embodiments of the present disclosure, the composition comprises ribonucleic acid (RNA) and YP, and optionally a Bt product. In some embodiments, the RNA is encapsulated in the YP.

[0219] Methods for encapsulating polynucleotides (e.g., RNA) into YPs are known in the art (e.g., U.S. Patent Nos. 8,007,814 and 9,682,135, each of which is incorporated herein by reference). Negatively charged nucleic acids can be encapsulated in YPs with the aid of cationic polymers and / or cationic lipids and / or other organic or inorganic carriers. In some embodiments, the cationic polymer is polyethyleneimine (PEI) or poly-L-lysine (PLL). In some embodiments, complex core A (CCA) is used to encapsulate nucleic acids in YPs. In some embodiments, the composition further comprises one or more nuclease inhibitors. In some embodiments, the one or more nuclease inhibitors are selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), polyvinylsulfonic acid (PVSA), sodium hexametaphosphate (SHMP), and sodium tripolyphosphate (TPP).

[0220] In some embodiments, the pesticide is a chemical pesticide. Chemical pesticides are typically small molecules (e.g., small organic molecules) having a molecular weight of less than 1000 daltons.

[0221] In some embodiments, the pesticide is a ryanodine receptor modulator (IRAC class 28). The ryanodine receptor modulator (IRAC class 28) may comprise a diamide selected from the group consisting of chlorantraniliprole, tetraniliprole, cyclaniliprole, brofuranilide, cyantraniliprole, imidacloprid, or flubendiamide.

[0222] In some embodiments, the pesticide is a chitin biosynthesis inhibitor (IRAC Class 15) acting on CHS1. The chitin biosynthesis inhibitor (IRAC Class 15) acting on CHS1 may comprise a benzoyl urea selected from the group consisting of bistrifluron, chlorfluazuron, diflubenzuron, flucycloxuron, flufenoxuron, hexaflumuron, lufenuron, novaluron, noviflumuron, teflubenzuron, and triflumuron.

[0223] In some embodiments, the pesticide is a nicotinic acetylcholine receptor (nACHR) allosteric modulator-site 1 (IRAC class 5). The nACHR allosteric modulator-site 1 molecule can be a spinosyn. In some embodiments, the spinosyn is spinetoram or spinosad.

[0224] In some embodiments, the pesticide is a nACHR competitive modulator (IRAC class 4). The nACHR competitive modulator (IRAC class 4) may be a neonicotinoid. In some embodiments, the neonicotinoid is selected from the group consisting of acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, thiacloprid, and thiamethoxam.

[0225] In some embodiments, the pesticide is a sodium channel modulator (IRAC Class 3). The sodium channel modulator (IRAC Class 3) may be a pyrethroid or pyrethrin. In some embodiments, the pyrethroid or pyrethrin is acrinathrin, allethrin, d-cis-trans-allethrin, d-trans-allethrin, bifenthrin, bioallethrin, bioallethrin S-cyclopentenyl, bioresmethrin, cycloprothrin, cyfluthrin, beta-cyfluthrin, cyhalothrin, lambda-cyhalothrin, gamma-cyhalothrin, cypermethrin, alpha-cypermethrin, beta-cypermethrin, theta-cypermethrin, zeta-cypermethrin, cyphenothrin [(1R)-trans-isomer] , deltamethrin, empenthrin [(EZ)-(1R)-isomer], esfenvalerate, etofenprox, fenpropathrin, fenvalerate, flucythrinate, flumethrin, tau-fluvalinate, kadethrin, pyrethrins (pyrethram), halfenprox, fenothrin [(1R)-trans-isomer], prallethrin, resmethrin, silafluofen, tefluthrin, tetramethrin, tetramethrin [(1R)-isomer], tralomethrin, transfluthrin, and permethrin.

[0226] In some embodiments, the pesticide is an acetylcholinesterase (ACHE) inhibitor (IRAC Class 1). In some embodiments, the pesticide is a carbamate, optionally alanycarb, aldicarb, bendiocarb, benfuracarb, butocarboxim, butoxycarboxim, carbaryl, carbofuran, carbosulfan, ethiofencarb, fenobucarb, formetanate, furathiocarb, isoprocarb, methiocarb, methomyl, metolcarb, oxamyl, pirimicarb, propoxur, thiodicarb, thiofanox, triazamate, trimethacarb, XMC, or xylylcarb.In some embodiments, the pesticide is an organophosphate, and optionally the organophosphate is acephate; formothion; azamethiphos; azinphos-ethyl; azinphos-methyl; chlorpyrifos; chlorfenvinphos; cyanophos; maniphos; fensulfothion; tribufos; O,O-diethyl O-[6-methyl-2-(1-methylethyl)-4-pyrimidinyl]phosphorothioate; O,O-diethyl O-(2-isopropyl-6-methyl -4-Pyrimidinyl)phosphorothioate;Dicrotophos;Dimethoate;Dioxathion;Disulfoton;Endothion;Ethion;Fenitrothion;Ethoprop;Chlorethoxyphos;Iprobenfos;Isazophos;Isofenphos;Isoxathion;Vamidothion;S-[2-(Ethylsulfinyl)-1-methylethyl]-0,0-dimethylphosphorothioate;Methidathion;Methylparathion;2-Carbomethoxy-1-methionyl The alpha isomer of 2-methylvinyl dimethyl phosphate; the beta isomer of 2-carbomethoxy-1-methylvinyl dimethyl phosphate; morphothion; naled; fenamiphos; fosmetilan; pyridaphenthion; omethoate; parathion; fenkapton; fenthoate; folate; phosalone; phosmet; phosnichlor; phosphamidon; leptophos; phoxim; pirimiphos-methyl; pirimiphos-ethyl; profenofos; prothidathion; prothoate; piperophos; tolclofos-methyl; ronnel; cadusafos; sofamid; demeton, demeton I (thiono isomer); demeton II (thiolo isomer); oxydemeton-methyl; cyanthoate; tebupirimfos; terbufos; tetrachlorvinphos; thiometon; prothiofos; dialifos; trichlorfon; and combinations thereof.

[0227] In some embodiments, the pesticide is a GABA-gated chloride channel blocker (IRAC class 2). In some embodiments, the pesticide is a cyclodiene organochlorine, optionally chlordane or endosulfan. In some embodiments, the pesticide is fiprole, optionally ethiprole or fipronil.

[0228] In some embodiments, the pesticide is a glutamate-gated chloride channel (GLUCL) allosteric modulator (IRAC class 6). In some embodiments, the pesticide is an avermectin, optionally abamectin, emamectin benzoate, lepimectin, or milbemectin.

[0229] In some embodiments, the pesticide is a juvenile hormone receptor modulator (IRAC class 7). In some embodiments, the pesticide is hydroprene, kinoprene, methoprene, fenoxycarb, or pyriproxyfen.

[0230] In some embodiments, the pesticide is an other non-specific (multi-site) inhibitor (IRAC Class 8). In some embodiments, the pesticide is an alkyl halide, chloropicrin, fluoride (e.g., cryolite, sulfuryl fluoride), borate (e.g., borax, boric acid, disodium octaborate, sodium borate), tartar emetic, or methyl isothiocyanate generator.

[0231] In some embodiments, the pesticide is a chordotonal TRPV channel modulator (IRAC class 9). In some embodiments, the pesticide is pymetrozine, pyrifluquinazone, or afidopiropen.

[0232] In some embodiments, the pesticide is an acarid growth inhibitor (IRAC class 10) that acts on CHS1. In some embodiments, the pesticide is clofentezine, diflovidazine, hexythiazox, or etoxazole.

[0233] In some embodiments, the pesticide is a mitochondrial ATP biosynthesis enzyme inhibitor (IRAC class 12). In some embodiments, the pesticide is diafenthiuron, azocyclotin, cyhexatin, fenbutatin oxide, propargite, or tetradifon.

[0234] In some embodiments, the pesticide is an oxidative phosphorylation uncoupler (IRAC class 13) that disrupts the proton gradient. In some embodiments, the pesticide is chlorfenapyr, dinitrophenol (e.g., DNOC), or sulfluramide.

[0235] In some embodiments, the pesticide is a nicotinic acetylcholine receptor (nACHR) channel blocker (IRAC class 14). In some embodiments, the pesticide is bensultap, cartap hydrochloride, thiocyclam, or thiosultap sodium.

[0236] In some embodiments, the pesticide is a chitin biosynthesis inhibitor, type 1 (IRAC class 16). In some embodiments, the pesticide is buprofezin.

[0237] In some embodiments, the pesticide is a molting disruptor Diptera (IRAC Class 17). In some embodiments, the pesticide is cyromazine.

[0238] In some embodiments, the pesticide is an ecdysone receptor agonist (IRAC class 18). In some embodiments, the pesticide is a diacylhydrazine, optionally chromafenozide, halofenozide, methoxyfenozide, or tebufenozide.

[0239] In some embodiments, the pesticide is an octopamine receptor agonist (IRAC class 19). In some embodiments, the pesticide is amitraz.

[0240] In some embodiments, the pesticide is a mitochondrial complex III electron transport inhibitor QO site (IRAC class 20). In some embodiments, the pesticide is hydramethylnon, acequinocyl, fluacrypyrim, or bifenazate.

[0241] In some embodiments, the pesticide is a mitochondrial complex I electron transport inhibitor (IRAC class 21). In some embodiments, the pesticide is fenazaquin, fenpyroximate, pyrimidifen, pyridaben, tebufenpyrad, rotenone, or tolfenpyrad.

[0242] In some embodiments, the pesticide is a voltage-gated sodium channel blocker (IRAC class 22). In some embodiments, the pesticide is indoxacarb or metaflumizone.

[0243] In some embodiments, the pesticide is an acetyl-CoA carboxylase inhibitor (IRAC class 23). In some embodiments, the pesticide is a tetronic acid derivative, optionally spirodiclofen, spiromesifen, spiropydione, spirotetramat, or spidoxamat.

[0244] In some embodiments, the pesticide is a mitochondrial complex IV electron transport inhibitor (IRAC class 24). In some embodiments, the pesticide is a phosphide or a cyanide.

[0245] In some embodiments, the pesticide is a mitochondrial transport inhibitor (IRAC class 25). In some embodiments, the pesticide is cyenopyrafen, cyflumetofen, or piflubumid.

[0246] In some embodiments, the pesticide is a chordotonal nicotinamidase inhibitor (IRAC class 29). In some embodiments, the pesticide is flonicamid.

[0247] In some embodiments, the pesticide is a GABA-gated chloride channel allosteric modulator (IRAC class 30). In some embodiments, the pesticide is broflanilide, fluxametamide, or isocycloceram.

[0248] In some embodiments, the pesticide is a nicotinic acetylcholine receptor (NACHR) allosteric modulator-site II (IRAC class 32).

[0249] In some embodiments, the pesticide is a calcium-activated potassium channel (KCa2) modulator (IRAC class 33). In some embodiments, the pesticide is acinonapyr.

[0250] In some embodiments, the pesticide is a mitochondrial complex III electron transport inhibitor QI site (IRAC class 34). In some embodiments, the pesticide is flometoquine.

[0251] In some embodiments, the pesticide is a chordotonal organ modulator - target site undefined (IRAC class 36). In some embodiments, the pesticide is dinpropylidaz.

[0252] Yeast particles and polysaccharides for use in compositions In some embodiments, the biological adjuvant comprises a yeast particle (YP) (eg, a yeast particle comprising yeast cell wall components).

[0253] Yeast particles (YP) are generally about 3-5 μm, 2-6 μm, 4-6 μm, or 3-7 μm in size. In some embodiments, yeast particles are about 3-5 μm, 2-6 μm, 4-6 μm, or 3-7 μm in diameter. In some embodiments, yeast particles comprise yeast cell wall components and produce hollow and porous microspheres. Yeast particles, in some embodiments, comprise yeast cell wall components and can be produced from intact yeast. Yeast particles contain β-glucan and / or mannan oligosaccharides, among other structural elements (e.g., mannoproteins, chitin, and / or lipids).

[0254] Beta-glucans found in yeast particles can include (but are not limited to) β-1,3-branched glucans, β-1,6-branched glucans, and / or β-1,3-1,6-branched glucans. β-1,3-branched glucans are typically the most prevalent beta-glucans in yeast particles.

[0255] In some embodiments, the β-glucan present in the yeast particle comprises β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan. In some embodiments, the β-glucan present in the yeast particle comprises β-1,3-branched glucan, β-1,6-branched glucan, and / or β-1,3-1,6-branched glucan. In some embodiments, the β-glucan is a linear polysaccharide (unbranched polysaccharide). In some embodiments, the β-glucan can form random coil, single helix, triple helix, or parallel fiber aggregates. In some embodiments, the yeast particle further comprises chitin (N-acylated polyglucosamine).

[0256] In some embodiments, the dry weight of mannan oligosaccharides in the yeast particles is about 10-60%, 20-50%, 20-60%, 30-50%, 30-60%, 25-50%, 40-60%, or 15-45% of the total dry weight. In some embodiments, the dry weight of mannan oligosaccharides in the yeast particles is about 10%, 20%, 30%, 40%, 50%, or 60% of the total dry weight. In some embodiments, the dry weight of beta-glucan in the yeast particles is about 20-80%, 30-60%, 30-70%, 30-50%, 25-55%, 30-70%, or 40-70% of the total dry weight. In some embodiments, the dry weight of beta-glucan in the yeast particle is about 20%, about 30%, about 40%, about 50%, about 60%, or about 70% of the total dry weight.

[0257] A yeast particle may comprise an intact yeast cell wall, a partially intact yeast cell wall, or a fragmented cell wall. The degree of cell wall fragmentation within a yeast particle may also depend on the method by which the yeast particle itself is produced.

[0258] Yeast particles are not the same as yeast extract. The yeast particles for use in the present disclosure do not include (and do not consist of) yeast extract. Yeast extract generally consists of the soluble internal contents of yeast cells (e.g., yeast proteins, cytoplasm, yeast nucleic acids) (i.e., no yeast cell wall components). In some embodiments, yeast extract does not include yeast cell wall components.

[0259] Yeast particles may be obtained, for example, as a by-product of some food-grade baker's yeast (i.e., Saccharomyces cerevisiae) extract manufacturing processes. Yeast particles can be obtained from various sources, such as Biorigin and LeSaffre. YP can also be prepared by extracting yeast cells. Methods for preparing extracted yeast cell wall particles are known in the art and are described, for example, in U.S. Patent Nos. 4,992,540, 5,082,936, 5,028,703, 5,032,401, 5,322,841, 5,401,727, 5,504,079, 5,968,811, 6,444,448, 6,476,003, U.S. Patent Application Publication Nos. 2003 / 0216346 and 2004 / 0014715, and PCT published application WO 02 / 12348, which are incorporated herein by reference. In some embodiments, yeast particles can be produced using any one of the yeast particle preparation methodologies detailed in Example 1.

[0260] Yeast particles containing yeast cell wall components may be prepared from yeast cells by extraction and purification of the insoluble particle fraction from the soluble components of the yeast cells. Fungal cell walls can be produced from the insoluble by-products of yeast extract production. Furthermore, yeast cells can be treated with aqueous hydroxide to digest the proteins and intracellular parts of the cells without destroying the yeast cell walls, and the yeast cell wall components remain free of significant protein contamination and have substantially unchanged cell wall structure of β(1-6) and β(1-3) linked glucans. In some embodiments, the process for preparing yeast particles of the present disclosure is as described in U.S. Patent No. 4,810,646; U.S. Patent No. 6,242,594; U.S. Patent No. 5,401,727; or U.S. Patent No. 5,607,677, the entire contents of which are incorporated herein by reference. In some embodiments, the process for preparing yeast particles of the present disclosure involves preparing yeast glucan particles by alkaline extraction, acid extraction, and then extraction with an organic solvent, and finally drying.

[0261] In some embodiments, yeast particles containing yeast cell wall components are prepared using shear force methodology. In some embodiments, yeast particles are prepared using a ball mill or bead mill, which functions to suspend and agitate yeast cells with small abrasive particles (e.g., glass or ceramic beads). In some embodiments, yeast particles are prepared using ultrasonic treatment. Yeast cells are lysed and / or disrupted due to shear force, grinding between beads, and collision with beads.

[0262] In some embodiments, the glass or ceramic beads have a diameter of 0.10 to 2 mm. In some embodiments, the glass or ceramic beads have a diameter of 0.1 to 1 mm, 0.2 to 2 mm, 0.3 to 1 mm, 0.3 to 0.8 mm, 0.5 to 1 mm, or 0.5 to 0.7 mm. In some embodiments, the glass or ceramic beads are 0.65 mm ceramic beads.

[0263] In certain embodiments, the YP comprises yeast cell wall particles (YCWP), yeast glucan particles (YGP), yeast glucan mannan particles (YGMP), yeast chitin particles (YCP), yeast glucan chitin particles (YGCP), yeast glucan lipid particles (YGLP), or whole glucan particles (WGP).

[0264] In some embodiments, the yeast particles are 2-10, 2-5, 2-6, 3-10, 3-5, or 3-7 microns in diameter. In some embodiments, the yeast particles are less than 10 microns, less than 9 microns, less than 8 microns, less than 7 microns, less than 6 microns, less than 5 microns, or less than 4 microns in diameter. In some embodiments, the yeast particles are about 10 microns, about 9 microns, about 8 microns, about 7 microns, about 6 microns, about 5 microns, or about 4 microns in diameter.

[0265] The yeast cell wall particles (YCWP) may be extracted YCWP. In some embodiments, the YCWP is as described in Figueiredo et al., "Yeast cell wall particles: a promising class of nature-inspired microcarriers for multimodal imaging," Chem. Commun., 2011, 47, 10635-10637, the contents of which are incorporated herein by reference.

[0266] Yeast glucan particles (GPs) are generally yeast particles that contain a high amount or concentration of beta-glucan compared to other yeast particles. In some embodiments, GPs can be derived from the cell walls of baker's yeast (Saccharomyces cerevisiae). Yeast glucan mannan particles (YGMPs) are yeast particles containing beta-glucan and mannan oligosaccharides. Yeast chitin particles (YCPs) are yeast particles that contain a high amount or concentration of chitin compared to other yeast particles. Yeast glucan chitin particles (YGCPs) are yeast particles containing beta-glucan and chitin. Yeast glucan lipid particles (YGLPs) are yeast particles containing beta-glucan and lipids (e.g., phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), and / or cytidine diphosphate diacylglycerol (CDP-DAG)).

[0267] In some embodiments of the present disclosure, the YP is commercially available YP. Commercially available YP can be derived from baker's yeast, lactic acid yeast, and brewer's / distiller's yeast sources.

[0268] In some embodiments, YP comprises a mixture composed of insoluble β-glucans (e.g., >20-30%) and mannan oligosaccharides (e.g., >18%) and lipids (e.g., about 2%) as well as other insoluble yeast components.

[0269] In certain embodiments of the present invention, the biological adjuvant comprises a polysaccharide selected from the group consisting of β-glucan, mannan oligosaccharide, and laminarin. In some embodiments, the polysaccharide is derived from a yeast source. In other embodiments, the polysaccharide is derived from a source other than yeast. Such polysaccharides can be chemically produced or can be derived from bacteria, fungi, algae, lichen, or plants. For example, β-glucans useful as biological adjuvants of the present invention can be derived from the cell walls or capsules of bacteria, fungi, algae (e.g., Euglena gracilis), lichen, or plants. Beta-glucans for use in the present disclosure include (but are not limited to) β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan. In some embodiments, the β-glucan is β-1,3-branched glucan, β-1,6-branched glucan, and / or β-1,3-1,6-branched glucan. In some embodiments, the β-glucan is a linear polysaccharide (unbranched polysaccharide). In some embodiments, the β-glucan can form random coil, single helix, triple helix, or parallel fiber aggregates.

[0270] In another embodiment, the polysaccharide is a mannan oligosaccharide, and optionally the source of the mannan oligosaccharide is a plant. In another embodiment, the polysaccharide is laminarin, optionally derived from algae. In another embodiment, the polysaccharide comprises a glucan derived from a plant, optionally from barley or oats, and optionally is a cereal glucan (β1,3 / 1,4-linked β-glucan). In another embodiment, the polysaccharide is derived from a fungus, optionally derived from a mushroom.

[0271] In certain embodiments, the biological adjuvant comprises a glucan selected from the group consisting of glucans with n=2-15 β1,3 oligosaccharide branches, β1,3,6 branches, and β1,3 backbones.

[0272] One or more polysaccharides for use in the composition may be derived from yeast. In some embodiments, one or more polysaccharides for use in the composition may be derived from Saccharomyces cerevisiae, baker's yeast, lactic acid yeast, and brewer's / distiller's yeast sources.

[0273] One or more polysaccharides for use in the composition may be derived from sources other than yeast.In some embodiments, one or more polysaccharides for use in the composition may be derived from bacteria, fungi, algae, lichens or plants.In some embodiments, one or more polysaccharides for use in the composition may be derived from Euglena gracilis.

[0274] In certain exemplary embodiments, the biological adjuvant present with the pesticidal polynucleotide comprises a polysaccharide derived from a source other than yeast, including, for example, bacteria, fungi, algae, lichen, or plants. In some embodiments, the polysaccharide is a β-glucan derived from the cell wall or capsule of bacteria, fungi, algae, lichen, or plants. In another embodiment, the polysaccharide is a mannan oligosaccharide, optionally from a plant. In another embodiment, the polysaccharide is laminarin, optionally from algae. In another embodiment, the polysaccharide is a cereal glucan (β1,3 / 1,4-linked β-glucan). In certain embodiments, the biological adjuvant is a long n=2-15 β1,3 oligosaccharide-branched glucan or a short β1,3,6-branched glucan.

[0275] In some embodiments, yeast particles are formulated into a complex core A. The complex core A generally comprises (i) yeast particles and (ii) a nuclease inhibitor. In some embodiments, the complex core A comprises (i) a polysaccharide selected from beta-glucan, mannan oligosaccharide, and / or laminarin that forms a spherical particle, and (ii) a nuclease inhibitor. In some embodiments, the complex core A further comprises a cationic polymer. In some embodiments, the complex core A particles have a diameter of 2 to 10, 2 to 5, 2 to 6, 3 to 10, 3 to 5, or 3 to 7 microns. In some embodiments, the nuclease inhibitor is ethylenediaminetetraacetic acid (EDTA), polyvinylsulfonic acid (PVSA), sodium hexametaphosphate (SHMP), sodium tripolyphosphate (TPP), diethylpyrocarbonate, aurintricarboxylic acid (ATA), formamide, macaloid, proteinase K, heparin, hydroxylamine-oxygen-copper(II) ion, bentonite, ammonium sulfate, dithiothreitol (DTT), beta-mercaptoethanol, cysteine, dithioerythritol, or tris(2-carboxyethyl)phosphene hydrochloride. The cationic polymer may be any cationically (positively) charged polymer. In some embodiments, the cationic polymer is a cationic polypeptide. In some embodiments, the cationic polymer is polyethyleneimine (PEI) or poly-L-lysine (PLL). In some embodiments, the cationic polymer is cationic gelatin, cationic chitosan, cationic cellulose, or cationic dextran. In some embodiments, the cationic polymer is poly(2-N,N-dimethylaminoethyl methacrylate) or poly(amidoamine).

[0276] In some embodiments, a complex core A encapsulating a pesticide (e.g., a pesticidal polynucleotide) is prepared by combining a yeast particle of the present disclosure, a nuclease inhibitor, and a pesticide, and incubating the mixture for a period of time (e.g., 10 to 60 minutes).

[0277] In some embodiments, the complex core A encapsulating a pesticide (e.g., a pesticidal polynucleotide) is prepared by first sterilizing and homogenizing yeast (e.g., using a bead mill process or sonication for 1-10, 5-10, or 4-6 minutes). In some embodiments, a maltodextrin solution is then added to the homogenized yeast and vortexed to create a homogenous maltodextrin-YP suspension. In some embodiments, to synthesize YP-EDTA for dsRNA capture, a 500 mg / mL EDTA tetrasodium salt solution in RNAase-free water is added to sterilized maltodextrin-YP particles (SHMP can be substituted for EDTA). The materials are mixed and allowed to incubate at room temperature for 30 minutes. A volume of strong acid (e.g., 3 M sulfuric acid) is then added to the incubated particles and mixed, followed by incubation for 1 hour. The material may be washed (e.g., with water) to remove excess acid and EDTA, and subsequently resuspended (e.g., via a Polytron dispersion unit). The resuspended sample may be centrifuged for 1 hour and finally frozen.

[0278] In some embodiments, to use polymer formulation to encapsulate dsRNA, YP-EDTA particles are combined with a certain volume of dsRNA (volume varies based on dsRNA concentration).Materials can be mixed, frozen in liquid nitrogen, and lyophilized.This process can be repeated until desired dsRNA concentration is reached.A certain volume of polymer capture agent (for example, cationic polymer) can be added to the encapsulated dsRNA-YP particles and mixed.

[0279] In some embodiments, yeast particles (e.g., including yeast cell wall components) are prepared by combining whole yeast with an anti-caking mixture (e.g., an emulsifying oil mixture including methyl oleate, castor oil ethoxylate, and calcium dodecylbenzenesulfonate), an antifoaming agent, and water in a vessel for homogenization. A bead milling protocol may then be initiated by adding ceramic beads (e.g., 0.65 mm ceramic beads) to the vessel and circulating the beads (e.g., by shear force) until yeast particles of the desired size are formed. The desired size of the yeast particles may be less than 5 microns in diameter.

[0280] method Aspects and embodiments of the present disclosure provide methods of controlling pest (e.g., insect or fungal pathogen) infestations, comprising delivering a composition as described herein to a plant or pest.

[0281] Aspects of the present disclosure provide, in some embodiments, a method for controlling plant pest infestation, the method comprising delivering an effective amount of a composition described herein to a plant or plant pest. In some embodiments, the delivery method comprises applying a composition described herein to the surface of the plant or plant pest. In some embodiments, the composition is a solid or liquid (e.g., a solution, suspension, or emulsion). Non-limiting examples include emulsifiable concentrates, concentrated solutions, low-concentrate solutions, ultra-low volume concentrated solutions, water-soluble concentrated solutions, water-soluble liquid solutions, baits (pastes, gels, liquids, solids, or injectables), smoke, fog, inverse emulsions, flowables, aerosols, homogeneous and non-homogeneous mixtures, suspensions (water and oil-based), dusts, powders (wettable or soluble), granules (water-dispersible or dry-flowable), pellets, capsules, fumigants, encapsulated or microencapsulated formulations, or any combination thereof.

[0282] In some embodiments, the compositing may be applied as a concentrate, spray (diluted or concentrated), mist, infaloat, seed treatment, drench, drip, insect bait, bait, or any other form suitable for in-furrow application. The compositions described herein may be delivered to any part of a plant, including, but not limited to, leaves, stems, flowers, fruits, shoots, roots, seeds, tubers, anthers, stamens, and / or pollen. In some embodiments, the composition is delivered mechanically through a high-pressure spray or sandblasting. In some embodiments, the composition comprises at least one additive selected from an adjuvant, attractant, sterilant, growth regulator, carrier or diluent, and / or stabilizer. Non-pesticides may also be used (e.g., adjuvants such as antifoaming agents, buffers, compatibilizers, drift control additives, emulsifiers, bulking agents, invert emulsifiers, plant penetrants, safeners, spreading agents, adhesives, surfactants, thickeners, and wetting agents).

[0283] In some embodiments, the compositions described herein are provided in insect feed.For example, the compositions can be applied topically to plants or seeds (e.g., by dipping, coating, dusting or spraying).In some embodiments, the compositions comprising yeast particles or polysaccharides are delivered to the cells of transgenic plants that may be engineered to express biopesticides.The compositions can also be provided in separate food or water sources.

[0284] Delivering the composition to the plant (e.g., a part of the plant) and / or plant pest can include, for example, topically applying (e.g., dipping, coating, or dusting) the composition to any part of the plant (e.g., roots, tubers, stems, branches, leaves, flowers, etc.), the ground (e.g., soil, mud, grass, etc.), insects, and / or insect feeding sites.

[0285] An effective amount of pesticide in a composition is the amount of pesticide required to provide a beneficial effect (e.g., mortality, cessation of feeding, inhibition of growth, development, or reproduction) against plant pest infestation, alone or in combination with one or more other additives. Beneficial effects include, for example, a reduction in infestation of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% compared to a control. In some embodiments, the control is the absence of yeast particles or polysaccharides. In some embodiments, the control is the absence of pesticide and the absence of yeast particles or polysaccharides. In some embodiments, an effective amount of pesticide completely eliminates plant pest infestation.

[0286] In some embodiments, an effective amount of pesticide in the composition is determined based on the surface area of ​​a plant or ground (e.g., soil, mud, grass, etc.) per square centimeter (cm 2 ) or μg / cm 2 Thus, in some embodiments, the effective amount is 0.001 μg / cm 2 ~10μg / cm 2 In some embodiments, the effective amount is 0.001 μg / cm 2 ~9μg / cm 2 , 0.001 μg / cm 2 ~8μg / cm 2 , 0.001 μg / cm 2 ~7μg / cm 2 ,0.001μg / cm 2 ~6μg / cm 2 , 0.001 μg / cm 2 ~5μg / cm 2 , 0.001 μg / cm 2 ~4μg / cm 2 , 0.001 μg / cm 2 ~3μg / cm 2 , 0.001 μg / cm 2 ~2μg / cm 2 , 0.001 μg / cm 2 ~1μg / cm2 , 0.001 μg / cm 2 ~0.1μg / cm 2 , or 0.001 μg / cm 2 ~0.01μg / cm 2 In some embodiments, the effective amount is 0.01 μg / cm 2 ~10μg / cm 2 , 0.1 μg / cm 2 ~10μg / cm 2 , 1 μg / cm 2 ~10μg / cm 2 ,2μg / cm 2 ~10μg / cm 2 , 3 μg / cm 2 ~10μg / cm 2 , 4 μg / cm 2 ~10μg / cm 2 , 5 μg / cm 2 ~10μg / cm 2 , 6 μg / cm 2 ~10μg / cm 2 , 7 μg / cm 2 ~10μg / cm 2 , 8 μg / cm 2 ~10μg / cm 2 , or 9 μg / cm 2 ~10μg / cm 2 Includes:

[0287] In some embodiments, the effective amount of pesticide in the composition is expressed as grams (g) of pesticide per acre (ac.) of plant or ground (e.g., soil, dirt, grass, etc.) surface, i.e., g / ac. Thus, in some embodiments, the effective amount of pesticide comprises between 0.01 g / ac. and 100 g / ac. In some embodiments, an effective amount of pesticide comprises between 0.01 g / ac and 90 g / ac, between 0.01 g / ac and 80 g / ac, between 0.01 g / ac and 70 g / ac, between 0.01 g / ac and 60 g / ac, between 0.01 g / ac and 50 g / ac, between 0.01 g / ac and 40 g / ac, between 0.01 g / ac and 30 g / ac, between 0.01 g / ac and 20 g / ac, between 0.01 g / ac and 10 g / ac, between 0.01 g / ac and 1 g / ac, or between 0.01 g / ac and 0.1 g / ac. In some embodiments, an effective amount of pesticide comprises 0.1 g / ac to 100 g / ac, 1 g / ac to 100 g / ac, 10 g / ac to 100 g / ac, 20 g / ac to 100 g / ac, 30 g / ac to 100 g / ac, 40 g / ac to 100 g / ac, 50 g / ac to 100 g / ac, 60 g / ac to 100 g / ac, 70 g / ac to 100 g / ac, 80 g / ac to 100 g / ac, or 90 g / ac to 100 g / ac.

[0288] In some embodiments, the effectiveness of a composition for controlling plant pests can be determined by the ability of the composition to kill or cause the death of a plant pest or a population of plant pests. The percentage of death in a population of plant pests can be determined by percent mortality (e.g., percent mortality over time). Generally, the percent mortality of a population of plant pests reflects the percentage of plant pests in the population that died as a result of the composition (e.g., 75% mortality indicates that the composition killed 75% of the total population). In some embodiments, percent mortality is measured over time (e.g., over the course of multiple days of exposure of insects to the composition). In some embodiments, percent mortality is measured after at least 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 days of exposure. In some embodiments, the composition causes at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 100% percent mortality of the plant pest population. In some embodiments, at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 100% of the plant pest population is killed by the composition. In some embodiments, the percent mortality of the composition is compared to a control (e.g., a control molecule or an untreated condition). In some embodiments, the percent mortality of the composition is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, or 200% higher than the control (e.g., a control molecule or an untreated condition).

[0289] In some embodiments, the effectiveness of a composition for controlling plant pests can be determined by its ability to limit leaf disk consumption by a plant pest or a population of plant pests. Leaf disk consumption refers to the amount (e.g., percentage) of plant material (e.g., eggplant leaves) consumed or eaten by a plant pest. In some embodiments, the composition causes at least a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 100% reduction in leaf disk consumption by a plant pest or a population of plant pests. In some embodiments, the ability of a composition to reduce leaf disk consumption is compared to a control (e.g., a control molecule or an untreated condition). In some embodiments, leaf disk consumption is measured over time (e.g., over the course of multiple days of exposure of a plant pest to the composition). In some embodiments, leaf disk consumption is measured after 3, 4, 5, 6, 7, 8, 9, 10, or more days of exposure.

[0290] In some embodiments, the effectiveness of the composition for controlling plant pests can be determined by the ability of RNAi molecules to reduce the percentage of plant defoliation caused by plant pests or plant pest populations.Percentage plant defoliation refers to the percentage of plant material (e.g., eggplant leaves) that is destroyed (e.g., consumed) by plant pests or plant pest populations.In some embodiments, the composition causes at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98% or 100% reduction in the percentage of plant defoliation caused by plant pests or plant pest populations.In some embodiments, the composition reduces the percentage of plant defoliation to less than 40%, 30%, 25%, 20%, 15%, 10%, 5%, 3% or 1%. In some embodiments, percent plant defoliation remains less than 40%, 30%, 25%, 20%, 15%, 10%, 5%, 3%, or 1% for at least 5, 6, 7, 8, 9, 10, 15, or 20 days after exposing the plant pest to the composition. In some embodiments, the ability of the composition to reduce percent plant defoliation is compared to a control (e.g., a control molecule or an untreated condition). In some embodiments, percent plant defoliation is measured over time (e.g., over the course of multiple days of exposure of the plant pest to the composition). In some embodiments, percent plant defoliation is measured after 3, 4, 5, 6, 7, 8, 9, 10, or more days of exposure.

[0291] In some embodiments, the composition can be formulated into a solution (e.g., applied to the surface of the plant pest and / or its feed (e.g., ingested food and / or water), plant, or ground (e.g., soil, mud, grass, etc.)). In some embodiments, the effective amount of RNAi molecule in solution is expressed as nanograms (ng) or micrograms (μg) of RNAi molecule per milliliter (ml) of solution, i.e., ng / ml. Thus, in some embodiments, the solution comprises the composition at a concentration of 10 ng / ml to 100 μg / ml. In some embodiments, the solution comprises the composition at a concentration of 10 ng / ml to 100 μg / ml, 100 ng / ml to 100 μg / ml, 250 ng / ml to 100 μg / ml, 750 ng / ml to 100 μg / ml, 1000 ng / ml to 100 μg / ml, 10 μg / ml to 100 μg / ml, 25 μg / ml to 100 μg / ml, 50 μg / ml to 100 μg / ml, or 75 μg / ml to 100 μg / ml. In some embodiments, the solution comprises the composition at a concentration of 10 ng / ml to 100 μg / ml, 10 ng / ml to 75 μg / ml, 10 ng / ml to 50 μg / ml, 10 ng / ml to 25 μg / ml, 10 ng / ml to 10 μg / ml, 10 ng / ml to 1000 ng / ml, 10 ng / ml to 1000 ng / ml, 10 ng / ml to 750 ng / ml, 10 ng / ml to 500 ng / ml, 10 ng / ml to 250 ng / ml, 10 ng / ml to 100 ng / ml, 10 ng / ml to 75 ng / ml, 10 ng / ml to 50 ng / ml, or 10 ng / ml to 25 ng / ml.

[0292] In some embodiments, the solution comprises the composition and at least one additional additive (e.g., a pesticide, surfactant, or other non-pesticide). In some embodiments, such a mixture comprises the composition at a concentration of 0.0001 μg / ml to 10 μg / ml (e.g., applied to the surface of a plant and / or ground (e.g., soil, mud, grass, etc.)). In some embodiments, such mixtures comprise the composition at a concentration of 0.001 μg / ml to 10 μg / ml, 0.01 μg / ml to 10 μg / ml, 0.1 μg / ml to 10 μg / ml, 1 μg / ml to 10 μg / ml, 2 μg / ml to 10 μg / ml, 3 μg / ml to 10 μg / ml, 4 μg / ml to 10 μg / ml, 5 μg / ml to 10 μg / ml, 6 μg / ml to 10 μg / ml, 7 μg / ml to 10 μg / ml, 8 μg / ml to 10 μg / ml, or 9 μg / ml to 10 μg / ml. In some embodiments, such mixtures comprise the composition at a concentration of from 0.0001 μg / ml to 9 μg / ml, from 0.0001 μg / ml to 8 μg / ml, from 0.0001 μg / ml to 7 μg / ml, from 0.0001 μg / ml to 6 μg / ml, from 0.0001 μg / ml to 5 μg / ml, from 0.0001 μg / ml to 4 μg / ml, from 0.0001 μg / ml to 3 μg / ml, from 0.0001 μg / ml to 2 μg / ml, from 0.0001 μg / ml to 1 μg / ml, from 0.0001 μg / ml to 0.1 μg / ml, from 0.0001 μg / ml to 0.01 μg / ml, or from 0.0001 μg / ml to 0.001 μg / ml.

[0293] In some embodiments, the composition is provided in the diet of the plant pest. Thus, in some embodiments, the effective amount of the composition is expressed as micrograms (μg) of RNAi molecules per milliliter (ml) of diet of the plant pest, i.e., μg / ml. In some embodiments, the diet of the plant pest contains the composition at a concentration of 0.001 μg / ml to 10 μg / ml. In some embodiments, the plant pest diet comprises the composition at a concentration of from 0.001 μg / ml to 9 μg / ml, from 0.001 μg / ml to 8 μg / ml, from 0.001 μg / ml to 7 μg / ml, from 0.001 μg / ml to 6 μg / ml, from 0.001 μg / ml to 5 μg / ml, from 0.001 μg / ml to 4 μg / ml, from 0.001 μg / ml to 3 μg / ml, from 0.001 μg / ml to 2 μg / ml, from 0.001 μg / ml to 1 μg / ml, from 0.001 μg / ml to 0.1 μg / ml, or from 0.001 μg / ml to 0.01 μg / ml. In some embodiments, the plant pest diet comprises the composition at a concentration of 0.01 μg / ml to 10 μg / ml, 0.1 μg / ml to 10 μg / ml, 1 μg / ml to 10 μg / ml, 2 μg / ml to 10 μg / ml, 3 μg / ml to 10 μg / ml, 4 μg / ml to 10 μg / ml, 5 μg / ml to 10 μg / ml, 6 μg / ml to 10 μg / ml, 7 μg / ml to 10 μg / ml, 8 μg / ml to 10 μg / ml, or 9 μg / ml to 10 μg / ml.

[0294] The step of delivering the composition to any part of the plant (e.g., roots, tubers, stems, branches, leaves, flowers, etc.), ground (e.g., soil, mud, grass, etc.), plant pest and / or plant pest feeding may involve a single application (single contact) or multiple applications (multiple contacts) of the composition to the plant, ground (e.g., soil, mud, grass, etc.), plant pest and / or plant pest feeding. Delivery to the plant, plant pest and / or part of the plant pest feeding may be in the form of a spray (e.g., pressurized / aerosolized spray, pump), solid (e.g., powder, pellet, bait), or liquid (e.g., homogeneous mixtures such as solutions and heterogeneous mixtures such as suspensions (water and oil based), colloids, micelles, and emulsions). The contact period may vary. In some embodiments, delivering includes exposing the composition to a portion of the plant and / or plant pest for a period of time sufficient to reduce the growth, reproduction (e.g., fertility and / or fecundity), and / or feeding of the plant pest and / or death, if any, of the plant pest.

[0295] In some embodiments, delivery of the composition to the plant and / or plant pest is followed by ingestion and / or absorption of the composition by the plant and / or plant pest. In some embodiments, ingestion of the composition by the plant pest alters the biological function of the plant pest, thereby controlling infestation by the plant pest. Examples of altered biological function of the plant pest include, but are not limited to, reduced growth, reduced reproduction (e.g., fecundity and / or fertility), reduced feeding, reduced movement, reduced development, reduced cell repair, and / or increased mortality.

[0296] In some embodiments, delivering comprises applying the composition to a surface of the plant and / or a portion of a surface contacted by the plant pest (e.g., ground (e.g., soil, mud, grass, etc.)). In some embodiments, applying the composition to a portion of a surface comprises spraying, coating, and / or dusting the surface or portion thereof. In some embodiments, applying the composition to a portion of a surface comprises ground drench or application of the RNAi molecule as a granulated or powdered formulation to the soil adjacent to the roots of the plant.

[0297] The composition may be applied to any part of the plant (e.g., roots, tubers, stems, branches, leaves, flowers, etc.). In some embodiments, the composition is contacted with the above-ground parts of the plant (e.g., leaves) and / or the below-ground parts of the plant (e.g., roots) and may include at least one infalo formulation selected from the group consisting of powder, granules, pellets, capsules, soluble liquid concentrates, sprays (after dilution or concentration), fogs, infalo, seed treatments, plant pest baits, baits, drenches, drip irrigation, or any other form suitable for in-furrow application. Plant parts that may be contacted with the compositions described herein include, but are not limited to, leaves, stems, flowers, fruits, shoots, roots, seeds, tubers, anthers, stamens, or pollen. In some embodiments, the composition is delivered mechanically via high-pressure spraying or sandblasting.

[0298] In some embodiments, delivering comprises providing a composition for dietary uptake by the plant pest. In some embodiments, contacting comprises providing a composition that can be ingested or otherwise absorbed by the plant pest. In some embodiments, the composition is provided in the diet for dietary uptake by the plant pest. In some embodiments, the composition is provided in / on the plant or plant part or topically applied (e.g., dipping, coating, dusting) to the plant or plant part.

[0299] In some embodiments, delivering a composition comprising a pesticidal polynucleotide to a plant pest inhibits (reduces or inhibits) the expression of an endogenous complementary nucleotide sequence (e.g., an RNA sequence) in the plant pest. In some embodiments, the endogenous complementary nucleotide sequence is an endogenous IAP sequence.

[0300] The results of inhibition by the pesticidal polynucleotide can be confirmed by any suitable assay for evaluating one or more characteristics of the plant pest, or by biochemical techniques that evaluate molecules indicative of gene expression (e.g., RNA, protein). In some embodiments, the extent to which a composition comprising a pesticidal polynucleotide provided herein reduces the expression level of a target gene is assessed by comparing the expression level (e.g., mRNA level or protein level) of the target gene with the expression level of an appropriate control (e.g., the level of gene expression in a cell or cell population to which the composition has not been delivered or to which a negative control has been delivered). In some embodiments, the appropriate control level of gene expression may be a predetermined level or value so that the control level does not need to be measured every time. The predetermined level or value can take various forms. In some embodiments, the predetermined level or value can be a single cutoff value, such as a median or mean value.

[0301] In some embodiments, delivering a composition comprising a pesticidal polynucleotide as described herein results in a reduction in gene expression levels in cells of a plant pest. In some embodiments, the reduction in gene expression levels may be 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%, at least 98%, at least 99%, or 100% reduction compared to the control level. In some embodiments, the control level is the level of gene expression (or the average level among a population of cells) in similar plant pest cells that have not been contacted with a composition comprising a pesticidal polynucleotide. In some embodiments, the control level is the level of gene expression (or the average level among a population of cells) in similar plant pest cells that have been contacted with a composition that targets a gene not expressed by plant pest cells, such as green fluorescent protein (GFP).

[0302] In some embodiments, the effect of delivering the composition to cells or plant pests is evaluated after a finite period of time.For example, the effect can be determined in cells or plant pests at least 4 hours, 8 hours, 12 hours, 18 hours, 24 hours after delivering the composition to cells or plant pests; or at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or 14 days after delivering the composition to cells or plant pests.

[0303] In some embodiments, delivery of a composition as described herein results in a reduction in the level of growth, reproduction (e.g., fertility and / or fecundity), and / or feeding of a plant pest. In some embodiments, the reduction in the level of growth, reproduction (e.g., fertility and / or fecundity), and / or feeding may be 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%, at least 98%, at least 99%, or 100% reduction compared to the control level. In some embodiments, the control level is the level of growth, reproduction (e.g., fertility and / or fecundity), and / or feeding of a similar plant pest that has not been contacted with the composition. In some embodiments, the control level is the level of growth, reproduction (e.g., fertility and / or fecundity), and / or feeding of a similar plant pest that has been contacted with a composition that targets a gene not expressed by plant pest cells, such as green fluorescent protein (GFP).

[0304] In some embodiments, delivery of a composition as described herein results in an increase in mortality among a population of plant pests. In some embodiments, the increase in mortality level may be 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%, at least 98%, at least 99%, or 100% increase compared to a control. In some embodiments, the control is mortality among a population of plant pests that are not contacted with the composition. In some embodiments, the control is among a population of plant pests that are contacted with a composition that targets a gene that is not expressed by plant pest cells, such as green fluorescent protein (GFP).

[0305] Aspects of the present disclosure provide transgenic plants that express a biopesticide as described herein. In some embodiments, delivery of yeast particles or polysaccharides (e.g., beta-glucan, manna oligosaccharides, and / or laminarin) increases the efficacy of the biopesticide expressed by the transgenic plant.

[0306] In some embodiments, the method comprises delivering a composition comprising a biological adjuvant that enhances the efficacy of a biopesticide and / or a pesticidal polynucleotide. In some aspects and embodiments, the composition comprising a biological adjuvant further comprises a biopesticide and / or a pesticidal polynucleotide. For example, the composition may comprise a biopesticide such as Bt or a Bt protein together with a biological adjuvant such as YP. In another example, the composition comprises a pesticidal polynucleotide such as a pesticidal dsRNA together with a biological adjuvant such as YP, optionally with the dsRNA encapsulated within the YP. In another example, the dsRNA is not encapsulated within the YP. In another example, the composition comprises a biopesticide such as Bt, a pesticidal polynucleotide such as a dsRNA, and a biological adjuvant such as YP, optionally with the dsRNA encapsulated within the YP. Further embodiments include methods of controlling pest infestations in plants genetically modified to produce a biopesticide such as a Bt protein or engineered to produce a pesticidal polynucleotide such as an siRNA by delivering to such plants a biological adjuvant, such as yeast particles, that enhances the efficacy of the biopesticide or pesticidal polynucleotide expressed by the plant. In another example, a composition includes a chemical pesticide and a biological adjuvant, such as YP, and optionally YCWP, that enhances the efficacy of the chemical pesticide.

[0307] In one aspect, there is provided a method of controlling plant pests, the method comprising delivering to a plant or plant pest a composition according to any one of the preceding claims.

[0308] In one embodiment of the method, the composition is delivered to the plant through the leaves, stems, flowers, seeds, roots, or soil.

[0309] The plant may be any plant that is infested by coleopteran or lepidopteran insects.In some embodiments, the plant is a Solanaceae plant (for example, Solanaceae).Examples of Solanaceae plants include, but are not limited to, potato plant (Solanum tuberosum), nightshade plant (Solanum rostratum), eggplant plant (Solanum melongena), tomato plant (Solanum lycopersicum), tobacco plant (Nicotiana tabacum), pepper plant (Capsicum annum) and woody nightshade plant (Solanum dulcamara).

[0310] In some embodiments, the plant being treated is a plant of the Brassicaceae family. Examples of plants in the Brassicaceae family include, but are not limited to, many vegetables such as cabbage, broccoli, cauliflower, kale, Brussels sprouts, collard greens, savoy, kohlrabi, and Chinese broccoli (Brassica oleracea), turnip, Napa cabbage, pomme de théâtre, bok choy, and rapini (Brassica rapa), rocket salad / arugula (Eruca sativa), garden cress (Lepidium sativum), water cress (Nasturtium officinale), and radish (Raphanus), as well as a few species such as horseradish (Armoracia rusticana), Brassica oleracea, wasabi (Eutrema japonicum), white mustard, Indian mustard, and black mustard (Sinapis alba, Brassica juncea, and B. nigra, respectively).

[0311] In some embodiments, the plant to be treated is a plant of the Poaceae family. Examples of plants in the Poaceae family include, but are not limited to, corn, millet, oats, barley, rye, sugarcane, wheat, sorghum, and wild rice.

[0312] In some embodiments, the plant is a plant of the Cucurbitaceae family. Examples of plants in the Cucurbitaceae family include, but are not limited to, pumpkins and cucumbers.

[0313] In some embodiments, the plant is a legume (Fabaceae) plant. Examples of plants in the Fabaceae family include, but are not limited to, Glycine max (soybean), Phaseolus (bean), Pisum sativum (pea), Cicer arietinum (chickpea), Vicia faba (fava bean), Medicago sativa (alfalfa), Arachis hypogaea (peanut), Ceratonia siliqua (carob), and Glycyrrhiza glabra (licorice).

[0314] In some embodiments, the plant is a plant of the Apiaceae family. Examples of plants in the Apiaceae family include, but are not limited to, many important vegetables such as carrot (Daucus carota), coriander (Coriandrum sativum), and celery (Apium graveolens), as well as important medicinal plants including ginseng (Angelica sinensis), Peucedanum praeruptorum, and Angelica dahurica.

[0315] In some embodiments, the plant is a plant of the Poaceae family. Examples of plants in the Poaceae family include, but are not limited to, corn, millet, oats, barley, rye, sugarcane, wheat, sorghum, and wild rice.

[0316] In some embodiments, the plant is a plant of the Fagaceae, Asteraceae, Amaryllidaceae, or Umbelliferae families.

[0317] In some embodiments, the plant is a plant of the Malvaceae family. Examples of plants in the Malvaceae family include, but are not limited to, various species of cotton (e.g., Gossypium hirsute, Gossypium barbadense, Gossypium arboretum, and Gossypium herbaceum).

[0318] In some embodiments, the delivery method comprises applying the composition to the surface of the plant or insect pest.

[0319] In some embodiments, the composition is delivered to the plant through the leaves, stems, seeds, roots, or soil.

[0320] Insects, as used herein, refer to insects in any developmental stage. In some embodiments, the insect is an insect egg. In some embodiments, the insect is an insect larva. In some embodiments, the insect is an insect pupa. In some embodiments, the insect is an adult insect.

[0321] In some embodiments, the insect pest belongs to the order Coleoptera, Lepidoptera, Diptera, or Hemiptera.

[0322] Examples of insects in the order Coleoptera include, but are not limited to, Chrysomelidae (leaf beetles), Curculionidae (weevils), Meloidae (blister beetles), Tenebrionidae (tenebrionid beetles), Scarabaeidae (scarab beetles), Cerambycidae (pine beetles), and weevils. Curculionidae (Chinese White Pine Beetle), Nitidulidae (Small Hive Beetle), Chrysomelidae (Broad-Shouldered Leaf Beetle), Cerambycidae (Mulberry Longhorn Beetle), Phyllotreta (Flea Beetle), Diabrotica (Corn Rootworm), Chrysomelidae (Shrimp Beetle) a) (Cottonwood Leaf Beetle), Hypothenemus (Coffee Berry Borer), Sitophilus (Maize Weevil), Epitrix (Tobacco Flea Beetle), E. cucumeris (Potato Beetle), P. pusilla (Western Black Flea Beetle); Anthonomus (Pepper Weevil), Hemicr epidus (wireworm), Melanotus (wireworm), Ceutorhychus (cabbage pod weevil), Aeolus (wireworm), Horistonotus (sandworm), Sphenophorus (corn billbug), S. zea (timothy billbug), S. parvulus (bluegrass billbug), S.callosus (southern cornbill bug); Phyllophaga (white grub), Chaetocnema (corn flea beetle), Popillia (bean beetle), Epilachna (green lady beetle), Cerotoma (bean leaf beetle), Epicauta (blister beetle), Chrysomelidae (alligatorweed flea beetle), and any combination thereof.

[0323] In some embodiments, the insect pest belonging to the order Coleoptera is of a species selected from the group consisting of Leptinotarsa ​​spp., Phyllotreta spp., Cerotoma spp., Diabrotica spp., Tribolium spp., Anthonomus spp., and Alticini spp.

[0324] In certain embodiments, the coleopteran insect species is a Leptinotarsa ​​species.

[0325] In some embodiments, the coleopteran insect may be any species of the genus Leptinotarsa, including, but not limited to, Leptinotarsa ​​decemlineata (Colorado potato beetle), Leptinotarsa ​​behrensi, Leptinotarsa ​​collinsi, Leptinotarsa ​​defecta, Leptinotarsa ​​haldemani (Holdman's green potato beetle), Leptinotarsa ​​heydeni, Leptinotarsa ​​juncta (false potato beetle), Leptinotarsa ​​lineolata (bloeblush leaf beetle), Leptinotarsa ​​peninsularis, Leptinotarsa ​​rubiginosa, Leptinotarsa ​​texana, Leptinotarsa ​​tlascalana, Leptinotarsa ​​tumamoca, and Leptinotarsa ​​typographica. In exemplary embodiments, the coleopteran insect is in the family Chrysomelidae, and optionally the insect is a Phyllotreta species or a Psylloides species, and optionally the insect is Phyllotreta cruciferae (canola flea beetle), Phyllotreta striolata (striolata flea beetle), or Psylliodes chrysocephala (cabbage stem flea beetle).

[0326] In an exemplary embodiment, the Leptinotarsa ​​species insect is a Colorado potato beetle.

[0327] Examples of insects in the order Lepidoptera include, but are not limited to, Nymphalidae (Nymphalians), Danaidae (Monarchs), Pieridae (Pieridae and Eurema butterflies), Papilionidae (Swallowtails), Lycaenidae (Lycaenidae), Hesperiidae (Hesperiids), Tineid moths, and the like. ae (bur moths), Sesiidae (sease moths), Pyralidae (pyralid moths), Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawk moths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (looper moths, noctuids, and burrowing moths). In exemplary embodiments, the lepidopteran insect is an insect within the Spodoptera species, and optionally the insect is S. frugiperda (stalk fall armyworm), Spodoptera exigua (beet armyworm), Spodoptera litura (tobacco rootworm), Spodoptera litoralis (tomato moth), or Spodoptera exigua (beet armyworm). In another exemplary embodiment, the insect is a Plutella species, and optionally the insect is within the Plutellidae family, and optionally the insect is Plutella xylostolla (diamond moth).In another exemplary embodiment, the insect is within the Noctuidae family, and optionally the insect is a Chrysodeixis species, a Helicoverpa species, or a Trichoplusia species, and optionally the insect is a Chyrysodeixis includens (soybean looper), a Chyrysodeixis acuta (tomato semilooper), a Helicoverpa armigera (cotton bollworm), or a Helicoverpa zea (tomato fruitworm), and the insect is a Trichoplusia ni (cabbage looper). In another exemplary embodiment, the insect is within the Tortricidae family, and optionally the insect is a Cydia species or a Lobesia species, and optionally the insect is a Cydia pomonella (tooth moth) or a Lobesia botrana (European grapevine moth). In another exemplary embodiment, the insect is within the Pieridae family, optionally the insect is a Pieris species, and optionally the insect is Pieris rapae (imported caterpillar). In another exemplary embodiment, the insect is within the Gelechiidae family, optionally the insect is a Tuta species, and optionally the insect is Tuta absoluta (tomato tea moth). In another exemplary embodiment, the insect is within the Pyralidae family, optionally the insect is an Amyelois species, and optionally the insect is Amyelois transitella (tomato tea moth).

[0328] The dipteran insect may be any dipteran insect of the order Diptera. Examples of insects of the order Diptera include, but are not limited to, Culicidae (mosquitoes), Tabanidae (horse flies / deer flies), Simuliidae (black flies), Psychodidae (moth flies), Ceratopogonidae (punkies, no-see-ums), Muscidae (house flies), and Cecidomyiidae (gall flies). , Tephritidae (fruit flies), Agromyzidae (leaf miners), Anthomyiidae (maggots), Drosophilidae (fruit flies), Tipulidae (crane flies), Calliphoridae (blow flies), Chironomidae (midges), and Sarcophagidae (flesh flies).

[0329] The hemipteran insect may be a hemipteran insect of the order Hemiptera. Examples of insects in the order Hemiptera include, but are not limited to, Miridae (plant bugs), Lygaeidae (seed bugs), Tingidae (tail bugs), Coreidae (squash bugs and earwigs), Alydidae (broadhead bugs), Rhopalidae (lace bugs), Berytidae (silver bugs), Reduviidae (assassin bugs), Phymatidae (ambush bugs), Nabidiidae (shrimp bugs), and the like. ae) (Teadbugs), Anthocoridae (Shrimp Bugs), Corixidae (Water Bugs), Gerridae (Water Striders), Nepidae (Water Scorpions), Belostomatidae (Stag Bugs), Naucoridae (Bugs), Notonectidae (Cicadas), Cicadidae (Cicadas), Cicadellidae (Leafhoppers), Membracidae (Treehoppers), Cercopidae (Foxtail Bugs or Spit Bugs) Froghoppers), Fulgoridae (plant hoppers), Psyllidae (lice or jumping plant lice), Aleyrodidae (whiteflies), Aphididae (aphids, plant lice), Pentatomidae (stink bugs), and Coccidae (soft scale insects).

[0330] In some embodiments of the present disclosure, any of the compositions described herein may be formulated into a deliverable form suitable for a particular application. Deliverable forms that can be used in accordance with embodiments of the present disclosure include, but are not limited to, liquids (including homogeneous mixtures such as soluble liquid concentrates, and heterogeneous mixtures such as suspensions, colloids, micelles, and emulsions), emulsifiable concentrates, solids, oil dispersions, pastes, granules, wettable powders, wettable powder granules, dusts, fumigants, suspensions, sprays, encapsulated, or microencapsulated formulations in or on microbeads or other carrier particles, in films or coatings, or on or in matrices, or as leaf, seed, root, or stem treatments. Suitable deliverable forms can be selected and formulated by those skilled in the art using methods currently known in the art. Formulation ingredients for the compositions of the present disclosure may include any known in the art and can be selected and formulated by those skilled in the art using methods currently known in the art. Formulation ingredients may include, but are not limited to, acidifiers, buffers, antifoaming agents, anti-transpirants, dyes and brighteners, compatibilizers, crop oil concentrates, surfactants, deposition agents, drift reducing agents, dyes and brighteners, feeding stimulants, herbicide safeners, spreading agents, bulking agents, adhesives, suspending agents, gelling agents, synergists, wetting agents, emulsifiers, dispersing agents, penetrating agents, neutralizing agents, water-absorbing agents, and / or water softeners. Conventional agricultural carriers may be used, including, for example, silica, clay, activated carbon, cellulose, lactose, sucrose, ammonium sulfate, and urea-based resins. The compositions may also be provided in liquid, solid, or gel form in combination with agriculturally acceptable, food-acceptable, or pharmaceutically acceptable carriers or excipients. For solid compositions, suitable carriers include pharmaceutical or food grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, and magnesium carbonate.Suitably, the formulation is in the form of a tablet or pellet.Suitable carriers may also be human or animal food materials.In some embodiments, the composition is delivered by contacting with a plant surface. In some embodiments, the surface is a plant leaf, flower, or fruit. In such embodiments, application may be achieved by spraying the plant leaf, flower, or fruit. Contacting can also be in the form of a seed treatment. As known to those skilled in the art in pesticide formulation and seed treatment, suitable binders, inert carriers, surfactants, etc. can optionally be included in the composition. In some embodiments, contacting includes providing the polynucleotide in a composition further comprising one or more carrier agents and / or one or more surfactants (e.g., organosilicon, organosilicon surfactants). Compositions suitable for formulation with polynucleotides for RNAi are well known in the art, including, for example, those described in U.S. Patent Application Publication No. 2022 / 0372478, published November 24, 2022, entitled "Stabilization of RNA for Exogenous RNAi Agricultural Applications and Formulations," which is incorporated herein by reference.

[0331] As recognized by those skilled in the art, effective amount varies depending on specific plant, severity of infestation, duration of infestation, previous exposure to pesticides and similar factors within the knowledge and expertise of the expert.These factors are well known to those skilled in the art and can be handled only by routine experimentation.Generally, to increase efficiency and reduce cost, lower effective concentration is used, that is, the lowest concentration that provides insect control.

[0332] Kits and Articles of Manufacture Another aspect of the present disclosure relates to a kit comprising a composition of the present disclosure and optional instructional materials. The instructional materials may include publications, records, diagrams, or any other medium of expression used to communicate the usefulness of the composition of the present disclosure (e.g., controlling plant pests with a lower effective amount of pesticide to control the plant pest compared to a composition that does not include a biological adjuvant). The instructional materials may also describe, for example, the effective amount of the composition of the present disclosure. The instructional materials of the kit of the present disclosure can be, for example, attached to a container containing the composition of the present disclosure or shipped together with a container containing the pharmaceutical composition. Alternatively, the instructional materials can be shipped separately from the container, with the intention that the instructional materials and the pharmaceutical composition be used cooperatively by the recipient.

[0333] The present disclosure also encompasses kits comprising the compositions of the present disclosure and a delivery device. By way of example, the delivery device can be adapted or configured to deliver the composition to a plant or a pest. The kit can further include educational materials as described herein.

[0334] In certain embodiments, the kit may include two containers: a first container containing a biopesticide; and a second container containing a biological adjuvant. Examples of containers include spray bottles, boxes, bags, etc. Typically, the kit includes instructions for application of the separate components.

[0335] In order that this invention may be better understood, the following examples are set forth. These examples are for illustrative purposes only and should not be construed as limiting the scope of the invention in any way.

[0336] example The foregoing description of specific embodiments sufficiently reveals the general nature of the present disclosure so that others can readily modify and / or adapt such specific embodiments to various uses by applying knowledge within the skill of those skilled in the art without departing from the general concept of the disclosure and without undue experimentation. Such adaptations and modifications are therefore intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology used herein is intended to be descriptive rather than limiting, as the terminology or terminology used herein would be interpreted by one of ordinary skill in the art in light of the teaching and guidance.

[0337] Example 1: Preparation of yeast particles (YP) and mass delivery system Yeast particles are often 3-5 μm hollow and porous microparticles derived from baker's yeast, composed primarily of approximately 80% 1→6-β-branched, 1→3-β-glucan, 2-4% chitin, and 40% mannan w / w. Yeast particles are readily available, biodegradable, substantially spherical particles approximately 2-4 mm in diameter.

[0338] Methods for preparing extracted yeast cell wall particles are well known in the art and are described, for example, in U.S. Pat. Nos. 4,992,540, 5,082,936, 5,028,703, 5,032,401, 5,322,841, 5,401,727, 5,504,079, 5,968,811, 6,444,448, 6,476,003, U.S. Patent Application Publication Nos. 2003 / 0216346, 2004 / 0014715, and published PCT application WO 02 / 12348, the disclosures of which are incorporated herein by reference.

[0339] The morphology of extracted yeast cell wall particles, referred to as "whole glucan particles" or "WGP" (see U.S. Pat. Nos. 5,032,401 and 5,607,677), may be modified to facilitate improved retention and / or delivery of payload molecules. Such modifications feature trapping molecules and nanoparticles, and multiple trapping molecules and nanoparticles formulated in specific forms to achieve the desired improved delivery characteristics. As used herein, WGPs are typically whole glucan particles with >90% beta-glucan purity.

[0340] Commercial yeast particles (YP) Yeast particles (YP) were used commercially available from Biorigin (Louiseville, KY, USA) or LeSaffre (Marcq-en-Baroeul, France), examples of which are described below.

[0341] Preparation of glucan particles (GP) Glucan particles (GPs), also referred to herein as yeast glucan particles ("YGPs"), are purified hollow yeast cell "ghosts" containing abundant β-glucan spheres, typically 2-4 microns in diameter. Generally, glucan particles can be prepared from yeast cells by extraction and purification of the alkali-insoluble glucan fraction from the yeast cell wall. Yeast cells can be treated with aqueous hydroxide, which digests the cell's proteins and intracellular components without disrupting the yeast cell wall, leaving the glucan wall components devoid of significant protein contamination and with essentially the same unaltered cell wall structure of β(1-6) and β(1-3)-linked glucans. The 1,3-β-glucan outer shell provides receptor-mediated uptake by phagocytes, such as macrophages, that express the β-glucan receptor.

[0342] Glucan particles can be produced as follows: Yeast particles (S. cerevisiae), Biorigin MOS55, are suspended in 1 liter of 1 M NaOH and heated to 85°C. The cell suspension is vigorously stirred at this temperature for 1 hour. The insoluble material, containing the cell walls, is recovered by centrifugation. This material is then suspended in 1 M NaOH, heated, and vigorously stirred for 1 hour. The suspension is cooled to room temperature, and extraction is continued for an additional 16 hours. The insoluble residue is recovered by centrifugation. This material is finally extracted into water adjusted to pH 4.5 with HCl. The insoluble residue is recovered by centrifugation and washed three times with water, isopropanol, and acetone. The resulting slurry is poured into a glass tray and dried under reduced pressure to produce a fine white powder.

[0343] Preparation of glucan lipid particles (GLP) GLPs retain some of the lipid content of yeast cells, creating a more hydrophobic internal cavity ideal for loading hydrophobic payloads. GLPs are prepared by modifying the GP preparation method described above. To prepare GLPs, the isopropanol and acetone washes are eliminated; instead, the insoluble residue recovered by centrifugation is washed three times with water. The particles are then dried by freeze-drying or spray-drying.

[0344] Whole Glucan Particles (WGP)

[0345] A more detailed description of the process for preparing WPG can be found in U.S. Patent Nos. 4,810,646, 4,992,540, 5,028,703, 5,607,677, and 5,741,495 (incorporated herein by reference). For example, U.S. Patent No. 5,028,703 discloses that yeast WGP particles can be produced from yeast strain R4 cells during fermentation culture. The cells are harvested by batch centrifugation at 8000 rpm for 20 minutes in a Sorval RC2-B centrifuge. To prepare the cells for total glucan extraction, the cells are washed twice in distilled water. The first step involved resuspending the cell mass in 1 liter of 4% w / v NaOH and heating to 100°C. The cell suspension is vigorously stirred at this temperature for 1 hour. The insoluble material containing the cell walls is recovered by centrifugation at 2000 rpm for 15 minutes. This material is suspended in 2 liters of 3% w / v NaOH and heated to 75°C. The suspension is vigorously stirred at this temperature for 3 hours. The suspension is then cooled to room temperature, and extraction is allowed to continue for an additional 16 hours. The insoluble residue is recovered by centrifugation at 2000 rpm for 15 minutes. The material is finally extracted for 1 hour at 75°C in 2 liters of 3% w / v NaOH adjusted to pH 4.5 with HCl. The insoluble residue is recovered by centrifugation and washed three times with 200 milliliters of water, once with 200 milliliters of dehydrated ethanol, and twice with 200 milliliters of dehydrated ethyl ether. The resulting slurry is placed on a Petri dish and allowed to dry.

[0346] Varying degrees of purity of the glucan particles are achieved by modifying the extraction / purification process. Generally, these GPs are approximately 80-85% pure on a w / w basis of beta-glucan, with slightly lower "purity" levels achieved after the introduction of payloads, sequestrants, or other components. In exemplary embodiments, the GPs are <90% beta-glucan pure.

[0347] Preparation of YCP particles Yeast cells (Rhodotorula sp.) from cultures obtained from the American Type Culture Collection (ATCC, Manassas, VA) were grown aerobically to stationary phase in YPD at 30°C. Rhodotorula sp. cultures available from ATCC include numbers 886, 917, 9336, 18101, 20254, 20837, and 28983. Cells were harvested by batch centrifugation at 2000 rpm for 10 minutes. The cells were then washed once in distilled water and then resuspended in water adjusted to pH 4.5 with HCl at 75°C for 1 hour. Insoluble material containing cell walls was recovered by centrifugation. This material was then suspended in 1 liter of 1 M NaOH and heated to 90°C for 1 hour. The suspension was cooled to room temperature, and extraction was continued for an additional 16 hours. The insoluble residue was collected by centrifugation and washed twice with water, isopropanol, and acetone. The resulting slurry was poured into a glass tray and allowed to dry at room temperature to yield 2.7 g of a fine, light brown powder.

[0348] Preparation of GCMP particles Yeast cells (Rhodotorula sp.) from a culture obtained from the American Type Culture Collection (ATCC, Manassas, VA) were grown aerobically to stationary phase in 10 L of YPD at 30°C. Rhodotorula sp. cultures available from ATCC include numbers 886, 917, 9336, 18101, 20254, 20837, and 28983. Cells (10 L) were harvested by batch centrifugation at 2000 rpm for 10 minutes. The cell pellet was then washed once with 1 liter of distilled water and then resuspended in 1 liter of distilled water adjusted to pH 4.5 with HCl and heated to 75°C with stirring for 1 hour. The insoluble material containing the cell wall was recovered by centrifugation and suspended in 1 liter of 1 M NaOH and heated to 55°C. The cell suspension was mixed at this temperature for 1 hour. The insoluble material containing the cell wall was collected by centrifugation at 2000 rpm for 10 minutes. This material was then suspended in 1 liter of water. The insoluble residue was collected by centrifugation and washed three times with 1 L of water, four times with 200 milliliters of dehydrated isopropanol, and two times with 200 milliliters of acetone, and the powder was dried by solvent evaporation. This process produced 3.1 g of a pale pink powder.

[0349] Preparation of YGMP particles S. cerevisiae (100 g of Fleishman's Baker's Yeast) was suspended in 1 liter of 1 M NaOH and heated to 55°C. The cell suspension was mixed at this temperature for 1 hour. The insoluble material, containing cell walls, was recovered by centrifugation at 2000 rpm for 10 minutes. This material was then suspended in 1 liter of water and adjusted to pH 4-5 with HCl and incubated at 55°C for 1 hour. The insoluble residue was recovered by centrifugation and washed once with 1000 milliliters of water, four times with 200 milliliters of dehydrated isopropanol, and twice with 200 milliliters of acetone. The resulting slurry was poured into a glass tray and allowed to dry at room temperature to yield 12.4 g of a fine, slightly off-white powder.

[0350] Preparation of glucan mannan lipid particles (GMLPs) GMLP was prepared by the procedure described above for preparing YGLP, except without the steps requiring washing with isopropanol and acetone.

[0351] Example 2: Preparation of dsRNA Based on the possibility that genes are sensitive to RNAi and that RNAi has a phenotypic effect, candidate genes for testing were hypothesized.RNAi is a highly unpredictable technology, and it is impossible to reliably predict which gene targets are sensitive to RNAi, and if so, which gene targets, when knocked down, will suppress the amount of encoded protein and lead to a phenotype that will result in insect control.Target gene-specific dsRNA was synthesized by in vitro transcription using a commercial kit or GreenLight Biosciences' proprietary RNA synthesis process described in U.S. Patent No. 10,858,385, which is incorporated herein by reference.Candidate genes were tested for lethal phenotypes in insect cell-based assays or microinjection.For cell-based assays, Plutella xylostella (diamond moth; DBM) cells were seeded into 12-well plates at 4 x 10e5 cells / well. For each well, Cellfectin II-dsRNA complexes were made in a sterile 1.5 ml tube using 8 μg of dsRNA and 8 μl of Cellfectin II mixed with nuclease-free (NF) water for a final volume of 100 μl. The Cellfectin II-dsRNA complexes were delivered to DBM cells. Cellfectin II-GS4, which targets GFP, which is not expressed in DBM cells, was used as a negative control. Assays were performed at least three times in replicates.

[0352] For microinjection assays, target gene-specific dsRNA sequences were injected into third-instar P. xylostella larvae to test for knockdown of the target gene. The dsRNA sequences were diluted to a concentration of 6–7 g / L using nuclease-free (NF) water. GS4, which targets GFP, not expressed in P. xylostella, was used as a negative control. Using a Nanoliter 2020 microinjector equipped with a micropipette needle, 200 nL of diluted dsRNA (1.2–1.4 μg of dsRNA) was injected into the second proboscis of the larvae. After 10 minutes, the larvae recovered from the injection and were placed on artificial diet for 72 hours. Live larvae were collected into a 1.5 mL microcentrifuge tube containing three 2.3 mm stainless steel beads and 210 mL of lysis buffer (DNA / RNA Shield + Proteinase K). The tubes were immediately placed in a GenoGrinder to homogenize the samples, followed by qPCR. Assays were replicated at least twice. dsRNA sequences that produced significant levels of target gene knockdown compared to the GS4 negative control in two or more independent runs were considered "positive hits."

[0353] Example 3: Encapsulation of dsRNA into YP containing complex core A Step 1: Preparation of YP EDTA

[0354] Loading and encapsulation of Na4EDTA as ethylenediaminetetraacetic acid (EDTA-H4) to obtain a target YP:EDTA-H4 weight ratio of 1:1 was achieved by the following protocol. EDTA is primarily used for its nuclease inhibitory properties. Other nuclease inhibitors, such as SHMP (sodium hexametaphosphate), TPP (sodium tripolyphosphate), or PVSA (polyvinyl sulfonic acid), may also be used. EDTA was encapsulated in YP by adding 781 mL of 500 mg / mL tetrasodium EDTA (Na4EDTA) solution to 300 mg of lyophilized YP and mixing to a uniform suspension, followed by adding 719 mL of 3 M sulfuric acid solution and mixing to a uniform suspension. After 1 hour of incubation at room temperature, the YP-EDTA formulation was washed four times with 6 mL of water, resuspended in 6 mL of 2 mg / mL maltodextrin, frozen, and lyophilized.

[0355] Step 2: dsRNA loading (target loading of 50μg dsRNA / mg YP)

[0356] Loading of dsRNA into YP was achieved by the following protocol: 300 mg of YP-EDTA (150 mg of YP) was weighed into a 15 mL centrifuge tube. An RNA loading solution was prepared at 5 mg dsRNA / mL in RNase-free water. To achieve a target of 50 μg dsRNA / mg YP, RNA loading was performed twice. 750 μL of dsRNA solution was added to YP-EDTA (5 μL of loading solution per mg of YP), mixed with a blunt pipette tip, and centrifuged three times. The sample was lyophilized and frozen. The dsRNA loaded into the hollow center of the YP was washed by adding 375 μL of RNase-free water to the dried YP-EDTA dsRNA pellet. The sample was mixed, frozen, and lyophilized three times.

[0357] Step 3: Capture dsRNA as a CCA complex

[0358] The capture reaction was carried out to form the CCA formulation (3:1 poly-l-lysine:dsRNA ratio) according to the following protocol: A 15 mg / mL PLL solution was prepared in RNase-free water. PLL loading steps 8-11 were repeated twice to achieve the target of 150 μg PLL / mg YP (3:1 PLL:dsRNA ratio). 750 μL of PLL solution was added to the dried YP-EDTA-dsRNA, mixed using a blunt pipette tip, centrifuged three times, and incubated at room temperature for 30+ minutes. The resulting lyophilized sample was frozen.

[0359] The YP EDTA dsRNA-PLL was resuspended in 7.5 mL of RNase-free water to achieve a concentration of 1 mg dsRNA / mL and 20 mg YP / mL. A homogenous YP EDTA dsRNA PLL suspension (CCA formulation) was obtained using a Polytron (20-30 seconds) and a sonicator (20-30 seconds). The Polytron / sonication step was repeated three times. The suspension was flash-frozen in dry ice. Samples were stored at -20°C or -80°C until ready for use.

[0360] Example 4: Mortality of diamondback moth (DBM) larvae on Chinese cabbage leaves treated with naked dsRNA or CCA-encapsulated dsRNA Control (GS134) and insecticidal dsRNA were encapsulated in the CCA delivery system as described above. Chinese cabbage leaves were sprayed with either naked / unencapsulated dsRNA (2.5 g / L) or CCA-encapsulated dsRNA (0.5 g / L) and allowed to dry. 45 mm diameter leaf discs were placed in petri dishes and infested with newly hatched first-instar Plutella xylostella (diamond moth, DBM) larvae. Larvae were counted 5, 7, and 10 days after infestation. Figure 1 shows that after a 10-day exposure period, larvae on leaf discs treated with CCA-encapsulated dsRNA GS329 showed increased mortality in the CCA formulation.

[0361] Example 5: Mortality of diamondback moth (DBM) larvae on Chinese cabbage leaves treated with a mixture of Bacillus thuringiensis toxin (Bt) products and naked dsRNA or CCA-encapsulated dsRNA

[0362] The commercially available Bacillus thuringiensis preparation DIPEL® (Valent Biosciences) was used. DIPEL® is a bioinsecticide containing Bacillus thuringiensis subsp. kurstaki (Btk) strain ABTS-351, which contains multiple insecticidal proteins and a balance of Bt spores to maximize efficacy against lepidopteran pests. XENTARI® is a bioinsecticide containing a naturally potent strain (ABTS-1857) of the microorganism Bacillus thuringiensis subsp. aizawai (Bta). The unique profile of XENTARI®'s insecticidal proteins provides control of difficult-to-control pests such as armyworms and diamondback moths.

[0363] A control (GS134) targeting Nicotiana benthamiana phytoene desaturase and dsRNA targeting a hypothetical diamondback moth gene encapsulated in a CCA delivery system as described above in Example 1 were mixed with commercial Bacillus thuringiensis (Bt) products. Chinese cabbage leaves were sprayed with a mixture of commercial Bt products mixed with either naked / unencapsulated dsRNA or CCA-encapsulated dsRNA at a concentration of 0.5 mg / mL dsRNA and allowed to dry. 45 mm diameter leaf disks were placed in Petri dishes and infested with "freshly hatched" first-instar Plutella xylostella (diamondback moth, DBM) larvae. Larvae were counted 5, 7, and 10 days after infestation. Figure 2 shows that after a 10-day exposure period, up to 71% of larvae died on leaf discs sprayed with a mixture of Bt products and CCA-encapsulated dsRNA, compared to the control (treated with Bt products only). These results unexpectedly demonstrate that the CCA delivery system was acting to enhance Bt.

[0364] Example 6: Mortality of diamondback moth (DBM) larvae on Chinese cabbage leaves treated with empty YP or a mixture of YP and the Bt product DIPEL®

[0365] Empty YP without any encapsulated dsRNA was used in this experiment. Chinese cabbage leaves were sprayed with either DIPEL® (Valent Biosciences) alone, empty YP alone, or a mixture of empty YP and DIPEL®. YP concentrations of 0.1, 1, 3, and 10 mg / ml were used for YP alone and the YP + DIPEL® composition. A sublethal LC20% dose of DIPEL® was used in all instances. Treated leaves were cut into 45 mm diameter discs. The leaf discs were placed in Petri dishes and then infested with "freshly hatched" first-instar Plutella xylostella (diamond moth, DBM) larvae. Larvae were counted 5, 7, and 10 days after infestation. Tukey statistical analysis was performed based on the DIPEL®-only control. Figure 3 shows that sublethal concentrations of DIPEL® resulted in low mortality, as expected. However, a significant increase in mortality was observed after 7 days for test compositions combining YP at 3 mg / mL and 10 mg / mL with the same sublethal concentrations of DIPEL® used in the control group. By day 11, mortality increased to nearly 90% for compositions containing sublethal DIPEL® concentrations plus a 10 g / L YP concentration, and to over 60% for compositions containing sublethal DIPEL® concentrations plus a 3 mg / mL YP concentration. Mortality in these blank YP + DIPEL® treatments was significantly higher than the sublethal mortality of the individual treatments combined, indicating that YP significantly enhanced the efficacy of Bt.

[0366] Example 7: Mortality of Fall Armyworm Larvae on Soybean Leaves Treated with Blank YP or a Mixture of YP and the Bt Product XENTARI® Empty YP without any encapsulated dsRNA was used in this experiment. For the control group, a sublethal LC20 concentration of the Bt product, XENTARI® (Valent Biosciences), was used, and the test group combined the same concentration of XENTARI® as the control with YP concentrations of 0.1, 1, 3, and 10 mg / ml. An additional control group used YP alone at the same concentration. Soybean leaves were sprayed with the composition and allowed to dry. 45 mm diameter leaf discs were placed in Petri dishes and infested with "freshly hatched" first-instar Spodoptera frugiperda (Fallen Armyworm, FAW) larvae. Larvae were counted 5, 7, and 10 days after infestation. Tukey statistical analysis was performed based on the XENTARI®-only control. 4 shows that after a 10-day exposure period, mortality in the YP + XENTARI® composition increased by nearly 50% at the highest YP concentration (10 g / L) when compared to the sublethal XENTARI®-only control. Mortality in the blank YP + XENTARI® treatment was higher than that of the combined sublethal XENTARI® control and the YP-only control, indicating that YP significantly enhanced the effect of XENTARI®.

[0367] Example 8: Mortality of Bt-resistant diamondback moth (DBM) larvae on Chinese cabbage leaves treated with DIPEL® or a mixture of YP and the Bt product DIPEL® Experiments were conducted to establish that YP can enhance the effectiveness of Bt even against Bt-resistant insect populations. Specifically, the experiment used Plutella xylostella (Benzon Research) resistant to various Cry proteins of Bacillus thuringiensis subsp. kurstaki, such as those found in the Bt product DIPEL® (Valent Biosciences). Chinese cabbage leaves were sprayed with either a DIPEL® control or a mixture of blank YP and DIPEL® at a YP concentration of 10 mg / ml. The leaves were cut into 45 mm diameter discs. The leaf discs were placed in Petri dishes and then infested with "freshly hatched" first-instar Plutella xylostella (diamond moth, DBM) larvae. Larvae were counted 5, 7, and 11 days after infestation. The results are modeled in Figure 5 as a dose-response curve indicating predicted mortality by dose. The dose-response curve for YP + DIPEL® was shifted to the left relative to the curve for DIPEL® alone, indicating that lower doses were required to cause mortality with Bt + YP than with Bt alone. This enhanced effect of YP on the efficacy of Bt against resistant P. xylostella increased over time. The LC50 was 2, 3, and 13-fold lower than Bt alone at 5, 7, and 11 days of exposure, respectively, as shown in Figure 5. The effect of Bt + YP was greater at lower concentrations; after 11 days of exposure, the LC50 was 13-fold lower and the LC20 was 103-fold lower than Bt alone.

[0368] Example 9. Delivery of yeast particles into transgenic plants expressing Bacillus thuringiensis (Bt) crystal proteins Experiments were conducted to determine damage to corn cultivars—transgenic corn plants expressing the Bacillus thuringiensis (Bt) Cry1Ab protein (crystal protein)—and standard corn plants (i.e., corn plants that do not express Cry1Ab) after infestation with fall armyworm susceptible to the Bacillus thuringiensis (Bt) Cry1Ab protein (crystal protein) and treatment with yeast particles. The yeast particles tested here (including yeast cell wall particles) were intact or fragmented yeast cell wall particles formulated with certain samples and further containing various standard agronomic formulation ingredients for delivery via spraying. One sample (Yeast Particle 1 in Figure 6) contained intact YCWP in water. The second sample (yeast particle 2 in Figure 6) contained bead-milled YCWP formulated with a surfactant (e.g., sodium isopropyl naphthalene sulfonate, acrylic copolymer), biological preservative(s) / biocide(s), and antifoam (e.g., silicone antifoam). The third sample (yeast particle 3 in Figure 6) contained bead-milled YCWP formulated with a surfactant (e.g., methyl oleate / methyl linoleate ester, castor oil ethoxylate), biological preservative(s) / biocide(s), and antifoam (e.g., silicone antifoam).

[0369] Corn plants were infested with first-instar Bt-susceptible fall armyworm (FAW) larvae at five larvae per plant (infested using 1.5 microcentrifuge tubes). The corn was treated with a spray formulation of yeast particles (containing yeast cell wall components) at 10 grams of yeast per liter (gAi / L). The yeast particles were applied to the leaves of the corn plants using a CO2 backpack sprayer at a rate of 50 gallons per acre. The carrier for the yeast particles was water. Twelve days after treatment of the corn with yeast particles, data were collected on the plants to determine the rating scale (Figure 6).

[0370] The addition of yeast particles to standard corn plants (also called "non-transformed" plants) did not result in fall armyworm mortality, indicating that the yeast particles themselves were not capable of controlling the fall armyworm pest. However, the addition of yeast particles to transgenic corn plants (also called "transformed" plants) caused an increase in fall armyworm mortality compared to transgenic plants that were not treated with yeast particles. These data indicate that yeast particles enhance the efficacy of Bacillus thuringiensis (Bt) proteins expressed by transgenic plants (biopesticides expressed by transgenic plants).

[0371] Example 10. Delivery of yeast particles into transgenic plants expressing Bacillus thuringiensis (Bt) crystal proteins Experiments were conducted to determine damage to transgenic maize plants expressing the Bacillus thuringiensis (Bt) Cry1Ab protein following infestation by fall armyworm resistant to the Bt Cry1Fa protein (a protein similar to the Cry1Ab protein (crystal protein) with which cross-resistance is evident in Cry1Fa-resistant fall armyworm populations) and treatment with yeast particles (containing yeast cell wall components).

[0372] Corn plants were infested with first-instar Bt-resistant fall armyworm (FAW) larvae at five larvae per plant (infested using 1.5 microcentrifuge tubes). The corn plants were treated with a spray formulation of yeast particles (containing yeast cell wall components) at varying concentrations (100 grams of yeast per hectare (gAI / ha), 500 gAI / ha, or 1000 gAI / ha). Yeast Particle 1 in Figure 7 was the same as described as Yeast Particle 1 in Example 9. Yeast Particle 2 in Figure 7 was the same as described as Yeast Particle 3 in Example 9. A single treatment was sprayed on the first selection of plants; two treatments were sprayed on the second selection of plants. Yeast particles were applied to the leaves of the corn plants using a CO2 backpack sprayer at a rate of 50 gallons per acre. The carrier for the yeast particles was water. After 14 days, data was collected on the plants to determine the rating scale (Figure 7).

[0373] Addition of yeast particles to transgenic corn plants (also called "transformed" plants) caused a reduction in insect populations (i.e., increased mortality of fall armyworm) compared to transgenic plants that were not treated with yeast particles. This technical effect was observed for plants that received a single treatment or two treatments. These data indicate that yeast particles enhance the efficacy of the Bt Cry1Ab protein (a biopesticide expressed by transgenic plants) even when used to control Bacillus thuringiensis (Bt)-resistant insects, and indicate that the use of the yeast particle methods and compositions of the present invention can be used to control insect populations that have developed resistance to Bt transgenic plants or crops.

[0374] Example 11. Delivery of yeast particles and chlorantraniliprole to transgenic plants expressing Bacillus thuringiensis (Bt) crystal proteins Experiments were conducted to determine damage to transgenic corn plants expressing the Bacillus thuringiensis (Bt) Cry1Ab protein (crystal protein) following infestation by fall armyworm resistant to the Bt Cry1Ab protein (crystal protein) and treatment with yeast particles (containing yeast cell wall components). Certain plants were further treated with Coragen® (a formulation of chlorantraniliprole).

[0375] Corn plants were infested with first-instar Bt-resistant fall armyworm (FAW) larvae at five larvae per plant (infested using 1.5 microcentrifuge tubes). The corn plants were treated with a spray formulation of: (1) yeast particles (containing yeast cell wall components) at varying concentrations (100 grams of active ingredient per hectare (gAI / ha), 500 gAI / ha, or 1000 gAI / ha); (2) Coragen® alone (1 oz / acre or 5 oz / acre); or (3) Coragen® (1 oz / acre) and yeast particles. A single treatment was sprayed on a first selection of plants; two treatments (the second treatment was sprayed 7 days after the first) were sprayed on a second selection of plants. The yeast particles were applied to the leaves of the corn plants using a CO2 backpack sprayer at a rate of 50 gallons per acre. The carrier for the yeast particles was water. After 14 days, data were collected on the plants to determine the rating scale (Figure 8).

[0376] The addition of yeast particles to transgenic corn plants (also called "transformed" plants) caused a reduction in insect populations (i.e., increased mortality of fall armyworms) compared to plants that were not treated with yeast particles. This technical effect was observed for plants that received a single treatment or two treatments. Delivery of yeast particles and Coragen® (chlorantraniliprole) provided complete control of the pest population, with all insects dead at the end of the experiment. Notably, the addition of yeast particles to this composition (containing 1 oz / ac Coragen®) was as effective as delivery of five times more Coragen® (5 oz / ac) and significantly better than Coragen® alone (i.e., no yeast particles).

[0377] These data indicate that yeast particles enhance the efficacy of chemical pesticides such as chlorantraniliprole for controlling insects.

[0378] Example 12. Delivery of yeast particles and chlorantraniliprole to transgenic plants expressing Bacillus thuringiensis (Bt) crystal proteins Plants used in this assay type expressed the Cry1Ab Bt trait. At the time of the experiment, plants were in late V2 or early V3 stage, and two to three of the lowest leaves had visible collars. A randomized complete block experimental design with eight replications was used. A single preventative application of experimental treatment was applied using a handheld CO2 pressurized sprayer fitted with an Even Flat Fan Nozzle 8002 at a volume of 50 gallons per acre and a pressure of 46 PSI.

[0379] Once dry, treated plants were infested with two first-instar Bt-resistant armyworm larvae (Spodoptera frugiperda) each. Feeding damage was assessed at 7, 11, and 13 days after application (DAA) according to a widely used categorical rating scale from Davis et al. (1992). Insects were counted at 13 DAA. Defoliation data were analyzed in JMP® (Version 17.0.0. SAS Institute Inc., Cary, NC, USA, 1989–2023) using the Kruskall-Wallis test for nonparametric comparisons, followed by pairwise comparisons using the Wilcoxon method. General linear analysis was performed on insect counts by performing ANOVA with Student's t-test for multiple comparisons to assess treatment means.

[0380] High rates (5 oz / ac, labeled for field use) and low rates (1 oz / ac) of Coragen® (chlorantraniliprole) were tested along with a low rate of 1000 grams of active ingredient per hectare (gAI / ha) yeast granules.

[0381] As shown in Figure 9, the results demonstrate that yeast particles (containing yeast cell wall components) had a statistically significant positive impact on the efficacy of a chemical pesticide (Coragen®). The results show that the combination of yeast particles with a low application rate of Coragen® provides similar efficacy as a five-fold higher Coragen® application rate.

[0382] Example 13. Delivery of yeast particles and chlorantraniliprole to soybean plants Plants used in this assay type were 17-day-old soybeans. A randomized complete block experimental design was used with eight replications. A single preventative application of experimental treatment was applied using a handheld CO2 pressurized sprayer fitted with an Even Flat Fan Nozzle 8002 at a volume of 50 gallons per acre and a pressure of 46 PSI.

[0383] Treated plants were placed into 16-inch wide x 16-inch deep x 24-inch high cages with zipper closures, two plants per cage. The plants were then infested with five first-instar soybean looper larvae (Chrysodeixis includens) per plant (10 larvae per cage).

[0384] Defoliation was assessed visually using a graphical scale on days 6-7 and 11-12 after treatment. Each leaflet was assessed individually, and total plant defoliation values ​​were averaged for the two plants in each cage. Defoliation values ​​were reported as cage means. On days 11-12, insect numbers per cage were assessed. Defoliation and insect data were analyzed in JMP® (Version 17.0.0. SAS Institute Inc., Cary, NC, 1989-2023). General linear analysis was performed by ANOVA with Student's t-test for multiple comparisons to assess treatment means.

[0385] A high rate (5 oz / ac) and three low rates of Coragen® (chlorantraniliprole) were tested, with 1000 g Ai / ha of yeast particles (containing yeast cell wall components) added to the low rate.

[0386] As shown in Figure 10, the results demonstrate that yeast particles (containing yeast cell wall components) had a statistically significant positive impact on the efficacy of Coragen® at all application rates tested. The results also indicate that adding yeast particles to Coragen® not only reduced insect feeding, but actually contributed to insect mortality in the test. The combination of yeast particles with a low application rate of Coragen® provided efficacy similar to a five-fold higher application rate of Coragen®. These results demonstrate the increased efficacy against chlorantraniliprole provided by yeast particles across two different insect species and crops.

[0387] Example 14. Delivery of yeast particles and novaluron to soybean plants Plants used in this assay type were 17-day-old soybeans. A randomized complete block experimental design was used with eight replications. A single preventative application of experimental treatment was applied using a handheld CO2 pressurized sprayer fitted with an Even Flat Fan Nozzle 8002 at a volume of 50 gallons per acre and a pressure of 46 PSI.

[0388] Treated plants were placed into 16-inch wide x 16-inch deep x 24-inch high cages with zipper closures, two plants per cage. The plants were then infested with five first-instar soybean looper larvae (Chrysodeixis includens) per plant (10 larvae per cage).

[0389] Defoliation was assessed visually using a graphical scale on days 6-7 and 11-12 after treatment. Each leaflet was assessed individually, and total plant defoliation values ​​were averaged for the two plants in each cage. Defoliation values ​​were reported as cage means. On days 11-12, insect numbers per cage were assessed. Defoliation and insect data were analyzed in JMP® (Version 17.0.0. SAS Institute Inc., Cary, NC, 1989-2023). General linear analysis was performed by ANOVA with Student's t-test for multiple comparisons to assess treatment means.

[0390] Yeast granules (containing yeast cell wall components) at 1000 grams of active ingredient per hectare (gAI / ha) were tested in combination with two rates of the insect growth regulator Rimon (Novaluron). As shown in Figure 11, the results demonstrate a statistically significant increase in efficacy over Rimon when yeast granules were added at both Rimon rates. Efficacy is demonstrated through both reduced defoliation and increased insect mortality. These results demonstrate that yeast granules enhance the activity of insecticide chemicals with widely differing modes of action.

[0391] Example 15. Delivery of yeast particles and spinosad to soybean plants Plants used in this assay type were 17-day-old soybeans. A randomized complete block experimental design was used with eight replications. A single preventative application of experimental treatment was applied using a handheld CO2 pressurized sprayer fitted with an Even Flat Fan Nozzle 8002 at a volume of 50 gallons per acre and a pressure of 46 PSI.

[0392] Treated plants were placed into 16-inch wide x 16-inch deep x 24-inch high cages with zipper closures, two plants per cage. The plants were then infested with five first-instar soybean looper larvae (Chrysodeixis includens) per plant (10 larvae per cage).

[0393] Defoliation was assessed visually using a graphical scale on days 6-7 and 11-12 after treatment. Each leaflet was assessed individually, and total plant defoliation values ​​were averaged for the two plants in each cage. Defoliation values ​​were reported as cage means. On days 11-12, insect numbers per cage were assessed. Defoliation and insect data were analyzed in JMP® (Version 17.0.0. SAS Institute Inc., Cary, NC, 1989-2023). General linear analysis was performed by ANOVA with Student's t-test for multiple comparisons to assess treatment means.

[0394] Yeast particles (containing yeast cell wall components) at 1000 g Ai / ha were tested in combination with the spinosyn Entrust (spinosad) at two application rates. As shown in Figure 12, the results demonstrate a statistically significant increase in efficacy over Entrust when yeast particles were added at both Entrust application rates. Efficacy is demonstrated through both reduced defoliation and increased insect mortality. These results indicate that yeast particles enhance the activity of insecticide chemicals with widely differing modes of action.

[0395] Example 16. Delivery of yeast particles and diflubenzuron to cabbage plants Greenhouse trials established that the addition of yeast particles containing yeast cell wall components to Dimilin brand pesticide (active ingredient: diflubenzuron) resulted in less plant damage compared to Dimilin alone at various concentrations. The addition of yeast particles provided a stronger improvement than diflubenzuron, even when the pesticide dose was reduced by 75%.

[0396] To conduct these greenhouse trials, cabbages were transplanted from the seedbed into pots in the greenhouse and grown for approximately 2-4 weeks after transplanting. Four replicates of one plant per pot were used. On day -1, each plant was artificially infested with 15 Plutella xylostella (diamondback moth) larvae (L1-L2) using a paintbrush. Two days after infestation (i.e., day 0), plant test material was applied to the leaves using an air-compressed boom sprayer. Plants were observed and scored for % damage on days 0, 2, 6, 9, and 13.

[0397] Eight test groups were utilized: (1) untreated control; (2) Dimlin 16 grams active ingredient per hectare (gAI / ha); (3) Dimlin 8 gAI / ha; (4) Dimlin 4 gAI / ha; (5) Dimlin 8 gAI / ha + yeast particles 100 gAI / ha; (6) Dimlin 8 gAL / ha + yeast particles 500 gAI / ha; (7) Dimlin 8 gAL / ha + yeast particles 1000 gAI / ha; and (8)(5) Dimlin 4 gAL / ha + yeast particles 500 gAI / ha.

[0398] Results across all data collection points are shown in Figure 13A. Observations at day 9 are shown in Figure 13B. As shown, yeast particles improved the efficacy of Dimlin. All test samples with yeast particles outperformed the highest concentration tested, 16 g AL / ha Dimlin, without yeast particles. For example, even when the concentration of Dimlin was reduced from 16 g AL / ha to 4 g / AI / ha, the addition of yeast particles resulted in an improvement in pest control.

[0399] Example 17. Delivery of yeast particles and cypermethrin to cauliflower plants Greenhouse trials established that the addition of yeast particles containing yeast cell wall components to Upala brand pesticide (active ingredient cypermethrin) resulted in less plant damage compared to Upala alone at various concentrations. The addition of yeast particles provided a stronger improvement than cypermethrin alone, with similar results obtained for Upala at a concentration 75% of the concentration of Upala required without the yeast cell wall particles.

[0400] The test protocol here corresponds to that of Example 16 above, except that cauliflower plants were used. The results are shown in Figure 14. The most potent control was provided by Upala at 0.15 liters per hectare (L / ha), but a composition of Upala at one-quarter that rate (0.0375 L / ha) plus yeast particles at 500 g AI / ha provided similar results. In general, each test composition of Upala with yeast cell wall particles was superior to Upala alone at the same application rate.

[0401] Example 18. Yeast particles and delivery of Bt pesticides Greenhouse trials established that the addition of yeast particles containing yeast cell wall components to Leprotec brand biopesticide (Bt kurstaki strain EVB-113-19) resulted in less plant damage compared to Leprotec alone at various concentrations. The addition of yeast particles provided a stronger improvement than Leprotec, resulting in similar results at a concentration of Leprotec that was 75% of the concentration of Leprotec required without the yeast cell wall particles.

[0402] The test protocol here corresponds to that of Example 16 above, except that cauliflower plants were used. The results are shown in Figure 15. As an example, Leprotec at 1 g AI / ha with 500 g AI / ha yeast cell wall particles (Item 11) performed as well as Leprotec at 4 g AI / ha without yeast cell wall particles (Item 8). Other yeast cell wall particle compositions containing Leprotec at 1 g AI / ha (Item 14), 2 g AI / ha (Item 15), and 4 g AI / ha (Item 16) all outperformed Leprotec at 4 g AI / ha alone.

[0403] Example 19. Delivery of yeast particles and acetamiprid to soybean plants Yeast particles containing yeast cell wall components (Biocapsule PF-I-011) at 1000 g Ai / ha were tested in combination with three concentrations of the neonicotinoid insecticide ArVida (acetamiprid), which are equivalent to field-relevant concentrations of neonicotinoids.

[0404] The results demonstrate a statistically significant increase in efficacy against ArVida with the addition of our formulation at the two ArVida application rates tested. For this chemical, efficacy was again demonstrated through both reduced defoliation and increased insect mortality (Figure 16). These results confirm that our formulation enhances efficacy in widely used insecticide chemicals with different modes of action.

[0405] Example 20. Delivery of biopesticides to leaves of cabbage plants In this example, the ability of polysaccharides and yeast particles (containing yeast cell wall components) to enhance the efficacy of a biopesticide was evaluated. The biopesticide tested was DIPEL® (Bacillus thuringiensis subsp. kurstaki - Bacillus thuringiensis microorganism).

[0406] Leaves of 3-week-old Chinese cabbage plants (N=40) were sprayed (using an airbrush) with the composition applied at a concentration of 0.15 mL and then infested with four neonate diamondback moth larvae. Insect mortality and leaf disc consumption were observed 5, 7, and 11 days after insect infestation.

[0407] The compositions tested were: (1) DIPEL® Low (LC20 sensitivity); (2) DIPEL® High (LC50 sensitivity); (3) Laminarin; (4) Laminarin + DIPEL® Low; (5) Laminarin + DIPEL® High; (6) β-1,3-Glucan; (7) β-1,3-Glucan + DIPEL® Low; (8) β-1,3-Glucan + DIPEL® High; (9) Yeast particles containing yeast cell wall components; (10) Yeast particles containing yeast cell wall components + DIPEL® Low; and (11) Yeast particles containing yeast cell wall components + DIPEL® High. Control experiments involved the delivery of water alone. DIPEL® Low was 327.8 ng / mL; DIPEL® High was 1545 ng / mL. The laminarin was derived from Laminaria digitata; the β-1,3-glucan was derived from Euglena gracilis; and the yeast particles were yeast glucan particles derived from Saccharomyces cerevisiae.

[0408] As shown in Figure 17, the addition of laminarin or β-1,3-glucan provided an increase in mortality when delivering Bacillus thuringiensis microorganisms in a composition at the low concentrations of DIPEL® tested. Yeast cell wall particles provided an enhancement compared to both concentrations of DIPEL®. This increase in mortality is most pronounced at day 11. These results demonstrate that yeast particles, including yeast cell wall particles, enhance the efficacy of biopesticides; and that polysaccharides (laminarin and β-1,3-glucan) enhance the efficacy of biopesticides.

[0409] Example 21. Delivery of yeast particles and Bt biopesticides to control Bt-resistant diamondback moth Greenhouse trials demonstrated that application of insecticidal dsRNA encapsulated in yeast particles containing yeast cell wall components after application of a sprayable Bt pesticide provided significantly improved efficacy against Dipel-resistant diamondback moth at both label and sublethal doses of Bt pesticide. Application of yeast cell wall-encapsulated insecticidal dsRNA provided similar efficacy after sublethal doses of Bt pesticide compared to application of Bt pesticide at label doses without subsequent application of yeast cell wall or insecticidal dsRNA. Combination treatments provided significantly improved efficacy at label doses of Bt pesticide. These results demonstrate that the use of dsRNA encapsulated in yeast cell walls following application of a Bt biopesticide can help users obtain either greater efficacy at the same application rate or similar efficacy at a reduced use rate of the Bt biopesticide, with the potential benefit of delaying the development of Bt resistance in lepidopteran populations.

[0410] A total of nine conditions, including an untreated control and eight different treatments, were run with eight replicates: (1) untreated control; (2) a sublethal reduced dose of Dipel (based on a separate dose-ranging study conducted to determine this sublethal dose); (3) a label dose of commercially available Dipel brand Bt biopesticide; (4) encapsulated control dsRNA (GS4); (5) a sublethal dose of Dipel followed by encapsulated GS4 control dsRNA; (6) a label dose of Dipel followed by encapsulated GS4 control dsRNA; (7) encapsulated insecticidal dsRNA GS6370 (targeting the diamondback moth Plutella xylostella IAP gene); (8) a sublethal dose of Dipel followed by encapsulated GS6370; and (9) a sublethal dose of Dipel followed by encapsulated GS6370.

[0411] Encapsulated dsRNA was prepared according to CCA using SHMP as a nuclease inhibitor and branched PEI as a capture agent at a weight ratio of 0.75 PEI:1 SHMP:1 dsRNA.

[0412] Ten-day-old Chinese cabbage plants were sprayed with the relevant composition on day 0, first treated with Dipel or water (if applicable), followed by encapsulated dsRNA or water on day 3, if applicable. After the initial application, the plants were artificially infested with a well-characterized laboratory colony of commercially available Dipel-resistant diamondback moth (Plutella xylostella) at the neonate larval stage. dsRNA for this test was applied curatively at a rate of 500 g of active ingredient per hectare three days after the initial application. Approximately 10 insects per plant were used for infestation. Defoliation was measured on day 11 based on the percent remaining leaves.

[0413] The results are shown in Figure 18. As shown, yeast cell wall-encapsulated GS6370 applied after Dipel at the sublethal EC50 provided statistically the same amount of defoliation compared to Dipel alone at the label dose, while the combination of Dipel treatment at the label dose followed by yeast cell wall-encapsulated GS6370 treatment provided significantly less defoliation than any other combination.

[0414] Example 22. Confirmation of gene knockdown from encapsulated pesticidal polynucleotides To collect insects for targeted gene knockdown analysis, the greenhouse assay protocol described above was replicated using two treatments of interest: (1) a labeling dose of Dipel plus encapsulated GS4 and (2) a labeling dose of Dipel plus encapsulated GS6370. Nine replicates were collected per treatment, and within each replicate, two Dipel-resistant diamondback moth larvae were collected 6 days after infestation and 3 days after encapsulated dsRNA application. IAP expression levels were significantly lower in insects exposed to Dipel plus encapsulated GS6370 (p-value 0.1, Student's t-test). IAP was knocked down by 38% in isoform 1 (p-value 0.098) and 27% in isoform 2 (p-value 0.086).

[0415] Example 23. Encapsulated dsRNA performed similarly to biological and chemical standard treatments Field trials confirmed that encapsulated GS6370 dsRNA performed statistically similarly to the labeled application rate of the biological standard, the Bt pesticide XenTari. Encapsulated dsRNA was prepared as described in Example YY above. Eight replicates were run for five groups: (1) untreated control; (2) chemical standard Coragen (active ingredient chlorantraniliprole); (3) encapsulated GS6370 dsRNA (targeting an IAP gene in DBM); (4) Xentari biological standard; and (5) encapsulated GS4 control dsRNA. Collards were sprayed with the test material on days 0, 7, and 14, and damage per plant and number of DBM larvae per plant were assessed on days 3, 7, 14, 21, and 28. The results are shown in Figure 19. At 28 days, damage and larvae per plant were statistically identical for the biological standard, chemical standard, and encapsulated GS6370 dsRNA, and all were significantly improved over the untreated and dsRNA controls.

[0416] Additional Aspects Additional aspects of the present disclosure are incorporated by the following numbered paragraphs:

[0417] 1.i) Biopesticides, and ii) Biological adjuvants A composition comprising: wherein the biological adjuvant is selected from the group consisting of yeast particles (YP) and polysaccharides derived from a source other than yeast; Optionally, wherein the YP is selected from the group consisting of intact yeast cell wall particles of Saccharomyces cerevisiae (baker's yeast), extracted yeast cell wall particles (YCWP), yeast cell particles (YCP), yeast glucan particles (GP or YGP), yeast glucan mannan particles (GMP or YGMP), yeast glucan chitin particles (GCP or YGCP), yeast glucan chitin mannan particles (GCMP or YGCMP), yeast glucan lipid particles (GLP or YGLP), and whole glucan particles (WGP); or Optionally, wherein the polysaccharide derived from a source other than yeast is selected from the group consisting of beta-glucan, mannan oligosaccharide, and laminarin; and A composition wherein the biological adjuvant enhances the efficacy of the biopesticide.

[0418] 2. The composition of paragraph 1, wherein the biopesticide comprises a naturally occurring microorganism.

[0419] 3. The composition of paragraph 2, wherein the naturally occurring microorganism produces one or more proteins that target plant pests.

[0420] 4. The composition of paragraph 3, wherein the plant pest is a fungus.

[0421] 5. The composition of paragraph 3, wherein the plant pest is an insect.

[0422] 6. The composition of any one of paragraphs 3 to 5, wherein one or more proteins produced by a naturally occurring microorganism are activated in the insect gut.

[0423] 7. The composition of any one of paragraphs 3 to 6, wherein the one or more proteins produced by naturally occurring microorganisms are selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth.

[0424] 8. The composition of paragraph 7, wherein the one or more proteins comprise crystal proteins, optionally wherein the crystal proteins belong to the class of Cry proteins.

[0425] 9. The composition of any one of paragraphs 2 to 8, wherein the naturally occurring microorganism is Bacillus thuringiensis.

[0426] 10. The composition of paragraph 1, wherein the biopesticide comprises a commercially available bioinsecticide prepared using Bacillus thuringiensis, wherein the insecticide targets plant pests.

[0427] 11. The composition of paragraph 10, wherein the commercially available bioinsecticide is selected from the group consisting of DIPEL, XENTARI, BACTOSPEINE ES, BIOBIT HP, FLORBAC, COSTAR WG, JAVELIN WP, THURICIDE, TEKAR, BACTIMOS, VECTOLEX, NOVODOR, TRIDENT, LEPROTEC, and any combination thereof.

[0428] 12. The composition of paragraph 1, wherein the biopesticide comprises a recombinant microorganism.

[0429] 13. The composition of paragraph 12, wherein the recombinant microorganism produces one or more recombinant proteins that target plant pests.

[0430] 14. The composition of paragraph 13, wherein the one or more recombinant proteins are activated in the insect gut.

[0431] 15. The composition of paragraph 13 or 14, wherein the one or more recombinant proteins are selected from the group consisting of crystal proteins and vegetatively produced insecticidal proteins.

[0432] 16. The composition of paragraph 15, wherein the recombinant protein comprises a crystal protein, optionally wherein the crystal protein belongs to the class of Cry proteins.

[0433] 17. The composition of any one of paragraphs 12 to 16, wherein the recombinant microorganism is Bacillus thuringiensis.

[0434] 18. The composition of paragraph 1, wherein the biopesticide comprises isolated cellular components derived from a naturally occurring microorganism.

[0435] 19. The composition of paragraph 1, wherein the biopesticide comprises an isolated recombinant cell component derived from a recombinant microorganism.

[0436] 20. The composition of paragraph 18 or 19, wherein the cellular component is a protein that targets a plant pest.

[0437] 21. The composition according to paragraph 20, wherein the plant pest is a fungus.

[0438] 22. The composition according to paragraph 20, wherein the plant pest is an insect.

[0439] 23. The composition of paragraph 22, wherein the protein is activated in the insect gut.

[0440] 24. The composition of any one of paragraphs 20 to 23, wherein the protein is selected from the group consisting of a crystal protein and an insecticidal protein produced during vegetative growth.

[0441] 25. The composition of paragraph 24, wherein the protein is a crystal protein, optionally wherein the crystal protein belongs to the class of Cry proteins.

[0442] 26. The composition of paragraph 1, wherein the biopesticide comprises a Bt protein.

[0443] 27. The composition of paragraph 26, wherein the Bt protein is derived from a naturally occurring microorganism.

[0444] 28. The composition of paragraph 26, wherein the biopesticide is derived from a recombinant microorganism.

[0445] 29. The composition of any one of paragraphs 26 to 28, wherein the Bt protein targets a plant pest.

[0446] 30. The composition according to paragraph 29, wherein the plant pest is a fungus.

[0447] 31. The composition according to paragraph 29, wherein the plant pest is an insect.

[0448] 32. The composition of paragraph 31, wherein the protein is activated in the insect gut.

[0449] 33. The composition of any one of paragraphs 26 to 32, wherein the Bt protein is selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth.

[0450] 34. The composition of paragraph 33, wherein the Bt protein is a crystal protein, optionally wherein the crystal protein belongs to the class of Cry proteins.

[0451] 35. The composition of any one of paragraphs 1 to 34, wherein the biological adjuvant comprises yeast particles (YP).

[0452] 36. The composition of any one of paragraphs 1-35, wherein the biological adjuvant comprises a polysaccharide derived from a source other than yeast, optionally wherein the source other than yeast is a bacterium, a fungus, an algae, a lichen, or a plant, and optionally wherein the polysaccharide is selected from the group consisting of beta-glucan, mannan oligosaccharide, and laminarin.

[0453] 37. The composition of any one of paragraphs 1 to 36, wherein the biological adjuvant comprises a glucan molecule selected from the group consisting of long n=2-15 β1,3 oligosaccharide branched glucans and short β1,3,6 branched glucans.

[0454] 38. The composition of any one of paragraphs 35 to 37, wherein the YP comprises one or more selected from the group consisting of extracted yeast cell wall particles (YCWP), yeast glucan particles (YGP), yeast glucan mannan particles (YGMP), yeast chitin particles (YCP), yeast glucan chitin particles (YGCP), yeast glucan lipid particles (YGLP), or whole glucan particles (WGP).

[0455] 39. The composition of any one of paragraphs 35 or 38, wherein the YP is commercially available YP.

[0456] 40. The composition of any one of paragraphs 35 to 39, wherein the composition targets plant pests.

[0457] 41. The composition according to paragraph 40, wherein the plant pest is a fungus that is resistant to a biological pesticide.

[0458] 42. The composition of any one of paragraphs 1-41, wherein the composition is effective in controlling a plant pest, and wherein the plant pest is an insect pest.

[0459] 43. The composition according to paragraph 42, wherein the plant insect pest is a fungus that is resistant to a biological pesticide.

[0460] 44. The composition according to paragraph 42 or 43, wherein the plant insect pest belongs to the order Lepidoptera or Coleoptera.

[0461] 45. The composition of paragraph 44, wherein the plant insect pest is resistant to Bacillus thuringiensis (Bt) or a Bt protein.

[0462] 46. ​​The composition of paragraph 44, wherein the lepidopteran insect is resistant to Bacillus thuringiensis (Bt) or a Bt protein.

[0463] 47. Lepidoptera include Nymphalidae (Nymphalians), Danaidae (Monarchs), Pieridae (Pieridae and Eurema), Papilionidae (Swallowtails), Lycaenidae (Lycaenidae), Hesperiidae (Hesperiids), Tineidae (Moths), Sesiidae (Clearwig moths), and Pyralidae (Pyralids). 47. The composition of paragraph 44 or 46, wherein the moth is a species of a family selected from the group consisting of: Acanthidae (pyralid moths), Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawk moths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (inchworms, noctuids, and underwing moths), and Plutellidae (diamond moths).

[0464] 48. The composition of paragraph 47, wherein the lepidopteran insect species is a Plutella species.

[0465] 49. The composition of paragraph 48, wherein the Plutella species insect is Plutella xylostella.

[0466] 50. The composition of paragraph 44 or 46, wherein the lepidopteran insect species is a Spodoptera species.

[0467] 51. The composition of paragraph 50, wherein the Spodoptera species insect is Spodoptera frugiperda.

[0468] 52. The composition of paragraph 44, wherein the insect belongs to the order Coleoptera, and wherein the Coleopteran insect is of a species selected from the group consisting of Leptinotarsa ​​spp., Phyllotreta spp., Cerotoma spp., Diabrotica spp., Tribolium spp., Anthonomus spp. and Alticini spp.

[0469] 53. The composition of paragraph 52, wherein the coleopteran insect species is a Leptinotarsa ​​species.

[0470] 54. The composition of paragraph 53, wherein the Leptinotarsa ​​species insect is the Colorado potato beetle.

[0471] 55. The composition of any one of paragraphs 1 to 54, further comprising a pesticidal polynucleotide.

[0472] 56. The composition of paragraph 55, wherein the pesticidal polynucleotide is single-stranded RNA.

[0473] 57. The composition of paragraph 55, wherein the pesticidal polynucleotide is dsRNA.

[0474] 58. The composition of any one of paragraphs 55 to 57, wherein the biological adjuvant comprises YP, and optionally, wherein the pesticidal polynucleotide is encapsulated within the YP.

[0475] 59. The composition of any one of paragraphs 55 to 57, wherein the biological adjuvant comprises a polysaccharide derived from a source other than yeast, optionally wherein the source other than yeast is a bacterium, a fungus, an algae, a lichen, or a plant, and optionally wherein the polysaccharide is selected from the group consisting of beta-glucan, mannan oligosaccharide, and laminarin.

[0476] 60. The composition of any of paragraphs 55 to 57, wherein the biological adjuvant comprises a glucan molecule selected from the group consisting of long n=2-15 β1,3 oligosaccharide branched glucans and short β1,3,6 branched glucans.

[0477] 61. The composition of paragraph 60, wherein the composition further comprises a compound selected from the group consisting of a cationic polymer, a non-cationic polymer, and an organic carrier, optionally wherein the compound is a cationic polymer, optionally wherein the cationic polymer is selected from the group consisting of polyethyleneimine (PEI) or poly-L-lysine (PLL), and optionally wherein the pesticidal polynucleotide is encapsulated within the YP using Complex Core A (CCA).

[0478] 62. The composition of any one of paragraphs 55 to 61, further comprising one or more nuclease inhibitors, optionally wherein one or more nuclease inhibitors is a chelating agent selected from the group consisting of EDTA, SHMP, PVSA, and TTP, and optionally wherein one or more nuclease inhibitors comprises EDTA.

[0479] 63. The composition according to any one of paragraphs 55 to 62, wherein the pesticidal polynucleotide inhibits the expression of one or more target genes of a plant pest.

[0480] 64. The composition of any one of paragraphs 55 to 61, wherein the pesticidal polynucleotide comprises a strand that is identical to or complementary to a region of messenger RNA (mRNA) encoded by one or more target genes.

[0481] 65. The composition of paragraph 64, wherein the pesticidal polynucleotide is a dsRNA further comprising a second strand complementary to the first strand.

[0482] 66. The composition of paragraph 64 or 65, wherein the target gene comprises one or more genes selected from the group consisting of proteasome beta 5 (PSMB5) gene, vacuolar ATPase E (vATPase E) gene, calmodulin gene, inhibitor of apoptosis protein (IAP) gene, soluble NSF attachment protein (a-SNAP) gene, Ras opposite (ROP) gene, proteasome alpha type 2 (PTSA2) gene, secretion-associated Ras-associated GTPase 1 (SAR1) gene, PBAN, ATPase, and CP4S3_DROME, C12C1_DROME.

[0483] 67. A composition for controlling plant pests, comprising yeast particles (YP) and a pesticidal polynucleotide, wherein the YP enhances the efficacy of the polynucleotide, and optionally, wherein the polynucleotide is encapsulated within the YP.

[0484] 68. The composition of paragraph 67, wherein the pesticidal polynucleotide is single-stranded RNA.

[0485] 69. The composition of paragraph 67, wherein the pesticidal polynucleotide is dsRNA.

[0486] 70. The composition of any one of paragraphs 67 to 69, wherein the pesticidal polynucleotide inhibits the expression of one or more target genes of a plant pest.

[0487] 71. The composition of any one of paragraphs 67 to 70, wherein the pesticidal polynucleotide comprises a strand that is identical to or complementary to a region of messenger RNA (mRNA) encoded by one or more target genes.

[0488] 72. The composition of paragraph 71, wherein the pesticidal polynucleotide is a dsRNA further comprising a second strand complementary to the first strand.

[0489] 73. The composition of paragraphs 70 to 72, wherein the target gene comprises one or more genes selected from the group consisting of proteasome beta 5 (PSMB5) gene, vacuolar ATPase E (vATPase E) gene, calmodulin gene, inhibitor of apoptosis protein (IAP) gene, soluble NSF attachment protein (a-SNAP) gene, Ras opposite (ROP) gene, proteasome alpha type 2 (PTSA2) gene, secretion-associated Ras-associated GTPase 1 (SAR1) gene, PBAN, ATPase, CP4S3_DROME, C12C1_DROME, and any combination thereof.

[0490] 74. The composition of any one of paragraphs 67 to 73, further comprising a biopesticide.

[0491] 75. The composition of paragraph 74, wherein the biopesticide comprises a naturally occurring microorganism.

[0492] 76. The composition of paragraph 75, wherein the naturally occurring microorganism produces one or more proteins that target plant pests.

[0493] 77. The composition of paragraph 76, wherein one or more proteins produced by naturally occurring microorganisms are activated in the insect gut.

[0494] 78. The composition of paragraph 76 or 77, wherein the one or more proteins produced by naturally occurring microorganisms are selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth.

[0495] 79. The composition of paragraph 78, wherein the one or more proteins comprise crystal proteins, optionally wherein the crystal proteins belong to the class of Cry proteins.

[0496] 80. The composition of any one of paragraphs 75 to 79, wherein the naturally occurring microorganism is Bacillus thuringiensis.

[0497] 81. The composition of paragraph 74, wherein the biopesticide comprises a commercially available bioinsecticide prepared using Bacillus thuringiensis, and wherein the insecticide targets plant pests.

[0498] 82. The composition of paragraph 81, wherein the commercially available bioinsecticide is selected from the group consisting of DIPELO, XENTARIO, BACTOSPEINE ESO, BIOBIT HPO, FLORBACO, COSTAR WGO, JAVELIN WPO, THURICIDEO, TEKARO, BACTIMOSO, VECTOLEXO, NOVODORO, TRIDENTO, LEPROTECO, and any combination thereof.

[0499] 83. The composition of paragraph 74, wherein the biopesticide compound comprises a recombinant microorganism.

[0500] 84. The composition of paragraph 83, wherein the recombinant microorganism produces one or more recombinant proteins that target plant pests.

[0501] 85. The composition of paragraph 84, wherein the one or more recombinant proteins are activated in the insect gut.

[0502] 86. The composition of paragraph 84 or 85, wherein one or more recombinant proteins are selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth.

[0503] 87. The composition of paragraph 86, wherein one or more recombinant proteins comprise crystal proteins, optionally wherein the crystal proteins belong to the class of Cry proteins.

[0504] 88. The composition of any one of paragraphs 83 to 87, wherein the recombinant microorganism is Bacillus thuringiensis.

[0505] 89. The composition of paragraph 74, wherein the biopesticide comprises isolated cellular components derived from a naturally occurring microorganism.

[0506] 90. The composition of paragraph 74, wherein the biopesticide comprises an isolated recombinant cell component derived from a recombinant microorganism.

[0507] 91. The composition of paragraph 89 or 90, wherein the cellular component is a protein that targets a plant pest.

[0508] 92. The composition of paragraph 91, wherein the protein is activated in the insect gut.

[0509] 93. The composition of paragraph 91 or 92, wherein the protein is selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth.

[0510] 94. The composition of paragraph 93, wherein the protein is a crystal protein, optionally wherein the crystal protein belongs to the class of Cry proteins.

[0511] 95. The composition of paragraph 74, wherein the biopesticide comprises a Bt protein.

[0512] 96. The composition of paragraph 95, wherein the Bt protein is derived from a naturally occurring microorganism.

[0513] 97. The composition of paragraph 95, wherein the biopesticide is derived from a recombinant microorganism.

[0514] 98. The composition of any one of paragraphs 94 to 97, wherein the Bt protein targets a plant pest.

[0515] 99. The composition of paragraph 98, wherein the plant pest is a fungus.

[0516] 100. The composition according to paragraph 98, wherein the plant pest is an insect.

[0517] 101. The composition of paragraph 100, wherein the protein is activated in the insect gut.

[0518] 102. The composition of any one of paragraphs 95-101, wherein the Bt protein is selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth.

[0519] 103. The composition of paragraph 102, wherein the Bt protein is a crystal protein, optionally wherein the crystal protein belongs to the class of Cry proteins.

[0520] 104. The composition of any of paragraphs 1-103, further comprising one or more additional ingredients selected from the group consisting of a carrier agent, a surfactant, an organosilicone, an organosilicone surfactant, a polynucleotide herbicidal molecule, a non-polynucleotide herbicidal molecule, a polynucleotide pesticide, a non-polynucleotide pesticide, a polynucleotide fungicide, a non-polynucleotide fungicide, a polynucleotide insecticide, a non-polynucleotide insecticide, a safener, and a pathogen growth regulator.

[0521] 105. The composition of any of paragraphs 1-104, wherein the composition is formulated for application in a form selected from the group consisting of a sprayable solution, an emulsion, a tank mix, and a powder.

[0522] 106. A method for controlling plant pests, the method comprising delivering to a plant or contacting a plant pest with a composition described in any one of paragraphs 1-105.

[0523] 107. The method of paragraph 106, wherein delivering comprises topically applying to said plant a composition of any one of paragraphs 1 to 105.

[0524] 108. The method of any one of paragraphs 107, wherein topically applying comprises spraying the composition onto the leaves, stems, flowers, or fruits of the plant.

[0525] 109. The method of paragraph 106, wherein delivering to the plant comprises a seed treatment.

[0526] 110. The method of paragraph 106, wherein delivery comprises application to the roots or soil.

[0527] 111. The method of paragraphs 106-110, wherein the plant is selected from the group consisting of Solanaceae plants, Brassicaceae plants, Poaceae plants, Cucurbitaceae plants, Fabaceae plants, Fagaceae plants, Asteraceae plants, Amaryllidaceae plants, Umbelliferae plants, Apiaceae plants, Amranthaceae plants, and Malvaceae plants.

[0528] 112. The method of any one of paragraphs 106-111, wherein the plant pest is selected from the group consisting of insect pests and fungal pathogens.

[0529] 113. The method of paragraph 112, wherein the plant pest is an insect, optionally wherein the insect pest belongs to the order Lepidoptera, Coleoptera, Hemiptera, Diptera, or Acari.

[0530] 114. The method of paragraph 113, wherein the plant pest is an insect that is resistant to Bacillus thuringiensis (Bt) or a Bt protein.

[0531] 115. The plant pest is a lepidopteran insect, optionally wherein the family includes Nymphalidae (nymphalians), Danaidae (monarch butterflies), Pieridae (pierid and ceratopsian butterflies), Papilionidae (swallowtail butterflies), Lycaenidae (lycaenid butterflies), Hesperiidae (skippers), Tineidae (moths), Sesiidae (cleanseed moths), Methoptera 115. The method of paragraph 113 or 114, wherein the species is selected from the group consisting of any species within the family Pyralidae (pyralid moths), Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawkmoths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (inchworms, noctuids, and underwing moths), and Plutellidae (diamond moths).

[0532] 116. The method of any one of paragraphs 113 to 115, wherein the plant pest is a lepidopteran insect species of the genus Plutella.

[0533] 117. The method of paragraph 116, wherein the insect species is Plutella xylostella (diamond moth, DBM).

[0534] 118. The method of paragraph 115, wherein the lepidopteran insect species is a Spodoptera species.

[0535] 119. The method of paragraph 118, wherein the Spodoptera species insect is Spodoptera frugiperda (flea fall armyworm, FAW).

[0536] 120. Lepidoptera insects include the Chrysomelidae (leaf beetles, broad-shouldered beetles, and alligator weed flea beetles), Curculionidae (weevils), Meloidae (blister beetles), Tenebrionidae (meal beetles), Scarabaeidae (scarab beetles), and Cerambycidae (pine beetles). Caterpillars), Curculionidae (Chinese White Pine Beetles), Nitidulidae (Small Hive Beetles), Cerambycidae (Mulberry Longhorn Beetles), Phyllotreta (Flea Beetles), Diabrotica (Corn Rootworms), Chrysomela (Cottonwood Leaf Beetles) , Hypothenemus (coffee berry borer), Sitophilus (may weevils), Epitrix (tobacco flea beetle), E. cucumeris (potato flea beetle), P. pusilla (western black flea beetle); Anthonomus (pepper weevil), Hemicrepidus (wireworm), Melanotus (wireworm), Ceutorhychus (cabbage pod weevil), Aeolus (wireworm), Horistonotus (sandworm), Sphenophorus (corn billbug), S. zea (timothy billbug), S. parvulus (bluegrass billbug), S.114. The method of paragraph 113, wherein the insect is a coleopteran insect of a species in a family selected from the group consisting of: Phylophaga (white grub), Chaetocnema (corn flea beetle), Popillia (bean beetle), Epilachna (pea beetle), Cerotoma (bean leaf beetle), and Epicauta (blister beetle).

[0537] 121. The method of paragraph 120, wherein the coleopteran insect species is a Leptinotarsa ​​species.

[0538] 122. The method of paragraph 121, wherein the Leptinotarsa ​​species insect is a Colorado potato beetle.

[0539] 123. The method of any one of paragraphs 106-122, wherein percent mortality of the plant pest is increased by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the biological adjuvant, and optionally, mortality of the plant pest is increased by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0540] 124. The method of any one of paragraphs 106-122, wherein percent plant part consumption by plant pests is reduced by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the biological adjuvant, and optionally, percent plant part consumption by plant pests is reduced by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0541] 125. The method of any one of paragraphs 106-122, wherein the percent plant part surface area affected by the plant pest is reduced by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the biological adjuvant, and optionally, the percent plant part surface area affected by the plant pest is reduced by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0542] 126. The method of any one of paragraphs 106-125, further comprising delivering to the plant or pest a pesticide having a mode of action different from that of the composition.

[0543] 127. A method for controlling plant pests, the method comprising delivering to a plant a composition comprising a biological adjuvant, and wherein the plant is a transgenic plant engineered to express one or more molecules selected from the group consisting of biopesticides and insecticidal polynucleotides.

[0544] 128. The method of paragraph 127, wherein the biological adjuvant is YP, optionally wherein the YP comprises one or more particles selected from the group consisting of (YCWP), yeast glucan particles (YGP), yeast glucan mannan particles (YGMP), yeast chitin particles (YCP), yeast glucan chitin particles (YGCP), yeast glucan lipid particles (YGLP), or whole glucan particles (WGP).

[0545] 129. The method of paragraph 127, wherein the biological adjuvant comprises a polysaccharide derived from a source other than yeast, optionally wherein the source other than yeast is a bacterium, a fungus, an algae, a lichen, or a plant, and optionally wherein the polysaccharide is selected from the group consisting of beta-glucan, mannan oligosaccharide, and laminarin.

[0546] 130. The method of paragraph 127, wherein the biological adjuvant comprises a glucan molecule selected from the group consisting of long n=2-15 β1,3 oligosaccharide branched glucans and short β1,3,6 branched glucans.

[0547] 131. The method of any one of paragraphs 127-130, wherein the one or more molecules expressed by the transgenic plant comprise a biopesticide, optionally wherein the biopesticide is a Bt toxin, and optionally wherein the Bt toxin is selected from the group consisting of crystal protein and vip3.

[0548] 132. The method of any one of paragraphs 127-131, wherein the plant pest is resistant to a biological pesticide.

[0549] 133. The method of any one of paragraphs 127-131, wherein the plant pest is resistant to the pesticidal polynucleotide.

[0550] 134. The method of any one of paragraphs 127 to 133, wherein the plant pest is an insect of the order Lepidoptera or Coleoptera.

[0551] 135. The method of paragraph 134, wherein the plant pest is an insect of the order Lepidoptera.

[0552] 136. The method of paragraph 135, wherein the insect is selected from the group consisting of P. xylostella and S. frugiperda.

[0553] 137. The method of any one of paragraphs 127 to 134, wherein the plant pest is an insect of the order Coleoptera.

[0554] 138. The method of any one of paragraphs 127 to 137, wherein the plant is selected from the group consisting of Solanaceae plants, Brassicaceae plants, Poaceae plants, Cucurbitaceae plants, Fabaceae plants, Apiaceae plants, Amranthaceae plants, Malvaceae plants, Fagaceae plants, Asteraceae plants, Amaryllidaceae plants, and Umbelliferae plants.

[0555] 139. The method of any one of paragraphs 127-138, wherein percent mortality of the plant pest increases by at least 10% after delivery of the composition compared to a control comprising the plant under untreated conditions, and optionally, percent mortality increases by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0556] 140. The method of any one of paragraphs 127-138, wherein the percent plant part consumption by plant pests is reduced by at least 10% after delivery of the composition compared to a control comprising the plant under untreated conditions, and optionally, the percent plant part consumption by plant pests is reduced by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0557] 141. The method of any one of paragraphs 127-138, wherein the percent plant parts affected by the plant pest is reduced by at least 10% after delivery of the composition compared to a control comprising the plant under untreated conditions, and optionally, the percent plant parts affected by the plant pest is reduced by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

[0558] 142.i) One or more Bacillus thuringiensis (Bt) insecticide products, and ii) Empty yeast particles (YP) A composition comprising:

[0559] wherein the composition causes at least about 10% greater mortality in plant-infesting insect pests compared to the Bt insecticide product alone;

[0560] wherein the plant is selected from the group consisting of Solanaceae plants, Brassicaceae plants, Poaceae plants, Cucurbitaceae plants, Fabaceae plants, Apiaceae plants, Amranthaceae plants, Malvaceae plants, Fagaceae plants, Asteraceae plants, Amaryllidaceae plants, and Umbelliferae plants. Selected, optionally, wherein the composition causes at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100%, or at least about 150%, or at least about 200%, or at least about 250%, or at least about 300%, or at least about 350%, or at least about 400% more mortality in plant-infesting insect pests compared to the Bt insecticide product alone.

[0561] 143. The composition of paragraph 142, wherein the Bt insecticide comprises a commercially available Bt insecticide, optionally wherein the commercially available Bt insecticide is selected from the group consisting of DIPELO, XENTARIO, BACTOSPEINE ESO, BIOBIT HPO, FLORBACO, COSTAR WGO, JAVELIN WPO, THURICIDEO, TEKARO, BACTIMOSO, VECTOLEXO, NOVODORO, TRIDENTO, LEPROTECO, and any combination thereof.

[0562] 144. The composition of paragraph 142 or 143, wherein the YP comprises extracted yeast cell wall particles (YCWP), yeast glucan particles (YGP), yeast glucan mannan particles (YGMP), yeast chitin particles (YCP), yeast glucan chitin particles (YGCP), yeast glucan lipid particles (YGLP), or whole glucan particles (WGP).

[0563] 145. The composition of any one of paragraphs 142 to 144, wherein the insect is a lepidopteran or coleopteran insect.

[0564] 146. The composition of paragraph 145, wherein the insect is resistant to Bacillus thuringiensis (Bt) or a Bt protein.

[0565] 147. Insects include Nymphalidae (Nymphalians), Danaidae (Monarchs), Pieridae (Pieridae and Eurema butterflies), Papilionidae (Swallowtails), Lycaenidae (Lycaenidae), Hesperiidae (Hesperiids), Tineidae (Moths), Sesiidae (Clearwig moths), and Pyralidae. 147. The composition of paragraph 145 or 146, wherein the insect is a lepidopteran insect selected from the group consisting of any species within the families: Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawk moths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (inchworms, noctuids, and underwing moths), and Plutellidae (diamond moths).

[0566] 148. The composition of claim 147, wherein the lepidopteran insect is a species selected from species of the genus Plutella, optionally wherein the lepidopteran insect is Plutella xylostella.

[0567] 149. The composition of claim 147, wherein the lepidopteran insect is a species selected from the species of the genus Spodoptera, optionally wherein the lepidopteran insect is Spodoptera frugiperda.

[0568] 150. A method for controlling insect infestation of a plant, the method comprising delivering to the plant or insect a composition described in any one of paragraphs 1-105 or 142-149.

[0569] 151. A kit comprising a composition according to any one of paragraphs 1-105 or 142-149 and optional instructions for use.

[0570] 152. The composition of any one of paragraphs 1-105 or 142-149, wherein the YP is at a concentration of about 2 g / L to about 15 g / L, or about 5 g / L to about 12 g / L, or about 10 g / L.

[0571] Cited literature Janeway, CAJr Approaching the asymptote?Evolution and revolution in immunology.Cold Spring Harb.Symp.Quant.Biol.54,1-13(1989).

[0572] Tabashnik et al., Managing Resistance to Bacillus thuringiensis: Lessons from the Diamondback Moth(Lepidoptera: Plutellidae) J. Econ. Entomol. 84(1):49-55 (1991).

[0573] Tabashnik et al., Global Patterns of Insect Resistance to Transgenic Bt Crops: The First 25 Years, Journal of Econ. Entomol., 2023, 1-13.

[0574] Equivalence and Scope In the claims, articles such as "a," "an," and "the" can mean one or more than one, unless indicated to the contrary or otherwise clear from the context. Unless indicated to the contrary or otherwise clear from the context, a claim or description including "or" between one or more members of a group is considered to be satisfied if one, more than one, or all members of the group are present in, employed in, or otherwise relevant to a given product or process. The invention includes embodiments in which exactly one member of the group is present in, used in, or otherwise relevant to a given product or process. The invention includes embodiments in which more than one, or all, of the group members are present in, employed in, or otherwise relevant to a given product or process.

[0575] Furthermore, the present invention covers all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the enumerated claims are introduced into another claim. For example, any claim that depends on another claim can be modified to include one or more limitations found in any other claim that depends on the same base claim. Where elements are presented as enumerated in Markush group format, by way of example, each subgroup of elements is also disclosed, and any element(s) can be removed from the group. In general, when the invention, or embodiments of the invention, are described as including certain elements and / or features, it should be understood that a particular embodiment of the invention or an aspect of the invention consists of or consists essentially of such elements and / or features. For purposes of brevity, those embodiments have not been specifically recited in haec verba herein.

[0576] It should also be noted that the terms "comprising" and "containing" are intended to be open and permit the inclusion of additional elements or steps. Where ranges are given, the endpoints are included. Furthermore, unless otherwise indicated or otherwise apparent from the context and the understanding of one of ordinary skill in the art, values ​​expressed as ranges can assume any specific value, or subrange within the ranges set forth in different aspects of the invention, to one-tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0577] This application references various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. In the event of a conflict between any of the incorporated references and this specification, this specification shall control. In addition, any particular aspect of the present invention that falls within the prior art may be expressly excluded from any one or more of the claims. Because such aspects would be known to those of skill in the art, they may be excluded even if the exclusion is not expressly stated herein. Any particular aspect of the present invention may be excluded from any claim for any reason, whether or not related to the existence of prior art.

[0578] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein. The scope of the embodiments described herein is not intended to be limited to the above description, but rather is as set forth in the appended claims. Those skilled in the art will appreciate that various changes and modifications to this description can be made without departing from the spirit or scope of the invention, as defined in the following claims.

Claims

1. A composition comprising a pesticide and yeast particles comprising yeast cell wall components, wherein the pesticide is a biopesticide or a pesticidal polynucleotide.

2. 1. A composition comprising a yeast particle comprising a pesticide and a yeast cell wall component, wherein the pesticide is a chemical pesticide, and wherein the pesticide is selected from the group consisting of ryanodine receptor modulators (IRAC class 28), chitin biosynthesis inhibitors acting on CHS1 (IRAC class 15), nicotinic acetylcholine receptor (nACHR) allosteric modulators-site 1 (IRAC class 5), nACHR competitive modulators (IRAC class 4), sodium channel modulators (IRAC class 3), acetylcholinesterase (ACHE) inhibitors (IRAC class 1), GABA-gated chloride channel blockers (IRAC class 2), glutamic acid inhibitors (IRAC class 3), and the like. Glucose-gated chloride channel (GLUCL) allosteric modulators (IRAC class 6), juvenile hormone receptor modulators (IRAC class 7), other nonspecific (multi-site) inhibitors (IRAC class 8), chordotonal organ TRPV channel modulators (IRAC class 9), mite growth inhibitors acting on CHS1 (IRAC class 10), mitochondrial ATP biosynthesis enzyme inhibitors (IRAC class 12), oxidative phosphorylation uncouplers that disrupt the proton gradient (IRAC class 13), nicotinic acetylcholine receptor (nACHR) channel blockers (IRAC class 14), chitin biosynthesis inhibitors, type 1 (IRAC class 16), and molting disruptors.Diptera (IRAC class 17), ecdysone receptor agonists (IRAC class 18), and octopamine receptor agonists (IRAC class 19), mitochondrial complex III electron transport inhibitors QO site (IRAC class 20), mitochondrial complex I electron transport inhibitors (IRAC class 21), voltage-gated sodium channel blockers (IRAC class 22), acetyl-CoA carboxylase inhibitors (IRAC class 23), mitochondrial complex IV electron transport inhibitors (IRAC class 24), mitochondrial transport inhibitors (IRAC class 25), and mitochondrial complex IV electron transport inhibitors (IRAC class 26). Class 25), chordotonal organ nicotinamidase inhibitor (IRAC class 29), GABAergic chloride channel allosteric modulator (IRAC class 30), nicotinic acetylcholine receptor (NACHR) allosteric modulator - site II (IRAC class 32), calcium-activated potassium channel (KCa2) modulator (IRAC class 33), mitochondrial complex III electron transport inhibitor QI site (IRAC class 34), or chordotonal organ modulator - target site undefined (IRAC class 36).

3. 3. The composition of claim 2, wherein the chemical pesticide is selected from the group consisting of chlorantraniliprole, novaluron, spinosad, acetamiprid, cypermethrin, diflubenzuron, and bifenthrin.

4. 4. The composition of any one of claims 1 to 3, wherein the yeast particle is selected from the group consisting of yeast cell wall particle (YCWP), yeast cell particle (YCP), yeast glucan particle (GP or YGP), yeast glucan mannan particle (GMP or YGMP), yeast glucan chitin particle (GCP or YGCP), yeast glucan chitin mannan particle (GCMP or YGCMP), yeast glucan lipid particle (GLP or YGLP), whole glucan particle (WGP), and combinations thereof.

5. 5. The composition of any one of claims 1 to 4, wherein the yeast particles enhance the efficacy of the pesticide, and optionally the yeast particles enhance the ability of the pesticide to control plant pests by at least 5%, 10%, 20%, 30%, 40%, or 50% compared to a control composition that does not include the yeast particles.

6. The composition of any one of claims 1 to 5, wherein the yeast particles are commercially available yeast particles.

7. The composition of any one of claims 1 to 6, wherein the yeast particles comprise β-glucan and / or mannan oligosaccharides.

8. 8. The composition of any one of claims 1 to 7, wherein the yeast particles comprise β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan, optionally wherein the β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan is branched.

9. 9. The composition of any one of claims 1 to 8, wherein the yeast particles comprise intact or fragmented yeast cell walls.

10. the yeast particles contain β-1,6-glucan, β-1,3-glucan, mannan oligosaccharides, mannoproteins, chitin, and / or lipids; Optionally, the composition of any one of claims 1 to 9, wherein the lipid is phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), and / or cytidine diphosphate diacylglycerol (CDP-DAG).

11. 11. The composition of any one of claims 1 to 10, wherein the yeast particles do not include yeast extract.

12. 11. The composition of any one of claims 1 to 10, wherein the yeast particles do not consist of yeast extract.

13. 13. The composition of any one of claims 1 to 12, wherein the pesticide is not encapsulated in the yeast particle.

14. 13. The composition of any one of claims 1 to 12, wherein the pesticide is encapsulated in the yeast particle.

15. A composition comprising a pesticide and one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharides, and laminarin, wherein the pesticide is a biopesticide or a pesticidal polynucleotide.

16. A composition comprising a pesticide and one or more polysaccharides selected from the group consisting of β-glucan, mannan oligosaccharides, and laminarin, wherein the pesticide is a chemical pesticide and wherein the pesticide is selected from the group consisting of ryanodine receptor modulators (IRAC class 28), chitin biosynthesis inhibitors acting on CHS1 (IRAC class 15), nicotinic acetylcholine receptor (nACHR) allosteric modulators-site 1 (IRAC class 5), nACHR competitive modulators (IRAC class 4), sodium channel modulators (IRAC class 3), acetylcholinesterase (ACHE) inhibitors (IRAC class 1), GABA-gated chloride channel blockers (GABA-gated chloride channel blockers), and methicillin-resistant steroids (META). - (IRAC class 2), glutamate-gated chloride channel (GLUCL) allosteric modulators (IRAC class 6), juvenile hormone receptor modulators (IRAC class 7), other nonspecific (multi-site) inhibitors (IRAC class 8), chordotonal organ TRPV channel modulators (IRAC class 9), mite growth inhibitors acting on CHS1 (IRAC class 10), mitochondrial ATP biosynthesis enzyme inhibitors (IRAC class 12), oxidative phosphorylation uncouplers that disrupt the proton gradient (IRAC class 13), nicotinic acetylcholine receptor (nACHR) channel blockers (IRAC class 14), chitin biosynthesis inhibitors, type 1 (IRAC class 16), and molting disruptorsDiptera (IRAC class 17), ecdysone receptor agonists (IRAC class 18), and octopamine receptor agonists (IRAC class 19), mitochondrial complex III electron transport inhibitors QO site (IRAC class 20), mitochondrial complex I electron transport inhibitors (IRAC class 21), voltage-gated sodium channel blockers (IRAC class 22), acetyl-CoA carboxylase inhibitors (IRAC class 23), mitochondrial complex IV electron transport inhibitors (IRAC class 24), mitochondrial transport inhibitors (IRAC class 25), and mitochondrial complex IV electron transport inhibitors (IRAC class 26). Class 25), chordotonal organ nicotinamidase inhibitor (IRAC class 29), GABAergic chloride channel allosteric modulator (IRAC class 30), nicotinic acetylcholine receptor (NACHR) allosteric modulator - site II (IRAC class 32), calcium-activated potassium channel (KCa2) modulator (IRAC class 33), mitochondrial complex III electron transport inhibitor QI site (IRAC class 34), or chordotonal organ modulator - target site undefined (IRAC class 36).

17. 17. The composition of claim 16, wherein the chemical pesticide is selected from the group consisting of chlorantraniliprole, novaluron, spinosad, acetamiprid, cypermethrin, diflubenzuron, and bifenthrin.

18. 18. The composition of any one of claims 15 to 17, wherein the one or more polysaccharides enhance the efficacy of the pesticide, and optionally the one or more polysaccharides enhance the ability of the pesticide to control plant pests by at least 5%, 10%, 20%, 30%, 40%, or 50% compared to a control composition that does not include the one or more polysaccharides.

19. 19. The composition of any one of claims 15 to 18, wherein the one or more polysaccharides comprise β-1,6-glucan, β-1,3-glucan, β-1,3-1,6-glucan, and / or mannan oligosaccharides, optionally wherein the β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan is branched.

20. 20. The composition of any one of claims 15 to 19, wherein the one or more polysaccharides are derived from yeast cell walls.

21. 21. The composition of any one of claims 15 to 20, wherein the one or more polysaccharides comprise β-glucan derived from yeast cell walls, optionally β-glucan derived from Saccharomyces cerevisiae.

22. 20. The composition of any one of claims 15 to 19, wherein the one or more polysaccharides are derived from a source other than yeast, optionally wherein the source other than yeast is a bacterium, a fungus, an algae, a lichen, or a plant.

23. 23. The composition of any one of claims 15 to 19 or 22, wherein the one or more polysaccharides comprise β-1,3-glucan or laminarin derived from algae, optionally β-1,3-glucan derived from Euglena gracilis or laminarin derived from Laminaria digitata.

24. 24. The composition of any one of claims 1, 4-15, or 16-23, wherein the pesticide is a biopesticide.

25. 25. The composition of claim 24, wherein the biopesticide comprises a microorganism.

26. 26. The composition of claim 25, wherein the microorganism is a naturally occurring or engineered microorganism.

27. 27. The composition of claim 25 or 26, wherein the microorganism produces one or more proteins that control a plant pest, optionally wherein the plant pest is a fungus or an insect.

28. 28. The composition of claim 27, wherein the plant pest is an insect.

29. 29. The composition of claim 27 or 28, wherein the one or more proteins produced by the microorganism are activated in the insect gut.

30. 30. The composition of any one of claims 26 to 29, wherein the one or more proteins produced by a naturally occurring microorganism are selected from the group consisting of crystal proteins, insecticidal proteins produced during vegetative growth, and toxin complex proteins.

31. 31. The composition of any one of claims 27-30, wherein the one or more proteins produced by a naturally occurring microorganism are Bacillus thuringiensis (Bt) crystal proteins, optionally wherein the Bt crystal proteins are selected from the group consisting of Cry1F, Cry1AB, Cry1Ac, Cry1.A105, Cry2AB, Cry1B.868, Cry1C, Cry1Da7, Cry2Ae, and Cry1Fa, and optionally wherein the Bt crystal proteins are selected from the group consisting of Cry1AB, Cry1F, or Cry1Fa.

32. 32. The composition of any one of claims 25 to 31, wherein the microorganism is a Bacillus thuringiensis microorganism.

33. 33. The composition of any one of claims 24 to 32, wherein the biopesticide comprises a commercially available biopesticide, wherein the biopesticide controls a plant pest, and optionally wherein the plant pest is a fungus or an insect.

34. 25. The composition of claim 24, wherein the biopesticide comprises isolated cellular components derived from a microorganism, optionally wherein the microorganism is a naturally occurring or engineered microorganism.

35. 35. The composition of claim 34, wherein the isolated cellular components comprise one or more proteins or polynucleotides that control a plant pest, optionally wherein the plant pest is a fungus or an insect.

36. 36. The composition of claim 35, wherein the one or more proteins or polynucleotides are activated in the insect gut.

37. 37. The composition of claim 35 or 36, wherein the one or more proteins are selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth.

38. 38. The composition of claim 37, wherein the biological pesticide comprises a Bacillus thuringiensis (Bt) protein, optionally wherein the Bt protein is a Bt crystal protein, further optionally wherein the Bt protein is derived from a microorganism, further optionally wherein the Bt crystal protein is selected from the group consisting of Cry1F, Cry1AB, Cry1Ac, Cry1.A105, Cry2AB, Cry1B.868, Cry1C, Cry1Da7, Cry2Ae, and Cry1Fa, further optionally wherein the Bt crystal protein is selected from the group consisting of Cry1AB, Cry1Fm, and Cry1Fa.

39. 39. The composition of any one of claims 1 to 38, wherein the composition further comprises a pesticidal polynucleotide.

40. 24. The composition of any one of claims 1, 4-15, or 18-23, wherein the pesticide is a pesticidal polynucleotide.

41. 41. The composition of claim 39 or 40, wherein the pesticidal polynucleotide inhibits expression of a target gene in a plant pest, optionally wherein the plant pest is an insect.

42. 42. The composition of any one of claims 39 to 41, wherein the pesticidal polynucleotide is single-stranded RNA (ssRNA) or double-stranded RNA (dsRNA).

43. 43. The composition of any one of claims 39 to 42, wherein the composition comprises yeast particles and the pesticidal polynucleotide is encapsulated within the yeast particles.

44. 44. The composition of claim 43, wherein the composition further comprises a cationic polymer and / or a nuclease inhibitor, and wherein the pesticidal polynucleotide is encapsulated within the yeast particle.

45. 45. The composition of claim 43 or 44, wherein the encapsulation efficiency is greater than about 80%, and optionally, the encapsulation efficiency is greater than 90%.

46. 46. ​​The composition of any one of claims 40-45, wherein the composition further comprises a biopesticide, optionally wherein the biopesticide comprises a Bacillus thuringiensis (Bt) protein, and optionally wherein the Bt protein is a Bt crystal protein.

47. 47. The composition of any one of claims 39 to 46, wherein the pesticidal polynucleotide comprises a first strand that is identical to or complementary to a region of messenger RNA (mRNA) encoded by one or more target genes.

48. 48. The composition of claim 47, wherein the pesticidal polynucleotide is a dsRNA comprising a second strand that is complementary to the first strand.

49. 49. The composition of any one of claims 41 to 48, wherein the target gene comprises one or more genes selected from the group consisting of proteasome beta 5 (PSMB5) gene, vacuolar ATPase E (vATPase E) gene, calmodulin gene, inhibitor of apoptosis protein (IAP) gene, soluble NSF attachment protein (a-SNAP) gene, Ras opposite (ROP) gene, proteasome alpha type 2 (PTSA2) gene, secretion-associated Ras-associated GTPase 1 (SAR1) gene, PBAN, ATPase, wingsup A (wupA), CP4S3_DROME, and C12C1_DROME.

50. 50. The composition of claim 49, wherein the target gene is IAP.

51. 51. The composition of claim 50, wherein the target gene has the sequence of SEQ ID NO:

1.

52. 52. The composition of any one of claims 39 to 51, wherein the pesticidal polynucleotide comprises at least 20, 21, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 contiguous nucleotides that are complementary to or contain at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity to a segment of the target gene.

53. 53. The composition of claim 52, wherein the pesticidal polynucleotide comprises at least 20, 21, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 contiguous nucleotides that are complementary to, or comprise at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity to, a segment of a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-13.

54. 54. The composition of any one of claims 1, 4-15, or 18-53, wherein the pesticidal polynucleotide comprises at least 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 nucleotides.

55. 40. The composition of any one of claims 1, 4-15, or 18-39, wherein the pesticide comprises at least 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, or 1000 amino acids.

56. 24. The composition of any one of claims 2-14 or 16-23, wherein the ryanodine receptor modulator (IRAC class 28) comprises a diamide selected from the group consisting of chlorantraniliprole, tetraniliprole, cyclaniliprole, brofuranilide, cyantraniliprole, imidacloprid, or flubendiamide.

57. 24. The composition of any one of claims 2 to 14 or 16 to 23, wherein the chitin biosynthesis inhibitor (IRAC class 15) acting on CHS1 comprises a benzoyl urea selected from the group consisting of bistrifluron, chlorfluazuron, diflubenzuron, flucycloxuron, flufenoxuron, hexaflumuron, lufenuron, novaluron, noviflumuron, teflubenzuron, and triflumuron.

58. 24. The composition of any one of claims 2-14 or 16-23, wherein the nACHR allosteric modulator-site 1 (IRAC class 5) is a spinosyn, optionally wherein the spinosyn is spinetoram or spinosad.

59. 24. The composition of any one of claims 2-14 or 16-23, wherein the nACHR competitive modulator (IRAC class 4) is a neonicotinoid, optionally wherein the neonicotinoid is selected from the group consisting of acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, thiacloprid, and thiamethoxam.

60. The sodium channel modulator (IRAC class 3) is a pyrethroid or pyrethrin, optionally wherein the pyrethroid or pyrethrin is acrinathrin, allethrin, d-cis-trans-allethrin, d-trans-allethrin, bifenthrin, bioallethrin, bioallethrin S-cyclopentenyl, bioresmethrin, cycloprothrin, cyfluthrin, β-cyfluthrin, cyhalothrin, lambda-cyhalothrin, gamma-cyhalothrin, cypermethrin, alpha-cypermethrin, β-cypermethrin, theta-cypermethrin, zeta-cypermethrin, cyphenothrin [(1R)-trans- 24. The composition of any one of claims 2-14 or 16-23, wherein the benzophenone-3-one is selected from the group consisting of benzophenone-3-one (EZ)-(1R)-isomer, deltamethrin, empenthrin [(EZ)-(1R)-isomer], esfenvalerate, etofenprox, fenpropathrin, fenvalerate, flucythrinate, flumethrin, tau-fluvalinate, kadethrin, pyrethrins (pyrethrams), halfenprox, fenothrin [(1R)-trans-isomer], prallethrin, resmethrin, silafluofen, tefluthrin, tetramethrin, tetramethrin [(1R)-isomer], tralomethrin, transfluthrin, and permethrin.

61. 61. The composition of any one of claims 1 to 60, wherein the composition controls a plant pest, optionally wherein the plant pest is a fungus or an insect.

62. 59. The composition of claim 58, wherein the plant pest is an insect.

63. 63. The composition of claim 61 or 62, wherein the plant pest is resistant to the pesticide.

64. 64. The composition of any one of claims 61 to 63, wherein the plant pest is an insect belonging to the order Lepidoptera, Coleoptera, Hemiptera, Diptera, or Acari.

65. 65. The composition of any one of claims 61 to 64, wherein the plant pest is resistant to Bacillus thuringiensis (Bt) or a Bt protein.

66. Plant pests belonging to the order Lepidoptera include Nymphalidae (nymphalians), Danaidae (monarchs), Pieridae (pierids and ceratopsians), Papilionidae (swallowtails), Lycaenidae (lycaenids), Hesperiidae (skippers), Tineidae (moths), and Sesiidae (cleanse moths). ), Pyralidae (pyralid moths), Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawk moths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (inchworms, noctuids, and underwing moths), and Plutellidae (diamond moths).

67. 67. The composition of claim 66, wherein the lepidopteran insect species is a Plutella species, optionally wherein the Plutella species insect is Plutella xylostella (diamond moth).

68. 67. The composition of claim 66, wherein the lepidopteran insect species is a Spodoptera species, optionally wherein the Spodoptera species insect is Spodoptera frugiperda (spodoptera fall armyworm).

69. 67. The composition of claim 66, wherein the lepidopteran insect species is Chrysodeixis species, optionally wherein the Chrysodeixis species insect is Chyrysodeixis includens (soybean looper).

70. 67. The composition of claim 66, wherein the lepidopteran insect species is Helicoverpa species, optionally wherein the Helicoverpa species insect is Helicoverpa armigera (cotton boll worm).

71. 67. The composition of claim 66, wherein the lepidopteran insect species is Helicoverpa armigera.

72. 67. The composition of claim 66, wherein the lepidopteran insect species is Plutella xylostella.

73. 67. The composition of claim 66, wherein the lepidopteran insect species is Spodoptera frugiperda.

74. 67. The composition of claim 66, wherein the lepidopteran insect species is Chyrysodeixis includens.

75. 67. The composition of claim 66, wherein the lepidopteran insect species is Helicoverpa species, optionally wherein the Helicoverpa species insect is Helicoverpa zea (tomato fruit worm).

76. 67. The composition of claim 66, wherein the lepidopteran insect species is Cydia species, optionally wherein the Cydia species insect is Cydia pomonella (the fruit moth).

77. 67. The composition of claim 66, wherein the lepidopteran insect species is a Trichoplusia species, optionally wherein the Trichoplusia species insect is Trichoplusia ni (cabbage looper).

78. 67. The composition of claim 66, wherein the lepidopteran insect species is a Spodoptera species, optionally wherein the Spodoptera species insect is Spodoptera exigua (beet armyworm).

79. 67. The composition of claim 66, wherein the lepidopteran insect species is a Spodoptera species, optionally wherein the Spodoptera species insect is Spodoptera litura (tobacco rootworm).

80. 67. The composition of claim 66, wherein the lepidopteran insect species is a Spodoptera species, optionally wherein the Spodoptera species insect is Spodoptera litoralis (tomato moth).

81. 67. The composition of claim 66, wherein the lepidopteran insect species is Pieris species, optionally wherein the Pieris species insect is Pieris rapae (imported caterpillar).

82. 67. The composition of claim 66, wherein the lepidopteran insect species is a Tuta species, optionally wherein the Tuta species insect is Tuta absoluta (tomato tea moth).

83. 67. The composition of claim 66, wherein the lepidopteran insect species is a Grapholita species.

84. 67. The composition of claim 66, wherein the lepidopteran insect species is a Chrysodeixis species, optionally wherein the Chrysodeixis species insect is Chyrysodeixis acuta (tomato semilooper).

85. 67. The composition of claim 66, wherein the lepidopteran insect species is Paramyelois species, optionally wherein the Paramyelois species insect is Paramyelois transitella (navel orangeworm).

86. 67. The composition of claim 66, wherein the lepidopteran insect species is Lobesia species, optionally wherein the Lobesia species insect is Lobesia botrana (European grapevine moth).

87. 66. The composition of claim 64 or 65, wherein the insect belongs to the order Coleoptera, and wherein the Coleopteran insect is a species selected from the group consisting of Leptinotarsa ​​spp., Phyllotreta spp., Cerotoma spp., Diabrotica spp., Tribolium spp., Anthonomus spp., and Alticini spp.

88. 88. The composition of claim 87, wherein the insect is a Phyllotreta species, optionally wherein the Phyllotreta species insect is Phyllotreta cruciferae (canola flea beetle).

89. 88. The composition of claim 87, wherein the insect is a Phyllotreta species, optionally wherein the Phyllotreta species insect is Phyllotreta striolata (Phyllotreta striolata).

90. 88. The composition of claim 87, wherein the insect is a Psylliodes species, optionally wherein the Psylliodes species insect is Psylliodes chrysocephala (cabbage stem flea beetle).

91. 88. The composition of claim 87, wherein the insect is a Leptinotarsa ​​species, optionally wherein the Leptinotarsa ​​species is the Colorado potato beetle.

92. 92. The composition of any one of claims 1 to 91, wherein the composition further comprises one or more compounds selected from the group consisting of cationic lipids, cationic polymers, non-cationic polymers, organic carriers, nuclease inhibitors, surfactants, antifoaming agents, and biocides.

93. 92. The composition of any one of claims 1 to 91, wherein the composition further comprises a cationic polymer and a nuclease inhibitor.

94. 94. The composition of claim 92 or 93, wherein the cationic polymer is polyethyleneimine (PEI), poly-L-lysine (PLL), cationic gelatin, cationic chitosan, cationic cellulose, cationic dextran, poly(2-N,N-dimethylaminoethyl methacrylate), or poly(amidoamine).

95. 94. The composition of claim 92 or 93, wherein the nuclease inhibitor is ethylenediaminetetraacetic acid (EDTA), polyvinylsulfonic acid (PVSA), sodium hexametaphosphate (SHMP), sodium tripolyphosphate (TPP), diethylpyrocarbonate, aurintricarboxylic acid (ATA), formamide, macaloid, proteinase K, heparin, hydroxylamine-oxygen-copper(II) ion, bentonite, ammonium sulfate, dithiothreitol (DTT), beta-mercaptoethanol, cysteine, dithioerythritol, or tris(2-carboxyethyl)phosphene hydrochloride.

96. 96. The composition of any one of claims 1 to 95, wherein the composition further comprises one or more of an acidifying agent, a buffering agent, an antifoaming agent, an anti-transpirant, a biocide preservative, a dye and brightener, a compatibilizer, a crop oil concentrate, a surfactant, a deposition agent, a drift reducing agent, a feeding stimulant, a spreading agent, a weighting agent, an adhesive, a suspending agent, a gelling agent, a synergist, a wetting agent, an emulsifier, a dispersing agent, a penetrating agent, a neutralizing agent, a water absorbing agent, and / or a water softener.

97. 97. The composition of any one of claims 1 to 96, wherein the composition is formulated as a spray, a solution, an emulsifiable concentrate, a solid, a suspension, a colloid, a micelle, or an emulsion, a soluble liquid concentrate, a wettable powder, a water dispersible granule, an emulsion, an aerosol, a homogeneous mixture, or a heterogeneous mixture.

98. (i) one or more Bacillus thuringiensis (Bt) proteins, and (ii) yeast particles containing yeast cell wall components A composition comprising:

99. 99. The composition of claim 98, wherein the composition causes at least about 10% greater mortality in plant-infesting insect pests compared to the one or more Bt proteins alone.

100. The concentration of yeast particles or one or more polysaccharides is (a) about 2 g / L to about 15 g / L, about 5 g / L to about 12 g / L, or about 10 g / L; or (b) about 2% w:w to about 15% w:w, about 5% w:w, about 12% w:w, or about 10% w:w 100. The composition of any one of claims 1 to 99, wherein

101. 101. The composition of any one of claims 1-100, wherein less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, or less than 5% of the pesticide is encapsulated within the yeast particles or one or more polysaccharides.

102. 102. The composition of claim 101, wherein 1 to 25% or 1 to 10% of the pesticide is encapsulated within the yeast particle or one or more polysaccharides.

103. 103. A kit comprising the composition of any one of claims 1 to 102, optionally further comprising instructions for use.

104. 103. A method for controlling plant pests, the method comprising delivering a composition according to any one of claims 1 to 102 to a plant, soil, a plant pest, or the food of a plant pest.

105. 103. A method for controlling insect infestation of a plant, the method comprising delivering a composition of any one of claims 1 to a plant, soil, insect food, or insect.

106. 103. A method for controlling infestation of a plant by lepidopteran insects, the method comprising delivering a composition according to any one of claims 1 to 102 to the plant, the soil, the food of the insect, or the insect.

107. A method for controlling infestation of a plant by a plant pest, wherein the plant pest is an insect, the method comprising delivering a composition to the plant, soil, insect food, or insect, wherein the composition comprises yeast particles comprising a pesticidal polynucleotide and a yeast cell wall component, and optionally wherein the insect is a lepidopteran insect.

108. 108. The method of claim 107, wherein the pesticidal polynucleotide is encapsulated in a yeast cell wall particle.

109. 109. The method of claims 104-108, wherein the plant is a transgenic plant engineered to express a biopesticide or pesticidal polynucleotide.

110. 110. The method of claim 109, wherein the transgenic plant is engineered to express a biopesticide, wherein the biopesticide comprises one or more Bt proteins.

111. 1. A method for controlling plant pests, the method comprising delivering yeast particles comprising yeast cell wall components to a transgenic plant engineered to express a biopesticide or pesticidal polynucleotide.

112. 112. The method of any one of claims 107 to 111, wherein the yeast particle is selected from the group consisting of yeast cell wall particle (YCWP), yeast cell particle (YCP), yeast glucan particle (GP or YGP), yeast glucan mannan particle (GMP or YGMP), yeast glucan chitin particle (GCP or YGCP), yeast glucan chitin mannan particle (GCMP or YGCMP), yeast glucan lipid particle (GLP or YGLP), and whole glucan particle (WGP), and combinations thereof.

113. 113. The method of any one of claims 107 to 112, wherein the yeast particles are commercially available yeast particles.

114. 114. The method of any one of claims 107 to 113, wherein the yeast particles comprise β-glucan and / or mannan oligosaccharides.

115. 115. The method of any one of claims 107 to 114, wherein the yeast particles comprise β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan, optionally wherein the β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan is branched.

116. 116. The method of any one of claims 107 to 115, wherein the yeast particles comprise intact or fragmented cell walls.

117. the yeast particles contain β-1,6-glucan, β-1,3-glucan, mannan oligosaccharides, mannoproteins, chitin, and / or lipids; Optionally, the method of any one of claims 107 to 116, wherein the lipid is phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), and / or cytidine diphosphate diacylglycerol (CDP-DAG).

118. 118. The method of any one of claims 107 to 117, wherein the yeast particles do not consist of yeast extract.

119. 118. The method of any one of claims 107 to 117, wherein the yeast particles do not include yeast extract.

120. 120. The method of any one of claims 107-119, wherein the yeast particles are in a composition further comprising one or more of an acidifying agent, a buffering agent, an antifoaming agent, an anti-transpiration agent, a biocide preservative, a dye and brightener, a compatibilizer, a crop oil concentrate, a surfactant, a deposition agent, a drift reducing agent, a feeding stimulant, a spreading agent, a weighting agent, an adhesive, a suspending agent, a gelling agent, a synergist, a wetting agent, an emulsifier, a dispersing agent, a penetrating agent, a neutralizing agent, a water absorbing agent, and / or a water softener.

121. 121. The method of any one of claims 107 to 120, wherein the yeast particles are in a composition further comprising one or more of a surfactant, an antifoaming agent, and a biocide preservative, optionally wherein the composition comprises a surfactant, an antifoaming agent, and a biocide.

122. 122. The method of any one of claims 107 to 121, wherein the yeast particles are in a composition formulated as a spray, a liquid (including homogeneous mixtures such as soluble liquid concentrates, and heterogeneous mixtures such as suspensions, colloids, micelles, and emulsions), a wettable powder, a water dispersible granule, a suspension, an emulsion, an aerosol, an emulsifiable concentrate, or a solid.

123. 123. The method of any one of claims 107 to 122, wherein the yeast particles are delivered in a pesticide-free composition.

124. 123. The method of any one of claims 107 to 122, wherein the yeast particles are delivered in a composition that does not contain a Bt pesticide.

125. 123. The method of any one of claims 107 to 122, wherein the yeast particles are delivered in a composition that does not contain a biopesticide.

126. 123. The method of any one of claims 107 to 122, wherein the yeast particles are delivered in a composition that does not contain a chemical pesticide.

127. 127. The method of any one of claims 104 to 126, wherein the yeast particles are applied at a dosage of about 50 to about 300 g yeast particles / ha; or about 200 to about 300 g yeast particles / ha; or about 100 to about 500 g yeast particles / ha; or about 100 to about 1,000 g yeast particles / ha; or about 200 g yeast particles / ha; or about 250 g yeast particles / ha; or about 300 g yeast particles / ha; or about 350 g yeast particles / ha; or about 400 g yeast particles / ha; or about 450 g yeast particles / ha; or about 500 g yeast particles / ha; or about 550 g yeast particles / ha.

128. A method for controlling plant infestation by insects, the method comprising delivering a composition to the plant, soil, insect food, or insect, wherein the composition comprises a pesticidal polynucleotide and one or more polysaccharides, and optionally wherein the insect is a lepidopteran insect.

129. 1. A method for controlling plant pests, the method comprising delivering one or more polysaccharides to a transgenic plant engineered to express a biopesticide or pesticidal polynucleotide.

130. 130. The method of claim 128 or 129, wherein the one or more polysaccharides comprise β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan, optionally wherein the β-1,3-glucan, β-1,6-glucan, and / or β-1,3-1,6-glucan is branched.

131. 131. The method of any one of claims 128 to 130, wherein the one or more polysaccharides comprise a β-glucan derived from yeast, optionally a β-glucan derived from Saccharomyces cerevisiae.

132. 131. The method of any one of claims 128-130, wherein the one or more polysaccharides are derived from a source other than yeast, optionally wherein the source other than yeast is a bacterium, a fungus, an algae, a lichen, or a plant.

133. 133. The method of any one of claims 128-130 or 132, wherein the one or more polysaccharides comprise a polysaccharide selected from the group consisting of β-1,3-glucan or laminarin derived from algae, optionally β-1,3-glucan derived from Euglena gracilis or laminarin derived from Laminaria digitata.

134. 134. The method of any one of claims 109 to 133, wherein the transgenic plant expresses one or more proteins that control a plant pest, optionally wherein the plant pest is a fungus or an insect.

135. 135. The method of claim 134, wherein the plant pest is an insect.

136. 136. The method of claim 134 or 135, wherein one or more proteins expressed by the transgenic plant are activated in the insect gut.

137. 137. The method of any one of claims 134 to 136, wherein the one or more proteins expressed by the transgenic plant are selected from the group consisting of crystal proteins and insecticidal proteins produced during vegetative growth.

138. 138. The method of claim 137, wherein one or more proteins expressed by the transgenic plant is a Bacillus thuringiensis (Bt) crystal protein.

139. The method of claim 138, wherein one or more proteins expressed by the transgenic plant are selected from the group consisting of VIP3a, Cry1F, Cry1AB, Cry1Ac, Cry1.A105, Cry2AB, Cry1B.868, Cry1C, Cry1Da7, Cry2Ae, and Cry1Fa.

140. 139. The method of claim 138, wherein one or more proteins expressed by the transgenic plant are selected from the group consisting of Cry1F, Cry1AB, Cry1Ac, Cry1.A105, Cry2AB, Cry1B.868, Cry1C, Cry1Da7, Cry2Ae, and Cry1Fa.

141. 139. The method of claim 138, wherein the one or more proteins expressed by the transgenic plant are selected from the group consisting of Cry1F, Cry1Fa, and Cry1Ab.

142. 142. The method of any one of claims 109-141, wherein the transgenic plant is selected from the group consisting of a Solanaceae plant, a Brassicaceae plant, a Poaceae plant, a Cucurbitaceae plant, a Fabaceae plant, a Fagaceae plant, an Asteraceae plant, an Amaryllidaceae plant, a Umbelliferae plant, an Apiaceae plant, an Amranthaceae plant, and a Malvaceae plant; optionally, the plant is selected from the group consisting of maize, soybean, and cotton.

143. 143. The method of claim 142, wherein the transgenic plant is in the family Malvaceae.

144. 144. The method of claim 143, wherein the transgenic plant in the Malvaceae family is a cotton seed.

145. 143. The method of claim 142, wherein the transgenic plant is in the Poaceae family.

146. 146. The method of claim 145, wherein the transgenic plant in the Poaceae family is a maize seed.

147. 143. The method of claim 142, wherein the transgenic plant is a plant in the Fabaceae family.

148. 148. The method of claim 147, wherein the transgenic plant of the Fabaceae family is soybean.

149. 146. The method of claim 145, wherein the transgenic plant in the Poaceae family is a rice seed.

150. 150. The method of any one of claims 107 to 149, wherein the pesticidal polynucleotide inhibits expression of a target gene in a plant pest, optionally wherein the plant pest is an insect.

151. 151. The method of claim 150, wherein the pesticidal polynucleotide is single-stranded RNA (ssRNA) or double-stranded RNA (dsRNA).

152. 152. The method of any one of claims 107-151, wherein the pesticidal polynucleotide comprises a first strand that is identical to or complementary to a region of messenger RNA (mRNA) encoded by one or more target genes.

153. 152. The method of claim 151, wherein the pesticidal polynucleotide is a dsRNA comprising a second strand that is complementary to the first strand.

154. The method of any one of claims 150 to 153, wherein the target gene comprises one or more genes selected from the group consisting of proteasome beta 5 (PSMB5) gene, vacuolar ATPase E (vATPase E) gene, calmodulin gene, inhibitor of apoptosis protein (IAP) gene, soluble NSF attachment protein (a-SNAP) gene, Ras opposite (ROP) gene, proteasome alpha type 2 (PTSA2) gene, secretion-associated Ras-associated GTPase 1 (SAR1) gene, PBAN, ATPase, wingsup A (wupA), CP4S3_DROME, and C12C1_DROME.

155. 155. The method of claim 154, wherein the target gene is IAP.

156. 156. The method of claim 155, wherein the target gene has the sequence of SEQ ID NO:

1.

157. 157. The method of any one of claims 107-156, wherein the pesticidal polynucleotide comprises at least 20, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 contiguous nucleotides that are complementary to or contain at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity to a segment of the target gene.

158. 158. The method of claim 157, wherein the pesticidal polynucleotide comprises at least 20, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 550 contiguous nucleotides that are complementary to, or comprise at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity to, a segment of a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-13.

159. 159. The method of any one of claims 104-158, wherein the composition is delivered to the leaves, stems, branches, seeds, fruits, flowers, roots, or soil of the plant.

160. 160. The method of any one of claims 104 to 159, wherein delivering the composition comprises topically applying the composition to the plant or soil.

161. 161. The method of any one of claims 104-160, wherein delivering the composition comprises spraying the composition onto leaves, stems, branches, seeds, fruits, flowers, or roots of the plant, and optionally, delivering comprises spraying the composition onto the leaves of the plant.

162. 162. The method of any one of claims 104-161, wherein delivering the composition to the plant comprises a seed treatment.

163. 163. The method of any one of claims 104 to 162, wherein the plant pest is an insect pest belonging to the order Lepidoptera, Coleoptera, Hemiptera, Diptera, or Acari.

164. 163. The method of any one of claims 104-162, wherein the plant pest is an insect that is resistant to one or more biopesticides, optionally wherein the one or more biopesticides comprise one or more Bt proteins, optionally wherein the one or more Bt proteins comprise one or more Bt crystal proteins, optionally wherein the one or more Bt crystal proteins comprise Cry1AB, Cry1F, or Cry1Fa.

165. Plant pests belonging to the order Lepidoptera include Nymphalidae (Nymphalians), Danaidae (Monarchs), Pieridae (Pieridae and Eurema butterflies), Papilionidae (Swallowtails), Lycaenidae (Lycaenidae), Hesperiidae (Hesperiids), Tineidae (Moths), Sesiidae (Clearwig moths), and Pyralidae. dae (pyralid moths), Lasiocampidae (lappet moths), Saturniidae (saturniid moths), Sphingidae (hawk moths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (inchworms, noctuids, and underwing moths), Tortricidae (tortricid moths), and Plutellidae (diamond moths).

166. 166. The method of claim 165, wherein the lepidopteran insect species comprises one or more of Plutella species, Spodoptera species, Helicoverpa species, or Chyrysodeixis species.

167. 166. The method of claim 165, wherein the lepidopteran insect species is a Plutella species, optionally wherein the Plutella species insect is Plutella xylostella (diamond moth).

168. 166. The method of claim 165, wherein the lepidopteran insect species is a Spodoptera species, optionally wherein the Spodoptera species insect is Spodoptera frugiperda (false armyworm).

169. 166. The method of claim 165, wherein the lepidopteran insect species is a Chrysodeixis species, optionally wherein the Chrysodeixis species insect is Chyrysodeixis includens (soybean looper).

170. 166. The method of claim 165, wherein the lepidopteran insect species is Helicoverpa species, optionally wherein the Helicoverpa species insect is Helicoverpa zea (tomato fruitworm).

171. 166. The method of claim 165, wherein the lepidopteran insect species is a Cydia species, optionally wherein the Cydia species insect is Cydia pomonella (the cephalopod moth).

172. 166. The method of claim 165, wherein the lepidopteran insect species is a Trichoplusia species, optionally wherein the Trichoplusia species insect is Trichoplusia ni (cabbage looper).

173. 166. The method of claim 165, wherein the lepidopteran insect species is a Spodoptera species, optionally wherein the Spodoptera species insect is Spodoptera exigua (beet armyworm).

174. 166. The method of claim 165, wherein the lepidopteran insect species is a Spodoptera species, optionally wherein the Spodoptera species insect is Spodoptera litura (tobacco rootworm).

175. 166. The method of claim 165, wherein the lepidopteran insect species is a Spodoptera species, optionally wherein the Spodoptera species insect is Spodoptera litoralis (tomato moth).

176. 166. The method of claim 165, wherein the lepidopteran insect species is Pieris species, optionally wherein the Pieris species insect is Pieris rapae (imported caterpillar).

177. 166. The method of claim 165, wherein the lepidopteran insect species is a Tuta species, optionally wherein the Tuta species insect is Tuta absoluta (tomato moth).

178. 166. The method of claim 165, wherein the lepidopteran insect species is Helicoverpa species, optionally wherein the Helicoverpa species insect is Helicoverpa armigera (cotton boll worm).

179. 166. The method of claim 165, wherein the lepidopteran insect species is a Chrysodeixis species, optionally wherein the Chrysodeixis species insect is Chyrysodeixis acuta (tomato semilooper).

180. 166. The method of claim 165, wherein the lepidopteran insect species is Paramyelois species, and optionally, wherein the Paramyelois species insect is Paramyelois transitella (navel orange worm).

181. 166. The method of claim 165, wherein the lepidopteran insect species is Lobesia species, optionally wherein the Lobesia species insect is Lobesia botrana (European grapevine moth).

182. 165. The method of claim 163 or 164, wherein the insect belongs to the order Coleoptera, and optionally the insect is of a species selected from the group consisting of Leptinotarsa ​​spp., Phyllotreta spp., Cerotoma spp., Diabrotica spp., Tribolium spp., Anthonomus spp. and Alticini spp.

183. 183. The method of claim 182, wherein the insect is a Phyllotreta species, optionally wherein the Phyllotreta species insect is Phyllotreta cruciferae (canola flea beetle).

184. 183. The method of claim 182, wherein the insect is a Phyllotreta species, optionally wherein the Phyllotreta species insect is Phyllotreta striolata (Phyllotreta striolata).

185. 183. The method of claim 182, wherein the insect is a Psylliodes species, optionally wherein the Psylliodes species insect is Psylliodes chrysocephala (cabbage stem flea beetle).

186. 186. The method of any one of claims 104 to 185, wherein the percent mortality of the plant pest is increased by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the yeast particles, and optionally the mortality of the plant pest is increased by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

187. 187. The method of any one of claims 104-186, wherein the percent plant part consumption by plant pests is reduced by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the yeast particles, and optionally the percent plant part consumption by plant pests is reduced by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

188. 188. The method of any one of claims 104 to 187, wherein the percent plant part surface area affected by the plant pest is reduced by at least 10% after delivery compared to the same treatment method comprising the same composition but lacking the yeast particles, and optionally the percent plant part surface area affected by the plant pest is reduced by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100%.

189. 189. The method of any one of claims 104-188, wherein the effective amount of pesticide required to produce a desired outcome is reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 60% compared to a control method, optionally wherein the control method does not include yeast particles, beta-glucan, mannan oligosaccharides, and / or laminarin.

190. The desired outcome is (a) an increase in mortality of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 75% of plant pests after delivery; (b) a reduction in plant part consumption by plant pests after delivery of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 75%; and / or (c) a reduction in the plant portion surface area affected by the plant pest by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 75% after delivery. The method of claim 189,

191. 191. The method of any one of claims 104 to 190, wherein the method comprises delivery of an effective amount of a pesticide.

192. The method of claim 191, wherein the effective amount of the pesticide required to control plant pests in a composition comprising yeast particles or one or more polysaccharides is lower than the effective amount of the pesticide in a composition that does not comprise yeast particles or one or more polysaccharides.

193. 193. The method of any one of claims 104 to 192, further comprising delivering a pesticide having a mode of action different from that of the composition to the plant, the ground, the plant pest, or the food of the plant pest.

194. 194. The method of any one of claims 104 to 193, wherein the plant is selected from the group consisting of Solanaceae plants, Brassicaceae plants, Poaceae plants, Cucurbitaceae plants, Fabaceae plants, Fagaceae plants, Asteraceae plants, Amaryllidaceae plants, Umbelliferae plants, Apiaceae plants, Amranthaceae plants, and Malvaceae plants; optionally, the plant is selected from the group consisting of corn, soybean, and cotton.

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