Antifungal peptides to control plant pathogens

Abstract

The present disclosure relates to methods of increasing plant immunity or resistance to plant pathogens or pests in a plant, or inhibiting a plant pathogen or pest by providing an antimicrobial or antifungal oligopeptide. Also provided herein are antimicrobial or antifungal oligopeptides, and nucleic acids encoding said oligopeptides. Also provided herein are compositions, kits and cells comprising antimicrobial or antifungal oligopeptides.

Description

ANTIFUNGAL PEPTIDES TO CONTROL PLANT PATHOGENSREFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0001] The contents ofthe electronic sequence listing (185952000940SEQLIST.xml; Size: 19,752 bytes; and Date of Creation: February 7, 2024) is herein incorporated by reference in its entirety.FIELD OF INVENTION

[0002] The present invention relates to oligopeptides and their use for increasing plant immunity and inhibiting plant pathogen or pests.CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Patent Application Nos. 63 / 486,652, 63 / 486,653, and 63 / 486,654, all of which were filed on February 23, 2023, and the entire contents of each of which are incorporated herein by reference.BACKGROUND

[0004] Fungal plant pathogens are major threats to food security worldwide. For example, infection by the fungus Botrytis cinerea causes yearly several 100 millions of US dollars crop losses worldwide (Bolton et al., 2006; Dean et al., 2012). Genetic modification approaches, such as the use of genetically- modified organisms (GMOs), and chemical pesticides are widely used to control wide range of plant pathogens, including fungal infections. However, these approaches have limited applicability and can have a significant environmental impact. As such, there is a need for natural and sustainable solutions for controlling plant pathogens.

[0005] Accordingly, there is a need for agents that regulate plant immunity and control infection by common plant pathogen, such as fungi.SUMMARY OF INVENTION

[0006] The disclosure is directed to multiple aspects including, without limitation, Embodiment Al: A method of increasing immunity in a plant comprising providing to the plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, or the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with one, two, three, or four amino acid substitutions, insertions, or deletions thereof.

[0007] Embodiment A2: The method of Embodiment Al, wherein the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0008] Embodiment A3: A method of increasing resistance to a plant pathogen or pest in a plant comprising providing to the plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 or the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 with one, two, three, or four amino acid substitutions, insertions, or deletions.

[0009] Embodiment A4: A method of inhibiting a plant pathogen or pest on a plant comprising providing to the plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 or the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 with one, two, three, or four amino acid substitutions, insertions, or deletions.

[0010] Embodiment A5: The method of Embodiment A3 or A4, wherein the plant pathogen or pest is selected from fungal pathogens or pests, bacterial pathogens or pests, and viral pathogens or pests.

[0011] Embodiment A6: The method of Embodiment A3 or A4, wherein the plant pathogen or pest is a fungal pathogen or pest selected from Cercospora spp., Mycosphaerella spp., Glomerella spp., Cladosporium spp., Diplodia maydis, Fusarium oxysporum, Fusarium graminearum, Fusarium monilforme, Fusarium verticillioides, Cochliobolus sativus, Collectotrichum graminicola, Stagonospora nodorum, Stagonospora avenae, Stenocarpella maydis, Sclerotinia minor, Sclerotinia sclerotiorum, Sclerotinia sp., Altemaria spp., Phytophthora spp., Botrytis spp., Pyrenophora fritici-repentis, Phytophthora parasitica, Phytophthora megasperma fsp. glycinea, Macrophomina phaseolina, Rhizoctonia solani, Sclerotinia sclerotiorum, Fusarium oxysporum, Diaporthe phaseolorum var. sojae (Phomopsis sojae), Diaporthe phaseolorum var. caulivora, Sclerotium rolfsii, Cercospora kikuchii, Cercospora sojina, Peronospora manshurica, Colletotrichum dematium (Colletotichum truncatum), Corynespora cassiicola, Septoria glycines, Phyllosticta sojicola, Altemaria altemata, Pseudomonas syringae p.v. glycinea, Xanthomonas campestris p.v. phaseoli, Microsphaera diffusa, Fusarium semitectum, Phialophora gregata, Glomerella glycines, Phakopsora pachyrhizi, Pythium aphanidermatum, Pythium ultimum, Pythium debaryanum, Fusarium solani; Canola: Albugo Candida, Altenaria brassicae, Leptosphaeria maculans, Rhizoctonia solani, Sclerotinia sclerotiorum, Mycosphaerella brassicicola, Pythium ultimum, Peronospora parasitica, Fusarium roseum, Altemaria altemata; Alfalfa: Clavibacter michiganese subsp. insidiosum, Pythium ultimum, Pythium irregulare, Pythium splendens, Pythium debaryanum, Pythium aphanidermatum, Phytophthora megasperma, Peronospora trifoliorum, Phoma medicaginis var. medicaginis, Cercospora medicaginis, Pseudopeziza medicaginis, Leptotrochila medicaginis, Fusarium oxysporum, Verticillium albo-atrum, Xanthomonas campestris p.v. alfalfae, Aphanomyces euteiches, Stemphylium herbarum, Stemphylium alfalfae, Colletotrichum trifolii,Leptosphaerulina briosiana, Uromyces striatus, Sclerotinia trifoliorum, Stagonospora meliloti, Stemphylium botryosum, Leptotrichila medicaginis; Pseudomonas syringae p.v. atrofaciens, Urocystis agropyri, Xanthomonas campestris p.v. translucens, Pseudomonas syringae p.v. syringae, Altemaria altemata, Cladosporium herbarum, Fusarium graminearum, Fusarium avenaceum, Fusarium culmorum, Ustilago tritici, Ascochyta tritici, Cephalosporium gramineum, Collotetrichum graminicola, Erysiphe graminis f.sp. tritici, Puccinia graminis f.sp. tritici, Puccinia recondite f.sp. tritici, Puccinia striiformis, Pyrenophora tritici-repentis, Septoria nodorum, Septoria tritici, Septoria avenae, Pseudocercosporella herpotrichoides, Rhizoctonia solani, Rhizoctonia cerealis, Gaeumannomyces graminis var. tritici, Pythium aphanidermatum, Pythium arrhenomanes, Pythium ultimum, Bipolaris sorokiniana, Claviceps purpurea, Tilletia tritici, Tilletia laevis, Ustilago tritici, Tilletia indica, Rhizoctonia solani, Pythium arrhenomannes, Pythium gramicola, Pythium aphanidermatum, Plasmopora halstedii, Sclerotinia sclerotiorum, Septoria helianthi, Phomopsis helianthi, Altemaria helianthi, Altemaria zinniae, Botrytis cinerea, Phoma macdonaldii, Macrophomina phaseolina, Erysiphe cichoracearum, Rhizopus oryzae, Rhizopus arrhizus, Rhizopus stolonifer, Puccinia helianthi, Verticillium dahliae, Erwinia carotovorum pv. carotovora, Cephalosporium acremonium, Phytophthora cryptogea, Albugo tragopogonis; Com: Colletotrichum graminicola, Fusarium verticillioides var. subglutinans, Erwinia stewartii, F. verticillioides, Gibberella zeae (Fusarium graminearum), Stenocarpella maydi (Diplodia maydis), Pythium irregulare, Pythium debaryanum, Pythium graminicola, Pythium splendens, Pythium ultimum, Pythium aphanidermatum, Aspergillus flavus, Bipolaris maydis O, T (Cochliobolus heterostrophus), Helminthosporium carbonum I, II & III (Cochliobolus carbonum), Exserohilum turcicum I, II & III, Helminthosporium pedicellatum, Physoderma maydis, Phyllosticta maydis, Kabatiella maydis, Cercospora sorghi, Ustilago maydis, Puccinia sorghi, Puccinia polysora, Macrophomina phaseolina, Penicillium oxalicum, Nigrospora oryzae, Cladosporium herbarum, Curvularia lunata, Curvularia inaequalis, Curvularia pallescens, Clavibacter michiganense subsp. nebraskense, Trichoderma viride, Claviceps sorghi, Pseudomonas avenae, Erwinia chrysanthemi pv. zea, Erwinia carotovora, Com stunt spiroplasma, Diplodia macrospora, Sclerophthora macrospora, Peronosclerospora sorghi, Peronosclerospora philippinensis, Peronosclerospora maydis, Peronosclerospora sacchari, Sphacelotheca reiliana, Physopella zeae, Cephalosporium maydis, Cephalosporium acremonium, Exserohilum turcicum, C. sublineolum, Cercospora sorghi, Gloeocercospora sorghi, Ascochyta sorghina, Pseudomonas syringae p.v. syringae, Xanthomonas campestris p.v. holcicola, Pseudomonas andropogonis, Puccinia purpurea, Macrophomina phaseolina, Perconia circinata, Fusarium verticillioides, Altemaria altemata, Bipolaris sorghicola, Helminthosporium sorghicola, Curvularia lunata, Phoma insidiosa, Pseudomonas avenae (Pseudomonas alboprecipitans), Ramulispora sorghi, Ramulispora sorghicola, Phyllachara sacchari, Sporisorium reilianum (Sphacelotheca reiliana), Sphacelotheca cruenta, Sporisorium sorghi, Clavicepssorghi, Rhizoctonia solani, Acremonium strictum, Sclerophthona macrospora, Peronosclerospora sorghi, Peronosclerospora philippinensis, Sclerospora graminicola, Fusarium graminearum, Fusarium oxysporum, Pythium arrhenomanes, and Pythium graminicola.

[0012] Embodiment A7: The method of Embodiment A3 or A4, wherein the plant pathogen or pest is a bacterial pathogen or pest selected from Pseudomonas spp., Pantoua spp., and Erwinia spp.

[0013] Embodiment A8: The method of Embodiment A3 or A4, wherein the plant pathogen or pest is a viral pathogen or pest selected from cucumber mosaic, tobacco mosaic, and barley yellow dwarf virus, alfalfa mosaic virus (Alfamovirus), Apple chlorotic leaf spot virus (Trichovirus), Apple scar skin viroid (Viroids), Arabis mosaic virus (Nepovirus), Barley mild mosaic virus (Bymovirus), Barley stripe mosaic virus (Hordeivirus), Barley yellow mosaic virus (Bymovirus), Bean common mosaic virus (Potyvirus), Bean yellow mosaic virus (Potyvirus), Beet necrotic yellow vein virus (Furovirus), Blackeye cowpea mosaic virus (Potyvirus), Bean common mosaic virus (Potyvirus), Broad bean wilt virus (Fabavirus), Butterbur mosaic virus (Carlavirus), Carnation mottle virus (Carmovirus), Carnation vein mottle virus (Potyvirus), Cauliflower mosaic virus (Caulimovirus), Chrysanthemum mild mottle virus (Cucumovirus), Tomato aspermy virus (Cucumovirus), Chrysanthemum stunt viroid (Viroids), Citrus mosaic virus, Citrus tristeza virus (Closterovirus), Clover yellow vein virus (Potyvirus), Cocksfoot mottle virus (Sobemovirus), Cucumber green mottle mosaic virus (Tobamovirus), Cucumber mosaic virus (Cucumovirus), Cycas necrotic stunt virus (Nepovirus), Dasheen mosaic virus (Potyvirus), Grapevine Algerian latent virus (Tombusvirus), Konjac mosaic virus (Potyvirus), Melon necrotic spot virus (Carmovirus), Mulberry ringspot virus (Nepovirus), and Narcissus mosaic virus (Potexvirus). Plant viruses are viruses affecting plants. Additional examples of viruses affecting plants include Odontoglossum ringspot virus (Tobamovirus), Papaya ringspot virus (Potyvirus), Peach latent mosaic viroid, Peanut mottle virus (Potyvirus), Peanut stripe virus (Potyvirus), Bean common mosaic virus (Potyvirus), Peanut stunt virus (Cucumovirus), Potato virus A (Potyvirus), Potato virus M (Carlavirus), Potato virus S (Carlavirus), Potato virus X (Potexvirus), Potato virus Y (Potyvirus), Prune dwarf virus (Ilarvirus), Prunus necrotic ringspot virus (Ilarvirus), Radish mosaic virus (Comovirus), Rice black streaked dwarf virus (Fijivirus), Rice dwarf virus (Reovirus), Rice grassy stunt virus (Tenuivirus), Rice stripe virus (Tenuivirus), Rice tungro spherical virus (Sequivirus), Rice waika virus, Rice tungro spherical virus (Sequivirus), Ryegrass mottle virus, Satsuma dwarf virus (Nepovirus), Soil -borne wheat mosaic virus (Furovirus), Southern bean mosaic virus (Sobemovirus), Soybean mosaic virus (Potyvirus), Soybean stunt virus (Cucumovirus), Cucumber mosaic virus (Cucumovirus), Tobacco mosaic virus (Tobamovirus), Tobacco mosaic virus (Tobamovirus), Tomato mosaic virus (Tobamovirus), Tobacco necrosis virus (Necrovirus), Tobacco rattle virus (Tobravirus), Tobacco ringspot virus (Nepovirus), Tomato aspermy virus (Cucumovirus), Tomato black ring virus (Nepovirus), Tomato mosaic virus (Tobamovirus), Tomato ringspot virus (Nepovirus),Tomato spotted wilt virus (Tospovirus), Turnip mosaic virus (Potyvirus), Watermelon mosaic virus 1 (Potyvirus), Papaya ringspot virus (Potyvirus), Watermelon mosaic virus 2 (Potyvirus), Wheat yellow mosaic virus (Bymovirus), and Zucchini yellow mosaic virus (Potyvirus).

[0014] Embodiment A9: The method of Embodiment A3 or A4, wherein the plant pathogen or pest is selected from the group consisting of Septoria tritici, Botrytis cinerea, Altemaria solani, Rhizoctonia solani anastomosis, Phytophthora capsici, and Phytophthora infestans.

[0015] Embodiment A10: The method of any one of Embodiments A1-A9, wherein the oligopeptide is provided by expressing a nucleic acid encoding the oligopeptide.

[0016] Embodiment Al l: The method of Embodiment A 10, wherein the nucleic acid is operably linked to a heterologous promoter.

[0017] Embodiment A 12 : The method of Embodiment A 10 or A 11 , wherein the heterologous promoter is a constitutive promoter, a tissue specific promoter, or an inducible promoter.

[0018] Embodiment A 13: The method of Embodiment A 12, wherein the inducible promoter is induced by fungal infection, optionally a promoter associated with a gene involved in phenylpropanoid metabolism (e.g., phenylalanine ammonia lyase, chaicone synthase promoters), a gene that modifies plant cell walls (e.g., hydroxyproline-rich glycoprotein, glycine-rich protein, and peroxidase promoters), a gene encoding an enzyme that degrade fungal cell walls (e.g., chitinase or glucanase promoters), or a gene encoding a thaumatin-like protein, or optionally a maize Misl promoter or a flax Fisl promoter.

[0019] Embodiment A 14: The method of any one of Embodiments A1-A9, wherein the oligopeptide is provided by application to the plant or a part thereof.

[0020] Embodiment A15: The method of Embodiment A14, wherein the plant part is a leaf, a bud, a root, a shoot, a floral part, or a seed.

[0021] Embodiment A 16 : The method of Embodiment A 14 or A 15 , wherein the oligopeptide is applied as a coating to the seed prior to planting.

[0022] Embodiment A 17: The method of any one of Embodiments A1-A9, wherein the oligopeptide is provided by application to the soil in which the plant is planted.

[0023] Embodiment A 18: The method of any one of Embodiments A1-A9, wherein the oligopeptide is provided by addition to water provided to the plant.

[0024] Embodiment A 19: The method of any one of Embodiments A1-A9, wherein the oligopeptide is provided following harvest to seeds, fruits, and / or plant parts.

[0025] Embodiment A20: The method of any one of Embodiments A14-A19, wherein the application is by a spreader, a power duster, a boom sprayer, a hand sprayers, a spray dusters, or a granular applicator.

[0026] Embodiment A21: The method of any one of Embodiments A1-A20, wherein the miPEP comprises a secretion tag, a tag that promotes entry into a plant cell, or a nuclear localization tag.

[0027] Embodiment A22: The method of Embodiment A21, wherein the tag that promotes entry into a plant cell comprises a cell penetrating peptide linked to the amino acid sequence.

[0028] Embodiment A23: The method of Embodiment A22, wherein the cell penetrating peptide comprises a protein transduction domain, an amphipathic peptide, a synthetic cationic polypeptide, optionally polylysine, polyhistidine, or polyarginine, a dendrimeric polycationic molecule, a peptide vascular endothelial-cadherin cell penetrating peptide, a transportan cell penetrating peptide, a monomer or dimer of HIV- 1 TAT basic domain cell penetrating peptide, a penetratin cell penetrating peptide, a synthetic cationic homoarginine oligopeptide cell penetrating peptide, a gamma zein cell penetrating peptide, a Zea mays knottedl cell penetrating peptide, a Saccharomyces pombe TP10 cell penetrating peptide, a Candida albicans Zebra cell penetrating peptide, an Antennapedia sequence, a TAT sequence , an Antp-3A (Antp mutant) sequence, a Buforin II sequence, a K-FGF sequence, a Ku70 sequence, a prion sequence, a pVEC sequence, a SynBl sequence, a Pep-7 sequence, a HN-1 sequence, a BGSC (Bis- Guanidinium-Spermidine-Cholesterol) sequence, or a BGTC (Bis-Guanidinium-Tren-Cholesterol) sequence.

[0029] Embodiment Bl: An isolated oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21 or the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21 comprising one, two, three, or four amino acid substitutions, insertions, or deletions.

[0030] Embodiment B2: The oligopeptide of Embodiment Bl, wherein the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0031] Embodiment B3: The oligopeptide of Embodiment Bl or B2, comprising the one, two, three, or four amino acid substitutions, insertions, or deletions.

[0032] Embodiment B4: The oligopeptide of any one of Embodiments B 1-B3, wherein the oligopeptide comprises a proline at its N-terminus, an aspartic acid at the C-terminus, or both.

[0033] Embodiment B5: The oligopeptide of any one of Embodiments B 1-B4, wherein the oligopeptide comprises one or more D-amino acids.

[0034] Embodiment B6: The oligopeptide of Embodiment B5, wherein the one or more D-amino acids enhance stability of the oligopeptide as compared to an oligopeptide of the same amino acid sequence comprising all L-amino acids.

[0035] Embodiment B7: The oligopeptide of any one of Embodiments B 1-B6, wherein the oligopeptide comprises a secretion tag, a tag that promotes entry into a plant cell, or a nuclear localization tag.

[0036] Embodiment B8 : The oligopeptide of Embodiment B7, wherein the tag that promotes entry into a plant cell comprises a cell penetrating peptide linked to the amino acid sequence.

[0037] Embodiment B9: The oligopeptide of Embodiment B8, wherein the cell penetrating peptide comprises a protein transduction domain, an amphipathic peptide, a synthetic cationic polypeptide, optionally polylysine, polyhistidine, or polyarginine, a dendrimeric polycationic molecule, a peptide vascular endothelial-cadherin cell penetrating peptide, a transportan cell penetrating peptide, a monomer or dimer of HIV- 1 TAT basic domain cell penetrating peptide, a penetratin cell penetrating peptide, a synthetic cationic homoarginine oligopeptide cell penetrating peptide, a gamma zein cell penetrating peptide, a Zea mays knottedl cell penetrating peptide, a Saccharomyces pombe TP10 cell penetrating peptide, a Candida albicans Zebra cell penetrating peptide, an Antennapedia sequence, a TAT sequence , an Antp-3A (Antp mutant) sequence, a Buforin II sequence, a K-FGF sequence, a Ku70 sequence, a prion sequence, a pVEC sequence, a SynBl sequence, a Pep-7 sequence, a HN-1 sequence, a BGSC (Bis- Guanidinium-Spermidine-Cholesterol) sequence, or a BGTC (Bis-Guanidinium-Tren-Cholesterol) sequence.

[0038] Embodiment C 1 : A composition comprising the oligopeptide of any one of Embodiments B1-B9 with an agriculturally acceptable formulant that can include water, organic solvents, paraffinic oils, vegetable oils, dispersants, emulsifiers, wetting agents, buffering agents, hydrotrope agents, rheology modifiers, antifoam agents and defoamers, antifreeze agents, biocides, dyes, polymer walls, catalysts, thermosetting materials, cross-polymerizing agents, UV protectants, antioxidants, and chelating agents.

[0039] Embodiment C2: A composition comprising the oligopeptide of any one of Embodiments B1-B9 with an agriculturally acceptable carrier, diluent, or excipient.

[0040] Embodiment C3: The composition of Embodiment C2, wherein the agriculturally acceptable carrier, diluent, or excipient comprises a buffer and the pH is in the range of from about 3.0 to about 9.0, or from about 4.5 to about 8.0.

[0041] Embodiment C4: The composition of any one of Embodiments C 1-C3, wherein the oligopeptide is in a concentration range of from about 0. 1 pg / ml to about 100 mg / ml, or from about 5 pg / ml to about 5 mg / ml.

[0042] Embodiment C5: The composition of any one of Embodiments C1-C4, wherein the composition is in the form of a finely-divided particulate solid, granules, pellets, wettable powders, dust, an aqueous suspension, a dispersion, a gel, or an emulsion.

[0043] Embodiment C6: The composition of any one of Embodiments C1-C5, further comprising one or more other active agents selected from the group consisting of a pesticide, a fertilizer, an insecticide, anattractant, a sterilizing agent, an acaricide, a nematocide, a herbicide, a biostimulant, a biological, and a growth regulator.

[0044] Embodiment D 1 : A nucleic acid encoding the oligopeptide of any one of Embodiments B1-B9.

[0045] Embodiment D2: A nucleic acid construct comprising the nucleic acid of Embodiment DI, operably linked to a promoter.

[0046] Embodiment D3: The nucleic acid construct of Embodiment D2, wherein the promoter is a heterologous promoter.

[0047] Embodiment D4: The nucleic acid construct of Embodiment D2 or D3, wherein the promoter is a constitutive promoter, a tissue specific promoter, a development stage specific promoter, or an inducible promoter.

[0048] Embodiment D5: The nucleic acid construct of Embodiment D4, wherein the inducible promoter is induced by fungal infection, optionally a promoter associated with a gene involved in phenylpropanoid metabolism (e.g., phenylalanine ammonia lyase, chaicone synthase promoters), a gene that modifies plant cell walls (e.g., hydroxyproline-rich glycoprotein, glycine-rich protein, and peroxidase promoters), a gene encoding an enzyme that degrade fungal cell walls (e.g., chitinase or glucanase promoters), or a gene encoding a thaumatin-like protein, or a maize Misl promotor or a flax Fisl promoter.

[0049] Embodiment E: A cell comprising the nucleic acid of Embodiment DI or the nucleic acid construct of any one of Embodiments D2-D5.

[0050] Embodiment F: The method of any one of Embodiments A1-A23, oligopeptide of any one of Embodiments B1-B9, composition of any one of Embodiments C1-C6, nucleic acid construct of any one of Embodiments DI -D4, or the cell of Embodiment E, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 19 or SEQ ID NO: 16, or wherein the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NO: 19 or SEQ ID NO: 16, with one, two, three, or four amino acid substitutions, insertions, or deletions thereof.

[0051] Embodiment A24: The method of any one of Embodiments A2-A23 and F, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 19 or wherein the amino acid sequence set forth in SEQ ID NO: 19 with one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and wherein the plant pathogen or pest selected from Phytium ultimum, Rhizoctonia solani, Zymoseptoria tritici, Phythopthora cactorum, Botrytis cinerea, Microdochium nivale, Alternaria alternata, Pyrenophora tritici-repenti, and Fusarium graminearum.

[0052] Embodiment A25: The method of any one of Embodiments A2-A23 and F, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 16 or wherein the amino acid sequence set forth in SEQ ID NO: 16 with one, two, three, or four amino acid substitutions, insertions, ordeletions thereof, and wherein the plant pathogen or pest selected from Zymoseptoria tritici, Phythopthora cactorum, Botrytis cinerea,Microdochium nivale, and Alternaria alternata.

[0053] Embodiment A26: The method of any one of Embodiments A2-A23 and F, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 19 or wherein the amino acid sequence set forth in SEQ ID NO: 19 with one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and wherein the plant pathogen or pest selected from Phytium ultimum, Rhizoctonia solani, Zymoseptoria tritici, Botrytis cinerea, Alternaria alternata, Pyrenophora tritici-repenti, and Fusarium graminearum.

[0054] Embodiment A27: The method of any one of Embodiments A2-A23 and F, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 16 or wherein the amino acid sequence set forth in SEQ ID NO: 16 with one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and wherein the plant pathogen or pest selected from Zymoseptoria tritici, Botrytis cinerea, and Alternaria alternata.BRIEF DESCRIPTION OF THE DRAWINGS

[0055] FIG. 1 shows the results of applying the oligopeptide MP 18279 to tomato leaves or tomato fruits infected with Botrytis cinerea (right) compared to a control (left). The images shown are 4 days post inoculation (“dpi”) for the tomato leaves, and 7 days post inoculation for the fruit. The black circles indicate typical B. cinerea symptoms, including brown lesions and grey mycelium. Leaves or fruits were treated either with MP 18279 or water (solvent for MP 18279) on the site of infection. The arrows indicate sites of infection without visible symptoms on the MP18279-treated leaves or fruits.

[0056] FIG. 2 provides the results of a germination assay of B. cinerea spores treated with MP 18279 (right), compared to a non-freated control (left). The images were taken after either 0 hours (top), 24 hours (middle), or 6 days (bottom) following peptide administration, as indicated at left.

[0057] FIG. 3 shows the results of germination assays of filamentous pathogen spores treated with MP 18279 (gray bars) compared to a control peptide (black bars). The x-axis shows the filamentous pathogen tested, including, from left to right, Septoria tritici, B. cinerea, Alternaria solani, Rhizoctonia, and Phytophthora capsici. The y-axis shows the EC50 of preventing spore germination in mg / L. The results shown are normalized according to the level of germination in the control peptide-freated samples, as indicated by the dotted black line.

[0058] FIG. 4 shows the results of germination assays of filamentous pathogen spores treated with MP 18279 compared to a scrambled control peptide (“sc control”). B. cinerea (left plot), S. tritici (centerplot), and Phytophthora capsid (right plot) were treated with MP 18279 (left three bars of each plot) or the scrambled control peptide (right three bars of each plot). The y-axis shows the EC50 of preventing spore germination in mg / L. Three replicates for each combination of peptide and species are shown, as indicated by the shading of the bars.

[0059] FIGS. 5A-5C show the results of germination assays of filamentous pathogen spores treated with MP18279, truncations of MP18279, variants of MP18279, or a scrambled control peptide (“sc”). In FIG. 5A, the truncations of MP18279 were generated by removing amino acids from the C-terminus of the peptide. B. cinerea (left plot), S. tritid (center plot), and P. capsici (right plot) were treated with the full-length, 10 amino acid MP18279, truncations of MP18279 that were 5-9 amino acids in length, or the scrambled control peptide. The x-axis of each plot indicates the peptide that was used including, from left to right, the scrambled control, the full- length, 10 ammo acid MP18279, and the 9-, 8-, 7-, 6-, and 5-amino acid truncations. The y-axis of each plot shows the EC50 of preventing spore germination in mg / L. Truncations that were active in preventing spore germination are indicated, and truncations that were not active are indicated. The dotted line indicates the level of germination in the full-length MP18279-treated samples. In FIG. 5B, the truncations of MP18279 were generated by removing amino acids from the N-terminus of the peptide. B. cinerea (left plot), S. tritid (center plot), and P. capsid (right plot) were treated with the full-length, 10 amino acid MP18279, truncations of MP18279 that were 5-9 amino acids in length, or the scrambled control peptide. The x-axis of each plot indicates the peptide that was used including, from left to right, the scrambled control, the full-length, 10 amino acid MP18279, and the 9-, 8-, 7-, 6-, and 5-amino acid truncations. The y-axis of each plot shows the EC50 of preventing spore germination in mg / L. Truncations that were active in preventing spore germination are indicated, and truncations that were not active are indicated. The dotted line indicates the level of germination in the full-length MP18279-treated samples. In FIG. 5C, S. tritici spores were treated with variants of MP 18279 with amino-acids additions and / or changes or scrambled control peptide. The y-axis of each plot shows the EC50 of preventing spore germination in mg / L.

[0060] FIG. 6 shows the level of B. cinerea control on tomato leaves achieved by various concentrations of MP 18279. As indicated on the x-axis, 2.5 x 105spores in 1 ml were incubated with 1 pM, 25 pM, 5 OpM, 75pM, or lOOpM, of MP18279, as compared to a control sample (“Ctrl”). The spores and peptides were added to tomato leaves as droplets, and lesion size was measured after two days. The y-axis shows the10RECTIFIED SHEET (RULE 91) ISA / EPpercentage of B. cinerea control. Below the histogram, representative images showing the phenotypes of B. cinerea lesions two days after peptide application are shown. The circled regions in the images are B. cinerea lesions, “n.s ” indicates not significant.

[0061] FIGS. 7A-7B show the results of spray application of MP 18279 to tomato plants infected with B. cinerea (Tomato Grey Mold). FIG. 7A shows representative images of a tomato plant infected with B. cinerea (left), and a tomato plant infected with B. cinerea and, one hour after infection, sprayed with 200 g / ha MP 18279 (right). FIG. 7B shows the percentage of disease control in tomato plants infected with B. cinerea and, one hour after infection, sprayed with 200-0.2 grams per hectare (“g / ha”) MP 18279. The level of disease control shown is 4 days after infection (“dai”). The x-axis indicates the sample, including, from left to right, an infected control (0% disease control) and treatment with 200 g / ha (40 % disease control), 20 g / ha (39% disease control), 2 g / ha (31% disease control), or 0.2 g / ha (13% disease control) MP 18279. The y-axis indicates the percentage of disease control.

[0062] FIGS. 8A-8B show the results of spray application of MP 18279 to tomato or wheat plants subsequently inoculated with pathogens. FIG. 8A shows the percentage of disease control in tomato plants sprayed with 3 grams per hectare (“g / ha”) MP 18279 and, two hours after MP 18279 application, inoculated with Phytophthora infestans (Tomato Late Blight). The x-axis indicates the sample, including, from left to right, an infected control (0% disease control) and treatment with 3 g / ha MP 18279 (32% disease control). The y-axis indicates the percentage of disease control. FIG. 8B shows the percentage of disease control in wheat plants sprayed with 300 grams per hectare (“g / ha”) MP 18279 and, two hours after MP 18279 application, inoculated with S. tritici (Wheat Brown Rust). The x-axis indicates the sample, including, from left to right, an infected control (0% disease control) and treatment with 300 g / ha MP18279 (38% disease control). The y-axis indicates the percentage of disease control.

[0063] FIGS. 9A-9C show the results of experiments analyzing the stability of MP 18279, compared to a variant of MP18279 with D-amino acid residues at the N- and C-terminus (“MP18279D1D10”). FIG. 9A shows the level of peptide stability over time in tomato leaf wash off solutions. The x-axis shows the storage time in hours, and the y-axis shows the percentage of peptide purity. Results are shown for MP18279 (light gray line) and MP18279D1D10 (black line), and the 20% difference in stability between the two peptides is indicated. FIG. 9B shows the level of B. cinerea control on tomato leaves achieved by MP18279 and MP18279D1D10. The x-axis indicates the sample, including, from left to right, an infected control, a scrambled peptide control, MP18279, and MP18279D1D10. The y-axis shows the percentage of B. cinerea control. FIG. 9C shows the percentage of disease control in tomato plants sprayed with 30 g / ha MP18279 or MP18279D1D10 and subsequently infected with P. infestans (Tomato Late Blight). The x-axis indicates the sample, including, from left to right, an infected control, MP18279(24% disease control), and MP18279D1D10 (22% disease control). The y-axis shows the percentage of P. infestans control.

[0064] FIG. 10 shows the abundance of MP 18279 present over time in tomato leaf wash off solutions compared to the 7 amino acid variant of MP18279 (“MP18865”). The x-axis indicates the sample, including, from left to right, MP18279 stored in the solution for 17-20 hours (901% peptide abundance), MP18865 stored in the solution for 17-20 hours (97% peptide abundance), MP18279 stored in the solution for 1 week (0% peptide abundance), and MP 18865 stored in the solution for 1 week (12% peptide abundance). The y-axis indicates the relative level amount of peptide.

[0065] FIG. 11 shows bar plot results of testing antimicrobial activity of peptides MP 19919, MP 19594, and a negative control peptide against seven pathogens (plotted along the horizontal axis). Half maximal effective concentration (EC50) in mg / L is plotted along the vertical axis to the left. Dotted horizontal lines correspond to demarcations of activity, which are plotted along the vertical axis to the right. The striped bars show activity of a negative control peptide, the black bars show activity of MP 19594, and light dotted bars show activity of MP 19919, and each measurement of MP 19594 and MP 19919 activity is listed (in mg / L) above each bar.

[0066] FIG. 12 shows bar plot results of measuring radial growth of 3 pathogen strains against peptides MP19919, MP19594, and a negative control peptide. The % control of pathogen relative to the colony growth (mm) of the negative control (unamended agar) is plotted along the vertical axis. The tested peptides MP 19919, MP 19594, negative control peptide, and positive control Amistar® commercial fungicide are plotted along the horizontal axis. % control is also listed above each bar. The relevant pathogen strain is above each panel Pythium ulimum for left panel, Rhizoctonia solani for middle panel, and Sclerotinia sclerotiorum for right panel).

[0067] FIG. 13 shows bar plot results of measuring each of 3 pathogen strains’ colony diameter (mm, vertical axis) when tested against peptides MP19919, MP19594, a negative control peptide, positive control Amistar® commercial fungicide, and a negative control of unamended agar (all plotted along the horizontal axis). Each measurement of colony diameter (mm) is also listed above each bar. The relevant pathogen strain is above each panel Pythium ulimum for left panel, Rhizoctonia solani for middle panel, and Sclerotinia sclerotiorum for right panel).

[0068] FIG. 14 shows bar plot results of measuring radial growth of 3 pathogen strains against peptides MP 19919, MP 19594, a negative control peptide, a negative control of unamended agar, and a positive control of Amistar® commercial fungicide (plotted along the horizontal axis). The % colony growth relative to the unamended agar is plotted along the vertical axis. % control is also listed above each bar.The relevant pathogen strain is above each panel (Pythium ulimum for left panel, Rhizoctonia solani for middle panel, and Sclerotinia sclerotiorum for right panel).DETAILED DESCRIPTION

[0069] Provided herein are methods of increasing plant immunity, methods of increasing resistance to a plant pathogen or pest in a plant, and methods of inhibiting a plant pathogen or pest. Also provided herein are antimicrobial oligopeptides, nucleic acids encoding said oligopeptides, and compositions comprising said oligopeptides.

[0070] The methods and oligopeptides of the disclosure are based, at least in part, on Applicant’s surprising discovery of an oligopeptide that can be used to regulate immunity and resistance to pathogens in plants. In particular, Applicants found that application of an oligopeptide to plants abolished infection by a broad spectrum of fungal pathogens. Moreover, the oligopeptide was effective at promoting immunity in both monocots and dicots. Furthermore, the oligopeptide was capable of inhibiting fungal pathogens in vitro, and higher stability variants of the oligopeptide also exhibited the antifungal activity.

[0071] As used herein, an “amino acid” or “amino acid residue” refers to any naturally occurring amino acid, any non-naturally occurring amino acid, any modified amino acid, including derivatized amino acids, or any amino acid mimetic known in the art. In some embodiments, the amino acids are D-amino acids. In some embodiments, the amino acids are L-amino acids. The amino acid may be referred by both their common three letter abbreviation and single letter abbreviation.

[0072] As used herein, the terms “peptide” or “oligopeptide” refer to any peptide structure comprising or consisting of two or more amino acids, including chemical modifications and derivatives of amino acids. In some embodiments, the oligopeptide comprises an amino acid sequence capable inducing increased immunity, or increased resistance to a plant pathogen or pest, in a plant exposed to said amino acid sequence. In some embodiments, the oligopeptide comprises an amino acid sequence that is capable of inhibiting a plant pathogen or pest. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide. In some embodiments, the oligopeptide is 4 to 50 or 6 to 30 amino acids in length.

[0073] As used herein, the term “purified” molecule refers to biological or synthetic molecules that are removed from their natural environment and are isolated or separated and are free from other components with which they are naturally associated.

[0074] The term "isolated" in reference to a molecule including a nucleic acid, construct, vector, etc., may refer to a molecule that is not found in nature and / or is present in a context in which it is not found in nature. The term “isolated” may also refer to a molecule that has undergone at least one step towardsbeing isolated or concentrated or enriched from a more complex solution or source. The term “isolated,” however, is in no way intended to limit the molecule to a particular location or state. For example, an isolated nucleic acid molecule includes the nucleic acid molecule introduced into the genome of a cell in a position where it is not found in nature or when it is resident in progeny of cells into which the nucleic acid molecule has been introduced into its genome in a position where it is not found in nature.

[0075] The term “sequence identity” refers to the degree of similarity between two nucleic acid sequences, or two amino acid sequences, is expressed in terms of the similarity between the sequences, and otherwise referred to as sequence identity. Sequence identity is frequently measured in terms of percentage identity (or similarity or homology); the higher the percentage, the more similar the two sequences are.I. Methods

[0076] Provided herein are methods of increasing immunity in a plant. Also provided herein are methods of increasing resistance to a plant pathogen or pest in a plant, and methods of inhibiting a plant pathogen or pest.Peptides

[0077] In some embodiments, the method comprises providing an oligopeptide to a plant.

[0078] In some embodiments, the method comprises providing to a plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21. In some embodiments, the oligopeptide comprises an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions as compared to any of SEQ ID NOS: 1-5 and 14-21. In some embodiments, the oligopeptide comprises amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any of SEQ ID NO: 1-5 and 14-21. In some embodiments, the oligopeptide is 4-50 or 5-30 amino acids in length. In some embodiments, the oligopeptide comprises a proline at its N-terminus, an aspartic acid at the C-terminus, or both. In some embodiments, the oligopeptide comprises one or more D-amino acids. In some embodiments, the one or more D-amino acids enhance stability of the oligopeptide as compared to an oligopeptide of the same amino acid sequence comprising all L-amino acids. In some embodiments, the oligopeptide is an antimicrobial or antifungal oligopeptide.Oligopeptide Application

[0079] In some embodiments, the method comprises providing an oligopeptide to a plant. In some embodiments, the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5and 14-21, or an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0080] The oligopeptide may be provided to a plant by any suitable method known in the art. In some embodiments, the method comprises providing an oligopeptide to a plant by foliar spray, foliar drench, drip irrigation, coating, mixing, pouring, dusting, atomizing, soil irrigation, fumigation, soil injection, seepage irrigation, sprinklers or manual irrigation. In some embodiments, the method comprises providing an oligopeptide to a plant using a spreader, a power duster, a boom sprayer, a hand sprayers, a spray dusters, or a granular applicator. In some embodiments, the method comprises providing an oligopeptide to a plant by foliar spray. In some embodiments, the method comprises providing to the plant an oligopeptide at a concentration of 2 g / ha to 300 g / ha. In some embodiments, the oligopeptide is provided at a concentration of 2 g / ha, 5 g / ha, 10 g / ha , 15 g / ha, 20 g / ha, 25 g / ha, 30 g / ha, 35 g / ha, 40 g / ha, 45 g / ha, 50 g / ha, 55 g / ha, 60 g / ha, 65 g / ha, 70 g / ha, 75 g / ha, 80 g / ha, 85 g / ha, 90 g / ha, 95 g / ha, 100 g / ha, 110 g / ha, 120 g / ha, 130 g / ha, 140 g / ha, 150 g / ha, 160 g / ha, 170 g / ha, 180 g / ha, 190 g / ha, 200 g / ha, 220 g / ha, 240 g / ha, 260 g / ha, 280 g / ha, or 300 g / ha. In some embodiments, the oligopeptide is provided to one or more of a leaf, a bud, a root, a shoot, a floral part, or a seed. In some embodiments, the oligopeptide is provided to a plant or part thereof after harvest. In some embodiments, the oligopeptide is provided to a plant or part thereof prior to harvest. In some embodiments, the oligopeptide is provided to the plant prior, during, or after an infection with a pathogen or pest. In some embodiments, the oligopeptide is provided to a plant by application to the soil in which the plant is planted. In some embodiments, the oligopeptide is provided to a plant by addition of water provided to the plant. In some embodiments, the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, or an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0081] In some embodiments, the method comprises preparing a composition comprising the oligopeptide. In some embodiments, the composition is an agriculturally acceptable composition. In some embodiments, the composition comprises an oligopeptide and an agriculturally acceptable formulant. In some embodiments, the agriculturally acceptable formulant comprises one or more of a water, organic solvents, paraffinic oils, vegetable oils, dispersants, emulsifiers, wetting agents, buffering agents, hydrotrope agents, rheology modifiers, antifoam agents and defoamers, antifreeze agents, biocides, dyes, polymer walls, catalysts, thermosetting materials, cross-polymerizing agents, UV protectants, antioxidants, and chelating agents. In some embodiments, the composition comprises an oligopeptide at a concentration range of from about 0.1 pg / ml to about 100 mg / ml, or from about 5 pg / ml to about 5 mg / ml. In some embodiments, the composition further comprises one or more other active agents selectedfrom the group consisting of a pesticide, a fertilizer, an insecticide, an attractant, a sterilizing agent, an acaricide, a nematocide, a herbicide, a biostimulant, a biological, and a growth regulator. In some embodiments, the composition is formulated as a liquid, a gel, an emulsion, a suspension, an encapsulation, a solid, a powder, an aerosol, a paste, a coating, a spray, a soil drench, a microcapsule, an emulsifiable concentrate, or as granules. In some embodiments, the composition is in the form of a finely-divided particulate solid, granules, pellets, wettable powders, dust, an aqueous suspension, a dispersion, a gel, or an emulsion. In some embodiments, the agriculturally acceptable composition is formulated as a seed treatment, a foliar spray, a foliar drench, a Ready-To-Use (RTU) formulation, a produce coating, a suspension concentrate, a tank-mix, an aerosol, a root dip, a soil treatment, a dipping formulation, an irrigation formulation, or a sprinkler formulation. In some embodiments, the agriculturally acceptable composition comprising an oligopeptide is a foliar spray composition. In some embodiments, the composition comprises an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21, or an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0082] In some embodiments, the method comprises preparing composition comprising a oligopeptide and an agriculturally acceptable carrier, diluent or excipient. In some embodiments, the composition comprises an oligopeptide and an agriculturally acceptable carrier, diluent or excipient. In some embodiments, the agriculturally acceptable carrier, diluent, or excipient comprises a buffer and the pH is in the range of from about 3.0 to about 9.0 or about 4.5 to about 8.0. In some embodiments, the agriculturally acceptable carrier comprises a solid carrier, a liquid carrier, a gel carrier, a suspension, or an emulsion. In some embodiments, the agriculturally acceptable carrier comprises an adjuvant, an inert component, a dispersant, a surfactant, a humectant, an emulsifier, a thickener, a wetting agent, a fertilizer, a mineral, a solvent, a tackifier, a binder, or a stabilizer. In some embodiments, the agriculturally acceptable composition comprising an oligopeptide and a surfactant or humectant. In some embodiments, the composition comprises an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, or an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and an agriculturally acceptable carrier, diluent or excipient. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0083] In some embodiments, the oligopeptide is provided by application to a plant or part thereof. In some embodiments, the plant part is a fruit, leaf, a bud, a root, a shoot, a floral part, or a seed. In some embodiments, the oligopeptide is provided to a plant or part thereof after harvest. In some embodiments, the oligopeptide is provided to a plant or part thereof prior to harvest. In some embodiments, the oligopeptide is provided to a plant by application to the soil in which the plant is planted. In someembodiments, the oligopeptide is provided to a plant by addition of water provided to the plant. In some embodiments, the oligopeptide is applied to a plant by a spreader, a power duster, a boom sprayer, a hand sprayers, a spray dusters, or a granular applicator. In some embodiments, the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0084] In some embodiments, the method comprises providing an oligopeptide to a plant by expressing a nucleic acid encoding the oligopeptide. In some embodiments, the nucleic acid is operably linked to a heterologous promoter. In some embodiments, the heterologous promoter is a constitutive promoter, a tissue specific promoter, a development stage specific promoter, or an inducible promoter. In some embodiments, the nucleic acid is operably linked to an inducible promoter. In some embodiments, the inducible promoter is induced by fungal infection, optionally a promoter associated with a gene involved in phenylpropanoid metabolism (e.g., phenylalanine ammonia lyase, chaicone synthase promoters), a gene that modifies plant cell walls (e.g., hydroxyproline-rich glycoprotein, glycine-rich protein, and peroxidase promoters), a gene encoding an enzyme that degrade fungal cell walls (e.g., chitinase or glucanase promoters), a gene encoding a thaumatin-like protein, or a maize or flax promoters. For instance, maize or flax promoters, such as Misl and Fisl promoters, respectively, are promoters induced by fungal infections in plants which may be used with the methods of the disclosure (US Patent Appl. Pub. No. 20020115849). In some embodiments, the nucleic acid expresses an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0085] In some embodiments, the method comprises providing a nucleic acid encoding an oligopeptide to a plant. In some embodiments, the nucleic acid encoding an oligopeptide is provided to a plant by transformation. In some embodiment, transformation comprises Agrobacterium-mediated transformation, micro-projectile-mediated transformation, sonication, electroporation, or liposome- or spheroplast- mediated vector delivery. In some embodiments, the nucleic acids encoding the peptide are added to a plant by genome editing. Gene editing may be performed by any method known in the art, including, but not limited to using zinc finger-nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), oligonucleotide-directed mutagenesis (ODM), a clustered regularly interspaced short palindromic repeats (CRISPR) / Cas, or by gene writing (see, e.g., PCT Patent Application Publication W02020 / 047124). In some embodiments, the nucleic acid encoding the oligopeptide are added to a plant by crossing a first plant comprising the nucleic acid encoding the oligopeptide to a second plant. In some embodiments, a plant comprising the nucleic acid expresses the oligopeptide encode by said nucleic acid.In some embodiments, the nucleic acid encodes an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.Plant Immunity

[0086] In some embodiments, the method comprises increasing immunity in a plant. In some embodiments, the method comprises providing to the plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof.

[0087] As used herein, “immunity in a plant” and “plant immunity” refer the ability of a plant to recognize triggers, signatures and / or patterns associated with exposure to a pathogen or pest, and respond accordingly (e.g., through gene induction) to ward off and / or recover from the pathogen.

[0088] Any suitable method of determining plant immunity known in the art may be used. For example, plant immunity may be measure as a reduction in pest number on the plant when compared to an untreated plant, or as a reduction in physical damage to the plant when compared to an untreated plant. Physical damage includes feeding damage and boring damage, and may manifest in a variety of plant phenotypes, including, but not limited to, chewed or ragged leaves, missing leaves, tunnels in leaves, holes in stems, leaf distortion, leaf discoloration, leaf spotting, wilting, stunted growth, girdled or dead stems, yellowing, breakage damage, or root damage. Convex hull analysis may be used to examine the physical damage to an infected plant.

[0089] In some embodiments, increasing plant immunity comprises increasing disease control in a plant by providing an antimicrobial oligopeptide to the plant. The term "disease control," as used herein, means killing, reducing in numbers, and / or reducing growth, feeding or normal physiological development of any or all life stages of a plant pathogen or pest, and / or reduction of the effects of a plant pest infection and / or infestation. In some embodiments, increasing immunity in a plant comprises increasing disease control in a plant by 10% to 90% as compared to an untreated infected plant. In some embodiments, increasing immunity in a plant comprises increasing disease control in a plant by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, or at least about 90% as compared to an untreated infected plant. In some embodiments, increasing immunity in a plant comprises increasing disease control in a plant by 2- to 20-fold as compared to an untreated infected plant. In some embodiments, increasing immunity in a plant comprises increasingdisease control in a plant by 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 12-fold, 14-fold, 16-fold, 18-fold, or 20-fold as compared to an untreated infected plant. In some embodiments, the antimicrobial oligopeptide is an oligopeptide comprising the amino acid sequence set forth in SEQ ID NO: 1-5 and 14-21, or an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions thereof.Plants

[0090] In some embodiments, the method comprises providing an antimicrobial oligopeptide to a plant. In some embodiments, the antimicrobial oligopeptide is an oligopeptide comprising the amino acid sequence set forth in in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof.

[0091] In some embodiment, the plant is a dicot or monocot plant. In some embodiments, the plant is a crop plant. In some embodiments, the plant is a row crop plant, a fruit-producing plant, a tree, a vine, a vegetable, or an ornamental plants (e.g. ornamental flowers, trees, shrubs, groundcovers, and turf grasses). In some embodiments, the plant is Solanum spp or Triticum spp. Other exemplary types of types of plants include, without limitiation, Medicago sativa, Prunus dulcis, Arabidopsis spp., Phaseolus spp., Fragaria spp., Malus domestica, Prunus spp., Asparagus officinalis, Arabidopsis spp., Musa spp., Hordeum vulgare, Phaseolus spp., Vaccinium spp., Theobroma cacao, Brassica spp., Dianthus caryophyllus, Daucus carota sativus, Manihot esculentum, Prunus avium, Cicer arietinum, Cichorium intybus, Capsicum spp., Chrysanthemum spp., Cocos nucifera, Coffea spp., Gossypium hirsutum L, Vigna spp., Vicia faba, Cucumis sativus, Ribes spp., Phoenix dactylifera, Solanum spp., Eucalyptus spp., Linum usitatissumum L), Pelargonium spp., Vitus spp., Psidium guajava, Humulus lupulus, Cannabis sativa and Cannabis spp., Iris spp. / Lactuca sativa, Citrus spp., Zea mays L, Mangifera indie), Garcinia mangostan), melon Cucumis mel), (Setaria spp., Echinochloa spp., Eleusine spp., Panicum spp., Pennisetum spp. / Avena sativ), Ellis quineensis, Olea europaea, Allium spp., Carica papaya, Prunus persic), Pyrus spp. / Pisa sativum, Arachis hypogaea, Paeonia spp., Petunia spp., Ananas comosus, Musa spp., Prunus domestica, (Euphorbia pulcherrima, Populus spp Cucurbita spp., Oryza sativa L, Rosa spp., Hevea brasiliensis, Secale cereale, Carthamus tinctorius L, Sesame indium, Sorghum bicolor, Glycine max L, Fragaria pp., Beta vulgari), Saccharum spp., Helianthus annuus, Ipomoea batatas, Camellia sinensis, Nicotiana tabacum L., Lycopersicon esculentum, Tulipa spp., Juglans spp. L, Citrulus lanatus, Triticum aestivum, and Discorea spp. In some embodiments, the plant is a wild plant variety. In some embodiments, the plant is a domesticated plant variety. In some embodiments, the plant is a hybrid plant variety. In some embodiments, the plant is a genetically modified plant and / or a gene-edited plant. Plant Pathogens and Pests

[0092] Provided herein are methods of increasing resistance to a plant pathogen or pest in a plant. As used herein, “resistance” or “increased resistance” in a plant to a pest or pathogen is an indication that the plant is more able to reduce the effect of the pest or pathogen than a non-resistant or less resistant plant.

[0093] Also provided herein are methods of inhibiting a plant pathogen or pest on a plant. As used herein, “inhibiting a plant pathogen or pest” means killing, disabling, immobilizing, reducing the numbers of, or otherwise rendering a plant pathogen or pest substantially incapable of causing harm to a plant.

[0094] In some embodiments, the method comprises providing to a plant an oligopeptide comprising the amino acid sequence set forth in in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide. In some embodiments, the plant pathogen or pest is a fungus, bacteria, virus, or a eukaryote.

[0095] In some embodiments, the plant pathogen or pest is a fungus. Exemplary plant fungal pathogens or pests that may be targeted by the present methods include, without limitation, Cercospora spp., Mycosphaerella spp., Glomerella spp., Cladosporium spp., Diplodia maydis, Fusarium oxysporum, Fusarium graminearum, Fusarium monilforme, Fusarium verticillioides, Cochliobolus sativus, Collectotrichum graminicola, Stagonospora nodorum, Stagonospora avenae, Stenocarpella maydis, Sclerotinia minor, Sclerotinia sclerotiorum, Sclerotinia sp., Alternaria spp., Phytophthora spp., Botrytis spp., Pyrenophora tritici-repentis, and , Phytophthora parasitica. Specific pathogens for major crops include: Soybeans: Phytophthora megasperma fsp. glycinea, Macrophomina phaseolina, Rhizoctonia solani, Sclerotinia sclerotiorum, Fusarium oxysporum, Diaporthe phaseolorum var. sojae (Phomopsis sojae), Diaporthe phaseolorum var. caulivora, Sclerotium rolfsii, Cercospora kikuchii, Cercospora sojina, Peronospora manshurica, Colletotrichum dematium (Colletotichum truncatum), Corynespora cassiicola, Septoria glycines, Phyllosticta sojicola, Alternaria alternata, Pseudomonas syringae p.v. glycinea, Xanthomonas campestris p.v. phaseoli, Microsphaera diffusa, Fusarium semitectum, Phialophora gregata, Glomerella glycines, Phakopsora pachyrhizi, Pythium aphanidermatum, Pythium ultimum, Pythium debaryanum, Fusarium solani; Canola: Albugo Candida, Altenaria brassicae, Leptosphaeria maculans, Rhizoctonia solani, Sclerotinia sclerotiorum, Mycosphaerella brassicicola, Pythium ultimum, Peronospora parasitica, Fusarium roseum, Alternaria alternata; Alfalfa: Clavibacter michiganese subsp. insidiosum, Pythium ultimum, Pythium irregulare, Pythium splendens, Pythium debaryanum, Pythium aphanidermatum, Phytophthora megasperma, Peronospora trifoliorum, Phoma medicaginis var. medicaginis, Cercospora medicaginis, Pseudopeziza medicaginis, Leptotrochila medicaginis, Fusarium oxysporum, Verticillium albo-atrum, Xanthomonas campestris p.v. alfalfae, Aphanomyces euteiches, Stemphylium herbarum, Stemphylium alfalfae, Colletotrichum trifolii, Leptosphaerulina briosiana,Uromyces striatus, Sclerotinia trifoliorum, Stagonospora meliloti, Stemphylium botryosum, Leptotrichila medicaginis; Whca / : Pseudomonas syringae p.v. atrofaciens, Urocystis agropyri, Xanthomonas campestris p.v. translucens, Pseudomonas syringae p.v. syringae, Alternaria altemata, Cladosporium herbarum, Fusarium graminearum, Fusarium avenaceum, Fusarium culmorum, Ustilago tritici, Ascochyta tritici, Cephalosporium gramineum, Collotetrichum graminicola, Erysiphe graminis f.sp. tritici, Puccinia graminis f.sp. tritici, Puccinia recondite f.sp. tritici, Puccinia striiformis, Pyrenophora tritici-repentis, Septoria nodorum, Septoria tritici, Septoria avenae, P seudocercosporella herpotrichoides, Rhizoctonia solani, Rhizoctonia cerealis, Gaeumannomyces graminis var. tritici, Pythium aphanidermatum, Pythium arrhenomanes, Pythium ultimum, Bipolaris sorokiniana, Claviceps purpurea, Tilletia tritici, Tilletia laevis, Ustilago tritici, Tilletia indica, Rhizoctonia solani, Pythium arrhenomannes, Pythium gramicola, Pythium aphanidermatum, Sunflower: Plasmopora halstedii, SclerotiniaSorghum: Exserohilum turcicum, C. sublineolum, Cercospora sorghi, Gloeocercospora sorghi, Ascochyta sorghina, Pseudomonas syringae p.v. syringae, Xanthomonas campestris p.v. holcicola, Pseudomonas andropogonis, Puccinia purpurea, Macrophomina phaseolina, Perconia circinata, Fusarium verticillioides, Alternaria altemata, Bipolaris sorghicola, Helminthosporium sorghicola, Curvularia lunata, Phoma insidiosa, Pseudomonas avenae (Pseudomonas alboprecipitans), Ramulispora sorghi, Ramulispora sorghicola, Phyllachara sacchari, Sporisorium reilianum (Sphacelotheca reiliana),Sphacelotheca cruenta, Sporisorium sorghi, Claviceps sorghi, Rhizoctonia solani, Acremonium strictum, Sclerophthona macrospora, Peronosclerospora sorghi, Peronosclerospora philippinensis, Sclerospora graminicola, Fusarium graminearum, Fusarium oxysporum, Pythium arrhenomanes, Pythium graminicola, etc.

[0096] In some embodiments, the plant pathogen or pest is a bacteria. Examples of plant pathogenic bacteria include those belonging to Pseudomonas spp., Pantoua spp., and Erwinia spp. Additional plant pathogenic bacteria are described in Robert W. Jackson, Plant Pathogenic Bacteria: Genomics and Molecular Biology, published by Horizon Scientific Press, 2009, ISBN 1904455379, 9781904455370; Samuel S. Gnanamanickam, Plant-Associated Bacteria, published by Springer, 2007, ISBN 1402045379, 9781402045370; Martin Dworkin et al., The Prokaryotes: a handbook on the biology of bacteria, Published by Springer, 2006, ISBN 0387254927, 9780387254920; George N. Agrios, Plant pathology, published by Academic Press, 2005, ISBN 0120445654, 9780120445653; and David W. Parry, Plant pathology in agriculture, published by CUP Archive, 1990, ISBN 0521368901, 9780521368902.

[0097] In some embodiments, the plant pathogen or pest is a virus. Examples of viruses that may cause infections in plants plant include, without limitation, cucumber mosaic, tobacco mosaic, and barley yellow dwarf virus, alfalfa mosaic virus (Alfamovirus), Apple chlorotic leaf spot virus (Tricho virus), Apple scar skin viroid (Viroids), Arabis mosaic virus (Nepovirus), Barley mild mosaic virus (Bymovirus), Barley stripe mosaic virus (Hordeivirus), Barley yellow mosaic virus (Bymovirus), Bean common mosaic virus (Potyvirus), Bean yellow mosaic virus (Potyvirus), Beet necrotic yellow vein virus (Furovirus), Blackeye cowpea mosaic virus (Potyvirus), Bean common mosaic virus (Potyvirus), Broad bean wilt virus (Fabavirus), Butterbur mosaic virus (Carlavirus), Carnation mottle virus (Carmovirus), Carnation vein mottle virus (Potyvirus), Cauliflower mosaic virus (Caulimovirus), Chrysanthemum mild mottle virus (Cucumovirus), Tomato aspermy virus (Cucumovirus), Chrysanthemum stunt viroid (Viroids), Citrus mosaic virus, Citrus tristeza virus (Closterovirus), Clover yellow vein virus (Potyvirus), Cocksfoot mottle virus (Sobemovirus), Cucumber green mottle mosaic virus (Tobamo virus), Cucumber mosaic virus (Cucumovirus), Cycas necrotic stunt virus (Nepovirus), Dasheen mosaic virus (Potyvirus), Grapevine Algerian latent virus (Tombusvirus), Konjac mosaic virus (Potyvirus), Melon necrotic spot virus (Carmovirus), Mulberry ringspot virus (Nepovirus), and Narcissus mosaic virus (Potexvirus). Plant viruses are viruses affecting plants. Additional examples of viruses affecting plants include Odontoglossum ringspot virus (Tobamovirus), Papaya ringspot virus (Potyvirus), Peach latent mosaic viroid, Peanut mottle virus (Potyvirus), Peanut stripe virus (Potyvirus), Bean common mosaic virus (Potyvirus), Peanut stunt virus (Cucumovirus), Potato virus A (Potyvirus), Potato virus M (Carlavirus), Potato virus S (Carlavirus), Potato virus X (Potexvirus), Potato virus Y (Potyvirus), Prune dwarf virus (liarvirus), Prunus necrotic ringspot virus (liarvirus), Radish mosaic virus (Comovirus), Rice blackstreaked dwarf virus (Fijivirus), Rice dwarf virus (Reovirus), Rice grassy stunt virus (Tenuivirus), Rice stripe virus (Tenuivirus), Rice tungro spherical virus (Sequivirus), Rice waika virus, Rice tungro spherical virus (Sequivirus), Ryegrass mottle virus, Satsuma dwarf virus (Nepovirus), Soil-borne wheat mosaic virus (Furovirus), Southern bean mosaic virus (Sobemovirus), Soybean mosaic virus (Potyvirus), Soybean stunt virus (Cucumovirus), Cucumber mosaic virus (Cucumovirus), Tobacco mosaic virus (Tobamovirus), Tobacco mosaic virus (Tobamovirus), Tomato mosaic virus (Tobamovirus), Tobacco necrosis virus (Necrovirus), Tobacco rattle virus (Tobravirus), Tobacco ringspot virus (Nepovirus), Tomato aspermy virus (Cucumovirus), Tomato black ring virus (Nepovirus), Tomato mosaic virus (Tobamovirus), Tomato ringspot virus (Nepovirus), Tomato spotted wilt virus (Tospovirus), Turnip mosaic virus (Potyvirus), Watermelon mosaic virus 1 (Potyvirus), Papaya ringspot virus (Potyvirus), Watermelon mosaic virus 2 (Potyvirus), Wheat yellow mosaic virus (Bymovirus), and Zucchini yellow mosaic virus (Potyvirus). More plant viruses have been described in F. C. Bawden, Plant Viruses and Virus Diseases, Publisher Biotech Books, 2002, ISBN 8176220647, 9788176220644.

[0098] In some embodiments, the plant pathogen or pest is a eukaryote. In some embodiments, the eukaryote is an insect or a nematode. Agricultural insect pests can be classified into: chewing insects, sucking insects, and soil insects. Common chewing insects are, for example, beet armyworm (Spodoptera exigua), diamondback moth (Plutei la xylostelld), com earworm (Heliolhis zea, a.k.a. boll worm and tomato fruitworm), blister beetles (Epicauta and others), carrot weevils (Listronotus oregonensis, Hyperodes texana), cabbage looper (Trichopulsia ni), grasshopper (several species), flea beetles (e.g., tobacco fleabeetle (Epitrix hirtipennis), eggplant fleabeetle (E. fuscula), potato fleabeetle (E. cucumeri) and other species), fall armyworm (Spodoptera frugiperda), Lesser cornstalk borer (Elasmopalpus lignosellus), Texas leafcutting ant (Atta texana), citrus leafminer (Phyllocnistis citrella), leafminers (Liiriomyza spp.), yellowstriped armyworm (Spodoptera ornithogalli). Common sucking insects are, for example, stink bugs (e.g. Nezara viridula and other species), sharpshooters (Homalodisca spp. and Oncopmetopia spp.), whiteflies (e.g. sliverleaf whitefly, greenhouse whitefly, sweetpotato whitefly (Bemisia tabaci , greenhouse whitefly (Trialeuroides vaporariorum), psyllid (e.g. Asian citrus psyllid), squash bug (Anasa tristis), leaffooted bugs (Leptoglossus spp.), leafhoppers (e.g., bean leafhopper, Empoasca solana, aster leafhopper, Macrosteles fascifrons, western potato leafhopper, Empoasca abrupta, grape leafhopper, variegated leafhopper, beet leafhopper, Circulifer tenellus), aphids (Aphidoidea, e.g. green peach aphid, turnip aphid, melon aphid, potato aphid, rosy apple aphid, spirea aphid,). Common rasping insects include, but are not limited to, thrips (e.g. citrus thrips, western flower thrips (Frankliniella occidentalis), onion thrips (Thrips tabaci), melon thrips, and chili thrips). Common soil insects are, for example, granulate cutworm (Feltia subterranea), mole crickets (e.g. northern mole cricket, Neocurtilla hexadactyla, southern moire cricket Scapteriscus acletus), com rootworm (e.g.Diabrotica undecimpunctata howardi), pillbugs and sowbugs (several species), sweetpotato weevil (Cylas formicarius elegantulus), white grubs (Pyllophaga spp.), wireworms (several species). Exemplary nematodes that act as plant pathogens or pest can be classified into parasitic nematodes, such as root-knot nematodes including Heterodera spp., Meloidogyne spp., and Globodera spp.; cyst nematodes, which include Heterodera spp., Meloidogyne spp. (cotton cyst nematode) and Globodera spp.; and lesion nematodes, which include Pratylenchus spp.

[0099] In some embodiments, the pathogen is a pathogen of Solanum spp. or Triticum spp. In some embodiments, the plant pathogen or pest is a fungi. In some embodiments, the fungi is In some embodiments, the plant pathogen or pest is Septoria tritici, Botrytis cinerea, Alternaria solani, Rhizoctonia spp., Phytophthora capsici, Phytophthora capsicum, or Phytophthora infestans.II. Oligopeptides

[0100] Also provided herein are antimicrobial and antifungal oligopeptides.

[0101] In some embodiments, the method comprises providing to a plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21. In some embodiments, the oligopeptide comprises an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions as compared to any of SEQ ID NOS: 1-5 and 14-21. In some embodiments, the In some embodiments, the oligopeptide comprises amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any of SEQ ID NO: 1-5 and 14-21. In some embodiments, the oligopeptide is 4-50 or 5-30 amino acids in length. In some embodiments, the oligopeptide comprises a proline at its N-terminus, an aspartic acid at the C-terminus, or both. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0102] In some embodiments, the oligopeptide comprises a protease-resistant amino acid sequence. In some embodiments, the oligopeptide comprises one or more modifications compared to a natural oligopeptide. In some embodiments, the oligopeptide comprises one or more modifications that increase stability and / or half-life. In some embodiments, the oligopeptide comprises one or more D-amino acids. In some embodiments, the one or more D-amino acids enhance the stability of the oligopeptide in a cell. In some embodiments, the one or more D-amino acids enhance stability of the oligopeptide as compared to an oligopeptide of the same amino acid sequence comprising all L-amino acids. In some embodiments, the oligopeptide comprises the sequence set forth in ONE OF SEQ ID NO: 1-5 and 14-21, with one or more D-amino acids. In some embodiments, the oligopeptide comprises an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions as compared to any of SEQ ID NOS : 1 -5 and 14-21, with one or more D-amino acids. In some embodiments, the oligopeptide comprising the one or more D-amino acids is an antimicrobial oligopeptide or an antifungal oligopeptide. In some embodiments, the oligopeptide comprises a D amino acid and the N-terminus. In some embodiments, the oligopeptide comprises a D-amino acid at the C-terminus. In some embodiments, the oligopeptide comprises a D-amino acid at the N-terminus and at the C-terminus.

[0103] In some embodiments, the oligopeptide comprises a tag. In some embodiments, the tag is a secretion tag, a tag that promotes entry into a plant cell, or a nuclear localization tag. In some embodiments, the oligopeptide comprises a tag at the N-terminus, the C-terminus, or both. In some embodiments, the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and a tag.

[0104] In some embodiments, the oligopeptide further comprises a cell penetrating peptide linked to the amino acid sequence of the oligopeptide. In some embodiments, the cell penetrating peptide increases cellular delivery and activity in a cell. Cell penetrating peptides useful in the present methods are a class of short peptides with a property to translocate across cell membranes and act as nano-carriers for protein delivery into plant cells. Exemplary cell penetrating peptide families include, but are not limited to, cell penetrating peptides derived from protein transduction domains, amphipathic peptides, and synthetic cationic polypeptides, such as polylysine, polyhistidine, and polyarginine, or dendrimeric polycationic molecules. Cell penetrating peptides that may be used with the oligopeptides of the disclosure include, but are not limited to, the peptide vascular endothelial-cadherin cell penetrating peptide, the transportan cell penetrating peptide, the monomer and dimer of HIV- 1 TAT basic domain cell penetrating peptide, the penetratin cell penetrating peptide, synthetic cationic homoarginine oligopeptide cell penetrating peptides (see Eudes and Chugh. (2008) Plant Signal Behav. 3:549-550), the gamma zein cell penetrating peptide (see U.S. Pat. No. 8,581,036, incorporated herein by reference in its entirety). Additional nonlimiting examples of cell penetrating peptides, and other internalization molecules, include the Zea mays knotted 1 cell penetrating peptide, the Saccharomyces pombe TP 10 cell penetrating peptide, the Candida albicans Zebra cell penetrating peptide, Antennapedia sequences, TAT, Antp-3A (Antp mutant), Buforin II, MAP (model amphipathic peptide), K-FGF, Ku70, Prion, pVEC, SynBl, Pep-7, HN-1, BGSC (Bis- Guanidinium-Spermidine-Cholesterol), BGTC (Bis-Guanidinium-Tren-Cholesterol), . Additional cell penetrating peptides are described in U.S. Pat. No. 9,757,473, WO / 2014 / 086835A1, US2017 / 0057997, US2020 / 0263189, US20200399647, and WO / 2011 / 084061, the entireties of each of which are incorporated by reference for these peptides. In some embodiments, the cell penetrating peptide comprises a C-terminal modification, an N-terminal modification, or both. In some embodiments, the oligopeptide comprises a cell penetrating peptide linked to the amino acid sequence set forth in one of SEQ ID NOs: 1- 5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof.

[0105] In some embodiments, the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and an N-terminus and / or C-terminus residues. In some embodiments, the oligopeptide comprises one or more proline residues at the N-or C-terminus. In some embodiments, the N-terminus and / or C-terminus residues correspond to a cleavage tag. For instance, the residue may be part of a peptide bond that is cleaved by chemical (e.g. acidic cleavage), or enzymatic reactions. In some embodiments, the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and an N-terminal proline, methionine, tryptophan, glycine or cysteine, and / or a C-terminal methionine, aspartic acid, cysteine, asparagine, or tryptophan. In some embodiments, the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and an N-terminal proline and / or a C-terminal aspartic acid. In some embodiments, the oligopeptide is 4-50 or 5-30 amino acids long. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0106] In some embodiments, the oligopeptides provided herein are produced by cleavage of a concatamer of multiple oligopeptides that are linked together and expressed as a single polypeptide chain. In some embodiments, cleavage results in a short (one to two) amino acid tag at the N and / or C terminus of the oligopeptide. In some embodiments, the oligopeptide cis produced by a method comprising expressing a fiision polypeptide comprising a polypeptide that forms inclusion bodies in a cell operably linked to two or more oligopeptides. In some embodiments, the oligopeptide is produced by a method comprising expressing a fusion polypeptide comprising an carrier polypeptide operably linked to two or more oligopeptides by a peptide bond. In some embodiments, the oligopeptide is produced by a method comprising expressing a fusion polypeptide comprising an carrier polypeptide operably linked to two or more oligopeptides by a peptide bond, and releasing the two or more oligopeptide from the carrier polypeptide. In some embodiments, the two or more oligopeptides in the fusion polypeptide are released from the carrier polypeptide and / or from each other by sequence-specific chemical cleavage of the peptide bond. In some embodiments, the peptide bond is an Asp-Pro bond and the sequence-specific cleavage is performed using acetic acid. In some embodiments, releasing the two or more oligopeptides from the carrier polypeptide is performed in the absence of a chaotropic agent. In some embodiments, releasing the two or more oligopeptides from the carrier polypeptide is performed without a column-based purification step. In some embodiments, the two or more oligopeptides are 4-50 amino acids long. In some embodiments, the two or more oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the released oligopeptides comprise an N-terminal proline and / or a C-terminal aspartic acid. In some embodiments, the released oligopeptides comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and anN-terminal proline residue and a C-terminal aspartic acid. In some embodiments, the two or more oligopeptides are antimicrobial oligopeptides or antifungal oligopeptides.Oligopeptide Synthesis

[0107] In some embodiments, the method comprises synthetizing an oligopeptide by chemical synthesis. Various approaches been developed for chemical synthesis of peptides, such as for example, by solidphase methods. Solid phase peptide synthesis is a method for chemically synthesizing peptides on a solid support. In solid phase peptide synthesis, amino acids or peptides are typically attached to a solid support via the C-terminus. The new amino acid is added to the bound amino acid or peptide by a coupling reaction. Translation-based approaches for peptide synthesis have also been developed, whereby a peptide is produced from an encoding transcript by in vitro translation.

[0108] In some embodiments, the method comprises synthetizing an oligopeptide by biological synthesis. Any suitable method of biological synthesis of peptides may be used. Biological synthesis may comprise expressing a peptide in a cell and purifying the peptide from the cell.

[0109] In some embodiments, synthetizing an oligopeptide comprises expressing a fusion polypeptide comprising a polypeptide that forms inclusion bodies in a cell operably linked to two or more oligopeptides. In some embodiments, synthetizing an oligopeptide comprises expressing a fusion polypeptide comprising an carrier polypeptide operably linked to two or more oligopeptides by a peptide bond. In some embodiments, synthetizing an oligopeptide comprises expressing a fusion polypeptide comprising an carrier polypeptide operably linked to two or more oligopeptides by a peptide bond, and releasing the two or more oligopeptide from the carrier polypeptide. In some embodiments, synthetizing an oligopeptide comprises expressing a fusion polypeptide comprising an carrier polypeptide operably linked to two or more oligopeptides by a peptide bond, and releasing the two or more oligopeptides from the carrier polypeptide. In some embodiments, the two or more oligopeptides in the fusion polypeptide are released from the carrier polypeptide and / or from each other by sequence-specific chemical cleavage of the peptide bond. In some embodiments, the peptide bond is an Asp-Pro bond and the sequence-specific cleavage is performed using acetic acid. In some embodiments, releasing the two or more oligopeptides from the carrier polypeptide is performed in the absence of a chaotropic agent. In some embodiments, releasing the two or more oligopeptides from the carrier polypeptide is performed without a column-based purification step. In some embodiments, the two or more oligopeptides are 4-50 amino acids long. In some embodiments, the two or more oligopeptides comprise an N-terminal proline, a C-terminal aspartic acid,or both. In some embodiments, the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and a tag. In some embodiments, the two or more oligopeptides are antimicrobial oligopeptides or antifungal oligopeptides.

[0110] In some embodiments, the method comprises synthetizing a nucleic acid encoding an oligopeptide. Synthetic nucleic acids (DNA, RNA or their analogues) may be prepared using column-based synthesizers, or produced from an existing nucleic acid by PCR or in vitro transcription. Other methods of nucleic synthesis are described for example in U.S. Pat. No. 6,586,211 Bl, in PCT / EP2004 / 013131, in WO 00 / 13017 A2, in S. Rayner et al., PCR Methods and Applications 8 (7), 741-747, 1998, in WO 90 / 00626 Al, in EP 385 410 A2, in WO 94 / 12632 Al, in WO 95 / 17413 Al, in EP 316 018 A2, in EP 022 242 A2, in L. E. Sindelar and J. M. Jaklevic, Nucl. Acids Res. 23 (6), 982-987, 1995, in D. A. Lashkari, Proc. Nat. Acad. Sci. USA 92 (17), 7912-7915, 1995, and in WO 99 / 14318 Al. Nucleic acids may also be synthetized by biological synthesis such as, for example, by introducing a nucleic acid into a cell (e.g., a bacteria) and allowing the nucleic acid to be replicated before isolating the newly synthetized nucleic acids.III. Compositions[oni] Also provided are compositions comprising an oligopeptide. In some embodiments, the composition comprises an isolated oligopeptide. In some embodiments, the composition comprises a purified oligopeptide. In some embodiments the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21

[0112] In some embodiments, the composition comprises an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21. In some embodiments, the composition comprises an oligopeptide with an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions as compared to one of SEQ ID NOs: 1-5 and 14-21. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide. In some embodiments, the oligopeptide comprises a proline at its N-terminus, an aspartic acid at the C-terminus, or both. In some embodiments, the oligopeptide comprises one or more D-amino acids. In some embodiments, the one or more D-amino acids enhance stability of the oligopeptide as compared to an oligopeptide of the same amino acid sequence comprising all L-amino acids. In some embodiments, the oligopeptide further comprises a cell penetrating peptide linked to the amino acid sequence of the oligopeptide.

[0113] In some embodiments, the composition comprises an oligopeptide and an agriculturally acceptable formulant. In some embodiments, the agriculturally acceptable formulant comprises one or more of a water, organic solvents, paraffinic oils, vegetable oils, dispersants, emulsifiers, wetting agents, buffering agents, hydrotrope agents, rheology modifiers, antifoam agents and defoamers, antifreeze agents, biocides, dyes, polymer walls, catalysts, thermosetting materials, cross-polymerizing agents, UV protectants, antioxidants, and chelating agents.

[0114] In some embodiments, the composition comprises an oligopeptide at a concentration range of from about 0. 1 pg / ml to about 100 mg / ml, or from about 5 pg / ml to about 5 mg / ml. In some embodiments, the composition comprises an oligopeptide at a concentration of about 0. 1 pg / ml, 0.5 pg / ml, 1 pg / ml, 5 pg / ml, 10 pg / ml, 20 pg / ml, 30 pg / ml, 40 pg / ml, 50 pg / ml, 60 pg / ml, 70 pg / ml, 80 pg / ml, 90 pg / ml, 1 mg / ml, 5 mg / ml, 10 mg / ml, 20 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, or 100 mg / ml.

[0115] In some embodiments, the composition further comprises one or more other active agents selected from the group consisting of a pesticide, a fertilizer, an insecticide, an attractant, a sterilizing agent, an acaricide, a nematocide, a herbicide, a biostimulant, a biological, and a growth regulator. In some embodiments, the composition is formulated as a liquid, a gel, an emulsion, a suspension, an encapsulation, a solid, a powder, an aerosol, a paste, a coating, a spray, a soil drench, a microcapsule, an emulsifiable concentrate, or as granules. In some embodiments, the composition is in the form of a finely-divided particulate solid, granules, pellets, wettable powders, dust, an aqueous suspension, a dispersion, a gel, or an emulsion. In some embodiments, the agriculturally acceptable composition is formulated as a seed treatment, a foliar spray, a foliar drench, a Ready-To-Use (RTU) formulation, a produce coating, a suspension concentrate, a tank-mix, an aerosol, a root dip, a soil treatment, a dipping formulation, an irrigation formulation, or a sprinkler formulation. In some embodiments, the composition is formulated for application to a plant or part thereof after harvest. In some embodiments, the composition is formulated for application to a plant or part thereof prior to harvest. In some embodiments, the plant part is a fruit leaf, a bud, a root, a shoot, a floral part, or a seed. In some embodiments, the composition is formulated for application to a plant prior, during, or after an infection with a pathogen or pest. In some embodiments, the composition is formulated for application to the soil in which the plant is planted. In some embodiments, the composition is formulated for application to a plant by addition of water provided to the plant.

[0116] In some embodiments, the composition comprises an oligopeptide and an agriculturally acceptable carrier, diluent or excipient. In some embodiments, the agriculturally acceptable carrier, diluent, or excipient comprises a buffer and the pH is in the range of from about 3.0 to about 9.0. In someembodiments, the agriculturally acceptable carrier, diluent, or excipient comprises a buffer and the pH is in the range of from about 3.0 to about 4.0, from about 4.0 to about 5.0, from about 5.0 to about 6.0, from about 6.0 to about 7.0, from about 7.0 to about 8.0, or from about 8.0 to about 9.0. In some embodiments, the agriculturally acceptable carrier, diluent, or excipient comprises a buffer and the pH is in the range of from about 4.5 to about 8.0. In some embodiments, the agriculturally acceptable carrier comprises a solid carrier, a liquid carrier, a gel carrier, a suspension, or an emulsion. In some embodiments, the agriculturally acceptable carrier comprises an adjuvant, an inert component, a dispersant, a surfactant, a humectant, an emulsifier, a thickener, a wetting agent, a fertilizer, a mineral, a solvent, a tackifier, a binder, or a stabilizer. In some embodiments, the agriculturally acceptable composition comprising an oligopeptide and a surfactant or humectant.IV. Nucleic acids

[0117] Also provided are nucleic acids encoding an oligopeptide.

[0118] In some embodiments, the nucleic acids encodes an oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial or antifungal oligopeptide. In some embodiments, the oligopeptide comprises an amino acid sequence with one, two, three, or four amino acid substitutions, insertions, or deletions as compared to any of SEQ ID NOS: 1-5 and 14-21. In some embodiments, the oligopeptide comprises amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any of SEQ ID NOS: 1-5 and 14-21. In some embodiments, the oligopeptide is 4-50 or 5-30 amino acids in length. In some embodiments, the oligopeptide further comprises an N-terminal proline and a C-terminal aspartate. In some embodiments, the oligopeptide is an antimicrobial oligopeptide, or an antifungal oligopeptide.

[0119] In some embodiments, the nucleic acid encoding the oligopeptide is operably linked to a promoter, for example in a nucleic acid construct. In some embodiments, the promoter may be a heterologous promoter. In some embodiments, the promoter is a constitutive promoter, a tissue specific promoter, a development stage specific promoter, or an inducible promoter. In some embodiments, the promoter is an inducible promoter. In some embodiments, the inducible promoter is induced by fungal infection, optionally a promoter associated with a gene involved in phenylpropanoid metabolism (e.g., phenylalanine ammonia lyase, chaicone synthase promoters), a gene that modifies plant cell walls (e.g, hydroxyproline-rich glycoprotein, glycine-rich protein, and peroxidase promoters), a gene encoding an enzyme that degrade fungal cell walls (e.g., chitinase or glucanase promoters), a gene encoding a thaumatin-like protein, or a maize or flax promoters, designated as Misl and Fisl.

[0120] In some embodiments, the nucleic acid encodes a fusion polypeptide comprising a polypeptide that forms inclusion bodies in a cell operably linked to two or more oligopeptides. In some embodiments, the nucleic acid encodes a fusion polypeptide comprising an carrier polypeptide operably linked to two or more oligopeptides. In some embodiments, the operable linkage comprises a peptide bond susceptible to sequence-specific chemical cleavage. In some embodiments, the operable linkage is an Asp-Pro bond and the sequence-specific cleavage is acetic acid cleavage. In some embodiments, the oligopeptide is 4- 50 amino acids long. In some embodiments, the oligopeptide comprises an N-terminal proline and a C- terminal aspartic acid. In some embodiments, the nucleic acid operably linked to a promoter encodes an oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial or antifungal oligopeptide.

[0121] In some embodiments, the nucleic acid encodes an oligopeptide further comprising a tag. In some embodiments, the tag is a secretion tag, a tag that promotes entry into a plant cell, or a nuclear localization tag.

[0122] The nucleic acids of the disclosure may be prepared using any technique known in the art. Without limitation, these may include cloning, DNA isolation, amplification and purification, enzymatic reactions involving DNA ligase, DNA polymerase, restriction endonucleases and the like, and various separation techniques, such as gel electrophoresis and chromatography. A number of standard techniques are described in Ausubel et al. (1992) Current Protocols in Molecular Biology, Green / Wiley, New York, N.Y.; Sambrook et al. (1989) Molecular Cloning, Second Edition, Cold Spring Harbor Laboratory, Plainview, N.Y.; Maniatis et al. (1982) Molecular Cloning, Cold Spring Harbor Laboratory, Plainview, N.Y.; Wu (ed.) (1993) Meth. Enzymol. 218, Part I; Wu (ed.) (1979) Meth. Enzymol. 68; Wu et al. (eds.) (1983) Meth. Enzymol. 100 and 101; Grossman and Moldave (eds.) Meth. Enzymol. 65; Miller (ed.) (1972) Experiments in Molecular Genetics, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.; Old and Primrose (1981) Principles of Gene Manipulation, University of California Press, Berkeley; Schleif and Wensink (1982) Practical Methods in Molecular Biology; Glover (ed.) (1985) DNA Cloning Vol. I and II, IRL Press, Oxford, UK; Hames and Higgins (eds.) (1985) Nucleic Acid Hybridization, IRL Press, Oxford, UK; Setlow and Hollaender (1979) Genetic Engineering: Principles and Methods, Vols. 1-4, Plenum Press, New York; and Ausubel et al. (1992) Current Protocols in Molecular Biology, Greene / Wiley, New York, N.Y. Abbreviations and nomenclature, where employed, are deemed standard in the field and commonly used in professional journals such as those cited herein.Vectors

[0123] Also provided herein are vectors comprising nucleic acids encoding any of the fusion polypeptides, carrier polypeptides or oligopeptides disclosed herein.

[0124] A “vector” is a nucleic acid that is capable of transporting another nucleic acid. Vectors may be, for example, plasmids, viruses, cosmids or phage. An “expression vector” is a vector that is capable of directing expression of a protein encoded by one or more genes carried by the vector when it is present in the appropriate environment. Examples of vectors are those that can autonomously replicate and express structural gene products present in the DNA segments to which they are operatively linked. Vectors, therefore, can contain the replicons and selectable markers described earlier. Vectors include, but are not necessarily limited to, expression vectors.

[0125] In some embodiments, the vector is a bacterial vector. In some embodiments, the vector is a yeast vector. Examples of expression vectors that can be used in prokaryotic host cells include those derived from commercially available plasmids such as the cloning vector pET plasmids (Novagen, Madison, Wis., USA) or pBR322 (ATCC 37017). The pBR322 vector contains genes for ampicillin and tetracycline resistance and thus provides simple means for identifying transformed cells. To construct an expression vector using pBR322, an appropriate promoter and a DNA sequence encoding one or more of the polypeptides of the invention are inserted into the pBR322 vector. Other commercially available vectors include, for example, pKK223-3 (Pharmacia Fine Chemicals, Uppsala, Sweden) and pGEM-1 (Promega Biotec, Madison, Wis., USA). Other commercially available vectors include those that are specifically designed for the expression of proteins; these would include pMAL-p2 and pMAL-c2 vectors that are used for the expression of proteins fused to maltose binding protein (New England Biolabs, Beverly, Mass., USA).

[0126] In bacterial systems a number of expression vectors may be advantageously selected depending upon the use intended for the variant squalene synthase enzyme expressed, and whether it is desired to isolate the enzyme and in what state of purity. For example, when large quantities are to be produced, vectors which direct the expression of high levels of fusion protein products that are readily purified may be desirable.

[0127] In yeast, a number of vectors containing constitutive or inducible promoters may be used. For a review, see Current Protocols in Molecular Biology, Vol. 2, Ed. Ausubel et al., Greene Publish. Assoc. & Wiley Interscience, Ch. 13 (1988); Bitter et al., Expression and Secretion Vectors for Yeast, in Methods in Enzymology, Eds. Wu & Grossman, 31987, Acad. Press, N.Y., Vol. 153, pp. 516-544 (1987); Glover, DNA Cloning, Vol. II, IRL Press, Wash., D.C., Ch. 3 (1986); Bitter, Heterologous Gene Expression in Yeast, Methods in Enzymology, Eds. Berger & Kimmel, Acad. Press, N.Y., Vol. 152, pp. 673-684 1987); and The Molecular Biology of the Yeast Saccharomyces, Eds. Strathem et al., Cold Spring Harbor Press,Vols. I and II (1982). A constitutive yeast promoter such as ADH1 or LEU2 or an inducible promoter such as GAL4 may be used (Cloning in Yeast, Ch. 3, R. Rothstein In: DNA Cloning Vol. 11, A Practical Approach, Ed. D M Glover, IRL Press, Wash., D.C. (1986)). Alternatively, vectors may be used which promote integration of foreign DNA sequences into the yeast or bacterial chromosome.

[0128] Promoter sequences commonly used for recombinant prokaryotic host cell expression vectors include the bacteriophage T7 promoter (Studier and Moffatt, J. Mol. Biol. 189: 113 (1986)), [3-lactamase (penicillinase), lactose promoter system (Chang et al., Nature 275:615, 1978; Goeddel et al., Nature 281:544 (1979)), tryptophan (tap) promoter system (Goeddel et al., Nucl. Acids Res. 8:4057 (1980); EP- A-36776), and tac promoter (Maniatis, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory; p. 412 (1982)). A particularly useful prokaryotic host cell expression system employs a phagePL promoter and a cl857ts thermolabile repressor sequence. Plasmid vectors available from the American Type Culture Collection (ATCC), which incorporate derivatives of the PL promoter, include plasmid pHUB2 (resident in E. coli strain JMB9 (ATCC 37092)) and pPLc28 (resident in E. coli RR1 (ATCC 53082)).V. Cells

[0129] Also provided are cells comprising antimicrobial or antifungal oligopeptides. In some embodiment, the cell comprises an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: l-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the cell is a host cell.

[0130] In some embodiments, the cell is a plant cell. In some embodiments, the cell is a cell of the plant. In some embodiments, the plant cell is a dicot or monocot plant cell. In some embodiment, the plant cell is a cell from a crop plant. In some embodiments, the plant cell is a cell from a Solarium spp or a Triticum spp. plant. In some embodiments, the cell is a transgenic cell. In some embodiments, the cell expresses an antimicrobial oligopeptide or an antifiingal oligopeptide.

[0131] In some embodiments, the cell is a bacterial cell. In some embodiments, the bacteria is an E. coli strain. In some embodiments, the cells is a cell that is capable of forming inclusion bodies. In some embodiments, the cell is a prokaryotic cell. In some embodiments, the cell is a eukaryotic cell. In some embodiments, the cell is a fiingal cell. In some embodiments, the cell is an algal cell. In some embodiments, the cell is an animal cell. In some embodiments, the animal cell is a mammalian cell or an insect cell.

[0132] Microorganism host cells usefill in the present invention may include, but are not limited to, bacteria, such as the enteric bacteria {Escherichia and Salmonella for example) as well as Bacillus,Acinetobacter, Streptomyces, Methylobacter, Rhodococcus and Pseudomonas,' Cyanobacteria, such as Rhodobacter and Synechocystis,' yeasts, such as Saccharomyces, Zygosaccharomyces, Kluyveromyces, Candida, Hansenula, Debaryomyces, Mucor, Pichia, Yarrowia, and Torulopsis,' and filamentous fungi such as Aspergillus and Arthrobotrys, and algae for example.

[0133] In some embodiments, the host cell comprises at least one copy of a nucleic acid sequence encoding an oligopeptide. The at least one copy of the nucleic acid sequence encoding an oligopeptide can be present in the chromosome of a prokaryotic (bacterial) cell or in one chromosome of a eukaryotic cell. Alternatively, the at least one copy of the nucleic acid sequence encoding an oligopeptide can be present in a vector or plasmid that is present in the cell. The host cell, as described above, can be a prokaryotic or eukaryotic cell. If it is a prokaryotic cell, it can be a bacterial cell. If it is a eukaryotic cell, it can be a yeast cell, a plant cell, or an animal cell. Suitable host cells are described herein.

[0134] In some embodiments, the cell comprises an oligopeptide, or a nucleic acid encoding an oligopeptide, comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide comprises amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any of SEQ ID NO: 1-5 and 14-21. In some embodiments, the oligopeptide is 4-50 or 5-30 amino acids in length. In some embodiments, the oligopeptide further comprises an N-terminal proline and a C-terminal aspartate. In some embodiments, the oligopeptide comprises a secretion tag, a tag that promotes entry into a plant cell, or a nuclear localization tag. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.VI. Kits

[0135] Some aspects of the present disclosure provide for kits comprising an oligopeptide and a package insert comprising instructions for providing the oligopeptide to a plant. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0136] In some embodiments, the kit comprises an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and a package insert comprising instruction for application of the oligopeptide to a plant. In some embodiments, the oligopeptide comprises amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any of SEQ ID NOS: 1-5 and 14-21, and a package insert comprising instruction for application of the oligopeptide to aplant. In some embodiments, the oligopeptide is 4-50 or 5-30 amino acids in length. In some embodiments, the oligopeptide further comprises an N-terminal proline and a C-terminal aspartate. In some embodiments, the oligopeptide comprises a secretion tag, a tag that promotes entry into a plant cell, or a nuclear localization tag. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0137] In some embodiments, the kit comprises a composition comprising an oligopeptide and, and a package insert comprising instructions for application of the composition. In some embodiments, the composition comprises an oligopeptide and an agriculturally acceptable formulant. In some embodiments, the agriculturally acceptable formulant comprises one or more of a water, organic solvents, paraffinic oils, vegetable oils, dispersants, emulsifiers, wetting agents, buffering agents, hydrotrope agents, rheology modifiers, antifoam agents and defoamers, antifreeze agents, biocides, dyes, polymer walls, catalysts, thermosetting materials, cross-polymerizing agents, UV protectants, antioxidants, and chelating agents. In some embodiments, the composition is in the form of a finely-divided particulate solid, granules, pellets, wettable powders, dust, an aqueous suspension, a dispersion, a gel, or an emulsion. In some embodiments, the composition further comprises one or more other active agents selected from the group consisting of a pesticide, a fertilizer, an insecticide, an attractant, a sterilizing agent, an acaricide, a nematocide, a herbicide, a biostimulant, a biological, and a growth regulator.. In some embodiments, the kit comprises an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

[0138] In some embodiments, the kit comprises a composition comprising an oligopeptide and an agriculturally acceptable carrier, diluent or excipient. In some embodiments, the kit comprises a composition comprising an oligopeptide and a package insert comprising instructions for applying the oligopeptide to a plant or part thereof. In some embodiments, the plant part is a leaf, a bud, a root, a shoot, a floral part, or a seed. In some embodiments, the package insert comprises instructions for providing the composition to a plant by application to the soil in which the plant is planted. In some embodiments, the package insert comprises instructions for providing the composition by addition of water provided to the plant. In some embodiments, the package insert comprises instructions for applying the composition by a spreader, a power duster, a boom sprayer, a hand sprayers, a spray dusters, or a granular applicator. In some embodiments, the kit comprises an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and a package insert comprising instruction for application of the oligopeptide to a plant.

[0139] Also provided herein are kits comprising cells comprising an oligopeptide, or a nucleic acid encoding the oligopeptide, and a package insert comprising instructions for application of the cells to a plant. In some embodiments, the cells comprise an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with zero, one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and a package insert comprising instruction for application of the oligopeptide to a plant. In some embodiments, the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.EXAMPLES

[0140] The present disclosure is described in further detail in the following examples, which are not in any way intended to limit the scope of the disclosure as claimed. The attached figures are meant to be considered as integral parts of the specification and description of the disclosure. The following examples are offered to illustrate, but not to limit the claimed disclosure.Example 1: Screen for antimicrobial oligopeptides targeting members of the Slmirl69 family, identification of an antimicrobial oligopeptide that clears Botrytis infection

[0141] The follow example describes a screen that resulted in the identification of MP 18279, an oligopeptide with anti-fungal activity.Materials and MethodsScreen

[0142] Botrytis cinerea was grown on potato dextrose agar (PDA) in the dark at 25° C. Actively growing mycelium was collected from the plates, dissolved in water and filtered through a Medifab cloth. Spores present in the filtrate were counted and adjusted to a final concentration of 2.5xl05spores / ml in potato dextrose broth (PDB).

[0143] Leaves from 3-week-old tomato plants were inoculated with 5pl of spore solution at six sites of the mature leaf. 22 peptides were chemically synthesized (SB-Peptide, Grenoble, France) and tested for activity against Botrytis cinerea. Peptides were diluted in Milli-Q water to a concentration of lOOpM. Each peptide was added daily (3 preventive treatments) on the site of future Botrytis inoculation. Spores were placed as droplet as described above on a leaf and plants were kept in humid environment to allow start of infection. At 24 and 48 hours after infection, 5 pl of lOOpM peptides were added to the site of infection. After 72h lesion area was measured and compared to control treatments, for which Milli-Q water was applied at the same timings as peptides.Application of MP 18279 to Botrytis cinerea-infected tomato

[0144] MP 18279 (SEQ ID NO: 1) was applied to Botrytis cinerea infections on tomato leaves and fruits. 100 pM of MP 18279 was applied directly on the infection sites daily for two (for leaves) or five (for fruits) consecutive applications (see FIG. 1).ResultsMP 18279 completely abolishes Botrytis infection on tomato leaves and fruits

[0145] A total of 22 candidate antimicrobial oligopeptides were analyzed. The activity of the peptides was assessed by applying the peptides to lesions provoked by B. cinerea infection on tomato plants. Typically, infected tomato plants showed brownish symptoms at the site of infection, caused by the spread of pathogen in the tissue.

[0146] Application of the 10 amino acid peptide MP18279 (SEQ ID NO: 1) led to complete lack of symptoms. MP18279 was identified in the primary transcripts of Solanum lycopersicum .SVi lR 169b. Solanum tuberosum StwmlR 169c, and Capsicum annuum mIR169 gene family. The plant tissue with MP18279 applied stayed healthy and green compared to the highly infected control plants (FIG. 1). The infected control tomato fruits depicted typical B. cinerea symptoms like the development of grey mycelium at the site of infection, whereas the peptide treated tomato plants did not develop any visible symptoms in the observed time period (FIG. 1). Accordingly, MP 18279 completely abolished B. cinerea infection on tomato leaves and fruits.Example 2: MP18279 prevents Botrytis cinerea germination

[0147] The follow example describes experiments investigating the effects of MP 18279 on B. cinerea germination.Materials and MethodsGermination Assay

[0148] 400 spores of Botrytis cinerea \\c c incubated with either water (control) or 100 pM MP 18279 (SEQ ID NO: 1) for 24h in the dark and analysed by microscopy. The spores were then deposited onto Potato Glucose Agar (PGA) medium, and their development (spore germination and mycelium growth) was analyzed after six days (see FIG. 2).ResultsMP 18279 prevents germination of Botrytis cinerea

[0149] To understand the mode of action of the anti-fungal effects of MP 18279, spores of B. cinerea were incubated with MP 18279 and spore germination and mycelium growth were monitored. In control conditions, the spores germinated after 24 hours. In contrast, no germination could be observed in the MP18279-treated spores. After six days, grey mycelium grew on control plates, while no growth couldbe observed on the MP18279-treated plate (FIG. 2). Therefore, MP18279 directly inhibited the germination of B. cinerea spores.Example 3: MP18279 blocks germination of various fungal and oomycete pathogens

[0150] The example describes experiments testing the ability of MP 18279 to prevent germination of the fungal pathogens B. cinerea, Septaria tritici, Alternaria solani, and Rhizoctonia solani anastomosis, and the oomycete Phytophthora capsici.Materials and MethodsGermination Assays

[0151] For the initial germination assay, B. cinerea, S. tritici, A. solani, Rhizoctonia solani anastomosis and P. capsici were grown on PDA agar plates, spores were harvested and diluted into 1% PDA. A 96- well plate was prepared containing 90 pL of same number of spores of the respective pathogen in each well. Peptides were prepared by re-suspension in 2-(N-morpholino)ethanesulfonic acid buffer (MES) 100 mM, and lOpL of the peptide solution was added to each well to reach a final concentration of 300, 100, 33 or 10 mg / L. A peptide with no known anti-fungal effect MP18021 (SEQ ID NO:3), was used as the control peptide. Blank (MES) and proline standards were also used as negative and positive controls, respectively. The final assessment was done after 2-7 days depending pathogen growth. The amount of fungal growth was compared to the control treatment and scored to a defined key, and the resulting growth reduction per part per millions (ppm) was determined as the EC50 (mg / L).

[0152] Subsequently, B. cinerea, S. tritici, and P. capsici spores were incubated with MP18279 or a scrambled control peptide at 300, 100, 30, or 10 mg / L. The scrambled control peptide MP18753 (SEQ ID NO: 13) consisted of all of the amino acids of the initial MP 18279 sequence, but in random order. Spore germination was visually detected and the EC50 was calculated for the control peptide and MP 18279 (see FIG. 4)ResultsIn vitro activity of MP 18279 on spore germination on five different pathogens

[0153] The ability of MP 18279 to block spore germination was tested for the species B. cinerea (fungus, Grey Mold), S. tritici (fungus, Wheat Leaf Blotch), A. solani (fungus, Early Blight), Rhizoctonia (fungus, Root Rot), and P. capsici (oomycete, Pepper Blight). MP 18279 reduced germination of S. tritici, P. capsici, B. cinerea, and Rhizoctonia spores (FIG. 3). MP 18279 showed the largest effect in preventing spore germination of S. tritici and P. capsici. B. cinerea and Rhizoctonia were similarly affected after peptide incubation. Finally, MP 18279 did not show any impact on A. solani growth, which points to a selectivity towards certain pathogens (FIG. 3). As control, the spores were incubated with an unrelatedpeptide sequence at same concentrations. The control peptide did not impact the spore germination in the in vitro condition.

[0154] In summary, MP 18279 exhibited inhibitory activity against four out of five tested filamentous pathogens, including both fungi and an oomycete.Confirmation of in vitro activity of MP 18279 on three different pathogens

[0155] S. tritici, P. capsici, and B. cinerea were further investigated because spore germination in these species was significantly affected by presence of MP 18279. For all pathogens tested, the EC50 of MP 18279 was significantly lower than the EC50 of the scrambled control peptide (FIG. 4). The scrambled peptide did not show any effect on germination on B. cinerea or S. tritici, and only a showed slight effect on the growth of Phytophthora.

[0156] In conclusion, the ability of MP 18279 to inhibit spore formation by three filamentous pathogens (including both fungi and an oomycete) was shown to be dependent on the specific amino acid sequence of MP 18279, since the scrambled peptide control had no effect on spore germination.Example 4: Identification of a seven amino acid residue region of MP18279 that is necessary for anti-pathogen activity

[0157] The following example describes experiments defining a region of MP 18279 that is necessary for anti-fungal activity. Specifically, various truncations of MP 18279 were generated and tested for their ability to block fungal or oomycete germination. A seven amino acid residue sequence of MP 18279 was identified that is necessary for MP 18279 activity.Materials and MethodsGeneration of truncated versions of MP 18279Table 1. Amino acid sequences of MP18279 variantsGermination Assays

[0158] Germination assays were performed using B. cinerea, S. tritici, and P. capsici spores as described in Example 3, above (see FIG. 5A, FIG. 5B, and FIG. 5C)Results

[0159] On all tested pathogens the 9, 8 and 7 amino acid truncated versions of MP 18279 (truncated from C-terminus) were more active than the original 10 amino acid sequence (FIG. 5A, see black line). Reducing the peptide to 6 and 5 amino acids in length led to a loss in peptide activity.

[0160] The truncation of amino acids at the N-terminus led to an immediate loss of peptide activity, demonstrating that the N terminal amino acids 1-7 are essential and required for the peptide activity (FIG.5B)

[0161] On S. tritici MP18279 variants (with amino-acids addition and / or changes) were still active (FIG. 5C). These modifications did not disturb peptide activity.Example 5: Concentration dependent anti-fungal activity of MP18279 in planta

[0162] The following example describes experiments investigating the minimum concentration of peptide needed to affect Botrytis cinerea growth on tomato leaves.Materials and MethodsBotrytis cinerea - Tomato pathosystem

[0163] The B. cinerea and tomato pathosystem was used to investigate the minimum concentration of peptide needed to affect B. cinerea growth on tomato leaves. To do so, spores at 2.5 x 105 / ml were incubated with either peptides at respective concentrations lOOpM, 75pM, 50pM, 25 pM and IpM or with water (control) for 1 hour, and then the spore solution was applied as a droplet on tomato leaves. For each treatment, six leaves were applied with six droplets each, resulting in n=36 per treatment. The lesion size after two days was measured and compared to control infection (see FIG. 6).Results

[0164] The effect of MP 18279 activity at different concentrations was tested in planta on tomato leaves. Incubation with 100 or 75 pM of MP18279 provided almost 100% B. cinerea control, manifesting in no lesions or extremely small lesions (FIG. 6). 50 pM of peptide provided 80% B. cinerea control, but lowering the peptide concentration to 25 pM and below did not impact B. cinerea growth and resulted in lesions that were as large as control lesions, showing only 0-20% B. cinerea control (FIG. 6).

[0165] Accordingly, concentrations of MP18279 ranging from 50-100 pM were effective at controlling Botrytis infections on tomato leaves.Example 6: Spray application of MP18279 provides disease control for crop plants

[0166] The following example describes tests of MP 18279 applied as spray on different plant / pathogen systems in order to test its ability to control pathogens on crop plants.Materials and MethodsSpray application of MP 18279 to tomato plants infected with Tomato Grey Mold

[0167] Three weeks old tomato plants were spray infected with Botrytis cinerea (Tomato Grey Mold). One hour after infection, the plats were sprayed with MP 18279 (SEQ ID NO: 1) at different doses ranging from 200-0.2 grams per hectare (g / ha). The convex hull was analyzed daily, and the severity of disease symptoms was visually inspected four days after infection. The convex hull measurement was used to calculate the disease control between infected and peptide treated plants (FIG. 7A, FIG. 7B). Peptides are diluted to the respective final concentration in TankMix-006 (pH 7.0; 1.44g / L alkylpolyglucoside, 0.5 g / L polysorbate 20, 0.1 g / 1 polydimethylsiloxane oil, and 1.96 g / L 2-ethanesulfonic acid).Spray application of MP 18279 to tomato plants subsequently inoculated with Tomato Late Blight, and wheat plants subsequently inoculated with Wheat Brown Rust

[0168] Tomato plants were sprayed with 3 grams per hectare MP 18279, and wheat plants were sprayed with 300 grams per hectare MP 18279. Two hours following peptide application, tomato plants were inoculated with Phytophthora infestans (Tomato Late Blight), and wheat plants were inoculated with Septoria tritici (Wheat Brown Rust) (see FIG. 8A, FIG. 8B). As standard fungicidal control, inoculated tomato plants were sprayed with Revus SC and inoculated wheat plants were sprayed with Fluxapyroxad.ResultsSpray application of MP 18279 provides 40% disease control against Tomato Grey Mold

[0169] To further test the ability of MP 18279 to control pathogens on crop plants, MP 18279 was applied as spray on different plant / pathogen systems. A 40% disease control was observed against B. cinerea when tomato plants were treated with peptide at 200 and 20 grams per hectare (FIG. 7A). Further, there was a concentration dependency of the applied peptide, with a loss of activity of MP 18279 at 0.2 grams per hectare (FIG. 7B).

[0170] In summary, spray application of MP 18279 significantly improved disease control against Tomato Grey Mold.Broad spectrum activity of MP 18279: Spray application of MP 18279 provides disease control against Tomato Late Blight and Wheat Brown Rust

[0171] It was further tested whether MP 18279 could provide disease control against other filamentous pathogens like rust fungi or oomycetes. Specifically, spray application of MP18279 was tested on tomato plants subsequently inoculated with Tomato Late Blight (FIG. 8A), and wheat plants infected with Wheat Brown Rust (FIG. 8B).

[0172] The fungicides used as controls provided 100% control, whereas MP 18279 application led to more than 30% disease control based on percentage of infected leaf area compared to non-treated inoculated plants (FIG. 8A, FIG. 8B). Therefore, spray application of MP 18279 can be used against multiple filamentous pathogens.Example 7: Stability analysis of MP18279 and variants thereof

[0173] The following example describes experiments analyzing the stability of MP 18279, as well as the synthesis of a stable variant of MP 18279 with D-amino acids at the N- and C-terminus.Materials and MethodsSynthesis ofMP18279DlD10, a variant of MP 18279 with D-amino acid residues at theN- and C- terminus

[0174] Peptide MP182879D1D10 was chemically synthesized by SB-peptide (Grenoble, France).Tomato leaf wash off degradation assays

[0175] For the preparation of the tomato leaf wash-off solution, leaves from 2-week-old plants grown in growth chambers were cut and placed in 50mL falcon tubes. 2mL of purified water were then added to the tubes and vortexed. Afterwards, the solution was collected in a 1.5mL tube and centrifuged at 500 xg for 4 mins at 20°C.The supernatant was then added to the peptide stock solution.

[0176] For the preparation of analytical samples, the peptides were diluted directly in an analytical vial with a 200pL insert to reach a final concentration of 0.5 g / L. The 200pL sample volume was obtained from 20pL of peptide stock solution and 180pL of either tomato leaf wash-off solution or MiliQ water.

[0177] For the comparing biological stability of MP18279 and MP18279D1D10 in tomato leaf wash-off solution, the samples r were stored at 20°C for 6, 9 or 48 hours. Afterwards, 5-10pl was used for high performance liquid chromatography (HPLC) analysis. The resulting chromatograms were then compared and plotted on the same curve graph, and the percentage of relative amount of peptide over time was calculated.Application ofBotrytis cinerea spores and peptides to tomato leaves

[0178] MP18279 and MP18279D1D10 at lOOpM were incubated for 30 minutes with isolated cinerea spores at 2.5 xlO5 / mL and applied as droplets (5pl) on leaves (see FIG. 9B). As an infection control, spores were applied without further treatment. As a peptide control, spores were incubated with lOOpM of MP19075 (SEQ ID NO:4), which contains the same amino acids as MP18279 but with a scrambled sequence.Spray application of MP 18279 and MP 18279D1D10 to tomato plants infected with Tomato Late Blight

[0179] Tomato plants were sprayed with MP18279 (SEQ ID NO: 1) or MP18279D1D10 at dose of 30 grams per hectare. Two hours following spray application, the plants were inoculated with P. infestans (see FIG. 9C).ResultsVariant forms of MP 18279 show higher stability by similar antifungal activity

[0180] Experiments were performed to test whether the stability MP18279 could be improved. The stability of MP 18279 in a solution washed off of tomato leaves was tested, to represent the microorganisms present on the leaf surface. MP 18279 degraded rapidly after storage for 40 hours in this solution (FIG. 9A).

[0181] To overcome this biological degradation, a variant of MP 18279 with the first and the last amino acids in the sequence replaced with the D-form of the respective amino acids was chemically synthesized. As D-amino acids are not naturally-derived amino acids, without wishing to be bound by theory, it was anticipated that a variant of MP18279 with N- and C-terminal D-amino acids would be degraded by microorganisms to a lesser extent than the peptide with entirely L-amino acids. Indeed, the modified peptide (“MP18279D1D10”) showed a 20% improvement in stability when stored for the same time in the leaf wash off solution compared to MP 18279 (FIG. 9A).

[0182] Two independent tests were performed to further analyze the impact of the D-amino acids on the biological activity of MP 18279. In the first experiment, the peptides were incubated with isolated B.cinerea spores and applied as droplets on leaves. In control conditions containing either buffer or a scrambled sequence of MP 18279, necrotic lesions were observed 2 days after infection with the spores. In contrast, both peptides MP18279 and MP18279D1D10 were able to completely abolish disease symptoms when used at 100 pM (FIG. 9B). Further, the activity of MP18279D1D10 as a spray application was validated on tomato plants subsequently infected with Phytophthora infestans, and both peptides showed a similar level of disease control between 22-24% (FIG. 9C). Accordingly, MP18279D1D10 was less prone to degradation, but retained its full biological activity.

[0183] Finally, the stability of MP 18279 and its shorter 7 amino acid variant (“MP 18865”) was compared in a wash off solution of tomato leaves. After one week. MP 18279 was almost folly degraded, whereas MP18865 was still present at 12% (FIG. 10). Further, the disease control of MP18279 and MP18865 using Tomato Grey Mold or Tomato Late Blight assays was tested. No significant difference between the two peptides was observed (Table 2). Accordingly, the shorter variant was more stable and showed the same biological activity as the original peptides in the tested conditions.Table 2. Summary of disease control by MP18279 and MP18865 using Tomato Grey Mold orExample 8: Genotoxicity tests of MP18279

[0184] The following example describes the results of genotoxicity screenings performed to test whether MP 18279 induces genetic damage.Results

[0185] Genotoxicity screenings were performed for MP18279. The bacterial reverse mutation test (AMES; Table 3) and in vitro mammalian micronucleus test (MNT; Table 4) did not show any sign of observed genotoxicity for MP 18279.Table 3. Bacterial reverse mutation test (AMES) summaryTable 4. Mammalian micronucleus test (MNT) summary*An AC50 was calculated, but was greater than the maximum surviving concentration.Example 9: Antimicrobial selectivity of MP19919 and MP19594

[0186] The following example describes the results of antimicrobial screenings performed to test whether MP19919 (SEQ ID NO: 19) and / or MP19594 (SEQ ID NO: 16), exhibit antimicrobial activity against Phytium ultimum, Rhizoctonia solani, Sclerotonia sclerotiorum, Zymoseptoria tritici, Phythopthora cactorum, Botrytis cinerea, Microdochium nivale, Alternaria alternata, Pyrenophora tritici-repenti, and / or Fusarium graminearum compared to a positive control commercial fungicide and / or a negative control peptide. Pathogen classification and importance (relevance) is listed in Table 5. MP 19919 (SEQ ID NO: 19) demonstrated selective antimicrobial activity against all tested pathogens except Sclerotoniasclerotiorum. MP19594 (SEQ ID NO: 16) demonstrated selective antimicrobial activity against only Zymoseptoria tritici, Phythopthora cactorum, Botrytis cinerea, Microdochium nivale, and Alternaria altemata. The negative control peptide did not demonstrate antimicrobial activity against any tested pathogens.MethodsMeasuring Half Maximal Effective Concentration (EC50)

[0187] Potency of peptide antimicrobial activity of MP19919 and MP19594 was tested against seven microbial strains: Zymoseptoria tritci, Phythophora cactorum, Botrytis cinerea, Microdochium nivale, Alternaria alternata, Fusarium graminearum, and Pyrenophora tritici-repentis . The negative control peptide was 11 amino acids long with an N-terminal P residue and C-terminal D residue and otherwise no sequence identity to MP 19919 or MP 19594. The peptides were each tested in amounts of 300 ppm, 100 ppm, 30 ppm, and 10 ppm each poured directly into growth medium on 96 well plates. A no-peptide negative control (no added compound / peptide) and 2 positive controls comprising chemical fungicide Proline 275, one at 10 mg / L and one at 50mg / L. Two replicates were conducted for each dose of each peptide and the controls , with one measurement per well and two wells measured per concentration. EC50 was calculated as the amount of peptide needed to induce 50% growth control against each tested pathogen, and calculation was provided by Fera Science Ltd. (York, United Kingdom).Measuring Growth Control

[0188] Capability of controlling pathogen colony growth was tested for MP 19919, MP 19594, and a negative control peptide along with a blank negative control of unamended agar and a positive control of Amistar® commercial fungicide. The tested pathogens were Pythium ultimum, Rhizoctonia solani, and Sclerotinia sclerotiorum. 100 ppm (or 100 mg / L) of each tested peptide were applied to amended agar, and subsequent growth of each respective pathogen colony was measured as the diameter of colony in millimeters. Positive control was applied in 0.5 ppm (or 0.5 mg / L), and the negative control remained untreated.

[0189] Two replicates were performed for each peptide and control, and 4 measurements were made per sample.

[0190] Growth was calculated as each peptide’s corresponding colony growth relative to the negative control (unamended agar) colony growth. This may be calculated by dividing the radial growth of the peptide’s associated colony by the radial growth of the negative control’s associated colony.

[0191] Growth control was calculated as each peptide’s corresponding colony growth (mm) relative to the colony growth (mm) corresponding to the negative control (unamended agar). This may be calculated by subtracting the radial growth of the peptide’s corresponding colony (mm) from the radial growth ofthe negative control’s corresponding colony (mm), then dividing that quotient by the radial growth of the negative control’s corresponding colony (mm). This may also be calculated by subtracting the % growth from 100% (also equivalent to the % growth of the negative control).Table 5. Classification and importance of tested pathogens, and corresponding peptide antimicrobial activity.Results

[0192] Peptide MP19919 exhibited antimicrobial activity on all tested pathogens when EC50 was measured. It exhibited an EC ,1 of less than 50mg / L against Z tritici, P.cactorum, B.cinerea, and AT. nivale, and it exhibited an EC50 of less than lOOmg / L against A. alternata, F.graminearum, and P .tritici-repentis (FIG. 11, light dotted bars). Peptide MP19594 exhibited selective antimicrobial activity, with an EC50 less than 50mg / L against Z. tritici and P.cactorum and an EC50 of less than lOOmg / L against A. alternata, F.graminearum, P. tritici-repentis, B. cinerea, andA nivale (FIG. 11, black bars). The negative control peptide did not show significant antimicrobial activity against any tested pathogen in measuring EC50.

[0193] In testing % control, peptide MP19919 provided 28% control and 36% control against P. ultimum and R. solani respectively at 100 ppm in vitro, (FIG. 12), calculated from % growth shown in FIG. 14, which was calculated from radial growth shown in FIG. 13. MP 19594 provided 13% control against P. ultimum. The peptides did not provide significant control against S. sclerotiorum, and the negative control peptide and MP19594 did not provide significant control against any of the three tested pathogens.

[0194] Table 5 shows the pathogens against which peptides MP19919 and / or MP19594 exhibited antimicrobial activity (marked “Active”).

Claims

CLAIMS1. A method of increasing immunity in a plant comprising providing to the plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, or the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21, with one, two, three, or four amino acid substitutions, insertions, or deletions thereof.

2. The method of claim 1, wherein the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

3. A method of increasing resistance to a plant pathogen or pest in a plant comprising providing to the plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14- 21 or the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 with one, two, three, or four amino acid substitutions, insertions, or deletions.

4. A method of inhibiting a plant pathogen or pest on a plant comprising providing to the plant an oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 or the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 with one, two, three, or four amino acid substitutions, insertions, or deletions.

5. The method of claim 3 or 4, wherein the plant pathogen or pest is selected from fungal pathogens or pests, bacterial pathogens or pests, and viral pathogens or pests.

6. The method of claim 3 or 4, wherein the plant pathogen or pest is a fungal pathogen or pest selected from Cercospora spp., Mycosphaerella spp., Glomerella spp., Cladosporium spp., Diplodia maydis, Fusarium oxysporum, Fusarium graminearum, Fusarium monilforme, Fusarium verticillioides, Cochliobolus sativus, Collectotrichum graminicola, Stagonospora nodorum, Stagonospora avenae, Stenocarpella maydis, Sclerotinia minor, Sclerotinia sclerotiorum, Sclerotinia sp., Alternaria spp., Phytophthora spp., Botrytis spp., Pyrenophora tritici-repentis, Phytophthora parasitica, Phytophthora megasperma fsp. glycinea, Macrophomina phaseolina, Rhizoctonia solani, Sclerotinia sclerotiorum, Fusarium oxysporum, Diaporthe phaseolorum var. sojae (Phomopsis sojae), Diaporthe phaseolorum var. caulivora, Sclerotium rolfsii, Cercospora kikuchii, Cercospora sojina, Peronospora manshurica, Colletotrichum dematium (Colletotichum truncatum), Corynespora cassiicola, Septoria glycines, Phyllosticta sojicola, Alternaria alternata, Pseudomonas syringae p.v. glycinea, Xanthomonas campestris p.v. phaseoli, Microsphaera diffusa, Fusarium semitectum, Phialophora gregata, Glomerella glycines, Phakopsora pachyrhizi, Pythium aphanidermatum, Pythium ultimum, PythiumClaviceps sorghi, Pseudomonas avenae, Erwinia chrysanthemi pv. zea, Erwinia carotovora, Corn stunt spiroplasma, Diplodia macrospora, Sclerophthora macrospora, Peronosclerospora sorghi,Peronosclerospora philippinensis, Peronosclerospora maydis, Peronosclerospora sacchari, Sphacelotheca reiliana, Physopella zeae, Cephalosporium maydis, Cephalosporium acremonium, Exserohilum turcicum, C. sublineolum, Cercospora sorghi, Gloeocercospora sorghi, Ascochyta sorghina, Pseudomonas syringae p.v. syringae, Xanthomonas campestris p.v. holcicola, Pseudomonas andropogonis, Puccinia purpurea, Macrophomina phaseolina, Perconia circinata, Fusarium verticillioides, Altemaria altemata, Bipolaris sorghicola, Helminthosporium sorghicola, Curvularia lunata, Phoma insidiosa, Pseudomonas avenae (Pseudomonas alboprecipitans), Ramulispora sorghi, Ramulispora sorghicola, Phyllachara sacchari, Sporisorium reilianum (Sphacelotheca reiliana), Sphacelotheca cruenta, Sporisorium sorghi, Claviceps sorghi, Rhizoctonia solani, Acremonium strictum, Sclerophthona macrospora, Peronosclerospora sorghi, Peronosclerospora philippinensis, Sclerospora graminicola, Fusarium graminearum, Fusarium oxysporum, Pythium arrhenomanes, and Pythium graminicola.

7. The method of claim 3 or 4, wherein the plant pathogen or pest is a bacterial pathogen or pest selected from Pseudomonas spp., Pantoua spp., and Erwinia spp.

8. The method of claim 3 or 4, wherein the plant pathogen or pest is a viral pathogen or pest selected from cucumber mosaic, tobacco mosaic, and barley yellow dwarf virus, alfalfa mosaic virus (Alfamovirus), Apple chlorotic leaf spot virus (Trichovirus), Apple scar skin viroid (Viroids), Arabis mosaic virus (Nepovirus), Barley mild mosaic virus (Bymovirus), Barley stripe mosaic virus (Hordeivirus), Barley yellow mosaic virus (Bymovirus), Bean common mosaic virus (Potyvirus), Bean yellow mosaic virus (Potyvirus), Beet necrotic yellow vein virus (Furovirus), Blackeye cowpea mosaic virus (Potyvirus), Bean common mosaic virus (Potyvirus), Broad bean wilt virus (Fabavirus), Butterbur mosaic virus (Carlavirus), Carnation mottle virus (Carmovirus), Carnation vein mottle virus (Potyvirus), Cauliflower mosaic virus (Caulimovirus), Chrysanthemum mild mottle virus (Cucumovirus), Tomato aspermy virus (Cucumovirus), Chrysanthemum stunt viroid (Viroids), Citrus mosaic virus, Citrus fristeza virus (Closterovirus), Clover yellow vein virus (Potyvirus), Cocksfoot mottle virus (Sobemovirus), Cucumber green mottle mosaic virus (Tobamovirus), Cucumber mosaic virus (Cucumovirus), Cycas necrotic stunt virus (Nepovirus), Dasheen mosaic virus (Potyvirus), Grapevine Algerian latent virus (Tombusvirus), Konjac mosaic virus (Potyvirus), Melon necrotic spot virus (Carmovirus), Mulberry ringspot virus (Nepovirus), and Narcissus mosaic virus (Potexvirus). Plant viruses are viruses affecting plants. Additional examples of viruses affecting plants include Odontoglossum ringspot virus (Tobamovirus), Papaya ringspot virus (Potyvirus), Peach latent mosaic viroid, Peanut mottle virus (Potyvirus), Peanut stripe virus (Potyvirus), Bean common mosaic virus(Potyvirus), Peanut stunt virus (Cucumovirus), Potato virus A (Potyvirus), Potato virus M (Carlavirus), Potato virus S (Carlavirus), Potato virus X (Potexvirus), Potato virus Y (Potyvirus), Prune dwarf virus (Ilarvirus), Primus necrotic ringspot virus (Ilarvirus), Radish mosaic virus (Comovirus), Rice black streaked dwarf virus (Fijivirus), Rice dwarf virus (Reovirus), Rice grassy stunt virus (Tenuivirus), Rice stripe virus (Tenuivirus), Rice tungro spherical virus (Sequivirus), Rice waika virus, Rice tungro spherical virus (Sequivirus), Ryegrass mottle virus, Satsuma dwarf virus (Nepovirus), Soil-borne wheat mosaic virus (Furovirus), Southern bean mosaic virus (Sobemovirus), Soybean mosaic virus (Potyvirus), Soybean stunt virus (Cucumovirus), Cucumber mosaic virus (Cucumovirus), Tobacco mosaic virus (Tobamovirus), Tobacco mosaic virus (Tobamovirus), Tomato mosaic virus (Tobamovirus), Tobacco necrosis virus (Necrovirus), Tobacco rattle virus (Tobravirus), Tobacco ringspot virus (Nepovirus), Tomato aspermy virus (Cucumovirus), Tomato black ring virus (Nepovirus), Tomato mosaic virus (Tobamovirus), Tomato ringspot virus (Nepovirus), Tomato spotted wilt virus (Tospovirus), Turnip mosaic virus (Potyvirus), Watermelon mosaic virus 1 (Potyvirus), Papaya ringspot virus (Potyvirus), Watermelon mosaic virus 2 (Potyvirus), Wheat yellow mosaic virus (Bymovirus), and Zucchini yellow mosaic virus (Potyvirus).

9. The method of claim 3 or 4, wherein the plant pathogen or pest is selected from the group consisting of Septaria tritici, Botrytis cinerea, Alternaria solani, Rhizoctonia solani anastomosis, Phytophthora capsici, and Phytophthora infestans.

10. The method of any one of claims 1-9, wherein the oligopeptide is provided by expressing a nucleic acid encoding the oligopeptide.

11. The method of claim 10, wherein the nucleic acid is operably linked to a heterologous promoter.

12. The method of claim 10 or claim 11, wherein the heterologous promoter is a constitutive promoter, a tissue specific promoter, or an inducible promoter.

13. The method of claim 12, wherein the inducible promoter is induced by fungal infection, optionally a promoter associated with a gene involved in phenylpropanoid metabolism (e.g., phenylalanine ammonia lyase, chaicone synthase promoters), a gene that modifies plant cell walls (e.g., hydroxyproline-rich glycoprotein, glycine-rich protein, and peroxidase promoters), a gene encoding an enzyme that degrade fungal cell walls (e.g., chitinase or glucanase promoters), or a gene encoding a thaumatin-like protein, or optionally a maize Misl promoter or a flax Fisl promoter.

14. The method of any one of claims 1-9, wherein the oligopeptide is provided by application to the plant or a part thereof.

15. The method of claim 14, wherein the plant part is a leaf, a bud, a root, a shoot, a floral part, or a seed.

16. The method of claim 14 or 15, wherein the oligopeptide is applied as a coating to the seed prior to planting.

17. The method of any one of claims 1-9, wherein the oligopeptide is provided by application to the soil in which the plant is planted.

18. The method of any one of claims 1-9, wherein the oligopeptide is provided by addition to water provided to the plant.

19. The method of any one of claims 1-9, wherein the oligopeptide is provided following harvest to seeds, fruits, and / or plant parts.

20. The method of any one of claims 14-19, wherein the application is by a spreader, a power duster, a boom sprayer, a hand sprayers, a spray dusters, or a granular applicator.

21. The method of any one of claims 1-20, wherein the miPEP comprises a secretion tag, a tag that promotes entry into a plant cell, or a nuclear localization tag.

22. The method of claim 21, wherein the tag that promotes entry into a plant cell comprises a cell penetrating peptide linked to the amino acid sequence.

23. The method of claim 22, wherein the cell penetrating peptide comprises a protein transduction domain, an amphipathic peptide, a synthetic cationic polypeptide, optionally polylysine, polyhistidine, or polyarginine, a dendrimeric polycationic molecule, a peptide vascular endothelial-cadherin cell penetrating peptide, a fransportan cell penetrating peptide, a monomer or dimer of HIV- 1 TAT basic domain cell penetrating peptide, a penetratin cell penetrating peptide, a synthetic cationic homoarginine oligopeptide cell penetrating peptide, a gamma zein cell penetrating peptide, a Zea mays knotted 1 cell penetrating peptide, a Saccharomyces pombe TP 10 cell penetrating peptide, a Candida albicans Zebra cell penetrating peptide, an Antennapedia sequence, a TAT sequence , an Antp-3A (Antp mutant) sequence, a Buforin II sequence, a K-FGF sequence, a Ku70 sequence, a prion sequence, a pVECsequence, a SynBl sequence, a Pep-7 sequence, a HN-1 sequence, a BGSC (Bis-Guanidinium- Spermidine-Cholesterol) sequence, or a BGTC (Bis-Guanidinium-Tren-Cholesterol) sequence.

24. An isolated oligopeptide comprising the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 or the amino acid sequence set forth in one of SEQ ID NOs: 1-5 and 14-21 comprising one, two, three, or four amino acid substitutions, insertions, or deletions.

25. The oligopeptide of claim 24, wherein the oligopeptide is an antimicrobial oligopeptide or an antifungal oligopeptide.

26. The oligopeptide of claim 24 or claim 25, comprising the one, two, three, or four amino acid substitutions, insertions, or deletions.

1. The oligopeptide of any one of claims 24-26, wherein the oligopeptide comprises a proline at its N- terminus, an aspartic acid at the C-terminus, or both.

28. The oligopeptide of any one of claims 24-27, wherein the oligopeptide comprises one or more D- amino acids.

29. The oligopeptide of claim 28, wherein the one or more D-amino acids enhance stability of the oligopeptide as compared to an oligopeptide of the same amino acid sequence comprising all L-amino acids.

30. The oligopeptide of any one of claims 24-29, wherein the oligopeptide comprises a secretion tag, a tag that promotes entry into a plant cell, or a nuclear localization tag.

31. The oligopeptide of claim 30, wherein the tag that promotes entry into a plant cell comprises a cell penetrating peptide linked to the amino acid sequence.

32. The oligopeptide of claim 31, wherein the cell penetrating peptide comprises a protein transduction domain, an amphipathic peptide, a synthetic cationic polypeptide, optionally polylysine, polyhistidine, or polyarginine, a dendrimeric polycationic molecule, a peptide vascular endothelial-cadherin cell penetrating peptide, a transportan cell penetrating peptide, a monomer or dimer of HIV- 1 TAT basic domain cell penetrating peptide, a penetratin cell penetrating peptide, a synthetic cationic homoarginine oligopeptide cell penetrating peptide, a gamma zein cell penetrating peptide, a Zea mays knotted 1 cell penetrating peptide, a Saccharomyces pombe TP 10 cell penetrating peptide, a Candida albicans Zebra cell penetrating peptide, an Antennapedia sequence, a TAT sequence , an Antp-3A (Antp mutant)sequence, a Buforin II sequence, a K-FGF sequence, a Ku70 sequence, a prion sequence, a pVEC sequence, a SynBl sequence, a Pep-7 sequence, a HN-1 sequence, a BGSC (Bis-Guanidinium- Spermidine-Cholesterol) sequence, or a BGTC (Bis-Guanidinium-Tren-Cholesterol) sequence.

33. A composition comprising the oligopeptide of any one of claims 24-32 with an agriculturally acceptable formulant that can include water, organic solvents, paraffinic oils, vegetable oils, dispersants, emulsifiers, wetting agents, buffering agents, hydrotrope agents, rheology modifiers, antifoam agents and defoamers, antifreeze agents, biocides, dyes, polymer walls, catalysts, thermosetting materials, cross-polymerizing agents, UV protectants, antioxidants, and chelating agents.

34. A composition comprising the oligopeptide of any one of claims 24-32 with an agriculturally acceptable carrier, diluent, or excipient.

35. The composition of claim 34, wherein the agriculturally acceptable carrier, diluent, or excipient comprises a buffer and the pH is in the range of from about 3.0 to about 9.0, or from about 4.5 to about 8.0.

36. The composition of any one of claims 33-35, wherein the oligopeptide is in a concentration range of from about 0. 1 pg / ml to about 100 mg / ml, or from about 5 pg / ml to about 5 mg / ml.

37. The composition of any one of claims 33-36, wherein the composition is in the form of a finely- divided particulate solid, granules, pellets, wettable powders, dust, an aqueous suspension, a dispersion, a gel, or an emulsion.

38. The composition of any one of claims 33-37, fiirther comprising one or more other active agents selected from the group consisting of a pesticide, a fertilizer, an insecticide, an attractant, a sterilizing agent, an acaricide, a nematocide, a herbicide, a biostimulant, a biological, and a growth regulator.

39. A nucleic acid encoding the oligopeptide of any one of claims 24-32.

40. A nucleic acid construct comprising the nucleic acid of claim 39, operably linked to a promoter.

41. The nucleic acid construct of claim 40, wherein the promoter is a heterologous promoter.

42. The nucleic acid construct of claim 40 or claim 41, wherein the promoter is a constitutive promoter, a tissue specific promoter, a development stage specific promoter, or an inducible promoter.

43. The nucleic acid construct of claim 42, wherein the inducible promoter is induced by fungal infection, optionally a promoter associated with a gene involved in phenylpropanoid metabolism (e.g., phenylalanine ammonia lyase, chaicone synthase promoters), a gene that modifies plant cell walls (e.g., hydroxyproline-rich glycoprotein, glycine-rich protein, and peroxidase promoters), a gene encoding an enzyme that degrade fungal cell walls (e.g., chitinase or glucanase promoters), or a gene encoding a thaumatin-like protein, or a maize Misl promotor or a flax Fisl promoter.

44. A cell comprising the nucleic acid of claim 39 or the nucleic acid construct of any one of claims 40- 43.

45. The method of any one of claims 1-23, oligopeptide of any one of claims 25-32, composition of any one of claims 33-38, nucleic acid construct of any one of claims 39-43, or the cell of claim 44, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 19 or SEQ ID NO: 16, or wherein the oligopeptide comprises the amino acid sequence set forth in one of SEQ ID NO: 19 or SEQ ID NO: 16, with one, two, three, or four amino acid substitutions, insertions, or deletions thereof.

46. The method of any one of claims 2-23 and 45, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 19 or wherein the amino acid sequence set forth in SEQ ID NO: 19 with one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and wherein the plant pathogen or pest selected from Phytium ultimum, Rhizoctonia solani, Zymoseptoria tritici, Phythopthora cactorum, Botrytis cinerea, Microdochium nivale, Alternaria alternata, Pyrenophora tritici-repenti, and Fusarium graminearum.

47. The method of any one of claims 2-23 and 45, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 16 or wherein the amino acid sequence set forth in SEQ ID NO: 16 with one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and wherein the plant pathogen or pest selected from Zymoseptoria tritici, Phythopthora cactorum, Botrytis cinerea, Microdochium nivale, and Alternaria alternata.

48. The method of any one of claims 2-23 and 45, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 19 or wherein the amino acid sequence set forth in SEQ ID NO: 19 with one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and wherein the plant pathogen or pest selected from Phytium ultimum, Rhizoctonia solani, Zymoseptoria tritici, Botrytis cinerea, Alternaria alternata, Pyrenophora tritici-repenti, and Fusarium graminearum.

49. The method of any one of claims 2-23 and 45, wherein the oligopeptide comprises the amino acid sequence set forth in SEQ ID NO: 16 or wherein the amino acid sequence set forth in SEQ ID NO: 16 with one, two, three, or four amino acid substitutions, insertions, or deletions thereof, and wherein the plant pathogen or pest selected from Zymoseptoria tritici, Botrytis cinerea, and Alternaria alternata.

Images