Microbial compositions for use in plants to prevent or reduce fungal pathogens - Patents.com

Abstract

Disclosed herein are biocontrol compositions for plant fungal pathogens and methods for their use to prevent or reduce crop loss or food quality deterioration. The biocontrol compositions may contain at least one microorganism or at least one secondary metabolite of a microorganism with antifungal or antipathogenic activity. The methods and compositions disclosed herein can prevent or inhibit the growth of a variety of different pathogens, including Penicillium pathogens. The biocontrol compositions can be applied to plants, seeds, or agricultural products thereof, or to packaging materials used to transport or store agricultural products.

Description

[Technical Field]

[0001] cross reference This application claims priority to U.S. Provisional Application No. 62 / 885,114, filed August 9, 2019, which is incorporated herein by reference in its entirety. [Background technology]

[0002] background Fungal pathogens cause significant agricultural losses, resulting in crop losses, food waste, and economic losses.Microorganisms with antifungal properties have been developed as biocontrol agents to reduce both crop losses and food quality deterioration caused by these fungal pathogens.Commercially available products cannot demonstrate the desired plant or fungal specificity or effectiveness.In addition, there are limited options for post-harvest protection of agricultural products, especially organic agricultural products.Biological control compositions for preventing fungal growth can provide an alternative to currently available products. Summary of the Invention [Means for solving the problem]

[0003] Abstract In one aspect, the disclosure provides a biocontrol composition comprising (i) at least one microorganism, or a metabolite produced by at least one microorganism, and (ii) a carrier, wherein the at least one microorganism comprises a 16S rRNA sequence greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:1, SEQ ID NO:22, or SEQ ID NO:23, and wherein the biocontrol composition is capable of inhibiting the growth of Penicillium species relative to a control not exposed to the biocontrol composition. In some embodiments, the at least one microorganism comprises a 16S rRNA sequence greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:1. In some embodiments, the 16S rRNA sequence is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:22. In some embodiments, the 16S rRNA sequence is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:23.

[0004] In some aspects, the disclosure provides a biocontrol composition comprising (i) at least one microorganism, or a metabolite produced by at least one microorganism, and (ii) a carrier, wherein the at least one microorganism comprises a 16S rRNA sequence greater than 90% identical to the 16S rRNA sequence of SEQ ID NO:24, or the at least one microorganism comprises an internal transcribed spacer (ITS) sequence greater than 90% identical to the 16S rRNA sequence of SEQ ID NO:25, and the biocontrol composition is capable of inhibiting the growth of Penicillium species relative to a control not exposed to the biocontrol composition. In some embodiments, the at least one microorganism comprises a 16S rRNA sequence greater than 90% identical to the 16S rRNA sequence of SEQ ID NO:24. In some embodiments, the at least one microorganism comprises a 16S rRNA sequence greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:24. In some embodiments, the at least one microorganism comprises an internal transcribed spacer (ITS) sequence greater than 90% identical to the 16S rRNA sequence of SEQ ID NO:25. In some embodiments, the at least one microorganism comprises an internal transcribed spacer (ITS) sequence greater than 99% identical to the ITS sequence of SEQ ID NO: 25. In some embodiments, the at least one microorganism is at least two microorganisms, including a first microorganism comprising a 16S rRNA sequence greater than 90% identical to the 16S rRNA sequence of SEQ ID NO: 24 and a second microorganism comprising an ITS sequence greater than 90% identical to the ITS sequence of SEQ ID NO: 25. In some embodiments, growth inhibition of Penicillium species is indicated by a reduction in lesion size or tissue necrosis in produce exposed to the biocontrol composition compared to a control not exposed to the biocontrol composition. In some embodiments, the biocontrol composition is capable of inhibiting the growth of Penicillium species by 5% or more compared to a control not exposed to the biocontrol composition. In some embodiments, the biocontrol composition is capable of inhibiting the growth of Penicillium species by 25% or more compared to a control not exposed to the biocontrol composition. In some embodiments, the Penicillium is Penicillium expansum.In some embodiments, the Penicillium is Penicillium digitatum. In some embodiments, the biocontrol composition comprises vegetative cells. In some embodiments, the biocontrol composition comprises spores. In some embodiments, the carrier is selected from the group consisting of oil, water, wax, resin, kaolinite clay, diatomaceous earth, or flour. In some embodiments, the carrier is water. In some embodiments, the biocontrol composition is formulated in liquid form. In some embodiments, the biocontrol composition is formulated in liquid form. In some embodiments, the biocontrol composition is formulated in powder form.

[0005] In another aspect, the disclosure provides a method of reducing or preventing the growth of a pathogen in a plant, seed, flower, or produce thereof, comprising applying a biocontrol composition to the plant, seed, flower, or produce thereof.

[0006] In another aspect, the disclosure provides a method of reducing or preventing the growth of pathogens in a plant, seed, flower, or produce thereof, comprising applying a biocontrol composition to an object or area adjacent to the plant, seed, flower, or produce. In some embodiments, the applying step occurs before harvesting the plant, seed, flower, or produce. In some embodiments, the applying step occurs after harvesting the plant, seed, flower, or produce. In some embodiments, the area adjacent to the plant includes soil used to grow the plant, seed, flower, or produce. In some embodiments, the object adjacent to the plant includes packaging used to store or transport the plant, seed, flower, or produce. In some embodiments, the applying step occurs by spraying the biocontrol composition. In some embodiments, the applying step occurs by dipping the plant, seed, flower, or produce in the biocontrol composition. In some embodiments, the plant is selected from the group consisting of almonds, apricots, apples, artichokes, bananas, barley, beets, blackberries, blueberries, broccoli, Brussels sprouts, cabbage, hemp, rapeseed, peppers, carrots, celery, Swiss chard, cherries, citrus fruits, corn, gourds, dates, figs, flax, garlic, grapes, herbs, spices, kale, lettuce, mint, oil palm, olives, onions, peas, pears, peaches, peanuts, papayas, parsnips, pecans, persimmons, plums, pomegranates, potatoes, quince, radishes, raspberries, roses, rice, plums, sorghum, soybeans, spinach, strawberries, sweet potatoes, tobacco, tomatoes, turnip greens, walnuts, and wheat. In some embodiments, the plant is an apple. In some embodiments, the plant is a member of the genus Malus. In some embodiments, the plant is a member of the genus Citrus. In some embodiments, the citrus fruit comprises a mandarin, a lemon, a lime, a navel orange, a pomelo, or a hybrid thereof.

[0007] In another aspect, the disclosure provides a method of inhibiting the growth of a pathogen, the method comprising applying a biocontrol composition to an apple, wherein the biocontrol composition comprises (i) at least one microorganism, or a metabolite produced by the at least one microorganism, and (ii) a carrier, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:1, SEQ ID NO:22, or SEQ ID NO:23, and wherein the biocontrol composition is capable of inhibiting the growth of the species Penicillium expansum relative to a control not exposed to the biocontrol composition.

[0008] In another aspect, the disclosure provides a method of inhibiting the growth of a pathogen, the method comprising applying a biocontrol composition to an apple, wherein the biocontrol composition comprises (i) a first microorganism and a second microorganism, or a metabolite produced by the first microorganism or the second microorganism, and (ii) a carrier, wherein the first microorganism comprises a 16S rRNA sequence that is greater than 90% identical to the 16S rRNA sequence of SEQ ID NO:24, and the second microorganism comprises an ITS sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO:25, and the biocontrol composition is capable of inhibiting the growth of the species Penicillium expansum relative to a control that is not exposed to the biocontrol composition. In another aspect, the disclosure provides a method of inhibiting the growth of a pathogen, the method comprising applying a biocontrol composition to a citrus plant, wherein the biocontrol composition comprises (i) at least one microorganism, or a metabolite produced by the at least one microorganism, and (ii) a carrier, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:1, SEQ ID NO:22, or SEQ ID NO:23, and wherein the biocontrol composition is capable of inhibiting the growth of the species Penicillium expansum relative to a control not exposed to the biocontrol composition.

[0009] In another aspect, the disclosure provides a method of inhibiting the growth of a pathogen, the method comprising applying a biocontrol composition to a citrus plant, wherein the biocontrol composition comprises (i) a first microorganism and a second microorganism, or a metabolite produced by the first microorganism or the second microorganism, and (ii) a carrier, wherein the first microorganism comprises a 16S rRNA sequence greater than 90% identical to the 16S rRNA sequence of SEQ ID NO:24, and the second microorganism comprises an ITS sequence greater than 90% identical to the ITS sequence of SEQ ID NO:25, and the biocontrol composition is capable of inhibiting the growth of the species Penicillium expansum relative to a control not exposed to the biocontrol composition.

[0010] Another aspect of the present disclosure provides a non-transitory computer-readable medium containing machine-executable code that, when executed by one or more computer processors, implements any of the methods described above or elsewhere herein.

[0011] Another aspect of the present disclosure provides a system including one or more computer processors and a computer memory coupled thereto, the computer memory including machine-executable code that, when executed by the one or more computer processors, implements any of the methods described above or elsewhere herein.

[0012]

[0013] Further aspects and advantages of the present disclosure will become readily apparent to those skilled in the art from the following detailed description, wherein merely illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the present disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature and not as restrictive. INCORPORATION BY REFERENCE

[0013] All publications, patents, and patent applications mentioned herein are incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. In the event that the publications and patents or patent applications incorporated by reference conflict with the present disclosure contained herein, the present specification is intended to supersede and / or take precedence over any such conflicting material.

[0014] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also referred to herein as "Figure" and "Fig."). [Brief explanation of the drawings]

[0015] [Figure 1] FIG. 1 shows a schematic diagram of a method for using and producing a biocontrol composition.

[0016] [Figure 2] Figure 2 illustrates the average lesion size for treated and untreated Fuji and Gala apples.

[0017] [Figure 3] Figure 3 illustrates apple rot for treated and untreated Fuji and Gala apples.

[0018] [Figure 4] 4A-4B illustrate the mean lesion size and mean weight of necrosis in treated and untreated Fuji apples.

[0019] [Figure 5] Figure 5 illustrates Fuji apples 6 days after infection.

[0020] [Figure 6]6A-6B illustrate the mean lesion size and mean weight of necrosis in treated and untreated Gala apples.

[0021] [Figure 7] Figure 7 illustrates Gala apples 6-7 days after infection.

[0022] [Figure 8] Figure 8A shows a schematic diagram of the seeding location of the biocontrol composition, and Figure 8B shows a photograph of the growth of the biocontrol composition on a citrus media plate.

[0023] [Figure 9] FIG. 9 shows a photograph of the inhibition of P. digitatum on citrus media plates.

[0024] [Figure 10] FIG. 10 shows a photograph of the inhibition of P. digitatum on citrus media plates. DETAILED DESCRIPTION OF THE INVENTION

[0025] Detailed Description While various embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous modifications, changes, and substitutions may occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed.

[0026] Provided herein are compositions, formulations, and methods of use of microorganisms, microbial consortia, or microbial aggregates for use on plants to prevent or reduce pathogens. The compositions, formulations, and methods described herein also relate to supernatants or culture compositions produced from or containing microorganisms, microbial consortia, or microbial aggregates for use on plants to prevent or reduce pathogens. These compositions may be referred to as biocontrol compositions. In particular, the compositions and formulations, as well as their methods of use, may be effective against fungal pathogens. The fungal pathogen may be a member of the Penicillium genus. For example, the fungal pathogen may be Penicillium expansum, also known as blue mold. In another example, the fungal pathogen may be Penicillium digitatum. The fungal pathogen may be Botrytis cinerea.

[0027] The plant may be a flower, a seed or an agricultural product. The plant, flower, seed, or produce thereof can be almond, apricot, apple, artichoke, banana, barley, beet, blackberry, blueberry, broccoli, Brussels sprouts, cabbage, hemp, rapeseed, chili pepper, carrot, celery, Swiss chard, cherry, citrus fruit, corn, gourd, date, fig, flax, garlic, grapes, herbs, spices, kale, lettuce, mint, oil palm, olive, onion, pea, pear, peach, peanut, papaya, parsnip, pecan, persimmon, plum, pomegranate, potato, quince, radish, raspberry, rose, rice, plum, sorghum, soybean, spinach, strawberry, sweet potato, tobacco, tomato, turnip greens, walnut, or wheat. The plant may be a member of the Citrus or Malus genus. For example, the plant may be a mandarin, lemon, or navel orange. The plant may be an apple. The plant may be a particular cultivar. For example, the apple may be a Fuji apple.

[0028] Selection of microbial consortia

[0029] Methods for identifying or selecting biocontrol compositions containing microbial consortia can be used. For example, methods such as those disclosed in U.S. Patent Application Publication No. 20180127796 can be used to identify or select for microbial consortia. In some cases, multiple species of microorganisms can be grown together. In some cases, the method can include diluting a sample to form multiple dilutions, where one dilution in the multiple dilutions contains a subset of multiple species of microorganisms. The dilutions can allow for the generation of multiple subsets in which different microorganisms of the multiple species can interact. The subset of multiple species of microorganisms can be cultured, so that the microorganisms can grow. The subset can be subjected to a sequencing reaction, so that the sequence of the microorganism can be obtained. Species, strains, or other taxonomic information can be obtained from the sequencing reaction. Sequences for identifying specific microorganisms are discussed elsewhere herein. The subset can be cultured for various times and subjected to sequencing reactions at various times to monitor the presence and / or relative abundance of specific species, strains, or other taxonomic categories. By observing the presence and / or relative abundance changes of specific species, strains, or other taxonomic categories, the interaction between multiple microorganisms can be determined.For example, when a first microorganism is cultured with a second microorganism, it can have a higher relative abundance compared to its relative abundance when not cultured with the second microorganism.In this example, the first microorganism can interact with the second microorganism, thereby increasing the overall viability of the first microorganism.Each of the multiple dilutions can be subjected to sequencing reactions, thereby allowing the microorganisms in each dilution to be identified, and multiple dilutions can enable multiplexed high-throughput approaches.

[0030] The multiple microorganisms can be diluted so that subsets of the multiple microorganisms grow together. In some cases, serial dilutions of the multiple microorganisms can be performed to form multiple serial dilutions of the sample. The microorganisms in the multiple serial dilutions of the sample can be due to dispersion or chance. The multiple serial dilutions can vary in different implementations. In some embodiments, multiple serial dilutions of a sample may include dilutions of a sample at or about 1:10, 1:100, 1:1000, 1:10000, 1:100000, 1:1000000, 1:10000000, 1:100000000, 1:100000000, or a number or range between any two of these values. In some embodiments, the multiple serial dilutions of a sample may include at least or at most 1:10, 1:100, 1:1000, 1:10000, 1:10000, 1:100000, 1:1000000, 1:10000000, 1:100000000, or 1:1000000000 dilutions of the sample. For example, a sample can be diluted 10-fold to a 1:10 dilution of the sample, for example, using a buffer. The 1:10 dilution of the sample can be diluted 10-fold to a 1:100 dilution of the sample. The multiple serial dilutions may include a 1:10 dilution of the sample, a 1:100 dilution of the sample, and other similarly prepared dilutions of the sample. As another example, a sample can be diluted 10-fold to a 1:10 dilution of the sample, for example, using a buffer. A sample can be diluted 100-fold to a 1:100 dilution of the sample. The multiple serial dilutions may include a 1:10 dilution of the sample, a 1:100 dilution of the sample, and other dilutions of the sample prepared similarly.

[0031] In some embodiments, culturing multiple dilutions of the sample in the first culture conditions comprises culturing multiple dilutions of the sample in the first culture conditions for multiple time periods that may vary from as little as one minute to up to one year.

[0032] Multiple kinds of microorganisms can be subjected to sequencing reaction, and specific microorganisms can be identified.When subset is cultured for a certain period, the overall percentage representation of each microorganism in subset can change from the percentage at the beginning of culture.For example, the microorganisms that survive in other microorganisms after different culture periods can indicate the symbiotic relationship or interaction between the microorganisms in culture, and these microorganisms can form a microbial community.Microorganism community can be tested for its effectiveness in inhibiting the growth of fungal pathogens in a manner similar to that used to identify the effectiveness of at least one microorganism as described elsewhere herein.

[0033] Specific microorganisms can also be isolated for use in the methods or compositions described elsewhere herein. For example, multiple species of microorganisms can be subjected to serial dilution, resulting in the isolation of colonies of specific microorganisms. The serial dilutions can be cultured in liquid, semi-solid, or solid media, respectively. Multiple species of microorganisms can form colonies in semi-solid or solid media, such as agar plates. The colonies can be sufficiently dispersed so that they contain a single strain or species of microorganism. Specific microorganisms can also be isolated using physical separation methods, such as centrifugation. For example, multiple species of microorganisms can be cultured in liquid media and then centrifuged to isolate the microorganisms from the culture. Specific growth conditions can also be used to isolate specific microorganisms. For example, a specific microorganism may have a higher viability than another microorganism when cultured under anaerobic conditions. A specific microorganism may have a higher viability than another microorganism when cultured in a specific nutrient-rich medium.

[0034] Compositions for preventing or reducing crop loss and food quality deterioration Disclosed herein is a biocontrol composition that can prevent or reduce the growth of fungal pathogens in plants, seeds, or their agricultural products. The term "agricultural products" can be used herein to refer to edible parts of plants, such as leaves, stems, seeds, roots, flowers, or fruits. The term "plant" can be used herein to refer to any part of a plant, such as leaves, stems, seeds, roots, or fruits. Preventing or reducing the growth of fungal pathogens in plants, seeds, or their agricultural products can reduce the amount of crop loss and food quality loss before, during, or after harvesting the produce from the plant.

[0035] The at least one microorganism may be a bacterium or yeast. The at least one microorganism may comprise a microorganism of a genus selected from the group consisting of Bacillus, Burkholderia, Cutaneotrichosporon, Cyberlindnera, Gluconacetobacter, Gluconobacter, Hanseniaspora, Paraburkholderia, Pseudomonas, Torulaspora, and any combination thereof.

[0036] The at least one microorganism may comprise a microorganism selected from the group consisting of Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus velezensis, Cutaneotrichosporon jirovecii, Cutaneotrichosporon moniliiforme, Cutaneotrichosporon mucoides, Cyberlindnera mrakii, Cyberlindnera saturnus, Gluconacetobacter liquefaciens, Gluconobacter cerinus, Hanseniaspora uvarum, Paraburkholderia phytofirmans, Pseudomonas fluorescens, Pseudomonas frederiksbergensis, Pseudomonas lini, Pseudomonas migulae, Torulaspora delbrueckii, and any combination thereof.

[0037] At least one microorganism may be a microorganism of the genus Bacillus. At least one microorganism may be a microorganism of the genus Burkholderia. At least one microorganism may be a microorganism of the genus Cutaneotrichosporon. At least one microorganism may be a microorganism of the genus Cyberlindnera. At least one microorganism may be a microorganism of the genus Gluconacetobacter. At least one microorganism may be a microorganism of the genus Gluconobacter. At least one microorganism may be a microorganism of the genus Hanseniaspora. At least one microorganism may be a microorganism of the genus Paraburkholderia. At least one microorganism may be a microorganism of the genus Pseudomonas. At least one microorganism may be a microorganism of the genus Torulaspora.

[0038] At least one microorganism may be Bacillus amyloliquefaciens. At least one microorganism may be Bacillus subtilis. At least one microorganism may be Bacillus velezensis. At least one microorganism may be Cutaneotrichosporon jivrovecii. At least one microorganism may be Cutaneotrichosporon moniliiforme. At least one microorganism may be Cutaneotrichosporon mucoides. At least one microorganism may be Cyberlindnera mrakii. At least one microorganism may be Cyberlindnera saturnus. At least one microorganism may be Gluconacetobacter liquefaciens. At least one microorganism may be Gluconobacter cerinus. At least one microorganism may be Hanseniaspora uvarum. At least one microorganism may be Paraburkholderia phytofirmans. At least one microorganism can be Paraburkholderia fluroescens. At least one microorganism can be Paraburkholderia frederiksbergensis. At least one microorganism can be Pseudomonas lini. At least one microorganism can be Pseudomonas migulae. At least one microorganism can be Torulaspora delbrueckii.

[0039] The at least one microorganism is Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus velezensis, Cutaneotrichosporon jirovecii, Cutaneotrichosporon moniliiforme, Cutaneotrichosporon mucoides, Cyberlindnera mrakii, Cyberlindnera saturnus, Gluconacetobacter liquefaciens, Gluconobacter cerinus, Hanseniaspora uvarum, Paraburkholderia phytofirmans, Pseudomonas fluorescens, Pseudomonas frederiksbergensis, Pseudomonas lini, Pseudomonas migulae, Torulaspora The at least one microorganism may have at least about 70%, 75%, 80%, 85%, 87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity with the rRNA of a microorganism selected from the group consisting of: Saccharomyces cerevisiae, ...

[0040] The biocontrol composition may comprise (i) at least one microorganism or a secondary metabolite of at least one microorganism, and (ii) a carrier, wherein at least one microorganism has a 16S rRNA sequence that is greater than 98% identical to a 16S rRNA sequence selected from the group of SEQ ID NO:1 and SEQ ID NO:9, or at least one microorganism has an ITS sequence that is greater than 98% identical to an ITS sequence selected from the group of SEQ ID NO:17 and SEQ ID NO:20, or at least one microorganism has an ITS sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO:18.

[0041] The microorganism may comprise an RNA sequence having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, and SEQ ID NO:25.

[0042] The biocontrol composition may further comprise a second microorganism, wherein the second microorganism is not identical to at least one of the microorganisms. The second microorganism may comprise an RNA sequence having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, and SEQ ID NO:25. In some cases, the first microorganism and the second microorganism are of the same species. For example, both the first microorganism and the second microorganism may be Bacillus amyloliquefaciens. In a further non-limiting example, the biocontrol compositions disclosed herein may include a first microorganism and a second microorganism, and optionally more than two microorganisms, each of which is a different strain of the same species. In some cases, the first microorganism and the second microorganism are not the same species. For example, the first microorganism may be Gluconobacter cerinus, and the second microorganism may be Hanseniaspora uvarum. In some cases, the first microorganism and the second microorganism are not the same genus. In some cases, the first microorganism and the second microorganism are not in the same family. In some cases, the first microorganism and the second microorganism are not in the same order. In some cases, the first microorganism and the second microorganism are not in the same class. In some cases, the first microorganism and the second microorganism are not in the same phylum. In some cases, the first microorganism and the second microorganism are not in the same kingdom.

[0043] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity with an rRNA sequence of a Bacillus species. The Bacillus species can be Bacillus amyloliquefaciens, Bacillus subtilis, or Bacillus velezensis. The rRNA sequence can be a 16S sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity with SEQ ID NO:1 or SEQ ID NO:23.

[0044] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to an rRNA sequence of a Gluconacetobacter species. The Gluconacetobacter species can be Gluconacetobacter liquefaciens. The rRNA sequence can be a 16S sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, or SEQ ID NO:16.

[0045] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to an rRNA sequence of a Gluconobacter species. The Gluconobacter species can be Gluconobacter cerinus. The rRNA sequence can be a 16S sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO:24.

[0046] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to an rRNA sequence of a Burkholderia species or a Paraburkholderia species. The Paraburkholderia species can be Paraburkholderia phytofirmans. The rRNA sequence can be a 16S sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO:3, SEQ ID NO:7, or SEQ ID NO:9.

[0047] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to an rRNA sequence of a Pseudomonas species. The Pseudomonas species can be Pseudomonas fluorescens, Pseudomonas lini, Pseudomonas migulae, or Pseudomonas frederiksbergensis. The rRNA sequence can be a 16S sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO:6, SEQ ID NO:10, SEQ ID NO:15, or SEQ ID NO:22.

[0048] In one embodiment, the at least one microorganism includes at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO:8.

[0049] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to an rRNA sequence of a Cyberlindnera species. The Cyberlindnera species can be Cyberlinderna saturnus or Cyberlindera mrakkii. The rRNA sequence can be an ITS sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO: 17.

[0050] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to an rRNA sequence of a Hanseniaspora species. The Hanseniaspora species may be Hanseniaspora uvarum. The rRNA sequence may be an ITS sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO: 18 or SEQ ID NO: 25. In one embodiment, the at least one microorganism comprises at least one microorganism having at least 90% sequence identity to SEQ ID NO: 18 or SEQ ID NO: 25. In one embodiment, the at least one microorganism comprises at least one microorganism having at least 95% sequence identity to SEQ ID NO: 18 or SEQ ID NO: 25. In one embodiment, the at least one microorganism comprises at least one microorganism having at least 99% sequence identity to SEQ ID NO: 18 or SEQ ID NO: 25.

[0051] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to an rRNA sequence of a Torulaspora species. The Torulaspora species can be Torulaspora delbrueckii. The rRNA sequence can be an ITS sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO:19.

[0052] In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity with an rRNA sequence of a Cutaneotrichosporon species. The Cutaneotrichosporon species can be Cutaneotrichosporon moniliiforme, Cutaneotrichosporon jirovecii, or Cutaneotrichosporon mucoides. The rRNA sequence can be an ITS sequence. In one embodiment, the at least one microorganism comprises at least one microorganism having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity with SEQ ID NO:20 or SEQ ID NO:21.

[0053] The biocontrol composition may include a microbial consortium comprising a plurality of microorganisms. The plurality of microorganisms may be at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten microorganisms. Each microorganism in the plurality of microorganisms may be a different microorganism. The biocontrol composition may include secondary metabolites from a microbial consortium comprising a plurality of microorganisms, wherein the plurality of microorganisms is at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten microorganisms.

[0054] The at least two microorganisms may include at least two microorganisms selected from the group consisting of a microorganism having a 16S rRNA sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, and a microorganism having an ITS sequence selected from the group consisting of SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:25. The at least two microorganisms may include a first microorganism having a 16S rRNA sequence selected from SEQ ID NO:1 or SEQ ID NO:9, or the first microorganism has an ITS sequence that is greater than 98% identical to an ITS sequence selected from the group of SEQ ID NO:17 and SEQ ID NO:20, or the first microorganism has an ITS sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO:18. The at least two microorganisms may include a first microorganism having an ITS sequence greater than 90% identical to SEQ ID NO:18, and the second microorganism may be a Gluconacetobacter species. The Gluconacetobacter species may be Gluconacetobacter liquefaciens. The Gluconacetobacter species may be a Gluconacetobacter species having a 16S rRNA sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:16. The at least two microorganisms may include a first microorganism that is a Gluconobacter species and a second microorganism that is a Hanseniaspora species. The at least two microorganisms may include a first microorganism that is Gluconobacter cerinus and a second microorganism that is Hanseniaspora uvarum.

[0055] The at least two microorganisms may include a first microorganism having a 16S sequence that is greater than 90% identical to SEQ ID NO: 24 and a second microorganism having an ITS sequence that is greater than 90% identical to SEQ ID NO: 25. The at least two microorganisms may include a first microorganism having a 16S sequence that is greater than 95% identical to SEQ ID NO: 24 and a second microorganism having an ITS sequence that is greater than 95% identical to SEQ ID NO: 25. The at least two microorganisms may include a first microorganism having a 16S sequence that is greater than 98% identical to SEQ ID NO: 24 and a second microorganism having an ITS sequence that is greater than 98% identical to SEQ ID NO: 25.

[0056] The at least three microorganisms may include a first microorganism having a 16S rRNA sequence greater than 99% identical to SEQ ID NO:23, a second microorganism having a 16S rRNA sequence greater than 99% identical to SEQ ID NO:23, and a third microorganism having a 16S rRNA sequence greater than 99% identical to SEQ ID NO:23, wherein the first microorganism, the second microorganism, and the third microorganism comprise non-identical genomes. In some cases, the genomes may differ by single nucleotide polymorphisms (SNPs). In some cases, the genomes may differ by more than one SNP. In some cases, the genomes may differ in the number of genes in each genome. In some cases, the genomes may differ by rearrangements, such as insertions, deletions, reordering, refactoring, or by lysogenic or inactive phages, insertion sequences, repeated genomic sequences, or other different contents of genomic regions or genes. In some cases, the cellular DNA content may differ by the inclusion of one or more plasmids, which may vary between strains. In some cases, genomes can encode different isoforms of genes.For example, the protein expressed from a gene can contain point mutations, deletions, insertions that can affect the function of the protein.For example, the protein expressed from a gene can contain point mutations, deletions, insertions that can not affect the function of the protein or can not substantially affect the function of the protein.

[0057] The at least three microorganisms may include at least three microorganisms selected from the group consisting of microorganisms having a 16S rRNA sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24, and microorganisms having an ITS sequence selected from the group consisting of SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:25. The at least three microorganisms may include at least one microorganism having a 16S rRNA sequence selected from SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:23, or an ITS sequence selected from SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:20.

[0058] The at least four microorganisms may include at least four microorganisms selected from the group consisting of microorganisms having a 16S rRNA sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24, and microorganisms having an ITS sequence selected from the group consisting of SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:25. The at least four microorganisms may include at least one microorganism having a 16S rRNA sequence selected from SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:23, or an ITS sequence selected from SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:20.

[0059] The at least five microorganisms may include at least five microorganisms selected from the group consisting of microorganisms having a 16S rRNA sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24, and microorganisms having an ITS sequence selected from the group consisting of SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:25. The at least five microorganisms may include at least one microorganism having a 16S rRNA sequence selected from SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:23, or an ITS sequence selected from SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:20.

[0060] Table 1 sets forth the identifiers, putative microbial genus or species, and corresponding SEQ ID NOs for the microbial strains described herein. At least one microorganism can be a microorganism in Table 1. Table 2 sets forth the sequences corresponding to these SEQ ID NOs. [Table 1-1] [Table 1-2] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5]

[0061] At least one microorganism can be grown in culture. The at least one microorganism can be isolated and purified from the culture. The at least one microorganism purified from the culture can include vegetative cells or spores of at least one microorganism. The culture can be a solid or semi-solid medium. The culture can be a liquid medium. The culture can be a bioreactor. Any suitable bioreactor can be used. Examples of bioreactors include, but are not limited to, flasks, continuous stirred tank bioreactors (CSTRs), bubbleless bioreactors, airlift reactors, and membrane bioreactors. In some cases, the culture supernatant contains at least one secondary metabolite of the microorganism. The at least one secondary metabolite of the microorganism can be isolated and purified from the supernatant. In some cases, the supernatant can be applied as a biocontrol composition as described elsewhere herein.

[0062] At least one microorganism can be affected by other microorganisms. Microorganisms can behave synergistically when cultured together, thus improving antifungal properties when cultured together compared to when cultured individually. For example, at least one microorganism can have increased viability when cultured with another microorganism. At least one microorganism can have increased growth when cultured with another microorganism. At least one microorganism can use chemicals or metabolites produced by another microorganism. At least one microorganism can directly interact with another microorganism. For example, at least one microorganism and another microorganism can form a biofilm or multicellular structure. At least one microorganism can produce and / or secrete increased amounts of secondary metabolites when cultured with another microorganism. For example, at least one microorganism can produce an intermediate metabolite that is then processed by another microorganism to produce a secondary metabolite. Methods disclosed elsewhere herein can be used to identify microorganisms that may benefit from culturing with another microorganism, as well as to identify biocontrol compositions comprising a first microorganism and a second microorganism, where the second microorganism is not identical to the first microorganism.

[0063] The biocontrol composition may include one or more secondary metabolites of at least one microorganism. The one or more secondary metabolites may have antifungal properties alone. The one or more secondary metabolites may have antifungal properties in combination with other microorganisms in the biocontrol composition. The one or more secondary metabolites may be isolated from the culture supernatant of at least one microorganism. The one or more secondary metabolites may include lipopeptides, dipeptides, aminopolyols, proteins, siderophores, phenazine compounds, polyketides, or combinations thereof.

[0064] The lipopeptide may be a linear lipopeptide or a cyclic lipopeptide (CLP). Examples of lipopeptides include, but are not limited to, surfactin, fengycin, iturin, macetolide, amphycin, arthrofactin, tracin, syringopeptide, syringomycin, ptisorbin, bacillomycin, bacillopeptin, bacitracin, polymyxin, daptomycin, mycosubtilin, crustakin, tensin, plipastatin, viscocin, and echinocandin. The echinocandin may be echinocandin B (ECB). In some examples, the secondary metabolite is surfactin, fengycin, iturin, or a combination thereof.

[0065] The dipeptide can be bacilysin or chlorotetaine. The polyketide can be deficidin, macrolactin, bacillaene, butyrolactol A, soraphen A, hippolachnin A, or forazolin A. The secondary metabolite can be an aminopolyol. The aminopolyol can be zwittermicin A. The secondary metabolite can be a protein. The protein can be bacisbin, subtilisin, or fungisin.

[0066] The siderophore may be pyoverdine, thioquinolobactin, or pyochelin. The phenazine compound may be phenazine-1-carboxylic acid, 1-hydroxyphenazine, or phenazine-1-carboxamide. The secondary metabolite may be chitinase, cellulase, amylase, or glucanase. The secondary metabolite may be a volatile antifungal compound.

[0067] The biocontrol composition can be formulated as a liquid or dry formulation. Liquid formulations can be flowable or aqueous suspensions. Liquid formulations can include at least one microorganism or its secondary metabolite suspended in water, oil, or a mixture thereof (emulsifiable concentrate). Dry formulations can be wettable powders, dry flakes, dusts, or granules. Wettable powders can be applied to plants, seeds, flowers, or their agricultural products as suspensions. Dusts can be applied to plants, seeds, or their agricultural products, such as seeds or leaves, in a dry state. Granules can be applied dry or mixed with water to create a suspension. The at least one microorganism or its secondary metabolite can be formulated as a microencapsulation, where the at least one microorganism or its secondary metabolite has a protective inner layer. The protective inner layer can include any suitable polymer.

[0068] The biocontrol composition may further comprise additional compounds. The additional compounds may be carriers, surfactants, wetting agents, penetrants, emulsifiers, spreaders, stickers, stabilizers, nutrients, binders, desiccants, thickeners, dispersants, UV protectants, or combinations thereof. The carrier may be a liquid carrier, an inorganic carrier, or an organic carrier. Examples of liquid carriers include, but are not limited to, vegetable oil or water. Examples of inorganic carriers include, but are not limited to, kaolinite clay or diatomaceous earth. Examples of organic carriers include, but are not limited to, flour. The surfactant may be an anionic surfactant, a cationic surfactant, an amphoteric surfactant, or a nonionic surfactant. The surfactant may be Tween® 20 or Tween® 80. The wetting agent may include polyoxyethylene esters, ethoxy sulfates, or derivatives thereof. In some cases, the wetting agent is mixed with a nonionic surfactant. The penetrant may include a hydrocarbon. The spreading agent may include a fatty acid, latex, a fatty alcohol, a crop oil (e.g., cottonseed), or a mineral oil. The adhesive may include emulsified polyethylene, a polymerized resin, a fatty acid, a petroleum distillate, or pregelatinized corn flour. The oil may be coconut oil, palm oil, castor oil, or lanolin. The stabilizer may be lactose or sodium benzoate. The nutrient may be molasses or peptone. The binder may be gum arabic or carboxymethylcellulose. The desiccant may be silica gel or anhydrous salt. The thickener may include polyacrylamide, a polyethylene polymer, a polysaccharide, xanthan gum, or a vegetable oil. The dispersant may be microcrystalline cellulose. The UV protectant may be oxybenzone, blankophor BBH, or lignin.

[0069] The biocontrol composition may further include dipicolinic acid.

[0070] The at least one microorganism can comprise an effective amount of isolated and purified microorganisms isolated and purified from liquid culture.The at least one microorganism from liquid culture can be air-dried, freeze-dried, spray-dried or fluidized bed dried to produce a dry preparation.The dry preparation can be redissolved in liquid to produce a liquid preparation.

[0071] Biocontrol compositions can be formulated so that at least one microorganism can replicate when they are applied and / or delivered to a target habitat (e.g., soil, plants, seeds, and / or agricultural products).

[0072] The biocontrol composition may have a shelf life of at least 1 week, 1 month, 6 months, at least 1 year, at least 2 years, at least 3 years, at least 4 years, or at least 5 years. Shelf life may refer to the length of time during which the biocontrol composition maintains at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% of its antifungal properties. The biocontrol composition may be stored at room temperature, 4°C or below, 0°C or below, or -20°C or below.

[0073] The biocontrol composition may contain spores. The spore-containing composition can be applied by the methods described herein. The spore-containing composition can extend the shelf life of the biocontrol composition. The spore-containing composition can survive the low pH or low temperature of the target habitat. For example, the spore-containing composition can be applied to soil at lower temperatures (e.g., below 10°C) and have antifungal properties for seeds planted at higher temperatures (e.g., 20°C). The spores can become vegetative cells, providing them with any of the advantages of vegetative cells.

[0074] The biocontrol composition may contain vegetative cells. The vegetative cell-containing composition can be applied by the methods described herein. The vegetative cells can proliferate, increasing the effectiveness of the composition. For example, the vegetative cells in the biocontrol composition can proliferate after application, increasing the surface area of ​​the plant exposed to the biocontrol composition. In another example, the vegetative cells in the biocontrol composition can proliferate after application, increasing the amount of time the biocontrol composition survives, thus extending the period during which the biocontrol composition is effective. The vegetative cells can proliferate and compete with fungal pathogens for nutrients. The vegetative cells can actively produce one or more secondary metabolites with antifungal properties. The vegetative cells can spore, giving them any of the advantages of spores.

[0075] The biocontrol composition may have antifungal activity, such as preventing or reducing the growth of fungal pathogens on plants, seeds, or agricultural products thereof. The biocontrol composition may prevent the growth of fungal pathogens on plants, seeds, or agricultural products thereof for at least 1 day, at least 2 days, at least 3 days, at least 4 days, or at least 5 days. The biocontrol composition may prevent the growth of fungal pathogens on plants, seeds, or agricultural products thereof for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days. The biocontrol composition may prevent the growth of fungal pathogens on plants, seeds, or agricultural products thereof for longer than 10 days.

[0076] The biocontrol composition can reduce the growth of fungal pathogens on a plant, seed, or produce compared to the growth of fungal pathogens on a control plant, seed, flower, or produce that has not been exposed to the biocontrol composition. The control can be a plant, seed, or produce that has not been treated with an antifungal agent, or a plant, seed, flower, or produce that has been treated with a commercially available antifungal agent. Examples of commercially available antifungal agents include, but are not limited to, Bacillus subtilis strain QST713 (Serenade®), Bacillus subtilis strain GB02 (Kodiak®), Bacillus subtilis strain MBI 600 (Subtilex®), Bacillus pumilus strain GB34 (YieldShield), and Bacillus licheniformis strain SB3086 (EcoGuard®). The biocontrol composition may reduce the growth of fungal pathogens on plants, seeds, or produce thereof for at least 1 day, at least 2 days, at least 3 days, at least 4 days, or at least 5 days. The biocontrol composition may reduce the growth of fungal pathogens on plants, seeds, or produce thereof for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days. The biocontrol composition may reduce the growth of fungal pathogens on plants, seeds, or produce thereof for longer than 10 days. The biocontrol composition may reduce the growth of fungal pathogens by at least 25% compared to the growth of fungal pathogens in a control. The biocontrol composition may reduce the growth of fungal pathogens by at least 60% compared to the growth of fungal pathogens in a control. The biocontrol composition can reduce the growth of the fungal pathogen by at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more compared to the growth of the fungal pathogen in a control.

[0077] In various aspects disclosed herein, the biocontrol compositions can be used to reduce the growth of fungal pathogens in plants. The plants can be flowers, seeds, or produce. The plant, flower, seed, or produce thereof may be almond, apricot, apple, artichoke, banana, barley, beet, blackberry, blueberry, broccoli, Brussels sprouts, cabbage, hemp, chili pepper, carrot, celery, Swiss chard, cherry, citrus fruit, corn, gourd, date, fig, garlic, grape, herb, spice, kale, lettuce, oil palm, olive, onion, pea, pear, peach, peanut, papaya, parsnip, pecan, persimmon, plum, pomegranate, potato, quince, radish, raspberry, rose, rice, plum spinosa, sorghum, soybean, spinach, strawberry, sweet potato, tobacco, tomato, turnip greens, walnut, or wheat. The plant may be a member of the genus Citrus or Malus. For example, the plant can be a mandarin, lemon, navel orange, or hybrid thereof. The plant can be an apple. The plant can be a particular cultivar. For example, the apple can be a Fuji apple.

[0078] Methods and compositions for preventing or reducing food spoilage and food spoilage Treating the plants, seeds, flowers, or produce thereof with the biocontrol composition prior to harvest. The method may be effective in inhibiting the growth of fungal pathogens. The method may be capable of inhibiting or reducing the growth of fungal pathogens by 25% or more compared to a control not exposed to the biocontrol composition. The method may be capable of inhibiting or reducing the growth of fungal pathogens by 60% or more compared to a control not exposed to the biocontrol composition. The methods and compositions may be capable of inhibiting or reducing the growth of fungal pathogens by at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 17%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, 99.5% or more compared to a control not exposed to the biocontrol composition.

[0079] Methods for preventing or reducing the growth of fungal pathogens in plants, seeds, or produce thereof can include applying a biocontrol composition comprising at least one microorganism or one or more secondary metabolites thereof described herein and a carrier to the plant, seed, flower, or produce before it is harvested. Harvesting the produce can refer to harvesting the edible parts of the plant from the rest of the plant, or can refer to harvesting the whole plant followed by later harvesting of the edible parts.

[0080] Applying the biocontrol composition before harvest can include sprinkling, pouring, spraying, or brushing the biocontrol composition onto the plant, seed, or its produce. Applying the biocontrol composition can include adding the biocontrol composition to a drip line, irrigation system, chemical irrigation system, spray, or soak. In some cases, the biocontrol composition is applied to the roots of the plant, the seeds of the plant, the leaves of the plant, the soil around the plant, or the edible parts of the plant, also referred to herein as the produce of the plant.

[0081] The method may further include applying to the plant a fertilizer, herbicide, pesticide, other biocontrol agent, or combination thereof. In some examples, the fertilizer, herbicide, pesticide, other biocontrol agent, or combination thereof is applied before, after, or simultaneously with the biocontrol composition.

[0082] A method for preventing or reducing the growth of fungal pathogens can include applying a biocontrol composition to seeds, the biocontrol composition comprising at least one microorganism or its secondary metabolite and a carrier, as described herein. Applying the biocontrol composition to plant seeds can occur before planting, at planting, or after planting and before germination. For example, the biocontrol composition can be applied to the surface of seeds before planting. In some cases, seed treatment that occurs before planting can include adding a colorant or dye, a carrier, a binder, a sticking agent, an antifoaming agent, a lubricant, nutrients, or a combination thereof to the biocontrol composition.

[0083] A method for preventing or reducing the growth of fungal pathogens can include applying a biocontrol composition to soil, comprising at least one microorganism or its secondary metabolite and a carrier, as described herein. The biocontrol composition can be applied before, after, or during planting seeds in the soil, or before transplanting plants to a new location. In one example, a soil conditioner is added to the soil before planting, and the soil conditioner improves plant growth, and the soil conditioner comprises the biocontrol composition. In some cases, the soil conditioner further comprises a fertilizer.

[0084] A method for preventing or reducing the growth of fungal pathogens may include applying a biocontrol composition comprising at least one microorganism or its secondary metabolite and a carrier as described herein to the roots. The biocontrol composition may be applied directly to the roots. An example of direct application to the roots of a plant may include immersing the roots in a solution containing the biocontrol composition. The biocontrol composition may be applied indirectly to the roots. An example of indirect application to the roots of a plant may include spraying the biocontrol composition near the base of the plant, allowing the biocontrol composition to penetrate the soil and reach the roots.

[0085] Figure 1 shows a general schematic diagram of a method for using a biocontrol composition. Microorganisms can be grown for use in a biocontrol composition. The active ingredients of the biocontrol composition can be extracted as needed, or the microbial growth can be manipulated to allow different components to be used in the biocontrol composition. For example, the microbial growth can be centrifuged, and the supernatant and microbial pellet can be collected. The supernatant or the microorganisms can be used as the biocontrol composition. Additional compounds can be added to the biocontrol composition to produce a formulation. The formulation can, for example, extend shelf life or allow the biocontrol composition to be applied. In parallel, plants can be grown. The plants can be grown from seeds or grafts. The plant produce can be harvested, and the harvested produce can be transported. The formulated biocontrol composition can be applied to the plant at any stage in the plant growth process, or can be applied during the harvesting or transport process. For example, the biocontrol composition can be applied to the seeds or roots of the plant. In another example, the biocontrol composition can be applied to produce before, during, or after harvest, or can be applied to packaging used for transporting the produce. After application of the biocontrol composition, the plants can have enhanced resilience to pathogen infection or growth.

[0086] Treating the produce with a biocontrol composition after harvest. Methods for preventing or reducing the growth of fungal pathogens on agricultural produce can include applying to the produce before or after harvest a biocontrol composition comprising at least one microorganism or a secondary metabolite thereof described herein and a carrier.

[0087] Applying the biocontrol composition before or after harvest can include sprinkling, dipping, spreading, injecting, rubbing, spraying, or brushing the biocontrol composition onto the plant produce. The biocontrol composition can be applied to the produce just before harvest, just after harvest, or within 1, 2, 3, 4, 5, 6, or 1 week after harvest. In some cases, the biocontrol composition is applied by the harvesting entity, the entity packaging the produce, the entity transporting the produce, or the entity selling or commercially displaying the produce for consumers in the process of treating the produce just before or just after harvest.

[0088] Applying the biocontrol composition after harvest can further include incorporating the biocontrol composition into a process for treating the harvested produce. Produce can be treated immediately after harvest, for example, during one or more washes. The washes can include the use of water with added bleach (chlorine) and / or sodium bicarbonate or ozonated water. Produce can also be treated with oil, resin, or a structural or chemical matrix. The biocontrol composition can be mixed with the oil, resin, or structural or chemical matrix for application. Produce can be treated before or after drying the produce. For example, the biocontrol composition can be added to wax, gum arabic, or other coatings used to coat the produce. The biocontrol composition can be added at any time in the process, included in one of the washes as part of a new wash, or mixed with the wax, gum arabic, or other coating on the produce.

[0089] Potential Formulations of Biocontrol Compositions

[0090] Certain formulations can be used to extend the shelf life of biocontrol compositions and improve ease of application. Formulations can include sucrose, glycerol, carboxymethylcellulose (CMC), gum arabic, polyvinylpyrrolidone (PVP), alginate, agar, lambda and kappa carrageenan, pectin, chitosan, bean gum, nonfat milk, starch, or trehalose. The formulations can contain a given component in an amount up to 100% of the composition. The formulations can contain a specific amount of a given component. For example, a given component can comprise at least 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more of the composition. For example, a given ingredient may make up up to 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or less of the composition.

[0091] For example, carboxymethylcellulose (CMC) may be present in an amount of 1:5 w / v. In another example, gum arabic may be present in a concentration of up to 40% w / w. The formulation may be in various liquid or solid states, or may be aerosolizable. For example, the formulation may be a freeze-dried powder. For example, the formulation may be liquid, or originally liquid before being freeze-dried.

[0092] The following examples are given for the purpose of illustrating various embodiments of the present invention and are not intended to limit the invention in any way. The examples, along with the methods described herein, are presently representative of preferred embodiments and are exemplary and are not intended to limit the scope of the invention. Variations in the examples and other uses will occur to those skilled in the art that are encompassed within the spirit of the invention, as defined by the scope of the claims. [Example]

[0093] Example 1 Protection of apples against P. expansum infection by the use of BC8, BC16, BC17 and BC18

[0094] The wounded and inoculated apples were incubated in closed containers and assessed for disease incidence and severity 4, 6 and 8 days after pest attack. Disease severity was assessed by measuring the diameter of the visible infection extending from the inoculated wound.

[0095] Apples were first disinfected by immersing them in a 10% bleach solution for 8 minutes. The bleached apples were rinsed three times with distilled water and left to dry for 30 minutes. A sterile 4mm-wide cork borer was used to artificially wound the apples, creating 3mm-deep holes. The apples were sorted into sets of four apples and photographed. For each treatment, the fruit was inoculated with the treatment by immersing it in a 1 / 4 dilution of the respective BC composition (BC8, BC16, BC17, or BC18) for 1 minute. The treatment was allowed to dry for at least 3 hours. Containers for fruit storage were prepared by wiping the containers with a sterilizing wipe and allowed to dry for 20 minutes. 200 mL of deionized water was filled into the bottom of the container. One side of a Petri dish was used to hold the fruit inoculated side up, and the fruit was placed in a covered container. Penicillium expansum was applied to the wound site. Four apples were left uninoculated (25 μL of sterile water). 5 25 μL of solution containing spores / mL (absolute number of spores 2.5 x 10 4 ) was inoculated onto each apple. The progression of infection was assessed on days 4, 6, and 8 after storage. To analyze the infection, readings of the lesion diameter were taken and the apples were photographed. In addition, the degree of infection was determined by weighing the whole apple, excising the necrotic area, weighing the apple again, and assessing the weight of infected tissue in grams.

[0096] The experimental conditions are shown in Table 3 for six conditions performed with four replicates per condition for both Gala and Fuji apples: spores from BC8, BC16, spores from BC17, BC18, an untreated control that was not infected with the pathogen (pathogen-uninfected UTC), and an untreated control apple that was infected with the pathogen (pathogen-infected UTC).

[0097] [Table 3-1] [Table 3-2]

[0098] Figure 2 shows apple rot as measured by lesion diameter across six experiments involving Fuji (3) and Gala (3) apples. Negative controls (Control (-)) are uninoculated and untreated, while positive controls (Control (+)) are inoculated but untreated. Boxplots not connected by the same letter are significantly different (p=0.01). Measurements were taken 6 days after infection.

[0099] Figure 3 shows apple rot, measured by weight of rotten tissue, across six experiments involving Fuji (3) and Gala (3) apples. Negative controls were uninoculated and untreated, while positive controls were inoculated but untreated. Boxplots not connected by the same letter are significantly different (p=0.01). Measurements were taken 6 days after infection.

[0100] Figure 4A shows Fuji apple rot assessed by mean lesion diameter. Figure 4B shows Fuji apple rot assessed by mean necrosis weight. Negative controls were uninoculated and untreated, while positive controls were inoculated but untreated. Bars not connected by the same letter are significantly different (p=0.05). Measurements were taken 6 days after infection.

[0101] Apples treated with BC18 were shown to be significantly less prone to decay than untreated apples, as measured by either size or weight of infection. BC18-mediated protection from decay was observed in Fuji and Gala apples. BC18 treatment is shown to significantly protect Fuji apples from decay caused by P. expansum compared to untreated (+) controls. BC18 treatment also significantly protected Gala apples from decay caused by P. expansum. Treatment results in Gala apples are comparable to uninoculated (-) control apples. Additionally, no adverse effects were observed on apples treated with these candidates. No abnormal odors or discoloration of the apple surface were observed.

[0102] Figure 5 is a photograph of Fuji apples 6 days after infection, and Figure 7 is a photograph of Gala apples 6-7 days after infection. Figure 6A shows Gala apple rot assessed by mean lesion diameter. Figure 6B shows Gala apple rot assessed by mean necrosis weight. Negative controls are uninoculated and untreated, while positive controls are inoculated but untreated. Bars not connected by the same letter are significantly different (p=0.05). Measurements were taken 6 days after infection.

[0103] Example 2 Protection of apples from decay caused by P. expansum with BC18 strains.

[0104] The wounded and inoculated apples are incubated in a closed container. Disease incidence and severity are assessed 6 days after pest attack. Disease severity is assessed by measuring the diameter of the visible infection extending from the inoculated wound. The two strains that make up BC18 (Gluconobacter cerinus = BC18A, and Hanseniaspora uvarium = BC18B) are tested in the same way as the single strains. In addition, various ratios of the components of BC18 are tested. Table 4 shows the experimental conditions for Gala apples, with eight conditions performed with four replicates per condition: BC18, BC18 microorganism A (BC18A), BC18 microorganism B (BC18B), BC18 microorganism A and microorganism B at a first ratio of microorganism A and B (BC18A+B ratio 1), BC18 microorganism A and microorganism B at a second ratio of microorganism A and B (BC18A+B ratio 2), BC18 microorganism A and microorganism B at a third ratio of microorganism A and B (BC18A+B ratio 3), an untreated control apple that was not infected with a pathogen (pathogen-uninfected UTC), and an untreated control apple that was infected with a pathogen (pathogen-infected UTC). [Table 4]

[0105] The apples are first disinfected by immersing them in a 10% bleach solution for 8 minutes. The bleached apples are rinsed three times with distilled water and left to dry for 30 minutes. A sterile 4mm wide cork borer is used to artificially wound the apples, creating 3mm deep holes in the apples. The apples are sorted into sets of 4 apples and photographed. For each treatment, the fruit is inoculated with the treatment by immersing the fruit in a 1 / 4 dilution of the respective BC composition for 1 minute. The treatment is left to dry for at least 3 hours. Containers for storing the fruit are prepared by wiping the container with a sterilizing sheet and left to dry for 20 minutes. 200mL of deionized water is filled into the bottom of the container. One side of a Petri dish is used to hold the fruit inoculated side up and place the fruit in a container with a lid. Penicillium expansum is applied to the wound site. Four apples are left uninoculated (25μL of sterile water). For all treatments, 1.0 × 105 25 μL of solution containing spores / mL (absolute number of spores 2.5 x 10 4 ) into each apple. The progression of the infection is assessed on days 4, 6 and 8 after storage. To analyze the infection, readings of the lesion diameter are taken and the apples are photographed. In addition, the degree of infection is measured by weighing the whole apple, cutting out the necrotic area, weighing the apple again and assessing the weight of infected tissue in grams.

[0106] The optimal ratio of BC18A to BC18B was determined, as well as the relative contributions of BC18A and BC18B. Synergistic effects of the co-cultures could be observed and compared with strains grown separately. Synergistic effects could be observed due to the mutual stimulation of antifungal metabolites.

[0107] Example 3 Protection of strawberries from decay caused by Botrytis cinerea in 8 BC, 16 BC, 17 BC, and 18 BC

[0108] The wounded and inoculated strawberries are incubated in closed containers and assessed for disease incidence and severity 4, 6, and 8 days after pest attack. Disease severity is assessed by measuring the diameter of the visible infection extending from the inoculated wound. The experimental conditions are shown in Table 5 for six conditions on strawberries, with four replicates per condition: BC8 spores, BC16, BC17 spores, BC18 organisms, untreated controls that were not infected with the pathogen (pathogen-free UTC), and untreated control apples that were infected with the pathogen (pathogen-infected UTC). [Table 5]

[0109] Strawberries were first surface disinfected by immersing them in a 10% bleach solution for 8 minutes. The bleached strawberries were rinsed three times with distilled water and left to dry for 30 minutes. A sterile 4mm-wide cork borer was used to artificially wound the strawberries, creating 3mm-deep holes. The strawberries were sorted into sets of four strawberries and photographed. For each treatment, the fruits were inoculated with the treatment by immersing them in a 1 / 4 dilution of the respective BC composition for 1 minute. The treatments were left to dry for at least 3 hours. Containers for fruit storage were prepared by wiping the containers with a sterilizing sheet and left to dry for 20 minutes. 200 mL of deionized water was filled into the bottom of the container. One side of a Petri dish was used to hold the fruit inoculated side up, and the fruit was placed in a container with a lid. Botrytis cinerea was applied to the wound site. Four strawberries were left uninoculated (25 μL of sterile water). 5 25 μL of solution containing spores / mL (absolute number of spores 2.5 x 10 4 ) is inoculated onto each strawberry. The progression of the infection is assessed on days 4, 6 and 8 after storage. To analyze the infection, readings of the lesion diameter are taken and the apples are photographed. In addition, the degree of infection is measured by weighing the whole apple, cutting out the necrotic area, weighing the apple again and assessing the weight of infected tissue in grams.

[0110] Example 4 Inhibition of Penicillium digitatum by BC8, BC17, and BC18 in vitro on various citrus-based media

[0111] Citrus-based media were prepared from homogenized fruit tissue, water, and agar. Homogenized fruit tissue was prepared by blending each fruit tissue type for mandarin, lemon, or navel to produce the corresponding medium. Six types of media were prepared using a blender: mandarin peel, whole mandarin, lemon peel, whole lemon, navel peel, or whole navel. The whole medium contained both the peel and pulp of the fruit, while the peel medium was prepared using the peel but not the pulp. After autoclaving the citrus-based media, the citrus medium was poured into Petri dishes to produce solid medium plates.

[0112] To determine the growth ability of the BC8, BC17, and BC18 microbial consortia on citrus agar, 4 μL of each consortium was spotted onto solid citrus media plates as shown in Figure 8A. Cultures were grown at room temperature for 4 days. Photographs of the citrus media plates illustrating the growth of BC8, BC17, and BC18 after 4 days are shown in Figure 8B. BC8 and BC17 did not show significant visible colony growth on any of the plates, while BC18 showed colony growth on all citrus media types.

[0113] Microorganism BC8, which contains Bacillus amyloliquefaciens; microorganism BC17, which contains Bacillus sp.; and microbial consortium BC18, which contains Gluconobacter cerinus and Hanseniaspora uvarum, were tested for their ability to inhibit Penicillium digitatum (P. digitatum) growth in various citrus-based media prepared as described above.

[0114] A lawn of P. digitatum was spread onto plates at a concentration of 500 spores / plate or 5,000 spores / plate using 50 μL of spore suspension. After the lawn dried, a center plug was cut and placed on each plate. Candidate suspensions were prepared by picking a single colony from a working stock plate and inoculating it into 1.5 mL of filter-sterilized deionized water. 100 μL of the microbial consortium was inoculated into the center plug. The plates were left to incubate at room temperature for 4-5 days. The degree of inhibition of P. digitatum was assessed by measuring the exclusion zone on the lawn. A control plate without the microbial consortium was used to represent 0% inhibition.

[0115] Results are shown four days after inoculation of P. digitatum on plates at an inoculum concentration of 500 spores / plate (Figure 9) and 5,000 spores / plate (Figure 10). Results were comparable at both concentrations, but P. digitatum growth inhibition was more visible on plates using an inoculum concentration of 500 spores / plate (Figure 9). BC8 showed no inhibition of P. digitatum on any of the citrus agar plates tested. BC17 showed inhibition of P. digitatum on multiple media.

[0116] For BC17, there was strong inhibition on mandarin peel medium and minimal inhibition on whole mandarin medium. There was no visible inhibition on lemon peel medium and clear inhibition on whole lemon medium. There was no visible inhibition of P. digitatum on navel peel and whole navel medium (Figures 9 and 10).

[0117] BC18 showed little inhibition of P. digitatum on mandarin peel medium and clear inhibition on whole mandarin medium. There was minimal inhibition on lemon peel medium and no inhibition on whole lemon medium. On both navel peel medium and whole navel medium, BC18 showed clear inhibition of P. digitatum (Figures 9 and 10). Despite the lack of visible growth of BC8 and BC17 on citrus fruit medium, inhibition of the pathogen was still observed. This demonstrates the potential of metabolites of the biocontrol compositions to have inhibitory properties.

[0118] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous modifications, changes, and substitutions will readily occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments described herein can be employed. It is intended that the following claims define the scope of the invention, and that methods and structures within the scope of these claims and their equivalents be covered thereby. In certain embodiments, for example, the following items are provided: (Item 1) (i) at least one microorganism, or a metabolite produced by said at least one microorganism, and (ii) Carrier A biocontrol composition comprising: A biocontrol composition, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:1, SEQ ID NO:22 or SEQ ID NO:23, and the biocontrol composition is capable of inhibiting the growth of Penicillium species compared to a control that is not exposed to the biocontrol composition. (Item 2) Item 1, wherein at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 1. The biocontrol composition of item 1. (Item 3) 2. The biocontrol composition of claim 1, wherein the 16S rRNA sequence is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 22. (Item 4) Item 1, wherein the 16S rRNA sequence is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 23. (Item 5) (i) at least one microorganism, or a metabolite produced by said at least one microorganism, and (ii) Carrier A biocontrol composition comprising: A biocontrol composition, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 90% identical to the 16S rRNA sequence of SEQ ID NO: 24, or the at least one microorganism comprises an internal transcribed spacer (ITS) sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO: 25, and the biocontrol composition is capable of inhibiting the growth of Penicillium species compared to a control that is not exposed to the biocontrol composition. (Item 6) 6. The biocontrol composition of claim 5, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 90% identical to the 16S rRNA sequence of SEQ ID NO: 24. (Item 7) 6. The biocontrol composition of claim 5, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 24. (Item 8) 6. The biocontrol composition of item 5, wherein the at least one microorganism comprises an internal transcribed spacer (ITS) sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO: 25. (Item 9) 6. The biocontrol composition of item 5, wherein the at least one microorganism comprises an internal transcribed spacer (ITS) sequence that is greater than 99% identical to the ITS sequence of SEQ ID NO: 25. (Item 10) 6. The biocontrol composition of claim 5, wherein the at least one microorganism is at least two microorganisms, including a first microorganism comprising a 16S rRNA sequence that is greater than 90% identical to the 16S rRNA sequence of SEQ ID NO: 24, and a second microorganism comprising an ITS sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO: 25. (Item 11) 11. The biocontrol composition of any of items 1 to 10, wherein growth inhibition of the Penicillium species is indicated by a reduction in lesion size or tissue necrosis in produce exposed to the biocontrol composition compared to the control not exposed to the biocontrol composition. (Item 12) 12. The biocontrol composition of any one of items 1 to 11, which is capable of inhibiting the growth of the Penicillium species by 5% or more compared to a control not exposed to the biocontrol composition. (Item 13) 13. The biocontrol composition of any one of items 1 to 12, which is capable of inhibiting the growth of the Penicillium species by 10% or more compared to a control not exposed to the biocontrol composition. (Item 14) 14. The biocontrol composition of any one of items 1 to 13, which is capable of inhibiting the growth of the Penicillium species by 20% or more compared to a control not exposed to the biocontrol composition. (Item 15) 14. The biocontrol composition of any one of items 1 to 13, which is capable of inhibiting the growth of the Penicillium species by 25% or more compared to a control not exposed to the biocontrol composition. (Item 16) 14. The biocontrol composition of any of items 1 to 13, wherein the Penicillium is Penicillium expansum. (Item 17) 14. The biocontrol composition of any of items 1 to 13, wherein the Penicillium is Penicillium digitatum. (Item 18) 18. The biocontrol composition of any of items 1 to 17, comprising vegetative cells. (Item 19) 18. The biocontrol composition of any of items 1 to 17, comprising spores. (Item 20) 20. The biocontrol composition of any of the preceding items, wherein the carrier is selected from the group consisting of oil, water, wax, resin, kaolinite clay, diatomaceous earth, or flour. (Item 21) 20. The biocontrol composition according to any of the preceding items, wherein the carrier is water. (Item 22) 22. The biocontrol composition according to any of items 1 to 21, formulated in liquid form. (Item 23) 22. The biocontrol composition according to any of items 1 to 21, formulated in solid form. (Item 24) 22. The biocontrol composition according to any of items 1 to 21, formulated in powder form. (Item 25) 10. A method of reducing or preventing the growth of pathogens in a plant, seed, flower, or produce thereof, comprising applying to the plant, seed, flower, or produce thereof a biocontrol composition according to any preceding item. (Item 26) 10. A method of reducing or preventing the growth of pathogens in a plant, seed, flower, or produce thereof, comprising applying to an object or area adjacent to the plant, seed, flower, or produce thereof a biocontrol composition of any of the preceding items. (Item 27) 27. The method of any of items 25 to 26, wherein the applying step is carried out before harvesting the plant, seed, flower, or produce. (Item 28) 27. The method of any of items 25 to 26, wherein the applying step is carried out after harvesting the plant, seed, flower, or produce. (Item 29) 27. The method of claim 26, wherein the area adjacent to the plant comprises soil used to grow the plant, seed, flower, or produce thereof. (Item 30) 27. The method of claim 26, wherein the object adjacent to the plant comprises packaging used to store or transport the plant, seed, flower, or produce. (Item 31) 31. The method of any of items 25 to 30, wherein the applying step is carried out by spraying the biocontrol composition. (Item 32) 29. The method of any of items 25, 27 to 28, wherein the applying step is carried out by immersing plants, seeds, flowers, or agricultural products in the biocontrol composition. (Item 33) 33. The method of any of items 25 to 32, wherein the plant is selected from the group consisting of almonds, apricots, apples, artichokes, bananas, barley, beets, blackberries, blueberries, broccoli, Brussels sprouts, cabbage, hemp, rapeseed, chili peppers, carrots, celery, Swiss chard, cherries, citrus fruits, corn, gourds, dates, figs, flax, garlic, grapes, herbs, spices, kale, lettuce, mint, oil palm, olives, onions, peas, pears, peaches, peanuts, papayas, parsnips, pecans, persimmons, plums, pomegranates, potatoes, quince, radishes, raspberries, roses, rice, plums, sorghum, soybeans, spinach, strawberries, sweet potatoes, tobacco, tomatoes, turnip greens, walnuts, and wheat. (Item 34) 34. The method according to any one of items 25 to 33, wherein the plant is an apple. (Item 35) 33. The method according to any of items 25 to 32, wherein the plant is a member of the genus Malus. (Item 36) 33. The method according to any of items 25 to 32, wherein the plant is a member of the genus Citrus. (Item 37) Item 34. The method of item 33, wherein the Citrus comprises mandarin, lemon, lime, navel orange, pomelo, or a hybrid thereof. (Item 38) 1. A method for inhibiting the growth of a pathogen, comprising: applying a biocontrol composition to an apple, the biocontrol composition comprising: (i) at least one microorganism, or a metabolite produced by said at least one microorganism, and (ii) Carrier Including, The method, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:1, SEQ ID NO:22, or SEQ ID NO:23, and the biocontrol composition is capable of inhibiting the growth of Penicillium expansum compared to a control that is not exposed to the biocontrol composition. (Item 39) 39. The method of claim 38, wherein at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 1. (Item 40) 39. The method of claim 38, wherein the 16S rRNA sequence is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 22. (Item 41) 39. The method of claim 38, wherein the 16S rRNA sequence is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 23. (Item 42) 1. A method for inhibiting the growth of a pathogen, comprising: applying a biocontrol composition to an apple, the biocontrol composition comprising: (i) a first microorganism and a second microorganism, or a metabolite produced by the first microorganism or the second microorganism; and (ii) Carrier Including, The method of claim 1, wherein the first microorganism comprises a 16S rRNA sequence that is greater than 90% identical to the 16S rRNA sequence of SEQ ID NO: 24, the second microorganism comprises an ITS sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO: 25, and the biocontrol composition is capable of inhibiting the growth of Penicillium expansum relative to a control that is not exposed to the biocontrol composition. (Item 43) 43. The method of claim 42, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 90% identical to the 16S rRNA sequence of SEQ ID NO: 24. (Item 44) 43. The method of claim 42, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 24. (Item 45) 43. The method of claim 42, wherein the at least one microorganism comprises an internal transcribed spacer (ITS) sequence that is more than 90% identical to the ITS sequence of SEQ ID NO: 25. (Item 46) 43. The method of claim 42, wherein the at least one microorganism comprises an internal transcribed spacer (ITS) sequence that is greater than 99% identical to the ITS sequence of SEQ ID NO: 25. (Item 47) 1. A method for inhibiting the growth of a pathogen, comprising: applying a biocontrol composition to a citrus plant, wherein the biocontrol composition comprises: (i) at least one microorganism, or a metabolite produced by said at least one microorganism, and (ii) Carrier Including, The method, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO:1, SEQ ID NO:22, or SEQ ID NO:23, and the biocontrol composition is capable of inhibiting the growth of the species Penicillium expansum compared to a control that is not exposed to the biocontrol composition. (Item 48) 44. The method of claim 43, wherein at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 1. (Item 49) 44. The method of claim 43, wherein the 16S rRNA sequence is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 22. (Item 50) 44. The method of claim 43, wherein the 16S rRNA sequence is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 23. (Item 51) 1. A method for inhibiting the growth of a pathogen, comprising: applying a biocontrol composition to a citrus plant, wherein the biocontrol composition comprises: (i) a first microorganism and a second microorganism, or a metabolite produced by the first microorganism or the second microorganism; and (ii) Carrier Including, The method of claim 1, wherein the first microorganism comprises a 16S rRNA sequence that is greater than 90% identical to the 16S rRNA sequence of SEQ ID NO: 24, the second microorganism comprises an ITS sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO: 25, and the biocontrol composition is capable of inhibiting the growth of the species Penicillium expansum relative to a control that is not exposed to the biocontrol composition. (Item 52) 52. The method of claim 51, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 90% identical to the 16S rRNA sequence of SEQ ID NO: 24. (Item 53) 52. The method of claim 51, wherein the at least one microorganism comprises a 16S rRNA sequence that is greater than 99% identical to the 16S rRNA sequence of SEQ ID NO: 24. (Item 54) 52. The method of claim 51, wherein the at least one microorganism comprises an internal transcribed spacer (ITS) sequence that is greater than 90% identical to the ITS sequence of SEQ ID NO: 25. (Item 55) 52. The method of claim 51, wherein the at least one microorganism comprises an internal transcribed spacer (ITS) sequence that is greater than 99% identical to the ITS sequence of SEQ ID NO: 25.

Claims

1. (i) at least one microorganism, and (ii) Carrier A biocontrol composition comprising: the at least one microorganism comprises a first microorganism belonging to Gluconobacter cerinus, the first microorganism comprising a 16S rRNA sequence of SEQ ID NO: 24, and a second microorganism belonging to Hansoniaspora uvarum, the second microorganism comprising an internal transcribed spacer (ITS) sequence of SEQ ID NO: 25; A biocontrol composition, wherein the biocontrol composition inhibits the growth of a fungal pathogen belonging to the genus Penicillium compared to a control not exposed to the biocontrol composition.

2. 10. The biocontrol composition of claim 1, wherein the ratio of the first microorganism to the second microorganism is from 1:2 to 2:

1.

3. 3. The biocontrol composition of claim 1 or 2, wherein growth inhibition of a fungal pathogen is indicated by a reduction in lesion size or tissue necrosis in produce exposed to the biocontrol composition compared to the control not exposed to the biocontrol composition.

4. 3. The biocontrol composition of claim 1 or 2, which is capable of inhibiting growth of the fungal pathogen by 70% or more compared to a control not exposed to the biocontrol composition.

5. 3. The biocontrol composition of claim 1 or 2, wherein the fungal pathogen is one or more selected from Penicillium expansum and Penicillium digitatum.

6. The biocontrol composition of claim 1 or 2, comprising vegetative cells or spores.

7. 3. The biocontrol composition of claim 1 or 2, wherein the carrier is selected from the group consisting of oil, water, wax, resin, kaolinite clay, diatomaceous earth, or flour, such that the biocontrol composition is formulated in a liquid, solid, or powder form.

8. 3. The biocontrol composition of claim 1 or 2, wherein the at least one microorganism is diluted in the carrier in a ratio of at least 1:

4.

9. 10. A method for reducing, inhibiting or preventing the growth of a fungal pathogen belonging to the genus Penicillium in a plant, seed, flower, or produce thereof, comprising the step of applying the biocontrol composition of claim 1 or 2 to (i) the plant, seed, flower, or produce thereof, or (ii) an object or area adjacent to the plant, seed, flower, or produce thereof.

10. 10. The method of claim 9, wherein the applying step occurs before harvesting the plant, seed, flower, or produce and / or after harvesting the plant, seed, flower, or produce.

11. 10. The method of claim 9, wherein the area adjacent to the plant comprises soil used to grow the plant, seed, flower, or produce thereof, and / or the object adjacent to the plant comprises packaging used to store or transport the plant, seed, flower, or produce.

12. 10. The method of claim 9, wherein the applying step is carried out by spraying the biocontrol composition and / or by dipping plants, seeds, flowers, or agricultural products into the biocontrol composition.

13. 10. The method of claim 9, wherein the plant is selected from the group consisting of almonds, apricots, apples, artichokes, bananas, barley, beets, blackberries, blueberries, broccoli, Brussels sprouts, cabbage, hemp, rapeseed, chili peppers, carrots, celery, Swiss chard, cherries, citrus fruits, corn, gourds, dates, figs, flax, garlic, grapes, herbs, spices, kale, lettuce, mint, oil palm, olives, onions, peas, pears, peaches, peanuts, papayas, parsnips, pecans, persimmons, plums, pomegranates, potatoes, quince, radishes, raspberries, roses, rice, plums, sorghum, soybeans, spinach, strawberries, sweet potatoes, tobacco, tomatoes, turnip greens, walnuts, and wheat.

14. 10. The method of claim 9, wherein the fungal pathogen is one or more selected from Penicillium expansum and Penicillium digitatum.

15. 10. The method of claim 9, wherein the biocontrol composition reduces, inhibits, or prevents the growth of a fungal pathogen for at least 7 days.

16. 10. The method of claim 9, wherein growth inhibition of fungal pathogens is indicated by a reduction in lesion size or tissue necrosis in produce exposed to the biocontrol composition compared to a control not exposed to the biocontrol composition.

17. 10. The method of claim 9, wherein the biocontrol composition is capable of inhibiting the growth of the fungal pathogen by 70% or more compared to a control not exposed to the biocontrol composition.

18. 10. The method of claim 9, wherein the biocontrol composition comprises vegetative cells or spores.

19. 10. The method of claim 9, wherein the carrier is selected from the group consisting of oil, water, wax, resin, kaolinite clay, diatomaceous earth, or flour, such that the biocontrol composition is formulated in a liquid, solid, or powder form.

20. 10. The method of claim 9, wherein the at least one microorganism is diluted in the carrier in a ratio of at least 1:4.

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