Microbial compositions for preventing or reducing the growth of fungal pathogens in plants

Abstract

To provide microbial compositions for prevention or reduction of growth of fungal pathogens on plants.SOLUTION: Disclosed herein are biocontrol compositions against plant fungal pathogens, and methods of use thereof for the prevention or reduction of crop loss or food spoilage. The biocontrol compositions can comprise at least one microbe with anti-fungal activity, or a secondary metabolite of the at least one microbe. The methods can comprise application of the biocontrol composition to a plant, a seed or the produce thereof, or to a packaging material used to transport or store the produce.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of priority of U.S. Provisional Application No. 62 / 629,525, filed on February 12, 2018, which is hereby incorporated by reference in its entirety.

Background Art

[0002] 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 degradation caused by these fungal pathogens. Commercially available products cannot show the desired plant or fungal specificity or effectiveness. Furthermore, options for post - harvest protection of agricultural products, especially organic agricultural products, are limited. A biocontrol composition for preventing fungal growth can provide an alternative to currently available products.

Summary of the Invention

Means for Solving the Problems

[0003] In this specification, in certain embodiments, a biological control composition comprising (i) at least one microorganism and (ii) a carrier is described, wherein at least one microorganism has a 16S rRNA sequence that is more than 99% identical to a 16S rRNA sequence selected from the group of SEQ ID NOs: 1 and SEQ ID NOs: 9, or at least one microorganism has an ITS sequence that is more than 99% identical to an ITS sequence selected from the group of SEQ ID NOs: 17 and SEQ ID NOs: 20, or at least one microorganism has an ITS sequence that is more than 90% identical to the ITS sequence of SEQ ID NOs: 18. Furthermore, in certain embodiments described herein, a biological control composition comprising (i) at least one microorganism and (ii) a carrier is described, wherein at least one microorganism contains an rRNA sequence that is more than 99% identical to a sequence longer than 200 nucleotides that includes an rRNA sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 9, or at least one microorganism has an ITS sequence that is more than 99% identical to the ITS sequence of SEQ ID NO: 17, or at least one microorganism has an ITS sequence that is more than 90% identical to SEQ ID NO: 18. Furthermore, in certain embodiments herein, a biological control composition comprising (i) at least one microorganism and (ii) a carrier is described which inhibits the growth of Fusarium oxysporum by 25% or more compared to a control not exposed to the biological control composition, or inhibits the growth of Verticillium dahliae by 60% or more compared to a control not exposed to the biological control composition, as determined by measuring the survival of Fusarium oxysporum or Verticillium dahliae, respectively. Furthermore, in certain embodiments herein, a biological control composition comprising (i) at least one microorganism and (ii) a carrier is described in which at least one microorganism has a 16S rRNA sequence that is more than 99% identical to the 16S rRNA sequence of Sequence ID No. 22.Furthermore, in certain embodiments of this specification, a biological control composition comprising (i) at least one microorganism and (ii) a carrier is described, wherein at least one microorganism has a 16S rRNA sequence that is more than 99% identical to the 16S rRNA sequence of SEQ ID NO: 23. Furthermore, in certain embodiments of this specification, a biological control composition comprising (i) at least one microorganism and (ii) a carrier is described, wherein at least one microorganism has a 16S rRNA sequence that is more than 99% identical to a 16S rRNA sequence selected from the group of SEQ ID NO: 24, or at least one microorganism has an ITS sequence that is more than 99% identical to an ITS sequence selected from the group of SEQ ID NO: 25, or at least one microorganism has an ITS sequence that is more than 90% identical to the ITS sequence of SEQ ID NO: 25.

[0004] Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of Botrytis cineria by 25% or more compared to a control that has not been exposed to the biological control composition. Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of Monilinia vaccinii-corymbosi by 25% or more compared to a control that has not been exposed to the biological control composition. Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of Colletotrichum spaethanium by 25% or more compared to a control that has not been exposed to the biological control composition. Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of Puccinia sorghi by 25% or more compared to a control that has not been exposed to the biological control composition. Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of Plasmopara viticola by 25% or more compared to a control that has not been exposed to the biological control composition. Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of Erysiphe necator by 25% or more compared to a control that has not been exposed to the biological control composition.Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of Podasphaera macularis by 25% or more compared to a control that has not been exposed to the biological control composition. Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of organisms of the genus Pytium by 25% or more compared to a control that has not been exposed to the biological control composition. Furthermore, in certain embodiments, this specification describes a biological control composition comprising (i) at least one microorganism and (ii) a carrier, which is capable of inhibiting the growth of organisms of the genus Rhizopus by 25% or more compared to a control that has not been exposed to the biological control composition.

[0005] Furthermore, in certain embodiments of this specification, a biological control composition comprising (i) a secondary metabolite of at least one microorganism and (ii) a carrier is described, wherein at least one microorganism has an ITS sequence that is more than 99% identical to an ITS sequence selected from the group 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. Furthermore, in certain embodiments of this specification, a biological control composition comprising (i) a secondary metabolite of at least one microorganism and (ii) a carrier is described, wherein at least one microorganism has a 16S rRNA sequence that is more than 99% identical to a 16S rRNA sequence selected from the group SEQ ID NO: 1, SEQ ID NO: 9, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24.

[0006] In one embodiment, the biological control composition further comprises a second microorganism, the second microorganism being not identical to at least one other microorganism. The second microorganism may comprise an RNA sequence that is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 1 to 25. The second microorganism may comprise a 16S rRNA sequence that is at least 95% identical to a 16S rRNA sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 22, 23, and 24. The second microorganism may comprise an ITS sequence that is at least 95% identical to an internal transcription spacer (ITS) sequence selected from the group consisting of SEQ ID NOs: 17, 18, 19, 20, 21, and 25. The second microorganism may contain a 16S rRNA sequence that is at least 99% identical to a 16S rRNA sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 22, 23, and 24. The second microorganism may contain an ITS sequence that is at least 99% identical to an internal transcription spacer (ITS) sequence selected from the group consisting of SEQ ID NOs: 17, 18, 19, 20, 21, and 25. The second microorganism may contain a 16S rRNA sequence that is a 16S rRNA sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 22, 23, and 24. The second microorganism may contain an ITS sequence, which is an internal transcription spacer (ITS) sequence selected from the group consisting of sequence numbers 17, 18, 19, 20, 21, and 25.

[0007] In some embodiments, at least one microorganism contains a 16S rRNA sequence that is at least 99% identical to SEQ ID NO: 24, and the second microorganism contains an ITS sequence that is at least 99% identical to SEQ ID NO: 25.

[0008] In one embodiment, the biological control composition further comprises a third microorganism, the third microorganism being not identical to the second or at least one of the microorganisms. In some embodiments, at least one microorganism comprises a 16S rRNA sequence that is more than 99% identical to SEQ ID NO: 23, the second microorganism comprises a 16S rRNA sequence that is more than 99% identical to SEQ ID NO: 23, and the third microorganism comprises a 16S rRNA sequence that is more than 99% identical to SEQ ID NO: 23. In one embodiment, the biological control composition further comprises a fourth microorganism, the third microorganism being not identical to the third, second, or at least one of the microorganisms. In one embodiment, the biological control composition further comprises a fifth microorganism, the fifth microorganism being not identical to the fourth, third, second, or at least one of the microorganisms. Any of the microorganisms in the biological control composition may be isolated and purified microorganisms. The biological control compositions disclosed herein may comprise one or more isolated and purified microorganisms. The biological control compositions disclosed herein may comprise one or more, two or more, three or more, four or more, or five or more isolated and purified microorganisms. In some examples, the biological control compositions may comprise isolated and purified microorganisms of different strains derived from a single microbial species.

[0009] At least one microorganism may have an ITS sequence that is more 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 have 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.

[0010] In one embodiment, a secondary metabolite of at least one microorganism is isolated from the supernatant of a culture of at least one microorganism. The secondary metabolite of at least one microorganism may include a lipopeptide. The lipopeptide may be a cyclic lipopeptide selected from the group consisting of surfactant, phengycin, and iturin. The secondary metabolite of at least one microorganism may include a polyketide. The secondary metabolite of at least one microorganism may include a volatile antifungal compound.

[0011] In one embodiment, a control is exposed to Bacillus subtilis strain QST 713. In one embodiment, at least one microorganism is isolated and purified. In one embodiment, the biological control composition is a liquid or powder. In one embodiment, the biological control composition contains spores.

[0012] In this specification, in certain embodiments, a method for preventing or reducing the growth of fungal pathogens in plants, roots, seeds, soil or furrows to which seeds are added, or agricultural products thereof, comprising the step of applying a biological control composition described herein having antifungal activity to plants, roots, seeds, soil or furrows to which seeds or plants are added, or agricultural products thereof. Furthermore, in this specification, in certain embodiments, a method for preventing or reducing the growth of fungal pathogens in plants, seeds, roots, soil or furrows to which seeds or plants are added, or agricultural products thereof, comprising the step of applying a biological control composition described herein having antifungal activity to the soil. Furthermore, in this specification, in certain embodiments, a method for preventing or reducing the growth of fungal pathogens in agricultural products, comprising the step of spraying or otherwise treating agricultural products with a biological control composition described herein having antifungal activity before harvest. Furthermore, in certain embodiments of this specification, a method for preventing or reducing the growth of fungal pathogens in agricultural products is described, comprising the step of spraying, dipping, or otherwise treating agricultural products with a biological control composition described herein that has antifungal activity. Furthermore, in certain embodiments of this specification, a method for preventing or reducing the growth of fungal pathogens in agricultural products is described, comprising the step of applying a biological control composition described herein that has antifungal activity to packaging materials used for transporting or storing agricultural products. Furthermore, in certain embodiments of this specification, a method for preventing or reducing the growth of fungal pathogens in seeds or agricultural products is described, comprising the step of incorporating a biological control composition described herein into a process selected from the group consisting of washing agricultural products or seeds, coating agricultural products or seeds, and a combination thereof.

[0013] In one embodiment, the plant, seed, or agricultural product thereof is a plant or agricultural product of the Rosaceae family. The plant, seed, or agricultural product thereof may belong to the genera Rubus, Malus, Pyrus, Cydonia, Prunus, Rosa, or Fragaria. The plant, seed, or agricultural product thereof of the genus Rubus may be a raspberry or a blackberry. The plant, seed, or agricultural product thereof of the genus Fragaria may be a strawberry. The plant, seed, or agricultural product thereof of the genus Pyrus may be a pear. The plant, seed, or agricultural product thereof of the genus Cydonia may be a quince. The plant, seed, or agricultural product thereof of the genus Prunus may be an almond, peach, plum, apricot, cherry, or spinosa plum. The plant, seed, or agricultural product thereof of the genus Rosa may be a rose. The plant, seed, or agricultural product thereof of the genus Malus may be an apple.

[0014] In one embodiment, the plant, seed, or agricultural product thereof may be a plant or agricultural product of the Ericaceae family. The plant, seed, or agricultural product thereof may belong to the genus Vaccinium. The plant, seed, or agricultural product thereof of the genus Vaccinium may be a blueberry.

[0015] In one embodiment, the plant, seed, or agricultural product thereof may be a plant or agricultural product of the family Vitaceae. The plant, seed, or agricultural product thereof may belong to the genus Vitis. The plant, seed, or agricultural product thereof of the genus Vitis may be a grape.

[0016] In one embodiment, the step of applying the biological control composition includes sprinkling, dipping, rolling, injecting, rubbing, spraying, or brushing the biological control composition onto plants, seeds, or agricultural products. The step of applying the biological control composition to plants may include adding the biological control composition to a drip line, irrigation system, chemigation system, spray, or dipping agent.

[0017] The step of applying the biological control composition to a plant may include the step of applying the biological control composition to the roots of the plant. Application to the roots may be indirect. The biological control composition may be applied to agricultural products after they have been harvested from the plants. In one embodiment, the application step does not kill the plants. In one embodiment, the method further includes the step of applying a fertilizer, herbicide, pesticide, or a combination thereof to the plants. The fertilizer, herbicide, or pesticide may be applied before, after, or simultaneously with the biological control composition.

[0018] In one embodiment, the packaging material includes polyethylene terephthalate (PET), molded fibers, oriented polystyrene (OPS), polystyrene (PS) foam, polypropylene (PP), or a combination thereof. Steps applied to the packaging material may include washing or impregnating the packaging material.

[0019] In one embodiment, antifungal activity is the prevention of fungal pathogen growth for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days. Antifungal activity may be a reduction in the growth of fungal pathogens in plants, seeds, or agricultural products thereof compared to the growth of fungal pathogens in a control, which is a Rosaceae plant, seed, or agricultural product thereof not exposed to the biological control composition. The growth of fungal pathogens can be reduced for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days after exposure of the fungal pathogens to the biological control composition, compared to the growth of fungal pathogens in plants, seeds, or agricultural products thereof not exposed to the biological control composition. In one embodiment, the biological control composition has antifungal activity against filamentous or non-filamentous fungal pathogens. Filamentous or non-filamentous fungal pathogens include Albugo candida, Albugo occidentalis, Alternaria alternata, Alternaria cucumerina, Alternaria dauci, Alternaria solani Alternaria tenuis, Alternaria tenuissima, Alternaria tomatophila, Aphanomyces euteiches, Aphanomyces raphani, Armillaria mellea, Botrydia theobromae, Botrytis cinerea, Botrytinia fuckeliana, Bremia lactuca, Cercospora beticola, Cercosporella rubi, Cladosporium herbarum, Colletotrichum acutatum, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum, Colletotrichum musae, Colletotrichum spaethanium, Cordana musae, Corynespora cassiicola, Daktulosphaira vitifoliae, Didymella bryoniae, Elsinoeampelina, Elsinoe mangiferae, Elsinoe veneta, Erysiphe cichoracearum, Erysiphe necator, Eutypa lata, Fusarium oxysporum, Fusarium solani, Ganoderma boninense, Guignardia bidwellii, Gymnoconia peckiana, Helminthosporium solani, Leptosphaeria coniothyrium, Leptosphaeria maculans, Leveillula taurica, Macrophomina phaseolina, Microsphaera alni, Monilinia fructicola, Monilinia vaccinii-corymbosi, Mycosphaerella angulate, Mycosphaerella brassicicola, Mycosphaerella fragariae, Mycosphaerella fijiensis, Oidopsis taurica, Passalora fulva, Peronospora sparse, Peronospora farinosa, Phoma exigua, Phomopsis obscurans, Phomopsis vaccinia, Phomopsis viticola, Phytophthora capsica, Phytophthora erythroseptica, Phytophthora infestans, Phytophthora parasitica, Plasmopara viticola, Plasmodiophora brassicae, Podosphaera macularis, Polyscytalum pustulans, Pseudocercospora vitis, Puccinia allii, Puccinia sorghi, Pucciniastrum vaccinia, Pythium debaryanum, Pythium sulcatum, Pythium ultimum, Ralstonia solanacearum, Ramularia tulasneii, Rhizoctoniasolani, Rhizopus arrhizus, Rhizopus stoloniferz, Sclerotinia minor, Sclerotinia sclerotiorum, Sclerotium cepivorum, Sclerotium rolfsii, Sclerotinia minor, Sclerotinia sclerotiorum, Septoria apiicola, Septoria lactucae, Septoria lycopersici, Septoria petroelini, Sphaceloma perseae, Sphaerotheca macularis, Spongospora subterrannea, Stemphylium vesicarium, Synchytrium endobioticum, Thielaviopsis basicola, Uncinula necator, Uromyces appendiculatus, Uromyces betae, Verticillium albo-atrum, Verticillium dahliae, Verticillium theobromae, and any combination thereof. Filamentous fungal pathogens include Fusarium oxysporum, Verticillium dahliae, Botrytis cinerea, Colletotrichum spaethaniu, Erysiphe necator, Podosphaera macularis, Monilinia vaccinii-corymbosi, PucciniaThe group can be selected from sorghi and any combination thereof. Plants, seeds, or agricultural products thereof can be selected from the group consisting of almonds, apricots, apples, artichokes, bananas, barley, beets, blackberries, blueberries, broccoli, Brussels sprouts, cabbage, hemp, chili peppers, carrots, celery, Swiss chard, cherries, citrus fruits, corn, melons, dates, figs, garlic, grapes, herbs, spices, kale, lettuce, oil palm, olives, onions, peas, pears, peaches, peanuts, papayas, parsnips, pecans, persimmons, plums, pomegranates, potatoes, quince, radishes, raspberries, roses, rice, spinosa plums, sorghum, soybeans, spinach, strawberries, sweet potatoes, tobacco, tomatoes, turnip leaves, walnuts, and wheat.

[0020] Inclusion by reference All publications, patents, and patent applications referenced herein are incorporated by reference to the same extent that each individual publication, patent, or patent application is specifically and individually indicated as being incorporated by reference herein.

[0021] The novel features of the present invention are described in detail in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by referring to the following detailed description, which describes exemplary embodiments utilizing the principles of the present invention, and to the following appended drawings: [Brief explanation of the drawing]

[0022] [Figure 1] Figure 1 illustrates the percentage survival of Verticillium dahliae and Fusarium oxysporum on semi-solid agar after application of 14 candidate microorganisms, compared to a control (Serenade®).

[0023] [Figure 2] Figure 2 illustrates the phylogenetic relationship between 16S and ITS sequences in Table 1.

[0024] [Figure 3] Figure 3 illustrates the identification of candidate agents using an understanding of interspecies interactions in the environment. Hanseniaspora uvarum was identified as directly interacting with Fusarium oxysporum and causing growth inhibition of this fungus. The ability of Hanseniaspora uvarum to inhibit the growth of Fusarium oxysporum was confirmed, and Hanseniaspora uvarum was advanced as a product candidate. This figure shows the identified first - layer interaction between H. uvarum and F. oxysporum; the identification and isolation of H. uvarum; and the confirmation of the inhibition of F. oxysporum caused by H. uvarum.

[0025] [Figure 4] Figure 4 illustrates the percent surface area of raspberries infected with Botrytis cinerea after different treatments: Botrytis cinerea - infected (+) control, non - infected (-) control, and a sample infected with Botrytis cinerea but to which the culture supernatant of product candidate BC8 (Bacillus amyloliquefaciens; strain 28B) was applied.

[0026] [Figure 5-1] Figures 5A - 5C illustrate fungal growth in raspberries after different treatment regimens. Figure 5A illustrates fungal growth in a Botrytis cinerea - infected control. Figure 5B illustrates fungal growth in a non - infected control. Figure 5C illustrates fungal growth in a raspberry infected with Botrytis cinerea and to which the culture supernatant of product candidate BC8 (Bacillus amyloliquefaciens; strain 28B) was applied. [Figure 5-2]Figures 5A–5C illustrate fungal growth in raspberries after different treatment regimens. Figure 5C illustrates fungal growth in raspberries infected with Botrytis cinerea and treated with the culture supernatant of candidate product BC8 (Bacillus amyloliquefaciens; strain 28B).

[0027] [Figure 6] Figure 6 illustrates the nucleotide alignment of the 16S RNA sequences of strain BC8 and two B. velezensis FZB42 isolates.

[0028] [Figure 7] Figure 7 illustrates the incidence of Botrytis in treated and untreated blueberry shrubs.

[0029] [Figure 8] Figure 8 illustrates the percentage of blueberries infected with Botrytis in treated and untreated plants.

[0030] [Figure 9] Figure 9 illustrates the number of Botrytis flower blight cases per blueberry shrub in treated and untreated plants.

[0031] [Figure 10] Figure 10 illustrates the percentage of blueberries infected with Botrytis in treated and untreated plants.

[0032] [Figure 11] Figure 11 illustrates the number of Botrytis flower blight outbreaks per blueberry shrub in treated and untreated plants.

[0033] [Figure 12] Figure 12 illustrates the percentage of blueberries infected with Botrytis in treated and untreated plants.

[0034] [Figure 13-1] Figure 13A illustrates the number of diseased shootstrikes in treated and untreated blueberry shrubs. [Figure 13-2] Figure 13B illustrates the number of mummified fruits in treated and untreated blueberry shrubs.

[0035] [Figure 14-1] Figure 14A illustrates the number of diseased seedlings in treated and untreated blueberry shrubs. [Figure 14-2] Figure 14B illustrates the number of mummified fruits in treated and untreated blueberry shrubs.

[0036] [Figure 15] Figure 15 illustrates the percentage disease caused by maize rust in treated and untreated maize plants.

[0037] [Figure 16-1] Figure 16A illustrates the percentage disease caused by maize rust in treated and untreated maize plants. [Figure 16-2] Figure 16B illustrates the disease severity index for maize rust in treated and untreated maize plants.

[0038] [Figure 17-1] Figure 17A illustrates the percentage disease caused by maize rust in treated and untreated maize plants. [Figure 17-2] Figure 17B illustrates the disease severity index for maize rust in treated and untreated maize plants.

[0039] [Figure 18-1] Figure 18A illustrates the percentage incidence of downy mildew in treated and untreated grape leaves. [Figure 18-2]Figure 18B illustrates the percentage incidence of downy mildew in treated and untreated grape leaves.

[0040] [Figure 19-1] Figure 19A illustrates the percentage incidence of Botrytis in treated and untreated grape clusters. [Figure 19-2] Figure 19B illustrates the percentage disease index for Botrytis in treated and untreated grape clusters.

[0041] [Figure 20-1] Figure 20A illustrates the percentage incidence of powdery mildew in treated and untreated grape leaves. [Figure 20-2] Figure 20B illustrates the percentage susceptibility index for powdery mildew in treated and untreated grape leaves.

[0042] [Figure 21-1] Figure 21A illustrates the percentage incidence of downy mildew in treated and untreated grape leaves. [Figure 21-2] Figure 21B illustrates the percentage susceptibility index for downy mildew in treated and untreated grape leaves.

[0043] [Figure 22] Figure 22 illustrates the percentage incidence of Botrytis in treated and untreated grape clusters.

[0044] [Figure 23-1] Figure 23A illustrates the percentage incidence of downy mildew in treated and untreated grape leaves. [Figure 23-2] Figure 23B illustrates the percentage susceptibility index for downy mildew in treated and untreated grape leaves.

[0045] [Figure 24-1] Figure 24A illustrates the percentage incidence of downy mildew in treated and untreated grape leaves. [Figure 24-2] Figure 24B illustrates the percentage susceptibility index for downy mildew in treated and untreated grape leaves.

[0046] [Figure 25-1] Figure 25A illustrates the percentage incidence of Botrytis in treated and untreated grape clusters. [Figure 25-2] Figure 25B illustrates the percentage disease index of Botrytis in treated and untreated grape clusters.

[0047] [Figure 26-1] Figure 26A illustrates the percentage incidence of powdery mildew in treated and untreated grape leaves. [Figure 26-2] Figure 26B illustrates the percentage susceptibility index for powdery mildew in treated and untreated grape leaves.

[0048] [Figure 27-1] Figure 27A illustrates the percentage mean incidence of Botrytis in treated and untreated raspberry shrubs. [Figure 27-2] Figure 27B illustrates the percent mean disease index of Botrytis in treated and untreated raspberry shrubs.

[0049] [Figure 28-1] Figure 28A illustrates the percentage mean incidence of powdery mildew in treated and untreated raspberry leaves. [Figure 28-2] Figure 28B illustrates the percentage mean susceptibility index for powdery mildew in treated and untreated raspberry leaves.

[0050] [Figure 29-1] Figure 29A illustrates the mean percentage incidence of powdery mildew in treated and untreated raspberries. [Figure 29-2] Figure 29B illustrates the percent mean susceptibility index for powdery mildew in treated and untreated raspberries.

[0051] [Figure 30-1] Figure 30A illustrates the average percentage incidence of Botrytis in treated and untreated raspberry shrubs. [Figure 30-2] Figure 30B illustrates the percent mean disease index of Botrytis in treated and untreated raspberry shrubs.

[0052] [Figure 31-1] Figure 31A illustrates the percentage incidence of powdery mildew on treated and untreated raspberry leaves. [Figure 31-2] Figure 31B illustrates the percentage susceptibility index for powdery mildew on treated and untreated raspberry leaves.

[0053] [Figure 32-1] Figure 32A illustrates the percentage incidence of powdery mildew in treated and untreated raspberries. [Figure 32-2] Figure 32B illustrates the percentage susceptibility index for powdery mildew in treated and untreated raspberries.

[0054] [Figure 33-1] Figure 33A illustrates the percentage mean incidence of Botrytis in treated and untreated raspberry shrubs. [Figure 33-2] Figure 33B illustrates the percent mean disease index of Botrytis in treated and untreated raspberry shrubs.

[0055] [Figure 34-1] Figure 34A illustrates the percentage incidence of powdery mildew in treated and untreated raspberry leaves. [Figure 34-2] Figure 34B illustrates the percentage susceptibility index for powdery mildew on treated and untreated raspberry leaves.

[0056] [Figure 35-1]Figure 35A illustrates the percentage incidence of powdery mildew in treated and untreated raspberries. [Figure 35-2] Figure 35B illustrates the percentage susceptibility index for powdery mildew in treated and untreated raspberries.

[0057] [Figure 36] Figure 36 illustrates the number of rotten strawberries infected with Botrytis and Rhizopus in treated and untreated plants.

[0058] [Figure 37] Figure 37 illustrates the number of rotten strawberries infected with Botrytis and Rhizopus in treated and untreated plants.

[0059] [Figure 38] Figure 38 illustrates the crop density per meter of soybean plants infected with Pythium in treated and untreated plants.

[0060] [Figure 39] Figure 39 illustrates treated and untreated raspberries infected with Botrytis cineria.

[0061] [Figure 40] Figure 40 illustrates treated and untreated grapes infected with Botrytis cinerea.

[0062] [Figure 41] Figure 41 illustrates treated and untreated apples infected with Botrytis cinerea.

[0063] [Figure 42] Figure 42 illustrates a treated apple infected with Botrytis cinerea.

[0064] [Figure 43]Figure 43 illustrates the percentage of necrotic apples in treated and untreated apples infected with Botrytis cinerea.

[0065] [Figure 44] Figure 44 illustrates treated and untreated peaches infected with Botrytis cinerea. [Modes for carrying out the invention]

[0066] Numerous fungal pathogens can infect agriculturally important plants, leading to food spoilage and reduced food quality while the plants are in the field or after harvest. For example, gray mold, caused by the fungal pathogen Botrytis cinerea, can often be found on fruits such as strawberries and raspberries, both in the field and in grocery stores. Finding ways to reduce losses caused by fungal pathogens is highly desirable for those involved in food production and consumption, and chemical and biological control strategies have been developed to date. However, the use of chemical and biological mycicides in food crops, while effective, can offer unintended side effects (e.g., toxicity) in addition to being undesirable from a consumer's perspective. Furthermore, commercially available biological control compositions may not provide the desired pathogen or plant specificity or efficacy. Finally, there can be a considerable burden on farmers and producers regarding the recording and reporting of the application of cumbersome synthetic chemical pesticides.

[0067] The biological control compositions described herein may have antifungal activity against agriculturally important fungi and may be formulated for use at various points in the production process. For example, these biological control compositions may be formulated for use before harvest, for example, by incorporating the composition into irrigation lines or administering it together with fertilizers, and during processing, packaging, transport, storage, and commercial display of agricultural products after harvest, for example, by spraying the composition onto harvested agricultural products or by applying the composition to packaging materials used for storing or transporting agricultural products. Furthermore, these biological control compositions may exhibit improved efficacy compared to commercially available biological control compositions.

[0068] As used herein, the term “severity index” generally refers to a score representing the degree of visible disease symptoms in a plant. For example, a given severity index may have a specific number (or range of numbers) of lesions on a leaf that indicate disease. For example, a plant with more disease symptoms will have a higher severity index than a plant with a lower severity index. Different plant species may have different severity indices associated with them.

[0069] As used herein, the terms “severity,” “average severity,” or “percent average severity” generally refer to the degree of visible disease symptoms in a plant or plant population. Severity may be calculated as the percentage of plants covered with disease symptoms. Percent average severity may be calculated for a population by assessing the severity of each plant and averaging the severity of each plant.

[0070] As used herein, the terms “susceptibility index,” “average susceptibility index,” or “percentage average susceptibility index” generally refer to a score for a plant population that represents the degree of visible disease symptoms in that population. The susceptibility index may be calculated by multiplying the incidence rate by the severity of the disease. The average susceptibility index may be calculated based on the severity index or score of individual plants, the number of plants with that severity index, the total number of plants, the highest susceptibility index, and the percentage incidence rate, in order to obtain a weighted mean representing the average severity of the disease. In a non-limiting example, a general calculation of the percentage average susceptibility index may be performed as [Total (number of plants with a given score multiplied by that score)] / [(total number of plants multiplied by the highest score)] × 100.

[0071] Compositions for preventing or reducing crop loss and deterioration of food quality This specification discloses biological control compositions that can prevent or reduce the growth of fungal pathogens in plants, seeds, or their agricultural products. The term “agricultural product” may be used herein to refer to the edible part of a plant, such as leaves, stems, seeds, roots, flowers, or fruits. The term “plant” may 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 deterioration of food quality before, during, or after harvesting agricultural products from plants.

[0072] At least one microorganism may be a bacterium or a yeast. At least one microorganism may include a genus selected from the group consisting of Bacillus, Burkholderia, Cutaneotrichosporon, Cyberlindnera, Gluconacetobacter, Gluconobacter, Hanseniaspora, Paraburkholderia, Pseudomonas, Torulaspora, and any combination thereof.

[0073] At least one microorganism may be 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.

[0074] At least one microorganism may belong to the genus Bacillus. At least one microorganism may belong to the genus Burkholderia. At least one microorganism may belong to the genus Cutaneotrichosporon. At least one microorganism may belong to the genus Cyberlindnera. At least one microorganism may belong to the genus Gluconacetobacter. At least one microorganism may belong to the genus Gluconobacter. At least one microorganism may belong to the genus Hanseniaspora. At least one microorganism may belong to the genus Paraburkholderia. At least one microorganism may belong to the genus Pseudomonas. At least one microorganism may belong to the genus Torulaspora.

[0075] 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 may be Paraburkholderia fluroescens. At least one microorganism may be Paraburkholderia frederiksbergensis. At least one microorganism may be Pseudomonas lini. At least one microorganism may be Pseudomonas migulae. At least one microorganism may be Torulaspora delbrueckii.

[0076] 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 rRNA of a microorganism selected from the group consisting of delbrueckii and any combination thereof may include at least one microorganism having sequence identity of at least approximately 70%, 75%, 80%, 85%, 87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100%. The rRNA may be 16S rRNA, 23S rRNA, internal transcription spacer (ITS), or a combination thereof. The at least one microorganism may be a combination of microbial strains from one or more microbial species.

[0077] A biological control 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 is a 16S rRNA sequence that is more than 98% identical to a 16S rRNA sequence selected from the group of SEQ ID NOs: 1 and SEQ ID NOs: 9. The microorganism has an rRNA sequence, or at least one microorganism has an ITS sequence that is more than 98% identical to an ITS sequence selected from the group of SEQ ID NOs. 17 and SEQ ID NOs. 20, or at least one microorganism has an ITS sequence that is more than 90% identical to the ITS sequence of SEQ ID NOs. 18.

[0078] Microorganisms may contain RNA sequences that have at least approximately 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity with sequences selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.

[0079] The biological control composition may further include a second microorganism, the second microorganism being not identical to at least one other microorganism. The second microorganism may include an RNA sequence having at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25. In some cases, the first and second microorganisms are the same species. For example, both the first and second microorganisms may be Bacillus amyloliquefaciens. In more non-limiting examples, the biological control compositions disclosed herein may include a first microorganism and a second microorganism, as well as two or more microorganisms, each being a different strain of the same species, as may be specified. In some cases, the first microorganism and the second microorganism are not of 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 of the same genus. In some cases, the first microorganism and the second microorganism do not belong to the same family. In some cases, the first microorganism and the second microorganism do not belong to the same order. In some cases, the first microorganism and the second microorganism do not belong to the same class. In some cases, the first microorganism and the second microorganism do not belong to the same phylum. In some cases, the first microorganism and the second microorganism do not belong to the same kingdom.

[0080] In one embodiment, 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 the rRNA sequence of a Bacillus species. The Bacillus species may be Bacillus amyloliquefaciens, Bacillus subtilis, or Bacillus velezensis. The rRNA sequence may be a 16S sequence. In one embodiment, 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.

[0081] In one embodiment, 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 the rRNA sequence of a Gluconacetobacter species. The Gluconacetobacter species may be Gluconacetobacter liquefaciens. The rRNA sequence may be a 16S sequence. In one embodiment, 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: 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.

[0082] In one embodiment, 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 the rRNA sequence of a Gluconobacter species. The Gluconobacter species may be Gluconobacter cerinus. The rRNA sequence may be a 16S sequence. In one embodiment, 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: 24.

[0083] In one embodiment, 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 the rRNA sequence of a Burkholderia species or Paraburkholderia species. The Paraburkholderia species may be Paraburkholderia phytofirmans. The rRNA sequence may be a 16S sequence. In one embodiment, 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: 3, SEQ ID NO: 7, or SEQ ID NO: 9.

[0084] In one embodiment, the 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 the rRNA sequence of a Pseudomonas species. The Pseudomonas species may be Pseudomonas fluorescens, Pseudomonas lini, Pseudomonas migulae, or Pseudomonas frederiksbergensis. The rRNA sequence may be a 16S sequence. In one embodiment, the 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: 6, SEQ ID NO: 10, SEQ ID NO: 15, or SEQ ID NO: 22.

[0085] In one embodiment, at least one microorganism comprises at least one microorganism having sequence identity of at least about 85%, 87%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% with sequence number 8.

[0086] In one embodiment, 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 the rRNA sequence of a Cyberlindnera species. The Cyberlindnera species may be Cyberlinderna saturnus or Cyberlindera mrakkii. The rRNA sequence may be an ITS sequence. In one embodiment, 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: 17.

[0087] In one embodiment, 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 the rRNA sequence of a Hanseniaspora species. The Hanseniaspora species may be Hanseniaspora uvarum. The rRNA sequence may be an ITS sequence. In one embodiment, 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: 18 or SEQ ID NO: 25. In one embodiment, at least one microorganism comprises at least one microorganism having at least 90% sequence identity with SEQ ID NO: 18 or SEQ ID NO: 25. In one embodiment, at least one microorganism includes at least one microorganism having at least 95% sequence identity with SEQ ID NO: 18 or SEQ ID NO: 25. In one embodiment, at least one microorganism includes at least one microorganism having at least 99% sequence identity with SEQ ID NO: 18 or SEQ ID NO: 25.

[0088] In one embodiment, 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 the rRNA sequence of a Torulaspora species. The Torulaspora species may be Torulaspora delbrueckii. The rRNA sequence may be an ITS sequence. In one embodiment, 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: 19.

[0089] In one embodiment, 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 the rRNA sequence of a Cutaneotrichosporon species. The Cutaneotrichosporon species may be Cutaneotrichosporon moniliiforme, Cutaneotrichosporon jirovecii, or Cutaneotrichosporon mucoides. The rRNA sequence may be an ITS sequence. In one embodiment, 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.

[0090] A biological control composition may include a microbial community comprising multiple microorganisms. The multiple microorganisms may consist of 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 of the multiple microorganisms may be a different microorganism. A biological control composition may also include secondary metabolites from a microbial community comprising multiple microorganisms, where the multiple microorganisms consist of 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.

[0091] The group may include at least two 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 at least two 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 group may include at least two microorganisms having a first microorganism having a 16S rRNA sequence selected from SEQ ID NO: 1 or SEQ ID NO: 9, or the first microorganism having an ITS sequence that is more than 98% identical to an ITS sequence selected from the group consisting of SEQ ID NO: 17 and SEQ ID NO: 20, or the first microorganism having an ITS sequence that is more than 90% identical to the ITS sequence of SEQ ID NO: 18. At least two microorganisms may include a first microorganism having an ITS sequence that is more 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. At least two microorganisms may include a first microorganism that is a Gluconobacter species and a second microorganism that is a Hanseniaspora species. At least two microorganisms may be Gluconobacter It may include a first microorganism that is Cerinus and a second microorganism that is Hanseniaspora uvarum.

[0092] At least two microorganisms may include a first microorganism having a 16S sequence that is more than 90% identical to sequence number 24, and a second microorganism having an ITS sequence that is more than 90% identical to sequence number 25. At least two microorganisms may include a first microorganism having a 16S sequence that is more than 95% identical to sequence number 24, and a second microorganism having an ITS sequence that is more than 95% identical to sequence number 25. At least two microorganisms may include a first microorganism having a 16S sequence that is more than 98% identical to sequence number 24, and a second microorganism having an ITS sequence that is more than 98% identical to sequence number 25.

[0093] At least three microorganisms may include a first microorganism having a 16S rRNA sequence more than 99% identical to SEQ ID NO: 23, a second microorganism having a 16S rRNA sequence more than 99% identical to SEQ ID NO: 23, and a third microorganism having a 16S rRNA sequence more than 99% identical to SEQ ID NO: 23, wherein the first, second, and third microorganisms contain non-identical genomes. In some cases, genomes may differ by single nucleotide polymorphisms (SNPs). In some cases, genomes may differ by one or more SNPs. In some cases, genomes may differ in the number of genes in each genome. In some cases, genomes may differ by rearrangements, e.g., insertions, deletions, reordering, refactoring, or by lysogenic or inactive phages, inserted sequences, repeating genome sequences, or other different contents of genomic regions or genes. In some cases, cellular DNA content may differ by including one or more plasmids, which may vary by strain. In some cases, a genome can encode different isoforms of a gene. For example, a protein expressed from a gene may contain point mutations, deletions, or insertions that can affect the function of the protein. For example, a protein expressed from a gene may contain point mutations, deletions, or insertions that cannot affect the function of the protein, or that can only substantially affect the function of the protein.

[0094] The group of 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 group of 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.

[0095] The group of 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 group of 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.

[0096] The group of 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 group of 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.

[0097] Table 1 describes the identifiers, putative genus or species, and corresponding sequence numbers of the microbial strains described herein. At least one microorganism may be one of the microorganisms listed in Table 1. Some phylogenetic relationships of these strains are shown in Figure 2. Table 2 describes the sequences corresponding to these sequence numbers. [Table 1-1] [Table 1-2] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4]

[0098] At least one microorganism can be grown in culture. At least one microorganism can be isolated and purified from the culture. At least one microorganism purified from the culture may contain vegetative cells or spores of at least one microorganism. The culture may be a solid or semi-solid medium. The culture may be a liquid medium. The culture may 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 supernatant of the culture contains secondary metabolites of at least one microorganism. The secondary metabolites of at least one 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.

[0099] The biological control composition may contain one or more secondary metabolites of at least one microorganism. One or more secondary metabolites may have antifungal properties on their own. One or more secondary metabolites may have antifungal properties together with other microorganisms in the biological control composition. One or more secondary metabolites can be isolated from the culture supernatant of at least one microorganism. One or more secondary metabolites may include lipopeptides, dipeptides, aminopolyols, proteins, siderophores, phenazine compounds, polyketides, or combinations thereof.

[0100] Lipopeptides can be linear or cyclic lipopeptides (CLPs). Examples of lipopeptides include, but are not limited to, surfactin, fengisin, iturin, macetolid, amphysin, arsolofactin, tracine, syringopeptide, syringomycin, ptysorbin, basilomycin, basilopeptin, bacitracin, polymyxin, daptomycin, mycosubtilin, crustaquin, tensin, prepastatin, viscosine, and echinocandin. Echinocandin may be echinocandib B (ECB). In some examples, the secondary metabolites are surfactin, fengisin, iturin, or a combination thereof.

[0101] The dipeptide may be basilicin or chlorotetaine. The polyketide may be deficidine, macrolactin, basilaene, butyrolactol A, sorafen A, hipporacin A, or forazoline A. The secondary metabolite may be an aminopolyol. The aminopolyol may be tzwittermycin A. The secondary metabolite may be a protein. The protein may be bacisvin, subtilisin, or fungisin.

[0102] The siderophore may be pioverdin, thioquinolobactin, or pioquelin. The phenazine compound may be fenzine-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.

[0103] The biological control composition can be formulated as a liquid or dry formulation. The liquid formulation may be a fluid or an aqueous suspension. The liquid formulation may contain at least one microorganism or its secondary metabolite suspended in water, oil, or a mixture thereof (emulsion). The dry formulation may be a wettable powder, dry flakes, dust, or granules. The wettable powder can be applied as a suspension to plants, seeds, flowers, or their agricultural products. The dust can be applied in a dry state to plants, seeds, or their agricultural products, such as seeds or leaves. The granules can be applied in a dry state or mixed with water to make a suspension. At least one microorganism or its secondary metabolite can be formulated as microencapsulated, and at least one microorganism or its secondary metabolite has a protective inner layer. The protective inner layer may contain any suitable polymer.

[0104] The biological control composition may further contain additional compounds. These additional compounds may include carriers, surfactants, wetting agents, penetrating agents, emulsifiers, spreaders, adhesives, stabilizers, nutrients, binders, drying agents, 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, cereal flour. The surfactant may be anionic, cationic, amphoteric, or nonionic surfactants. The surfactant may be Tween® 20 or Tween® 80. The wetting agent may include polyoxyethylene esters, ethoxysulfates, or derivatives thereof. In some cases, the wetting agent is mixed with a nonionic surfactant. The penetrating agent may include hydrocarbons. The spreading agent may include fatty acids, latex, aliphatic alcohols, crop oil (e.g., cottonseed), or inorganic oils. The adhesive may include emulsified polyethylene, polymerized resin, fatty acids, petroleum distillates, or pregelatinized cornflour. 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, polyethylene polymer, polysaccharides, xanthan gum, or vegetable oil. The dispersant may be microcrystalline cellulose. The UV protectant may be oxybenzone, blankophor BBH, or lignin.

[0105] The biological control composition may further contain dipicolinic acid.

[0106] At least one microorganism may comprise an effective amount of isolated and purified microorganisms isolated and purified from the liquid culture. At least one microorganism from the liquid culture can be air-dried, freeze-dried, spray-dried, or fluid-bed-dried to produce a dried formulation. The dried formulation can be redissolved in liquid to produce a liquid formulation.

[0107] The biological control composition can be formulated so that at least one microorganism can replicate once it is applied to / delivered to a target habitat (e.g., soil, plants, seeds, and / or agricultural products).

[0108] The biological control composition may have a shelf life of at least one week, one month, six months, at least one year, at least two years, at least three years, at least four years, or at least five years. The shelf life may indicate the length of time during which the biological control composition maintains at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% of its antifungal properties. The biological control composition can be stored at room temperature, 4°C or below, 0°C or below, or -20°C or below.

[0109] The biological control composition may contain spores. The spore-containing composition may be applied by the method described herein. The spore-containing composition may extend the shelf life of the biological control composition. The spore-containing composition may survive low pH or low temperature of the target habitat. For example, the spore-containing composition may be applied to soil at lower temperatures (e.g., below 10°C) and may have antifungal properties with respect to seeds planted at higher temperatures (e.g., 20°C). The spores may become vegetative cells and give them any advantages of being vegetative cells.

[0110] A biological control composition may contain vegetative cells. A vegetative cell-containing composition may be applied by the method described herein. Vegetative cells may proliferate and increase the effectiveness of the composition. For example, vegetative cells in a biological control composition may proliferate after application to increase the surface area of ​​plants exposed to the biological control composition. In another example, vegetative cells in a biological control composition may proliferate after application to increase the amount of time the biological control composition survives, thus extending the period during which the biological control composition is effective. Vegetative cells may proliferate and compete with fungal pathogens for nutrients. Vegetative cells may actively produce one or more secondary metabolites having antifungal properties. Vegetative cells may become spores and give them any advantages of spores.

[0111] A biological control composition may have antifungal activity, for example, prevention or reduction of the growth of fungal pathogens in plants, seeds, or their agricultural products. A biological control composition can prevent the growth of fungal pathogens in plants, seeds, or their agricultural products for at least 1 day, at least 2 days, at least 3 days, at least 4 days, or at least 5 days. A biological control composition can prevent the growth of fungal pathogens in plants, seeds, or their agricultural products 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. A biological control composition can prevent the growth of fungal pathogens in plants, seeds, or their agricultural products for longer than 10 days.

[0112] A biological control composition can reduce the growth of fungal pathogens in plants, seeds, or agricultural products thereof compared to the growth of fungal pathogens in a control, which is a plant, seed, flower, or agricultural product thereof that has not been exposed to the biological control composition. The control may be a plant, seed, or agricultural product thereof that has not been treated with an antifungal agent, or a plant, seed, flower, or agricultural product thereof 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®). A biological control composition can reduce the growth of fungal pathogens in plants, seeds, or their agricultural products for at least 1 day, at least 2 days, at least 3 days, at least 4 days, or at least 5 days. A biological control composition can reduce the growth of fungal pathogens in plants, seeds, or their agricultural products 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. A biological control composition can reduce the growth of fungal pathogens in plants, seeds, or their agricultural products for longer than 10 days. A biological control composition can reduce the growth of fungal pathogens by at least 25% compared to the growth of fungal pathogens in a control. A biological control composition can reduce the growth of fungal pathogens by at least 60% compared to the growth of fungal pathogens in a control. The biological control composition can reduce the growth of fungal pathogens 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 fungal pathogens in the control.

[0113] Albugo, Alternaria, Aphanomyces, Armillaria, Aspergillus, Botrytis, Botrydiplodia, Botrytinia, Bremia, Cercospora, Cercosporella, Cladosporium, Colletotrichum, Cordana, Corynespora, Cylindroca rpon, Daktulosphaira, Didymella, Elsinoe, Erysiphe, Eutypa, Fusarium, Ganoderma, Guignardia, Gymnoconia, Helminthosporium, Leptosphaeria, Leveillula, Macrophomina, Microsphaera, Monolinia, Mycosphaere It may be a fungal pathogen of the genera lla, Oidopsis, Passalora, Peronospora, Phomopsis, Phytophthora, Peronospora, Phoma, Plasmodiophora, Plasmopara, Podosphaera, Polyscytalum, Pseudocercospora, Puccinia, Pucciniastrum, Pythium, Ralstonia, Ramularia, Rhizoctonia, Rhizopus, Septoria, Sclerotinia, Sclerotium, Sphaerotheca, Sphaceloma, Spongospora, Stemphylium, Synchytrium, Thielaviopsis, Uncinula, Uromyces, or Verticillium.The fungal pathogens are Albugo candida, Albugo occidentalis, Alternaria alternata, Alternaria cucumerina, Alternaria dauci, Alternaria solani, Alternaria tenuis, Alternaria tenuissima, Alternaria tomatophila, Aphanomyces euteiches, Aphanomyces raphani, Armillaria mellea, Botrydia theobromae, Botrytis cinerea, Botrytinia fuckeliana, Bremia lactuca, Cercospora beticola, Cercosporella rubi, Cladosporium herbarum, Colletotrichum acutatum, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum, Colletotrichum musae, Colletotrichum spaethanium, Cordana musae, Corynespora cassiicola, Daktulosphaira vitifoliae, Didymella. bryoniae, Elsinoe ampelina, Elsinoe mangiferae, Elsinoe veneta, Erysiphe cichoracearum, Erysiphe necator, Eutypa lata, Fusarium germinareum, Fusarium oxysporum, Fusarium solani, Ganoderma boninense, Guignardia bidwellii, Gymnoconia peckiana, Helminthosporium solani, Leptosphaeria coniothyrium, Leptosphaeria maculans, Leveillula taurica, Macrophomina phaseolina, Microsphaera alni, Monilinia fructicola, Monilinia vaccinii-corymbosi, Mycosphaerella angulate, Mycosphaerella brassicicola, Mycosphaerella fragariae, Mycosphaerella fijiensis, Oidopsis taurica, Passalora fulva, Peronospora sparse, Peronospora farinosa, Phoma exigua, Phomopsis obscurans, Phomopsis vaccinia, Phomopsis viticola, Phytophthora capsica, Phytophthora erythroseptica, Phytophthora infestans, Phytophthora parasitica, Plasmopara viticola, Plasmodiophora brassicae, Podosphaera macularis, Polyscytalum pustulans, Pseudocercospora vitis, Puccinia allii, Puccinia sorghi, Pucciniastrum vaccinia, Pythium debaryanum, Pythium sulcatum, Pythium ultimum, Ralstoniasolanacearum, Ramularia tulasneii, Rhizoctonia solani, Rhizopus arrhizus, Rhizopus stoloniferz, Sclerotinia minor, Sclerotinia sclerotiorum, Sclerotium cepivorum, Sclerotium rolfsii, Sclerotinia minor, Sclerotinia sclerotiorum, Septoria apiicola, Septoria lactucae, Septoria lycopersici, Septoria petroelini, Sphaceloma perseae, Sphaerotheca macularis, Spongospora subterranea, Stempphylium vesicarium, Synchytrium endobioticum, Thielaviopsis basicola, Uncinula necator, Uromyces appendiculatus, Uromyces betae, Verticillium albo-atrum, Verticillium dahliae, Verticillium It may be a theobromae or a combination thereof. The fungal pathogen may be Fusarium oxysporum or Verticillium dahliae. The fungal pathogen may be Botrytis cinerea. The fungal pathogen may be Colletotrichum spaethanium. The fungal pathogen may be Erysiphe necator. The fungal pathogen may be Peronospora farinosa. The fungal pathogen may be Podosphaera maculari. The fungal pathogen may be Monilinia vaccinii-corymbosi. The fungal pathogen may be Puccinia sorghi. The fungal pathogen may be a fungal pathogen that causes powdery mildew. The fungal pathogen may be a fungal pathogen that causes downy mildew. The fungal pathogen may be a fungal pathogen that causes mummy berry disease. The fungal pathogen may be a fungal pathogen that causes maize rust.

[0114] Plants, flowers, seeds, or agricultural products thereof may be of almonds, apricots, apples, artichokes, bananas, barley, beets, blackberries, blueberries, broccoli, Brussels sprouts, cabbage, hemp, peppers, carrots, celery, Swiss chard, cherries, citrus fruits, corn, melons, dates, figs, garlic, grapes, herbs, spices, kale, lettuce, oil palm, olives, onions, peas, pears, peaches, peanuts, papayas, parsnips, pecans, persimmons, plums, pomegranates, potatoes, quince, radishes, raspberries, roses, rice, spinosa plums, sorghum, soybeans, spinach, strawberries, sweet potatoes, tobacco, tomatoes, turnip leaves, walnuts, or wheat. Plants, seeds, flowers, or agricultural products thereof may be plants or agricultural products of the Rosaceae family. Plants, flowers, seeds, or agricultural products of the Rosaceae family may be derived from the genus Rubus, e.g., raspberry or blackberry; the genus Fragaria, e.g., strawberry; the genus Pyrus, e.g., pear; the genus Cydonia, e.g., quince; the genus Prunus, e.g., almond, peach, plum, apricot, cherry, or spinosa plum; the genus Rosa, e.g., rose; or the genus Malus, e.g., apple. Plants, seeds, flowers, or agricultural products thereof may be plants or agricultural products of the Ericaceae family. Plants, seeds, flowers, or agricultural products of the Ericaceae family may be derived from the genus Vaccinium, e.g., blueberry. Plants, seeds, flowers, or agricultural products thereof may be plants or agricultural products of the Ericaceae family. Plants, seeds, flowers, or agricultural products of the Ericaceae family may be derived from the genus Vaccinium, e.g., blueberry. The plants, seeds, flowers, or agricultural products thereof may belong to the Vitaceae family. The plants, seeds, flowers, or agricultural products thereof may originate from the genus Vitis, for example, grapes.

[0115] Method for identifying and isolating biological control compositions

[0116] Methods for identifying and / or selecting biological control compositions may include the step of culturing at least one microorganism alone or together with several other microorganisms and / or fungal pathogens. For example, culturing at least one microorganism together with a fungal pathogen can identify the effectiveness of the at least one microorganism in inhibiting the growth of the fungal pathogen. The effectiveness of the at least one microorganism in inhibiting the growth of the fungal pathogen can be determined by observing the growth parameters of the fungal pathogen. For example, the effectiveness of high inhibition may be determined by using the absence of viable fungal pathogens near at least one microorganism in a semi-solid or solid growth medium. The effectiveness of at least one microorganism may be identified using the optical concentration of a liquid medium containing at least one microorganism and a fungal pathogen.

[0117] At least one microorganism can be identified by various methods. At least one microorganism can be subjected to a sequencing reaction. The sequencing reaction may identify sequences of 16S rRNA, 12S rRNA, 18S rRNA, 28S rRNA, 13S rRNA and 23S rRNA, internal transcription spacers (ITS), ITS1, ITS2, cytochrome oxidase I (COI), cytochrome b, or any combination thereof. The sequencing reaction may identify 16S rRNA sequences, ITS sequences, or combinations thereof. The sequencing reaction may be used to identify at least one microbial species or strain.

[0118] At least one microorganism may be influenced by other microorganisms. Microorganisms may behave synergistically when cultured together such that their antifungal properties are improved when cultured together compared to when cultured individually. For example, at least one microorganism may have increased viability when cultured with another microorganism. At least one microorganism may have increased growth when cultured with another microorganism. At least one microorganism may utilize chemicals or metabolites produced by another microorganism. At least one microorganism may directly interact with another microorganism. For example, at least one microorganism and another microorganism may form a biofilm or multicellular structure. At least one microorganism may produce and / or secrete increased amounts of secondary metabolites when cultured with another microorganism. For example, at least one microorganism may produce an intermediate metabolite, which is then processed by another microorganism to yield a secondary metabolite. Microorganisms that may benefit from culture with other microorganisms can be identified using methods disclosed elsewhere herein, as can biocontrol compositions comprising a first microorganism and a second microorganism, where the second microorganism is not identical to the first microorganism.

[0119] In some cases, at least one microorganism may be affected by environmental conditions. At least one microorganism may grow at a specific pH or produce secondary metabolites. For example, the pH at which at least one microorganism grows may be 3.0, 4.0, 5.0, 6.0, 6.2, 6.4, 6.6, 6.8, 7.0, 7.2, 7.4, 7.6, 7.8, 8.0, 9.0, 10.0, or higher. For example, the pH at which at least one microorganism grows may be 3.0, 4.0, 5.0, 6.0, 6.2, 6.4, 6.6, 6.8, 7.0, 7.2, 7.4, 7.6, 7.8, 8.0, 9.0, 10.0, or lower. At least one microorganism may grow in the presence of salt or produce secondary metabolites. The salt may be a buffer salt. At least one microorganism may grow or produce secondary metabolites in the presence of sugars or carbohydrates. These sugars or carbohydrates may be glucose or glycerol.

[0120] The biological control composition can be cultured using a variety of media or substrates. At least one microorganism may be a culture on an agar dish. At least one microorganism may be cultured on a semi-solid agar dish. At least one microorganism may be cultured on a liquid medium.

[0121] Selection of Microbial Communities

[0122] Methods for identifying or selecting a biological control composition containing a microbial community can be used. For example, the method disclosed in U.S. Patent Publication US20180127796 can be used to identify or select a microbial community. In some cases, multiple microorganisms can be grown together. In some cases, the method may include the step of diluting a sample to form multiple dilutions, one of which contains a subset of multiple microorganisms. The dilutions may allow for the generation of multiple subsets of the multiple microorganisms from which different microorganisms can interact. The subsets of multiple microorganisms can be subjected to culture so that the microorganisms can grow. The subsets can be subjected to a sequencing reaction so that a sequence of microorganisms can be obtained. From the sequencing reaction, species, strain, or other taxonomic information can be obtained. Sequences for identifying specific microorganisms are discussed elsewhere herein. The subsets can be subjected to various culture times, thereby allowing them to be subjected to a sequencing reaction at various point in time to monitor the presence and / or relative abundance of a particular species, strain, or other taxonomic category. Interactions between multiple microorganisms can be determined by observing the presence and / or changes in the relative abundance of specific species, strains, or other taxonomic categories. For example, a first microorganism may have a higher relative abundance when cultured with a second microorganism compared to its relative abundance when not cultured with a second microorganism. In this example, the first microorganism may interact with the second microorganism in such a way that its overall viability increases. Multiple dilutions can each be subjected to sequencing reactions to identify the microorganisms in each dilution, enabling a multiplexed, high-throughput approach.

[0123] Multiple microorganisms can be diluted so that subsets of multiple microorganisms grow together. In some cases, serial dilutions of multiple microorganisms can be performed to form multiple serial dilutions of the sample. The microorganisms in the multiple serial dilutions of the sample may be due to dispersion or chance. The multiple serial dilutions may differ in different implementations. In some embodiments, a series of dilutions of the sample may include dilutions of the sample at ratios of 1:10, 1:100, 1:1000, 1:10000, 1:100000, 1:1000000, 1:10000000, 1:100000000, or approximately 1:10, 1:100, 1:1000, 1:10000, 1:100000, 1:1000000, 1:10000000, 1:100000000, or a number or range between any two of these values. In some embodiments, a series of dilutions of a sample may include at least or many 1:10, 1:100, 1:1000, 1:10000, 1:100000, 1:1000000, 1:10000000, 1:1000000000, or 1:10000000000 dilutions of the sample. For example, a sample can be diluted 10-fold to a 1:10 dilution of the sample using, for example, a buffer. A 1:10 dilution of the sample can then be diluted 10-fold to a 1:100 dilution of the sample. A series of 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 using, for example, a buffer. A sample can then be diluted 100-fold to a 1:100 dilution of the sample. Multiple sequential 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.

[0124] In some embodiments, culturing multiple dilutions of a sample under first culture conditions includes culturing multiple dilutions of a sample under first culture conditions for multiple periods that can range from as short as one minute to up to one year.

[0125] Multiple microorganisms can be subjected to sequencing reactions to identify specific microorganisms. When a subset is cultured for a certain period, the overall percentage representation of each microorganism in the subset may change from its percentage at the start of culture. For example, microorganisms that remain alive among other microorganisms after different culture periods may exhibit symbiotic relationships or interactions between microorganisms in the culture, and these microorganisms may form a microbial community. This microbial 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 in this specification.

[0126] The isolation of specific microorganisms may also be carried out for use in methods or compositions described elsewhere herein. For example, multiple microorganisms may be subjected to serial dilutions so that colonies of a particular microorganism can be isolated. Each serial dilution may be cultured in a liquid, semi-solid, or solid medium. Multiple microorganisms may form colonies on a semi-solid or solid medium such as an agar plate. The colonies may be dispersed so that each colony may contain a single strain or species of microorganism. The isolation of specific microorganisms may also be carried out using physical separation methods such as centrifugation. For example, multiple microorganisms may be cultured in a liquid medium and then centrifuged to isolate the microorganisms from the culture. Specific microorganisms may also be isolated using specific growth conditions. For example, a particular microorganism may have higher viability than another microorganism when cultured under anaerobic conditions. A particular microorganism may have higher viability than another microorganism when cultured in a specific nutrient-rich medium.

[0127] Methods for preventing or reducing food spoilage and deterioration of food quality The plants, seeds, flowers, or agricultural products thereof are treated with a biological control composition before harvest. A method for preventing or reducing the growth of fungal pathogens in plants, seeds, or agricultural products thereof may include the step of applying a biological control composition comprising at least one microorganism or one or more of its secondary metabolites and carriers described herein to plants, seeds, flowers, or agricultural products before harvesting. Harvesting of agricultural products may refer to taking the edible part of the plant from the rest of the plant, or to harvesting the whole plant and then taking the edible part at a later date.

[0128] The step of applying the biological control composition before harvest may include sprinkling, injecting, spraying, or brushing the biological control composition onto the plant, seeds, or its agricultural products. The step of applying the biological control composition may include adding the biological control composition to a drip line, irrigation system, chemical irrigation system, spray, or immersion agent. In some cases, the biological control composition is applied to the roots of the plant, the seeds of the plant, the leaves of the plant, the soil surrounding the plant, or the edible parts of the plant, also referred to herein as the agricultural products of the plant.

[0129] The method may further include the step of applying fertilizers, herbicides, pesticides, other biological control agents, or a combination thereof to the plants. In some examples, fertilizers, herbicides, pesticides, other biological control agents, or a combination thereof are applied before, after, or simultaneously with the biological control composition.

[0130] A method for preventing or reducing the growth of fungal pathogens may include the step of applying a biological control composition comprising at least one microorganism or its secondary metabolites and carrier as described herein to seeds. The step of applying the biological control composition to plant seeds may occur before planting, at planting, or after planting but before germination. For example, the biological control composition may be applied to the surface of seeds before planting. In some cases, seed treatment occurring before planting may include the step of adding a colorant or dye, carrier, binder, adhesive, defoamer, lubricant, nutrient, or a combination thereof to the biological control composition.

[0131] A method for preventing or reducing the growth of fungal pathogens may include the step of applying a biological control composition comprising at least one microorganism or its secondary metabolites and carrier as described herein to the soil. The biological control composition may 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, the soil conditioner resulting in improved plant growth, and the soil conditioner comprises the biological control composition. In some cases, the soil conditioner further comprises a fertilizer.

[0132] A method for preventing or reducing the growth of fungal pathogens may include the step of applying a biological control composition comprising at least one microorganism or its secondary metabolites and carrier as described herein to the roots. The biological control 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 biological control composition. The biological control composition may be applied indirectly to the roots. An example of indirect application to the roots of a plant may include spraying the biological control composition near the base of the plant, so that the biological control composition penetrates the soil and reaches the roots.

[0133] After harvest, the agricultural products are treated with a biological control composition. A method for preventing or reducing the growth of fungal pathogens in agricultural products may include the step of applying a biological control composition comprising at least one microorganism or its secondary metabolites and carrier as described herein to agricultural products before or after harvest.

[0134] The step of applying a biological control composition before or after harvest may include sprinkling, dipping, spreading, injecting, rubbing, spraying, or brushing the biological control composition onto the plant produce. The biological control composition may be applied to the produce immediately before or immediately after harvest, or within one, two, three, four, five, six, or one week after harvest. In some cases, the biological control composition may be applied by an entity that harvests, an entity that packages the produce, an entity that transports the produce, or an entity that sells or commercially displays the produce for consumers in the process of treating the produce immediately before or immediately after harvest.

[0135] The step of applying the biological control composition after harvest may further include the step of incorporating the biological control composition into a process for treating the crop after harvest. The crop may be treated immediately after harvest, for example, during one or more washes. One or more washes may include the use of water or ozonated water to which bleach (chlorine) and / or sodium bicarbonate has been added. The crop may also be treated with oil, resin, or structural or chemical matrix. The biological control composition may be mixed with oil, resin, or structural or chemical matrix for application. The crop may be treated before or after drying. For example, the biological control composition may be added to wax, gum arabic, or other coatings used to coat the crop. The biological control composition may be added at any point in the process, included in one of the washing solutions as part of a new washing solution, or mixed with wax, gum arabic, or other coatings of the crop.

[0136] Treatment of packaging materials with biological control compositions A method for preventing or reducing the growth of fungal pathogens in agricultural products may include the step of applying a biocontrol composition comprising at least one microorganism or its secondary metabolites and carrier as described herein to packaging materials used for transporting or storing agricultural products.

[0137] Packaging materials may include polyethylene terephthalate (PET), molded fibers, oriented polystyrene (OPS), polystyrene (PS) foam, polypropylene (PP), or a combination thereof. Packaging materials may include cardboard, rigid board, Styrofoam®, or molded pulp. Packaging materials may include a substrate, such as cellulose. Packaging materials may be horizontal flow (HFFS) packages, vertical flow (VFFS) packages, thermoformed packages, sealed trays, or stretchable films. Thermoformed packages may be clamshell packages. Packaging materials may be baskets, trays, or clamshells.

[0138] The packaging material treated with the biological control composition may be an insert. The insert may be a pad, a sheet, or a blanket. The sheet may be placed in or on a cage, tray, basket, or clamshell. The insert may contain cellulose or a cellulose derivative. The insert may contain at least one layer of a microporous polymer, e.g., polyethylene or polypropylene, and at least one layer of a superabsorbent polymer. In some cases, the insert includes an outer layer and an inner layer. The inner layer may be a water-absorbing layer. The inner layer may contain carboxymethylcellulose, cellulose ether, polyvinylpyrrolidone, starch, dextrose, gelatin, pectin, or a combination thereof. The outer layer may be a water-permeable layer.

[0139] The step of applying the biological control composition to the packaging material may include the step of washing, spraying, or impregnating the packaging material with the biological control composition.

[0140] The terms used herein are for the purpose of describing specific cases only and are not intended to be restrictive. The following terms are considered to illustrate the meaning of the terms used herein, in addition to the understanding of these terms by those skilled in the art. Where used herein and in the appended claims, the singular forms “one,” “an,” and “it” include plural nouns unless the context clearly indicates otherwise. It should be further noted that the claims may be drafted to exclude elements as needed. This statement is therefore intended to serve as a precedent for the use of exclusive terms such as “only,” “only,” or “negative” limitations in relation to the enumeration of elements in the claims.

[0141] A particular range is expressed herein as a numerical value following the term “approximately.” The term “approximately” is used herein to provide a literal basis for the exact number that follows it, as well as for numbers that are close to or approximate the number that follows the term. When determining whether a number is close to or approximates a specifically enumerated number, an unenumerated number that is close to or approximates may be a number that, in the context in which it is presented, provides a substantial equivalent of the specifically enumerated number. Where a range of values ​​is provided, each intervening value between the upper and lower limits of that range, up to one-tenth of the unit of its lower limit unless the context clearly indicates otherwise, and any other enumerated or intervening values ​​within that specified range, are understood to be included in the methods and compositions described herein. The upper and lower limits of these smaller ranges may independently be included in smaller ranges and are also included in the methods and compositions described herein, subject to any specifically excluded limits within the specified ranges. Where a specified range includes one or both limits, a range excluding either or both of the limits they include is also included in the methods and compositions described herein.

[0142] Unless otherwise defined, all scientific and technical terms used herein have the same meaning as those commonly understood by those skilled in the art of the methods and compositions described herein. Any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the methods and compositions described herein, but representative exemplary methods and materials are listed below. [Examples]

[0143] (Example 1: Screening of microorganisms for antifungal activity) Microorganisms were screened for their ability to inhibit the growth of the fungal pests Verticillium dahliae and Fusarium oxysporum. Fourteen candidate organisms were identified as active in the first layer screening and were able to form clear zones in the fungal community (see Figure 3 for the identification of Hanseniaspora uvarum, which inhibits Fusarium oxysporum, for example). These 14 candidate organisms were then screened in a second layer assay designed to mimic a structured soil environment using semi-solid agar. In this assay, the reduction in growth compared to untreated fungi was determined, with untreated fungal growth set at 100%. Serenade, a commercially available product containing the active ingredient of Bacillus subtilis QST 713 strain, was used as a control, and it reduced the growth of F. oxysporum by approximately 25% and the growth of V. dahliae by >99%. Of these candidates, 10 significantly reduced the growth of F. oxysporum compared to Serenade®, and 11 reduced the growth of V. dahliae to a level indistinguishable from Serenade (Figure 1).

[0144] The candidate strains tested are found in Table 3, along with the most closely related microbial species or genus identified. [Table 3-1] [Table 3-2]

[0145] (Example 2: Antifungal activity of Bacillus amyloliquefaciens (strain 28B; BC8) supernatant in raspberries) The antifungal activity of the culture supernatant of an isolated strain of Bacillus amyloliquefaciens (strain 28B; BC8) against Botrytis cinerea in raspberries was determined. After 120 hours, raspberries treated with the supernatant of the Bacillus amyloliquefaciens strain showed fungal growth similar to that of negative control raspberries (not infected with Botrytis cinerea), with fungal growth covering less than 5% of the raspberry surface area. In contrast, positive control raspberries (infected with Botrytis cinerea) showed approximately 90% of their surface area covered by fungal growth after 120 hours (Figure 4; Figures 5A-5C).

[0146] (Example 3: Evaluation of the effectiveness of BC8 against Botrytis cinerea and Colletotrichum spaethanium-induced flower blight in blueberry crops) The efficacy of BC8 against Botrytis cinerea and Colletotrichum spaethanium-induced flower blight in blueberries was evaluated. Blueberry shrubs were treated with either BC8 or a control treatment. As a control treatment, shrubs were left untreated or treated with Lifegard (Certis; active ingredient: Bacillus mycoides), a combination of Stargus (Marrone; active ingredient: Bacillus amyloliquefaciens strain F727) and Nufilm (Fertrell; mixture of terpene polymer and emulsifier), or sequential application of Bravo Weatherstik (Syngenta; active ingredient: Chlorothalonil (tetrachloroisophthalonitrile) 54%), Captevate (Arysta; active ingredients: fenhexamide, captan) and Pristine (BASF; active ingredients: pyraclostrobin, boscide). Standard commercial fertilizer and insecticide programs were also applied to all treated areas. The treatment product was mixed with 75 gallons of water (0.9 L / treatment or according to the label specifications) and applied to plants at regular intervals using a sprayer. The shrubs were treated at various growth stages, including early leaf buds (EGT), late leaf buds (LGT), pink buds (PB), flowering (BLM), flower fall (PF), green fruit (GRF), and 10% blue fruit (BLF). The shrubs were grown and maintained according to the grower's standard practices.

[0147] Botrytis cinerea infection was assessed by counting the number of flower blights per blueberry shrub. Four replicates per treatment were performed in a randomized plot design format.

[0148] Figure 7 shows the results, expressed as the incidence of B. cinerea-induced flower blight per shrub. BC8 was effective in reducing the incidence of flower blight by approximately 55% compared to the untreated control.

[0149] The data were analyzed using one-way analysis of variance (ANOVA), and the means were compared using Fisher's least significant difference (LSD). Box plots, which are represented by the same letters in each graph, are not significantly different (LSD p=0.05).

[0150] (Example 4: Evaluation of the effectiveness of BC8 against post-harvest rot caused by Botrytis cinerea and Colletotrichum spaethanium in blueberry crops) The effectiveness of BC8 against post-harvest rot caused by Botrytis cinerea or Colletotrichum sp. infection in blueberry crops was evaluated. Blueberry shrubs were treated with BC8 or a control treatment before harvest, and the blueberries obtained were observed after harvest. As a control treatment, shrubs were left untreated or treated with Lifegard (Certis), a combination of Stargus and Nufilm (Fertrell), or sequential treatment with Bravo Weatherstik (Syngenta), Captevate (Aresta), and Pristine (Bayer).

[0151] Post-harvest rot caused by B. cinerea and Colletrotrichum sp. was evaluated after harvesting 50 berries and placing them in a humid chamber at room temperature for 12–14 days. The results, expressed as % infected fruit, are shown in Figure 8. Treated berries in the harvest had a 10% berry rot incidence, while untreated berries had an 85% rot incidence (Figure 8). BC8 was as effective in reducing post-harvest rot in blueberry crops as the commercially available standard combination of Bravo Weatherstik (Syngenta), Captevate (Aresta), and Pristine (Bayer) (Figure 8).

[0152] The data were analyzed using one-way analysis of variance (ANOVA), and the means were compared using Fisher's least significant difference (LSD). Box plots, which are represented by the same letters in each graph, are not significantly different (LSD p=0.05).

[0153] (Example 5: Evaluation of the effectiveness of BC16 against Botrytis cinerea and Colletotrichum spaethanium-induced flower blight in blueberry crops) BC16 was evaluated for its efficacy against Botrytis cinerea and Colletotrichum spaethanium-induced flower blight in blueberries. Blueberry shrubs were treated with either BC16 or a control treatment. As a control treatment, shrubs were left untreated or treated with Lifegard (Certis), a combination of Stargus and Nufilm (Fertrell), or a sequential application of Bravo Weatherstik (Syngenta), Captevate (Aresta), and Pristine (Bayer). A standard commercial fertilizer and insecticide program was also applied to all treatment areas. The treatment product was mixed with 75 gallons of water (0.9 L / treatment or according to manufacturer's specifications) and applied to plants at regular intervals using a sprayer. The shrubs were treated at various growth stages, including early leaf bud (EGT), late leaf bud (LGT), pink bud (PB), flowering (BLM), flower drop (PF), green fruit (GRF), and 10% blue fruit (BLF). The shrubs were grown and maintained according to the grower's standard practices.

[0154] The incidence of Botrytis cinerea and Colletotrichum spaethanium infections was assessed by counting the number of flower blight cases per blueberry shrub. Four replicates were performed per treatment in a randomized plot design format. The results, expressed as the incidence of flower blight per shrub, are shown in Figure 9. BC16 was effective in reducing the incidence of flower blight by approximately 52% compared to the untreated control.

[0155] The data were analyzed using one-way analysis of variance (ANOVA), and the means were compared using Fisher's least significant difference (LSD). Box plots, which are represented by the same letters in each graph, are not significantly different (LSD p=0.05).

[0156] (Example 6: Evaluation of the effectiveness of BC16 against post-harvest rot caused by Botrytis cinerea and Colletotrichum spaethanium in blueberry crops) The effectiveness of BC16 against post-harvest rot caused by Botrytis cinerea or Colletrotrichum sp. infection in blueberry crops was evaluated. Blueberry shrubs were treated with BC16 or a control treatment before harvest, and the blueberries obtained were observed after harvest. As a control treatment, berries were left untreated or treated with Lifegard (Certis), a combination of Stargus and Nufilm (Fertrell), or sequential treatment with Bravo Weatherstik (Syngenta), Captevate (Aresta), and Pristine (Bayer). 50 berries were harvested and placed in a humid chamber at room temperature for 12–14 days, after which post-harvest rot caused by B. cinerea and Colletrotrichum sp. was evaluated. The results, expressed as % infected fruit, are shown in Figure 10. BC16-treated shrubs in harvest 1 had a 5% berry rot incidence (Figure 10), while untreated berries in harvest 1 had an 85% rot rate (Figure 10). BC16 was as effective as commercially available standards Bravo Weatherstik (Syngenta), Captevate (Aresta), and Pristine (Bayer) in reducing post-harvest rot in blueberry crops (Figure 10).

[0157] The data were analyzed using one-way analysis of variance (ANOVA), and the means were compared using Fisher's least significant difference (LSD). Box plots, which are represented by the same letters in each graph, are not significantly different (LSD p=0.05).

[0158] (Example 7: Evaluation of the effectiveness of BC17 against Botrytis cinerea and Colletotrichum spaethanium-induced flower blight in blueberry crops) BC17 was evaluated for its efficacy against Botrytis cinerea and Colletotrichum spaethanium-induced flower blight in blueberries. Blueberry shrubs were treated with either BC17 or a control product. As a control treatment, shrubs were left untreated or treated with Lifegard (Certis), a combination of Stargus and Nufilm (Fertrell), or a sequential application of Bravo Weatherstik (Syngenta), Captevate (Aresta), and Pristine (Bayer). Standard commercial fertilizer and insecticide programs were also applied to all treatment areas. The treatment product was mixed with 75 gallons of water at a rate of 0.9 L / treatment or according to the manufacturer's specifications and applied to plants at regular intervals using a sprayer. The shrubs were treated at various growth stages, including early leaf bud (EGT), late leaf bud (LGT), pink bud (PB), flowering (BLM), flower drop (PF), green fruit (GRF), and 10% blue fruit (BLF). The shrubs were grown and maintained according to the grower's standard practices.

[0159] Botrytis cinerea infection was assessed by counting the number of flower blights per blueberry shrub. Four replicates per treatment were performed using a randomized plot design format.

[0160] Figure 11 shows the results, expressed as the incidence of flower blight per shrub. BC17 was effective in reducing the incidence of flower blight by 80% compared to the untreated control.

[0161] The data were analyzed using one-way analysis of variance (ANOVA), and the means were compared using Fisher's least significant difference (LSD). Box plots, which are represented by the same letters in each graph, are not significantly different (LSD p=0.05).

[0162] (Example 8: Evaluation of the effectiveness of BC17 against post-harvest rot caused by Botrytis cinerea and Colletotrichum spaethanium in blueberry crops) The effectiveness of BC17 against post-harvest rot caused by Botrytis cinerea or Colletrotrichum sp. infection in blueberry crops was evaluated. Blueberry shrubs were treated with BC17 or a control treatment before harvest, and the blueberries obtained were observed after harvest. As a control treatment, berries were left untreated or treated with Lifegard (Certis), a combination of Stargus and Nufilm (Fertrell), or sequential treatment with Bravo Weatherstik (Syngenta), Captevate (Aresta), and Pristine (Bayer). Fifty berries were harvested and placed in a humid chamber at room temperature for 12–14 days, after which post-harvest rot caused by B. cinerea and Colletrotrichum sp. was evaluated.

[0163] The results, expressed as % infected fruit, are shown in Figure 12. BC17-treated shrubs had a 7% berry rot incidence (Figure 12), while untreated berries had an 85% rot rate (Figure 12). BC17 was as effective as commercially available standards Bravo Weatherstik (Syngenta), Captevate (Aresta), and Pristine (Bayer) in reducing post-harvest rot in blueberry crops (Figure 12).

[0164] The data were analyzed using one-way analysis of variance (ANOVA), and the means were compared using Fisher's least significant difference (LSD). Box plots, which are represented by the same letters in each graph, are not significantly different (LSD p=0.05).

[0165] (Example 9: Evaluation of the effectiveness of BC16 against Monilinia vaccinii-corymbosi-induced mummie berry disease in blueberry crops) BC16 was evaluated for its efficacy against Monilinia vaccinii-corynbosi-induced mummie berry disease in blueberry crops. As a control treatment, shrubs were left untreated or treated with a combination of Bravo Weatherstik (Syngenta), Indar 2F (Corteva Agriscience; active ingredient: fenbuconazole), and Pristine (Bayer). Standard commercial fertilizer and insecticide programs were also carried out in all treated areas. The treatment products were mixed with 75 gallons of water at a rate of 0.9 L / treatment or according to the manufacturer's specifications and applied to plants at regular intervals, approximately weekly, using a sprayer. Shrubs were treated at various growth stages, including early leaf bud (EGT), late leaf bud (LGT), pink bud (PB), flowering (BLM), flower drop (PF), green fruit (GRF), and 10% blue fruit (BLF). Shrubs were grown and maintained according to standard grower practices. Four replicates were performed per treatment using a randomized plot design format. Infection was assessed before fruit production and after berry production.

[0166] The incidence of mummie berry disease caused by Monilinia vaccinii-corymbosi infection was evaluated by the presence of blueberry seedlings infected with flower blight, commonly referred to as diseased seedlings. The results, expressed as the incidence of M. vaccinii-corymbosi-induced diseased seedlings (per shrub), are shown in Figure 13A. BC16 was effective in reducing the incidence of flower blight by approximately 52% compared to the untreated control.

[0167] The incidence was also assessed by the phenotype of mummified fruit, characterized by hardening and wilting of infected berries. Blueberries treated with BC16 were evaluated for the presence of mummified fruit 7 days after the crop reached the green fruit stage. The results, expressed as the incidence of mummified fruit, are shown in Figure 13B. BC16 was effective in reducing the number of mummified fruit by approximately 59% compared to the untreated control.

[0168] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0169] (Example 10: Evaluation of the effectiveness of BC17 against Monilinia vaccinii-corymbosi-induced mummie berry disease in blueberry crops) BC17 was evaluated for its effectiveness against Monilinia vaccinii-corynbosi-induced mummie berry disease in blueberry crops. Shrubs were treated with either BC17 or a control treatment. As a control, shrubs were either left untreated or treated with a combination of Bravo Weatherstik (Syngenta), Indar 2F (Corteva Agriscience), and Pristine (BASF). Standard commercial fertilizer and insecticide programs were also applied to all treated areas. The treatment product was mixed with 75 gallons of water at a concentration of 0.9 L / treatment or according to the manufacturer's specifications and applied to plants at regular intervals using a sprayer. Shrubs were treated at various growth stages, including early leaf bud (EGT), late leaf bud (LGT), pink bud (PB), flowering (BLM), flower drop (PF), green fruit (GRF), and 10% blue fruit (BLF). Shrubs were grown and maintained according to standard grower practices. Four replicates were performed per treatment using a randomized plot design format. Infection was assessed before fruit production and after berry production.

[0170] Mummy berry disease caused by Monilinia vaccinii-corymbosi infection was evaluated by the presence of blueberry seedlings infected with flower blight, commonly referred to as diseased seedlings. The results, expressed as the incidence of diseased seedlings per shrub, are shown in Figure 14A. BC17 was effective in reducing the incidence of flower blight by approximately 75% compared to the untreated control.

[0171] Disease incidence was also assessed by the phenotype of mummified fruit, characterized by hardening and wilting of infected berries. Blueberries treated with BC17 were evaluated for the presence of mummified fruit 7 days after the crop reached the green fruit stage. The results, expressed as the incidence of mummified fruit, are shown in Figure 14B. BC17 was effective in reducing the number of mummified fruit by approximately 80% compared to the untreated control.

[0172] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0173] (Example 11: Evaluation of the efficacy of BC8 against the fungal maize pathogen Puccinia sorghi) The efficacy of BC8 against maize rust caused by the fungal pathogen Puccinia sorghi in maize crops was evaluated. The BC8 treatment consisted of three applications at 7-10 day intervals at a rate of 40 qt / acre. Two rows, each 20 feet long, were treated with BC8, leaving one row as a buffer zone between the treated sections. Four repetitions were performed in each treatment protocol.

[0174] As a control treatment, rows were left untreated or treated with Daconil SDG (Syngenta; active ingredient: chlorothalonil). Standard commercial fertilizer and insecticide programs were also applied to all treated areas. Shrubs were grown and maintained according to grower's standard practices.

[0175] The treatment product was mixed with 20 gallons / acre of water and applied to plants at regular intervals using a napsack sprayer (7.5-foot boom with a 28 psi flat fan nozzle). Crops were treated at the start of the normal season (end of June) or at the first signs of disease (whichever came first).

[0176] BC8 was effective in inhibiting damage caused by maize rust and maize disease compared to untreated plots. Four days after the last treatment, the average disease rate in plots treated with BC8 was observed to be approximately 8.8%, compared to the untreated plots where disease incidence was approximately 20% (Figure 15). BC8 was more effective than commercially available treatments in inhibiting damage caused by maize rust in maize crops (Figure 15).

[0177] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0178] (Example 12: Evaluation of the efficacy of BC16 against the fungal maize pathogen Puccinia sorghi) The efficacy of BC16 against maize rust caused by the fungal pathogen Puccinia sorghi in maize crops was evaluated. BC16 treatment consisted of three applications at 7-10 day intervals at rates of 20 qt / acre or 40 qt / acre. Two rows, each 20 feet long, were treated with BC16, with one row left as a buffer zone between the treated sections. Four replicates were performed in each treatment protocol.

[0179] As a control measure, rows were left untreated or treated with Daconil SDG (Syngenta). Standard commercial fertilizer and insecticide programs were also applied to all treated areas. Maize crops were grown and maintained according to standard grower practices. The treatment product was mixed with 20 gallons / acre of water and applied to plants at regular intervals using a napsack sprayer (7.5-foot boom with 28 psi flat fan nozzle). Crops were treated at the start of the normal season (end of June) or at the first signs of disease (whichever came first).

[0180] BC16 was effective in inhibiting maize rust and disease-induced damage in maize crops compared to untreated plots. The average disease rate in plots treated with 20 qt / acre of BC16 was approximately 10% compared to untreated plots where approximately 20% disease was observed 4 days after the last treatment (Figure 16A). The average disease rate in plots treated with 20 qt / acre of BC16 was approximately 5% compared to untreated plots where approximately 20% disease was observed 4 days after the last treatment (Figure 16A). BC16 was more effective than standard commercial treatments in inhibiting damage caused by maize rust in maize crops (Figure 16A).

[0181] The disease severity index was measured after three applications. Compared to untreated plots and plots treated with commercially available standards, plots treated with BC16 at 20 qt / acre and 40 qt / acre showed reduced disease severity (Figure 16B).

[0182] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0183] (Example 13: Evaluation of the efficacy of BC17 against the fungal maize pathogen Puccinia sorghi) The efficacy of BC17 against maize rust caused by the fungal pathogen Puccinia sorghi in maize crops was evaluated. The BC17 treatment consisted of three applications at 7-10 day intervals at a rate of 20 qt / acre. Two rows, each 20 feet long, were treated with BC17, with one row left as a buffer zone between the treated sections. Four repetitions were performed in each treatment protocol.

[0184] As a control measure, shrubs were left untreated or treated with Daconil SDG (Syngenta). Standard commercial fertilizer and insecticide programs were also applied to all treated areas. Shrubs were grown and maintained according to standard grower practices. The treatment product was mixed with 20 gallons / acre of water and applied to plants at regular intervals using a napsack sprayer (7.5-foot boom with 28 psi flat fan nozzle). Crops were treated at the start of the normal season (end of June) or at the first signs of disease (whichever came first).

[0185] BC17 was effective in inhibiting maize rust and disease-induced damage in maize compared to untreated plots. The average disease level in plots treated with 20 qt / acre of BC17 was observed to be approximately 10% compared to untreated plots where approximately 20% disease was observed 4 days after the last treatment (Figure 17A). The average disease level in plots treated with 20 qt / acre of BC17 was observed to be approximately 13.8% compared to untreated plots where approximately 20% disease was observed 4 days after the last treatment (Figure 17A). BC17 was more effective than the standard commercially available treatment Daconil SDG (Syngenta) in inhibiting damage caused by maize rust in maize crops (Figure 17A).

[0186] The disease severity index was measured after three applications. Compared to untreated plots and commercially available standard treatments, plots treated with BC17 at 20 qt / acre showed reduced disease severity (Figure 17B).

[0187] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0188] (Example 14: Evaluation of the effectiveness of BC8 against Plasmopara viticola in Vignol grapes) BC8 was evaluated for its effectiveness against the progression of downy mildew caused by Plasmopara viticola in Vignol vines. Vineyards were treated with either BC8 or a control treatment. The BC8 treatment consisted of eight applications applied at intervals of 7 to 14 days, depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyards were left untreated or treated with a combination of RevusTop (Syngenta; active ingredients: madnipropamide, defenoconazole) and Intuity (Valent USA; active ingredient: mandestrobin) (referred to as Intuity in Figures 18A and 18B), or a combination of Manzate (Keystone Pest Solutions; active ingredient: mancozeb) and Pristine (Bayer) (referred to as a commercially available standard in Figures 18A and 18B). Standard commercial fertilizer and insecticide programs were also applied to all treatment areas. The vines were grown and maintained according to standard grower practices. Treatment products were mixed with water according to the manufacturer's specifications and applied to the shrubs using a spray blooming device. Four replicates were performed for each treatment. A randomized plot design was employed for the trial.

[0189] BC8 treatment resulted in effective control of downy mildew induced by Plasmopara viticola, as demonstrated by reductions in both disease severity (Figure 18A) and susceptibility index (Figure 18B) compared to untreated grapevine leaves.

[0190] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0191] (Example 15: Evaluation of the effectiveness of BC8 against rot caused by Botrytis cinerea in vineyard grapes) BC8 was evaluated for its effectiveness against rot caused by Botrytis cinerea in vineyard vines. Vineyard vines were treated with either BC8 or a control treatment. The BC8 treatment consisted of eight applications applied at intervals of 7–14 days depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyard vines were left untreated or treated with a combination of RevusTop (Syngenta) and Intuity (Valent USA) (referred to as Intuity in Figures 19A and 19B), or a combination of Manzate (Keystone Pest Solutions) and Pristine (Bayer) (referred to as a commercially available standard in Figures 19A and 19B). Standard commercial fertilizer and insecticide programs were also carried out in all treated areas. Vineyard vines were grown and maintained according to standard grower practices.

[0192] The treatment product was mixed with water according to the manufacturer's instructions and applied to the grapevines using a spray bloomer. Four replicates were performed for each treatment. A randomized plot design was employed for the study. In total, four replicates were performed for three grapevines per plot.

[0193] The BC8 treatment resulted in effective control of rot induced by Botrytis cinerea in grape clusters, as demonstrated by both a nearly 32% reduction in disease severity (Figure 19A) and a roughly 50% reduction in the disease index (Figure 19B) compared to untreated grape clusters.

[0194] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0195] (Example 16: Evaluation of the effectiveness of BC8 against powdery mildew induced by Erysiphe necator in Vignol grapes) BC8 was evaluated for its effectiveness against powdery mildew caused by Erysiphe necator in Vignol vines. Vineyards were treated with either BC8 or a control treatment. The BC8 treatment consisted of eight applications applied at intervals of 7–14 days depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyards were left untreated or treated with a combination of RevusTop (Syngenta) and Intuity (Valent USA) ("Intuity" in Figures 20A and 20B), or a combination of Manzate (Keystone Pest Solutions) and Pristine (Bayer) ("Commercial Standard" in Figures 20A and 20B). Standard commercial fertilizer and insecticide programs were also carried out in all treated areas. Vineyards were grown and maintained according to standard grower practices.

[0196] The treatment product was mixed with water according to the manufacturer's specifications and applied to the grapevines using a spray bloom device. Four replicates were performed for each treatment. A randomized plot design was employed for the trial.

[0197] BC8 treatment reduced powdery mildew induced by Erysiphe necator in grape leaves, as demonstrated by both a nearly 30% reduction in disease severity (Figure 20A) and a roughly 50% reduction in the disease index (Figure 20B) compared to untreated grape leaves.

[0198] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0199] (Example 17: Evaluation of the effectiveness of BC16 against Plasmopara viticola in Vignol grapes) BC16 was evaluated for its effectiveness against the progression of downy mildew caused by Plasmopara viticola in Vignol vines. Vineyards were treated with either BC16 or a control treatment. The BC16 treatment consisted of eight applications applied at intervals of 7–14 days depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyards were left untreated, or treated with a combination of RevusTop (Syngenta) and Intuity (Valent USA) ("Intuity" in Figures 21A and 21B), or Manzate (Keystone). The vines were treated with a combination of Pest Solutions and Pristine (Bayer) (referred to as "Commercial Standard" in Figures 21A and 21B). A standard commercial fertilizer and insecticide program was also applied to all treated areas. The vines were grown and maintained according to the grower's standard practices.

[0200] The treatment product was mixed with water according to the manufacturer's specifications and applied to the grapevines using a spray blooming device.

[0201] Four replicates were performed for each treatment. A randomized plot design was employed for the trial.

[0202] BC16 treatment reduced downy mildew on leaves induced by Plasmopara viticola, as demonstrated by reductions in both disease severity (Figure 21A) and susceptibility index (Figure 21B) compared to leaves of untreated grape plants.

[0203] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0204] (Example 18: Evaluation of the effectiveness of BC16 against rot caused by Botrytis cinerea in vineyard grapes) BC16 was evaluated for its effectiveness against rot caused by Botrytis cinerea in vineyard vines. Vineyard vines were treated with either BC16 or a control treatment. The BC16 treatment consisted of eight applications applied at intervals of 7–14 days depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyard vines were left untreated or treated with a combination of RevusTop (Syngenta) and Intuity (Valent USA) ("Intuity" in Figure 22), or a combination of Manzate (Keystone Pest Solutions) and Pristine (Bayer) ("Commercial Standard" in Figure 22). Standard commercial fertilizer and insecticide programs were also carried out in all treated areas. Vineyard vines were grown and maintained according to standard grower practices.

[0205] The treatment product was mixed with water according to the manufacturer's specifications and applied to the grapevines using a spray bloom device. Four replicates were performed for each treatment. A randomized plot design was employed for the trial.

[0206] BC16 treatment inhibited rot induced by Botrytis cinerea in grape clusters, as demonstrated by a nearly 32% reduction in disease severity compared to untreated grape clusters (Figure 22).

[0207] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0208] (Example 19: Evaluation of the effectiveness of BC16 against powdery mildew induced by Erysiphe necator in Vignol grapes) BC16 was evaluated for its effectiveness against powdery mildew caused by Erysiphe necator in Vignol vines. Vineyards were treated with either BC16 treatment or a control treatment. The BC16 treatment consisted of eight applications applied at intervals of 7–14 days depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyards were left untreated or treated with a combination of RevusTop (Syngenta) and Intuity (Valent USA) (referred to as "Intuity" in Figures 23A and 23B), or a combination of Manzate (Keystone Pest Solutions) and Pristine (Bayer) (referred to as "Commercial Standard" in Figures 23A and 23B). Standard commercial fertilizer and insecticide programs were also carried out in all treated areas. Vineyards were grown and maintained according to standard grower practices.

[0209] The treatment product was mixed with water according to the manufacturer's specifications and applied to the grapevines using a spray bloom device. Four replicates were performed for each treatment. A randomized plot design was employed for the trial.

[0210] BC16 treatment reduced the degree of Erysiphe necator-induced powdery mildew in grape leaves (Figure 23A), resulting in an approximately 30% reduction in the susceptibility index compared to untreated grape leaves (Figure 23B).

[0211] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0212] (Example 20: Evaluation of the effectiveness of BC18 against Plasmopara viticola in Vignol grapes) BC18 was evaluated for its effectiveness against the progression of downy mildew caused by Plasmopara viticola in Vignol vines. Vineyards were treated with either BC18 or a control treatment. The BC18 treatment consisted of eight applications applied at intervals of 7–14 days depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyards were left untreated or treated with a combination of RevusTop (Syngenta) and Intuity (Valent USA) ("Intuity" in Figures 24A and 24B), or a combination of Manzate (Keystone Pest Solutions) and Pristine (Bayer) ("Commercial Standard" in Figures 24A and 24B). Standard commercial fertilizer and insecticide programs were also carried out in all treated areas. Vineyards were grown and maintained according to standard grower practices.

[0213] The treatment product was mixed with water according to the manufacturer's specifications and applied to the grapevines using a spray bloom device. Four replicates were performed for each treatment. A randomized plot design was employed for the trial.

[0214] BC18 was as effective as commercially available standard treatments in controlling downy mildew on grape leaves (Figures 24A and 24B, respectively). BC18 treatment reduced downy mildew on leaves induced by Plasmopara viticola, as indicated by an approximately 71% reduction in disease severity (Figure 24A) and an approximately 80% reduction in the disease index (Figure 24B) compared to untreated grape leaves.

[0215] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0216] (Example 21: Evaluation of the effectiveness of BC18 against rot caused by Botrytis cinerea in vineyard grapes) BC18 was evaluated for its effectiveness against rot caused by Botrytis cinerea in vineyard vines. Vineyard vines were treated with either BC18 or a control treatment. The BC18 treatment consisted of eight applications applied at intervals of 7 to 14 days, depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyard vines were left untreated or treated with a combination of RevusTop (Syngenta) and Intuity (Valent USA) ("Intuity" in Figures 25A and 25B), or a combination of Manzate (Keystone Pest Solutions) and Pristine (Bayer) ("Commercial Standard" in Figures 25A and 25B). Standard commercial fertilizer and insecticide programs were also carried out in all treated areas. Vineyard vines were grown and maintained according to standard grower practices.

[0217] The treatment product was mixed with water according to the manufacturer's specifications and applied to the grapevines using a spray bloom device. Four replicates were performed for each treatment. A randomized plot design was employed for the trial.

[0218] BC18 was as effective as commercially available standard treatments in controlling Botrytis cinerea infection in grape clusters (Figures 25A and 25B, respectively). BC18 treatment inhibited Botrytis cinerea-induced rot in grape clusters, as demonstrated by a nearly 80% reduction in disease severity (Figure 25A) and an approximately 87% reduction in the morbidity index (Figure 25B) compared to untreated grape clusters.

[0219] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0220] (Example 22: Evaluation of the effectiveness of BC18 against powdery mildew induced by Erysiphe necator in Vignol grapes) The effectiveness of BC18 against powdery mildew caused by Erysiphe necator in Vignol vines was evaluated. Vineyards were treated with either BC18 or a control treatment. The BC18 treatment consisted of eight applications applied at intervals of 7–14 days depending on the growth stage. The first four applications were applied at a rate of 40 gallons / acre, and the last four applications were applied at a rate of 50 gallons / acre. As a control treatment, vineyards were left untreated, or treated with a combination of RevusTop (Syngenta) and Intuity (Valent USA) ("Intuity" in Figures 26A and 26B), or Manzate (Keystone Pest). The areas were treated with a combination of Solutions and Pristine (Bayer) (referred to as "Commercial Standard" in Figures 26A and 26B). A standard commercial fertilizer and insecticide program was also applied to all treated areas. The vines were grown and maintained according to the grower's standard practices.

[0221] The treatment product was mixed with water according to the manufacturer's specifications and applied to the grapevines using a spray bloom device. Four replicates were performed for each treatment. A randomized plot design was employed for the trial.

[0222] BC18 was as effective as commercially available standard and Intuity treatments in controlling powdery mildew in grape leaves (Figures 26A and 26B, respectively). BC18 treatment reduced the degree of Erysiphe necator-induced powdery mildew in grape leaves by approximately 80% compared to untreated grape leaves (Figure 26A) and resulted in an approximately 87% reduction in the susceptibility index (Figure 26B).

[0223] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0224] (Example 23: Evaluation of the effectiveness of BC8 against Botrytis cinerea infection in raspberries) BC8 was evaluated for its efficacy against powdery mildew caused by Botrytis cinerea and Podosphaera macularis in raspberries. Shrubs were treated with either BC8 or a control treatment. BC8 treatment was applied at 14-day or 7-day intervals depending on the growth stage. Treatment consisted of 5-6 applications in total at a rate of 39 gallons / acre. As a control treatment, shrubs were left untreated or treated with industry standards (a combination of Rally (Corteva Agriscience; active ingredient: mycobutanil), Pristine (BASF), Elevate (Arysta LifeScience; active ingredient: fenhexamide, and Switch (Syngenta)), or biological controls including Botector (Nufarm; active ingredient: Aureobasidium pullalans), Double Nickel (Certis; active ingredient: Bacillus amyloliquefaciens strain D747), or Stargus / NuFilm P. Standard commercial fertilizer and insecticide programs were also implemented in all treatment areas. Shrubs were grown and maintained according to standard grower practices.

[0225] The treatment product was mixed with water according to the manufacturer's specifications and applied to plants at regular intervals. Four replicates were performed for each treatment. A randomized plot protocol was employed for the trial, using 10' plots per treatment.

[0226] BC8 was as effective as commercial treatments in controlling Botrytis cinerea infection in raspberry bushes. The BC8 treatment reduced the extent of Botrytis cinerea infection in raspberry plants by approximately 75% compared to the untreated control (Figure 27A) and reduced the average disease index by more than 90% (Figure 27B).

[0227] All data were analyzed through one-way analysis of variance (ANOVA), and the means were compared using Fisher's least significant difference (LSD). Box-and-whisker plots denoted with the same letter within each graph were not significantly different (LSD p = 0.05).

[0228] (Example 24: Evaluation of the efficacy of BC8 against powdery mildew caused by Podosphaera macularis in raspberry bushes) BC8 was evaluated for its efficacy against powdery mildew caused by Podosphaera macularis in raspberry bushes. The bushes were treated with either BC8 or a control treatment. The BC8 treatment was applied at 14-day or 7-day intervals depending on the growth stage. The treatment consisted of 5 - 6 applications in total at a rate of 39 gallons / acre. As a control treatment, the bushes were left untreated or treated with a biological control containing the industry standards (a combination of Rally (Corteva Agriscience), Pristine (Bayer), Elevate (Arysta LifeScience), and Switch (Syngenta)), or Botector, Double Nickel, or Stargus / NuFilm P. A standard commercial fertilizer and pesticide program was also conducted in all treatment areas. The bushes were grown and maintained according to the producer's standard practices.

[0229] The treatment products were mixed with water according to the manufacturer's specifications and applied to the plants at regular intervals. Four replicates were performed for each treatment. A randomized plot protocol was employed for the test using 10 plots per treatment.

[0230] The effectiveness of BC8 against powdery mildew, measured by the reduction of the average disease incidence and average disease index in raspberry leaves, is shown in FIGS. 28A and 28B, respectively. The BC8 treatment reduced the degree of Podosphaera macularis infection in raspberry leaves by approximately 75% (FIG. 28A). The BC8 treatment was as effective as the commercial treatment and reduced the average disease index by approximately 70% compared to the untreated control (FIG. 28B).

[0231] The effectiveness of BC8 against powdery mildew, measured by the reduction of the average disease incidence and average disease index in raspberry berries, is shown in FIGS. 29A and 29B, respectively. The BC8 treatment reduced the degree of Podosphaera macularis infection in raspberry berries by approximately 70% (FIG. 29A). The BC8 treatment reduced the average disease index in raspberry berries by approximately 90% compared to the untreated control (FIG. 29B).

[0232] All data were analyzed through one-way analysis of variance (ANOVA), and the mean values were compared using Fisher's least significant difference (LSD). Box-and-whisker plots labeled with the same letter within each graph are not significantly different (LSD p = 0.05).

[0233] (Example 25: Evaluation of the effectiveness of BC16 against Botrytis cinerea infection in raspberries) BC16 was evaluated for its effectiveness against powdery mildew caused by Botrytis cinerea and Podosphaera macularis in raspberries. Shrubs were treated with either BC16 or a control treatment. BC16 treatment was applied at 14-day or 7-day intervals depending on the growth stage. Treatment consisted of 5-6 applications across a total area of ​​39 gallons / acre. As a control treatment, shrubs were left untreated or treated with industry standards (a combination of Rally (Corteva Agriscience), Pristine (Bayer), Elevate (Arysta LifeScience), and Switch (Syngenta)), or biological controls including Botector, Double Nickel, or Stargus / NuFilm P. Standard commercial fertilizer and insecticide programs were also applied to all treated areas. Shrubs were grown and maintained according to grower standard practices.

[0234] The treatment product was mixed with water according to the manufacturer's specifications and applied to plants at regular intervals. Four replicates were performed for each treatment. A randomized plot protocol was employed for the trial, using 10' plots per treatment.

[0235] BC16 was effective in controlling Botrytis cinerea infection in raspberry shrubs. Compared to untreated controls, BC16 treatment reduced the degree of Botrytis cinerea infection in raspberry plants by approximately 50% (Figure 30A) and reduced the mean morbidity index by more than 63% (Figure 30B).

[0236] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0237] (Example 26: Evaluation of the effectiveness of BC16 against powdery mildew caused by Podosphaera macularis in raspberry shrubs) BC16 was evaluated for its effectiveness against powdery mildew caused by Podosphaera macularis in raspberry shrubs. Shrubs were treated with either BC16 or a control treatment. BC16 treatment was applied at 14-day or 7-day intervals depending on the growth stage. Treatment consisted of 5-6 applications across a total area of ​​39 gallons / acre. As a control treatment, shrubs were either left untreated or treated with industry standards (a combination of Rally (Corteva Agriscience), Pristine (Bayer), Elevate (Arysta LifeScience), and Switch (Syngenta)), or biological controls including Botector, Double Nickel, or Stargus / NuFilm P. Standard commercial fertilizer and insecticide programs were also applied to all treated areas. Shrubs were grown and maintained according to grower standard practices.

[0238] The treatment product was mixed with water according to the manufacturer's specifications and applied to plants at regular intervals. Four replicates were performed for each treatment. A randomized plot protocol was employed for the trial, using 10' plots per treatment.

[0239] The effectiveness of BC16 against powdery mildew, as measured by the reduction in disease severity and morbidity index in raspberry leaves, is shown in Figures 31A and 31B, respectively. BC16 treatment reduced the degree of Podosphaera macularis infection in raspberry leaves by approximately 56% (Figure 31A). BC16 treatment reduced the mean morbidity index by approximately 70% compared to the untreated control (Figure 31B).

[0240] The effectiveness of BC16 against powdery mildew, as measured by the reduction in disease severity and morbidity index in raspberry berries, is shown in Figures 32A and 32B, respectively. BC16 treatment reduced the degree of Podosphaera macularis infection in raspberry berries by approximately 50% (Figure 32A). BC16 treatment reduced the mean morbidity index in raspberry berries by approximately 55% compared to the untreated control (Figure 32B).

[0241] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0242] (Example 27: Evaluation of the effectiveness of BC17 against Botrytis cinerea infection in raspberries) BC17 was evaluated for its effectiveness against powdery mildew caused by Botrytis cinerea and Podosphaera macularis in raspberries. Shrubs were treated with either BC17 or a control treatment. BC17 treatment was applied at 14-day or 7-day intervals depending on the growth stage. Treatment consisted of 5-6 applications across a total area of ​​39 gallons / acre. As a control treatment, shrubs were either left untreated or treated with industry standards (a combination of Rally (Corteva Agriscience), Pristine (Bayer), Elevate (Arysta LifeScience), and Switch (Syngenta)), or biological controls including Botector, Double Nickel, or Stargus / NuFilm P. Standard commercial fertilizer and insecticide programs were also applied to all treated areas. Shrubs were grown and maintained according to grower standard practices.

[0243] The treatment product was mixed with water according to the manufacturer's specifications and applied to plants at regular intervals. Four replicates were performed for each treatment. A randomized plot protocol was adopted for the study.

[0244] BC17 was effective in controlling Botrytis cinerea infection in raspberry shrubs. Compared to untreated controls, BC17 treatment reduced the degree of Botrytis cinerea infection in raspberry plants by approximately 50% (Figure 33A) and the mean morbidity index by more than 55% (Figure 33B).

[0245] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0246] (Example 28: Evaluation of the effectiveness of BC17 against powdery mildew caused by Podosphaera macularis in raspberry shrubs) BC17 was evaluated for its effectiveness against powdery mildew caused by Podosphaera macularis in raspberry shrubs. Shrubs were treated with either BC17 or a control treatment. BC17 treatment was applied at 14-day or 7-day intervals depending on the growth stage. Treatment consisted of 5-6 applications across a total area of ​​39 gallons / acre. As a control treatment, shrubs were either left untreated or treated with industry standards (a combination of Rally (Corteva Agriscience), Pristine (Bayer), Elevate (Arysta LifeScience), and Switch (Syngenta)), or biological controls including Botector, Double Nickel, or Stargus / NuFilm P. Standard commercial fertilizer and insecticide programs were also applied to all treated areas. Shrubs were grown and maintained according to grower standard practices.

[0247] The treatment product was mixed with water according to the manufacturer's specifications and applied to plants at regular intervals. Four replicates were performed for each treatment. A randomized plot protocol was adopted for the study.

[0248] The effectiveness of BC17 against powdery mildew, measured by the reduction of disease incidence and disease index in raspberry leaves, is shown in FIGS. 34A and 34B, respectively. The BC17 treatment reduced the degree of Podosphaera macularis infection in raspberry leaves by approximately 45% (FIG. 34A). The BC17 treatment reduced the average disease index by approximately 70% compared to the untreated control (FIG. 34B).

[0249] The effectiveness of BC17 against powdery mildew, measured by the reduction of disease incidence and disease index in raspberry berries, is shown in FIGS. 35A and 35B, respectively. The BC17 treatment reduced the degree of Podosphaera macularis infection in raspberry berries by approximately 50% (FIG. 35A). The BC17 treatment reduced the average disease index in raspberry berries by approximately 50% compared to the untreated control (FIG. 35B).

[0250] All data were analyzed through one-way analysis of variance (ANOVA), and the mean values were compared using Fisher's least significant difference (LSD). Box-and-whisker plots labeled with the same letter within each graph are not significantly different (LSD p = 0.05).

[0251] (Example 29: Evaluation of the effectiveness of BC16 against decay caused by Botrytis cinerea and Rhizopus spp. infections in strawberry fruits) BC16 was evaluated for its effectiveness against rot caused by Botrytis cinerea and Rhizopus spp. in strawberry fruit. Strawberry broadleaf herb plots were treated with either BC16 or a control treatment. The BC16 treatment consisted of five leaf applications, applied weekly at a rate of 40 quarts / acre. As a control treatment, plots were left untreated or treated with commercial standards including CAPTAN (Keystone Pest solutions) and Procidic (Greenspire Global Inc.; active ingredient: citrate), Aviv (Sym Agro; active ingredient: Bacillus subtilis strain IAB / BS03), Stk 73 (STK;), and Procidic (Greenspire Global Inc.). Standard commercial fertilizer and insecticide programs were also carried out in all treated areas. Broadleaf herbs were grown and maintained according to grower's standard practices.

[0252] The treatment product was mixed with 150 gallons of water per acre and applied to broadleaf herbaceous plants using a portable CO2 backpack sprayer with eight nozzles. Four replicates were performed for each treatment. A randomized plot design was employed for the study. The first harvest was taken the day after the last application, and the second harvest was taken 7 days after the last application. Data were collected from 32 ripe berries gathered from each plot and observed for 12–14 days to assess berry rot in the presence of Botrytis cinerea or Rhizopus spp.

[0253] Figure 36 shows the results, expressed as the number of rotten fruits. BC16-treated berries showed significantly less rot compared to untreated berries, and the results were comparable to those of the commercially available fungicide used in this study.

[0254] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0255] (Example 30: Evaluation of the effectiveness of BC17 against spoilage caused by infection with Botrytis cinerea and Rhizopus spp. in strawberry fruits) BC17 was evaluated for its effectiveness against rot caused by Botrytis cinerea and Rhizopus spp. in strawberry fruit. Strawberry broadleaf herb plots were treated with either BC17 or a control treatment. The BC17 treatment consisted of five leaf applications, applied weekly at a rate of 40 quarts / acre. As a control treatment, shrubs were left untreated or treated with CAPTAN (Keystone Pest solutions) and Procidic (Greenspire). The plants were treated with commercially available standards including Botrytis cinerea (Greenspire Global Inc.), Aviv (Sym Agro), Stk 73 (STK), and Procidic (Greenspire Global Inc.). Standard commercial fertilizer and insecticide programs were also applied to all treated areas. The shrubs were grown and maintained according to the grower's standard practices. The treatment product was mixed with 150 gallons of water per acre and applied to the shrubs using a portable CO2 backpack sprayer with 8 nozzles. Four replicates were performed for each treatment. A randomized plot design was employed for the trial. The first harvest was taken the day after the last application, and the second harvest was taken 7 days after the last application. Data were collected from 32 ripe berries collected from each plot and observed for 12–14 days to assess berry rot in the presence of Botrytis cinerea or Rhizopus spp.

[0256] Figure 37 shows the results, expressed as the number of rotten fruits. Berries treated with BC17 showed significantly less rot compared to untreated berries, and the results were comparable to those of the commercially available fungicide used in this study.

[0257] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0258] (Example 31: Evaluation of the effectiveness of BC17 against root rot caused by Pythium sp. in soybeans) BC17 was evaluated for its effectiveness against root rot caused by Pythium sp. in soybeans. Plots were treated with either BC17 or a control treatment. The BC17 treatment consisted of three applications to Credenz variety soybeans. The first application was at planting time, either in furrows or trenches, over seedlings (seed line) after planting. The second application in trenches was at 100% germination. The third application was carried out 7–10 days after the second application. The treatment consisted of two different application rates: 20 quarts / acre or 40 quarts / acre. As a control treatment, plots were left untreated or treated with the commercially available standard Daconil SDG (Syngenta) ("Commercial Standard" in Figure 38). Standard commercial fertilizer and insecticide programs were also applied to all treated areas. Plots were grown and maintained according to grower's standard practices.

[0259] The treatment product was mixed with 20 gallons / acre of water and applied to plants at normal times (end of June) or at the first signs of disease (whichever came first) using a napsack sprayer (0.5-foot boom with a 28 psi flat nozzle). Four replicates were performed for each treatment. Two rows, each 20 feet long, were treated with BC17, leaving one row as a buffer zone between the treatment sections. Four replicates were performed for each treatment protocol. A randomized complete block design was employed for this study.

[0260] Crop planting density was evaluated as a measure of plant health and emergence rate. The results, expressed as crop planting density (per meter), are shown in Figure 38. Crop planting density was evaluated at the time of the second application, before the third application, and 14 days after the third application. Treatment with BC17 at 40 qt / acre significantly increased soybean crop planting density compared to untreated soybeans. BC17 performed better than commercially available standards in increasing crop planting density.

[0261] All data were analyzed using one-way analysis of variance (ANOVA), and means were compared using Fisher's least significant difference (LSD). Box plots, indicated by the same lettering within each graph, were not significantly different (LSD p=0.05).

[0262] (Example 32: Efficacy of a biocontrol composition against infection by Botrytis cinerea on the shelf life of raspberries) Raspberries were harvested from the plants and placed in sterile containers. The raspberries showed no recognizable fungal infections. Raspberries were intentionally infected with Botrytis cinerea. Two groups of raspberries were evaluated: one group was treated with BC8 culture supernatant, and the other was left untreated. Treatment was applied by immersing the fruit in the treatment formulation, but it may also be incorporated into the packaging holding the raspberries, or applied as a spray or by other suitable methods as described elsewhere herein. After 3 days, visible fungal infection was observed in the untreated raspberries. On the other hand, the BC8-treated raspberries showed no infection even after 5 days. Treated and untreated raspberries are shown in Figure 39.

[0263] (Example 33: Efficacy of a biological control composition against infection by Botrytis cinerea on the shelf life of grapes) Grapes were harvested from the plants and placed in sterile containers. The grapes showed no recognizable fungal infections. Two groups of grapes were intentionally infected with Botrytis cinerea. Figure 40 shows the three groups of grapes evaluated. One group was not intentionally infected and is labeled (-)ctrl, one group was intentionally infected and treated with the biological control composition BC16 and is labeled BC16 product, and one group was intentionally infected but left untreated and is labeled (+)ctrl. Treatment was applied by immersing the fruit in the treatment formulation, but may also be incorporated into the packaging holding the grapes, or applied as a spray or by other suitable methods described elsewhere herein. The BC16-treated grapes showed no recognizable fungal infections.

[0264] (Example 34: Efficacy of a biological control composition against infection by Botrytis cinerea on the shelf life of apples) Apples were harvested from the plants and placed in sterile containers. The apples did not have any recognizable fungal infections. Two apples were intentionally infected with Botrytis cinerea. Figure 42 shows the three apples evaluated. One apple was not intentionally infected and is labeled (-)ctrl, one apple was intentionally infected and treated with the biological control composition BC16 and is labeled BC16 product, and one apple was intentionally infected but left untreated and is labeled (+)ctrl. Treatment was applied by immersing the fruit in the treatment formulation, but may also be incorporated into the packaging holding the apples, or applied as a spray or by other suitable methods described elsewhere herein. The BC16-treated apples showed a smaller area of ​​fungal infection compared to the untreated apples. Apples were also intentionally infected and treated with BC17 and labeled BC17 product. The apples treated with BC17 (Figure 42) showed a smaller area of ​​fungal infection compared to the untreated apples. Figure 43 shows the percentage of necrotic fruit in various apple varieties.

[0265] (Example 35: Efficacy of a biological control composition against infection by Botrytis cinerea on the shelf life of peaches) Peaches were harvested from the plants and placed in sterile containers. The peaches did not show any recognizable fungal infections. Peaches were intentionally infected with Botrytis cinerea. Figure 44 shows the three peaches evaluated. One peach was not intentionally infected and was labeled (-) control, one peach was intentionally infected and treated with the biological control composition BC17 and was labeled BC17, and one peach was intentionally infected but left untreated and was labeled (+) control. Treatment was applied by immersing the fruit in the treatment formulation, but may also be incorporated into the packaging holding the peaches, or applied as a spray or by other suitable methods as described elsewhere herein. The BC17-treated peaches did not show any recognizable fungal infections.

[0266] Preferred embodiments of the present invention have been shown and described herein, but it will be apparent to those skilled in the art that such embodiments are provided only as examples. Numerous modifications, changes, and substitutions will be conceivable to those skilled in the art without departing from the present invention. It should be understood that various substitutes for the embodiments of the present invention described herein may be used in the practice of the present invention. The following claims define the scope of the present invention, and the methods and structures within these claims, as well as their equivalents, are intended to be included therein. The present invention provides, for example, the following items. (Item 1) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, The at least one microorganism is selected from the group of Sequence ID No. 1 and Sequence ID No. 9. A biological control composition having a 16S rRNA sequence that is more than 99% identical to an rRNA sequence, or at least one microorganism having an ITS sequence that is more than 99% identical to an ITS sequence selected from the group of SEQ ID NOs. 17 and SEQ ID NOs. 20, or at least one microorganism having an ITS sequence that is more than 90% identical to the ITS sequence of SEQ ID NOs. 18. (Item 2) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition wherein at least one microorganism contains an rRNA sequence that is more than 99% identical to a sequence longer than 200 bases that includes an rRNA sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 9, or the at least one microorganism has an ITS sequence that is more than 99% identical to the ITS sequence of SEQ ID NO: 17, or the at least one microorganism has an ITS sequence that is more than 90% identical to SEQ ID NO: 18. (Item 3) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of Fusarium oxysporum by 25% or more compared to a control not exposed to the biological control composition, or inhibiting the growth of Verticillium dahliae by 60% or more compared to a control not exposed to the biological control composition, as determined by measuring the survival of Fusarium oxysporum or Verticillium dahliae, respectively. (Item 4) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition comprising at least one microorganism having a 16S rRNA sequence that is more than 99% identical to the 16S rRNA sequence of Sequence ID No. 22. (Item 5) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition comprising at least one microorganism having a 16S rRNA sequence that is more than 99% identical to a 16S rRNA sequence selected from the group of Sequence ID No. 23. (Item 6) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition in which at least one microorganism has a 16S rRNA sequence that is more than 99% identical to the 16S rRNA sequence of SEQ ID NO: 24, or at least one microorganism has an ITS sequence that is more than 99% identical to the ITS sequence of SEQ ID NO: 25, or at least one microorganism has an ITS sequence that is more than 90% identical to the ITS sequence of SEQ ID NO: 25. (Item 7) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of Botrytis cineria by 25% or more compared to a control that has not been exposed to the biological control composition. (Item 8) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of Monilinia vaccinii-corymbosi by 25% or more compared to a control that has not been exposed to the biological control composition. (Item 9) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, Compared to a control that was not exposed to the aforementioned biological control composition, Colletotrichum A biological control composition capable of inhibiting the growth of spaethanium by 25% or more. (Item 10) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of Puccinia sorghi by 25% or more compared to a control that has not been exposed to the biological control composition. (Item 11) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of Plasmopara viticola by 25% or more compared to a control that has not been exposed to the biological control composition. (Item 12) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of Erysiphe necator by 25% or more compared to a control that has not been exposed to the biological control composition. (Item 13) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of Podasphaera macularis by 25% or more compared to a control that has not been exposed to the biological control composition. (Item 14) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of organisms of the genus Pytium by 25% or more compared to a control that has not been exposed to the biological control composition. (Item 15) (i) at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition capable of inhibiting the growth of organisms of the genus Rhizopus by 25% or more compared to a control that has not been exposed to the biological control composition. (Item 16) (i) secondary metabolites of at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition wherein at least one microorganism has an ITS sequence that is more than 99% identical to an ITS sequence selected from the group of SEQ ID NOs: 17, 18, 19, 20, 21, and 25. (Item 17) (i) one or more secondary metabolites of at least one microorganism, and (ii) carrier A biological control composition comprising, A biological control composition wherein at least one microorganism has a 16S rRNA sequence that is more than 99% identical to a 16S rRNA sequence selected from the group of SEQ ID NOs: 1, 9, 22, 23, and 24. (Item 18) A biological control composition according to any one of items 1 to 17, wherein at least one of the aforementioned microorganisms is isolated and purified. (Item 19) The biological control composition according to any one of items 1 to 17, further comprising a second microorganism, wherein the second microorganism is not identical to the at least one microorganism. (Item 20) The biological control composition according to item 19, wherein the second microorganism is isolated and purified. (Item 21) The biological control composition according to item 18 or 20, wherein the second microorganism comprises an RNA sequence that is at least 95% identical to a sequence selected from the group consisting of Sequence IDs 1 to 25. (Item 22) The second microorganism is selected from the group consisting of SEQ ID NOs: 16S A biological control composition according to item 21, comprising a 16S rRNA sequence that is at least 95% identical to the rRNA sequence. (Item 23) The biological control composition according to item 21, wherein the second microorganism comprises an ITS sequence that is at least 95% identical to an internal transcription spacer (ITS) sequence selected from the group consisting of SEQ ID NOs: 17, 18, 19, 20, 21, and 25. (Item 24) The second microorganism is selected from the group consisting of SEQ ID NOs: 16S A biological control composition according to item 21, comprising a 16S rRNA sequence that is at least 99% identical to the rRNA sequence. (Item 25) The biological control composition according to item 21, wherein the second microorganism comprises an ITS sequence that is at least 99% identical to an internal transcription spacer (ITS) sequence selected from the group consisting of SEQ ID NOs: 17, 18, 19, 20, 21, and 25. (Item 26) The second microorganism is selected from the group consisting of SEQ ID NOs: 16S A biological control composition as described in item 21, comprising a 16S rRNA sequence, which is an rRNA sequence. (Item 27) The biological control composition according to item 21, wherein the second microorganism comprises an ITS sequence which is an internal transcription spacer (ITS) sequence selected from the group consisting of SEQ ID NOs: 17, 18, 19, 20, 21, and 25. (Item 28) The biological control composition according to item 21, wherein at least one microorganism contains a 16S rRNA sequence that is at least 99% identical to SEQ ID NO: 24, and the second microorganism contains an ITS sequence that is at least 99% identical to SEQ ID NO: 25. (Item 29) The biological control composition according to any one of items 21 to 27, wherein the biological control composition comprises a third microorganism, the third microorganism being not identical to the second microorganism or the at least one of the microorganisms. (Item 30) The biological control composition according to item 29, wherein the biological control composition comprises a fourth microorganism, and the third microorganism is not identical to the third microorganism, the second microorganism, or the at least one of the microorganisms. (Item 31) The biological control composition according to item 30, wherein the biological control composition comprises a fifth microorganism, the fifth microorganism being not identical to the fourth microorganism, the third microorganism, the second microorganism, or the at least one of the microorganisms. (Item 32) The biological control composition according to item 29, wherein at least one microorganism contains a 16S rRNA sequence that is more than 99% identical to SEQ ID NO: 23, the second microorganism contains a 16S rRNA sequence that is more than 99% identical to SEQ ID NO: 23, and the third microorganism contains a 16S rRNA sequence that is more than 99% identical to SEQ ID NO: 23. (Item 33) The biological control composition according to item 18, wherein at least one microorganism has an ITS sequence that is more than 90% identical to Sequence ID No. 25, and the second microorganism is a Gluconobacter species. (Item 34) The biological control composition according to item 33, wherein the Gluconobacter species is Gluconobacter cerinus. (Item 35) The aforementioned Gluconobacter is 16S, which is more than 99% identical to sequence number 24. A biological control composition according to item 33, having an rRNA sequence. (Item 36) The biological control composition according to item 18, wherein at least one microorganism has an ITS sequence that is more than 90% identical to sequence number 18, and the second microorganism is a Gluconacetobacter species. (Item 37) The biological control composition according to item 35, wherein the Gluconacetobacter species is Gluconacetobacter liquefaciens. (Item 38) The biological control composition according to item 35, wherein the Gluconacetobacter species has a 16S rRNA sequence selected from the group consisting of SEQ ID NOs. 2, SEQ ID NOs. 4, SEQ ID NOs. 5, SEQ ID NOs. 11, SEQ ID NOs. 12, SEQ ID NOs. 13, SEQ ID NOs. 14, and SEQ ID NOs. 16. (Item 39) The biological control composition according to item 16 or 17, wherein the secondary metabolite of the at least one microorganism is isolated from the culture supernatant of the at least one microorganism. (Item 40) The biological control composition according to item 16 or 17, wherein one or more secondary metabolites of at least one microorganism comprises a lipopeptide. (Item 41) The biological control composition according to item 40, wherein the lipopeptide is a cyclic lipopeptide selected from the group consisting of surfactant, phengycin, and iturin. (Item 42) The biological control composition according to item 16 or 17, wherein one or more secondary metabolites of at least one microorganism comprises a polyketide. (Item 43) The biological control composition according to item 16 or 17, wherein one or more secondary metabolites of at least one microorganism contain a volatile antifungal compound. (Item 44) A biological control composition according to any one of items 3 to 15, wherein the control is exposed to Bacillus subtilis strain QST 713. (Item 45) A biological control composition according to any one of items 1 to 15, wherein at least one of the aforementioned microorganisms is isolated and purified. (Item 46) A biological control composition, which is a liquid or powder, as described in any one of items 1 to 45. (Item 47) A biological control composition according to any one of items 1 to 46, comprising vegetative cells. (Item 48) A biological control composition containing spores, as described in any of items 1 to 46. (Item 49) A method for preventing or reducing the growth of fungal pathogens in plants, seeds, or agricultural products thereof, comprising the step of applying a biological control composition described in any one of items 1 to 46 to the plants, seeds, flowers, or agricultural products, wherein the biological control composition has antifungal activity. (Item 50) A method for preventing or reducing the growth of fungal pathogens in plants, seeds, flowers, or agricultural products thereof, comprising the step of applying a biological control composition described in any one of items 1 to 46 to the soil, wherein the biological control composition has antifungal activity. (Item 51) A method for preventing or reducing the growth of fungal pathogens in agricultural products, comprising the step of applying a biological control composition described in any one of items 1 to 46 to a packaging material used for transporting or storing the agricultural products, wherein the biological control composition has antifungal activity. (Item 52) A method for preventing or reducing the growth of fungal pathogens in seeds or agricultural products, comprising the step of incorporating a biological control composition described in any one of items 1 to 46 into a process selected from the group consisting of washing the agricultural product or seeds, coating the agricultural product or seeds, and a combination thereof. (Item 53) The method according to any one of items 49 to 52, wherein the plant, the seed, the flower, or the agricultural product thereof is a plant, seed, flower, or agricultural product thereof belonging to the family Ericaceae. (Item 54) The method according to item 53, wherein the plant, the seed, or the agricultural product thereof is a plant, seed, flower, or agricultural product thereof belonging to the Ericaceae family, which is of the genus Vaccinium. (Item 55) The method according to item 54, wherein the plant of the genus Vaccinium, the seeds, the flowers, or the agricultural product thereof is a blueberry. (Item 56) The method according to any one of items 49 to 52, wherein the plant, the seed, the flower, or the agricultural product thereof is a plant, seed, flower, or agricultural product thereof belonging to the family Vitaceae. (Item 57) The method according to item 56, wherein the plant, the seed, the flower, or the agricultural product thereof is a plant, seed, or agricultural product thereof of the family Vitaceae, which is of the genus Vitis. (Item 58) The method according to item 57, wherein the plant of the genus Vitis, the seeds, the flowers, or the agricultural product thereof is a grape. (Item 59) The method according to any one of items 49 to 52, wherein the plant, the seed, the flower, or the agricultural product thereof is a plant, seed, or agricultural product thereof belonging to the family Rosaceae. (Item 60) The method according to item 59, wherein the plant, seed, flower, or agricultural product of the family Rosaceae is of the genus Rubus, Malus, Pyrus, Cydonia, Prunus, Rosa, or Fragaria. (Item 61) The method according to item 60, wherein the plant of the genus Rubus, the seeds, the flowers, or the agricultural product thereof is a raspberry or a blackberry. (Item 62) The method according to item 60, wherein the plant of the genus Fragaria, the seeds, the flowers, or the agricultural product thereof is a strawberry. (Item 63) The method according to item 60, wherein the plant of the genus Pyrus, the seeds, the flowers, or the agricultural product thereof is a pear. (Item 64) The method according to item 60, wherein the plant of the genus Cydonia, the seeds, the flowers, or the agricultural product thereof is quince. (Item 65) The method according to item 60, wherein the plant of the genus Prunus, the seeds, the flowers, or the agricultural product thereof is an almond, peach, plum, apricot, cherry, or spinosa plum. (Item 66) The method according to item 60, wherein the plant of the genus Rosa, the seeds, the flowers, or the agricultural product thereof is a rose. (Item 67) The method according to item 60, wherein the plant of the genus Malus, the seeds, the flowers, or the agricultural product thereof is an apple. (Item 68) The method according to item 49, wherein the step of applying the biological control composition includes sprinkling, dipping, spreading, injecting, rubbing, spraying, or brushing the biological control composition onto the plant, the seed, the flower, or the agricultural product. (Item 69) The method according to item 49, wherein the step of applying the biological control composition to the plant includes the step of adding the biological control composition to a drip line, irrigation system, chemical irrigation system, spray, or immersion agent. (Item 70) The method according to item 49, wherein the step of applying the biological control composition to the plant includes the step of applying the biological control composition to the roots of the plant. (Item 71) The method according to item 70, wherein the step applied to the root is indirect. (Item 72) The method according to item 49, wherein the biological control composition is applied to the agricultural product after it has been harvested from the plant. (Item 73) The method according to item 49, wherein the step of application described above does not kill the plant. (Item 74) The method according to item 49, further comprising the step of applying a fertilizer, herbicide, pesticide, another biological control composition, or a combination thereof to the plant. (Item 75) The method according to item 49, wherein the fertilizer, the herbicide, the pesticide, or the other biological control composition is applied before, after, or simultaneously with the biological control composition. (Item 76) The method according to item 51, wherein the packaging material includes polyethylene terephthalate (PET), molded fibers, oriented polystyrene (OPS), polystyrene (PS) foam, polypropylene (PP), or a combination thereof. (Item 77) The method according to item 51, wherein the step applied to the packaging material includes the step of washing or impregnating the packaging material. (Item 78) The method according to any one of items 49 to 52, wherein the antifungal activity is prevention of the growth of the fungal pathogen for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days. (Item 79) The method according to any one of items 49 to 52, wherein the antifungal activity is a reduction in the growth of the fungal pathogen in the plant, the seed, the flower, or the agricultural product thereof, compared to the growth of the fungal pathogen in a control which is a Rosaceae plant or agricultural product thereof that has not been exposed to the biological control composition. (Item 80) The method according to item 79, wherein the growth of the fungal pathogen is reduced for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days after exposure of the fungal pathogen to the biological control composition, compared to the growth of the fungal pathogen in the plant, seed, flower, or agricultural product thereof that has not been exposed to the biological control composition. (Item 81) The method according to any one of items 49 to 52, wherein the biological control composition has antifungal activity against filamentous or non-filamentous fungal pathogens. (Item 82) The filamentous or non-filamentous fungal pathogens are Albugo candida, Albugo occidentalis, Alternaria alternata, Alternaria cucumerina, Alternaria dauci, Alternaria solani, Alternaria tenuis, Alternaria tenuissima, Alternaria tomatophila, Aphanomyces euteiches, Aphanomyces raphani, Armillaria mellea, Botrydia theobromae, Botrytis cinerea, Botrytinia fuckeliana, Bremia lactuca, Cercospora beticola, Cercosporella rubi, Cladosporium herbarum, Colletotrichum acutatum, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum, Colletotrichum musae, Colletotrichum spaethanium, Cordana musae, Corynespora cassiicola, Daktulosphaira vitifoliae, Didymella bryoniae, Elsinoe ampelina, Elsinoe mangiferae, Elsinoe veneta, Erysiphe cichoracearum, Erysiphe necator, Eutypa lata, Fusarium germinareum, Fusarium oxysporum, Fusarium solani, Ganoderma boninense, Guignardia bidwellii, Gymnoconia peckiana, Helminthosporium solani, Leptosphaeria coniothyrium, Leptosphaeria maculans, Leveillula taurica, Macrophomina phaseolina, Microsphaera alni, Monilinia fructicola, Monilinia vaccinii-corymbosi, Mycosphaerella angulate, Mycosphaerella brassicicola, Mycosphaerella fragariae, Mycosphaerella fijiensis, Oidopsis taurica, Passalora fulva, Peronospora sparse, Peronospora farinosa, Phoma exigua, Phomopsis obscurans, Phomopsis vaccinia, Phomopsis viticola, Phytophthora capsica, Phytophthora erythroseptica, Phytophthora infestans, Phytophthora parasitica, Plasmopara viticola, Plasmodiophora brassicae, Podosphaera macularis, Polyscytalum pustulans, Pseudocercospora vitis, Puccinia allii, Puccinia sorghi, Pucciniastrum vaccinia, Pythium debaryanum, Pythium sulcatum, Pythium ultimum, Ralstonia solanacearum, Ramularia tulasneii, Rhizoctonia solani, Rhizopus arrhizus, Rhizopus stoloniferz, Sclerotinia minor, Sclerotinia sclerotiorum, Sclerotium cepivorum, Sclerotium rolfsii, Sclerotinia minor, Sclerotinia sclerotiorum, Septoria apiicola, Septoria lactucae, Septoria lycopersici, Septoria petroelini, Sphaceloma perseae, Sphaerotheca macularis, Spongospora subterranea, Stemphylium vesicarium, Synchytrium endobioticum, Thielaviopsis basicola, Uncinula necator, Uromyces appendiculatus, Uromyces betae, Verticillium The method described in item 81, selected from the group consisting of albo-atrum, Verticillium dahliae, Verticillium theobromae, and any combination thereof. (Item 83) The method according to item 81, wherein the filamentous fungal pathogen is selected from the group consisting of Fusarium oxysporum, Verticillium dahliae, Botrytis cinerea, Colletotrichum spaethaniu, Erysiphe necator, Podosphaera macularis, Monilinia vaccinii-corymbosi, Puccinia sorghi, and any combination thereof. (Item 84) The method according to any one of items 49 to 52, wherein the plant, the seed, or the agricultural product thereof is selected from the group consisting of almonds, apricots, apples, artichokes, bananas, barley, beets, blackberries, blueberries, broccoli, Brussels sprouts, cabbage, hemp, peppers, carrots, celery, Swiss chard, cherries, citrus fruits, corn, melons, dates, figs, garlic, grapes, herbs, spices, kale, lettuce, oil palm, olives, onions, peas, pears, peaches, peanuts, papayas, parsnips, pecans, persimmons, plums, pomegranates, potatoes, quince, radishes, raspberries, roses, rice, spinosa plums, sorghum, soybeans, spinach, strawberries, sweet potatoes, tobacco, tomatoes, turnip leaves, walnuts, and wheat.

Claims

1. A composition for preventing or reducing the growth of fungal pathogens in plants, or in their seeds, flowers, or crops, The first microorganism or the spores of the first microorganism, A second microorganism or spores of a second microorganism, and Carrier Includes, The first microorganism is Gluconobacter cerinus, which contains a nucleic acid having a 16S rRNA sequence that is 99.5% or more identical to Sequence ID No.

24. A composition in which the second microorganism is Hanseniaspora uvarum, comprising a nucleic acid containing an ITS sequence having 99.5% or more identity with SEQ ID NO:

25.

2. The composition according to claim 1, wherein the first microorganism comprises a nucleic acid containing the 16S rRNA sequence of SEQ ID NO: 24, and the second microorganism comprises a nucleic acid containing the ITS sequence of SEQ ID NO:

25.

3. The composition according to claim 1, wherein the carrier comprises a liquid carrier, an inorganic carrier, or an organic carrier.

4. A method for preventing or reducing the growth of fungal pathogens in plants, or in their seeds, flowers, or crops, wherein the plant, or its seeds, flowers, or crops The first microorganism or the spores of the first microorganism, A second microorganism or spores of a second microorganism, and Carrier Includes, The first microorganism is Gluconobacter cerinus, which contains a nucleic acid having a 16S rRNA sequence that is 99.5% or more identical to Sequence ID No.

24. A method comprising contacting the second microorganism, which is Hanseniaspora uvarum, with a composition, wherein the second microorganism is Hanseniaspora uvarum, which contains a nucleic acid having an ITS sequence having 99.5% or more identity with SEQ ID NO:

25.

5. The aforementioned contact is (i) Applying the composition to the plant, or its seeds, flowers, or crops, (ii) Applying the composition to the soil surrounding the plant, (iii) Applying the composition to packaging materials used for transporting or storing the plant, or the seeds, flowers, or crops thereof, or (iv) Incorporating the composition into a process that includes washing or coating the plant, or the seeds, flowers, or crops thereof. The method according to claim 4, which includes any of the following.

6. Applying the composition may involve sprinkling, dipping, spreading, injecting, rubbing, spraying, or brushing the composition; or The method according to claim 5, wherein applying or incorporating the composition includes adding the composition to a drip line, an irrigation system, a chemical irrigation system, a spray agent, or a immersion agent.

7. The method according to claim 5, wherein the composition is applied to the plant or the crop before the crop is harvested from the plant.

8. The method according to claim 5, wherein the composition is applied to the crop after the crop has been harvested from the plant.

9. The method according to claim 4, further comprising applying a fertilizer, herbicide, pesticide, or other composition to the plant.

10. The method according to claim 4, wherein the contact prevents or reduces the growth of the fungal pathogen in the plant, or in its seeds, flowers, or crops.

11. The method according to claim 10, wherein the contact prevents or reduces the growth of the fungal pathogen for at least one day.

12. The method according to claim 10, wherein the contact reduces the growth of the fungal pathogen by at least 10% compared to a control.

13. The above fungal pathogens are Albugo candida, Albugo occidentalis, Alternaria alternata, Alternaria cucumerina, Alternaria dauci, Alternaria solani, Alternaria tenuis, Alternaria tenuissima, Alternaria tomatophila, Aphanomyces euteiches, Aphanomyces raphani, Armillaria mellea, Botrydia Theobromae, Botrytis cinerea, Botrytinia fuckeliana, Bremia lactuca, Cercospora beticola, Cercosporella rubi, Cladosporium herbarum, Colletotrichum acutatum, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum, Colletotrichum musae, Colletotrichum Spaethanium, Cordana musae, Corynespora cassiicola, Dactulosphaira vitifolia, Didymella bryoniae, Elsinoe ampelina, Elsinoe mangiferae, Elsinoe veneta, Erysiphe cichoracearum, Erysiphe necator, Eutypa lata, Fusarium germinarium, Fusarium oxysporum, Fusarium solani, Ganoderma boninense, Guignardia bidwellii, Gymnoconia peckiana, Helminthosporium solani, Leptosphaeria coniothyrium, Leptosphaeria maculans, Leveillula taurica, Macrophomina phaseolina, Microsphaera alni, Monilinia fructicola, Monilinia vaccinii-corymbosi,Mycosphaerella angulate, Mycosphaerella brassicicola, Mycosphaerella fragariae, Mycosphaerella fijiensis, Oidopsis taurica, Passalora fulva, Peronospora sparse, Peronospora farinosa, Phoma exigua, Phomopsis obscurans, Phomopsis vaccinia, Phomopsis viticola, Phytophthora capsica, Phytophthora erythroseptica, Phytophthora infestans, Phytophthora parasitica, Plasmopara viticola, Plasmodiophora brassicae, Podosphere macularis, Polyscytalum pustulans, Pseudocercospora vitis, Puccinia allii, Puccinia sorghi, Pucciniastrum vaccinia, Pythium debaryanum, Pythium sulcatum, Pythium ultimum, Ralstonia solanacearum, Ramularia tulasneii, Rhizoctonia solani, Rhizopus arrhizus, Rhizopus stoloniferz, Sclerotinia minor, Sclerotinia sclerotiorum, Sclerotium cepivorum, Sclerotinia rolfsii, Sclerotinia minor, Sclerotinia sclerotiorum, Septoria apicola, Septoria lactucae, Septoria lycopersici, Septoria petrolini, Spaceloma perseae, Sphaerotheca macularis, Spongospora subterranea, Stemphylium vesicarium, Syncytrium endobioticum,The method according to claim 10, comprising a fungal pathogen selected from the group consisting of Thielaviopsis basicola, Uncinula necator, Uromyces appendiculatus, Uromyces betae, Verticillium albo-atrum, Verticillium dahliae, and Verticillium theobromae.

14. The aforementioned plants, or their seeds, flowers, or crops, include almonds, apricots, apples, artichokes, bananas, barley, beans, beets, blackberries, blueberries, broccoli, Brussels sprouts, cabbage, hemp, chili peppers, carrots, celery, Swiss chard, cherries, citrus fruits, corn, melons, dates, figs, garlic, grapes, herbs, spices, kale, and lettuce. The method according to claim 4, selected from the group consisting of sorghum, oil palm, olive, onion, pea, pear, peach, peanut, papaya, parsnip, pecan, persimmon, plum, pomegranate, potato, Prunus, quince, radish, raspberry, rose, rice, spinosa plum, sorghum, soybean, spinach, strawberry, sweet potato, tobacco, tomato, turnip leaves, walnut, and wheat.

15. The fungal pathogens include fungal pathogens selected from the group consisting of Fusarium oxysporum, Mycosphaerella fijiensis, Botrytis cinerea, Monilinia, Monilinia fructicola, Monilinia vaccinii-corymbosi, Phomopsis viticola, Plasmopara viticola, Rhizopus, Rhizopus arrhizus, Rhizopus stoloniferz, Macrophonina phaseolina, and Verticillium dahlia; The method according to claim 4, wherein the plant, or its seeds, flowers, or crops, is selected from the group consisting of bananas, beans, peaches, plums, citrus fruits, grapes, raspberries, blueberries, strawberries, blackberries, spinach, lettuce, and rice.

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