TW201740812A - Bacillus amyloliquefaciens RTI301 compositions and methods of use for benefiting plant growth and treating plant disease - Google Patents

Abstract

Compositions and methods include a new strain of Bacillus amyloliquefaciens having growth promoting activity and activity against plant pathogens. The compositions are useful for benefiting plant growth and/or conferring protection against a pathogenic infection when applied to plant foliage, flowers, fruits, bark, roots, seeds, callus tissue, grafts, cuttings, surrounding soil or growth medium, and soil or growth medium concomitant with sowing seed and planting callus tissue, grafts, and cuttings. The compositions containing the Bacillus amyloliquefaciens RTI301 strain can be applied alone or in combination with other microbial, biological, or chemical insecticides, fungicides, nematicides, bacteriocides, herbicides, plant extracts, plant growth regulators, and fertilizers. In one example, the Bacillus amyloliquefaciens RTI301 strain can be delivered to the plant as part of an integrated pest management program, with other microbial or chemical insecticides, fungicides, nematicides, bacteriocides, herbicides, plant extracts, and plant growth regulators.

Description

Bacillus amyloliquefaciens RTI301 composition useful for plant growth and treatment of plant diseases and methods of use thereof Cross-reference to related applications

The present application claims the benefit of U.S. Provisional Application No. 62/097,203, filed on Dec. 29, 2014, the disclosure of which is hereby incorporated by reference.

The subject matter disclosed in the present invention relates to a composition comprising an isolate of Bacillus amyloliquefaciens RTI301 for application to plant leaves, plant fruits and flowers, plant seeds and roots, and soil around plants to benefit plant growth and treatment. Plant diseases.

A variety of microorganisms known to have beneficial effects on plant growth and health are present in the soil, particularly in combination with plants in the root zone (plant root growth promoting bacteria "PGPR"), or in the form of endoparasites in plants. It is beneficial to promote plant growth characteristics including nitrogen fixation; iron chelation; phosphate dissolution; inhibition of non-beneficial microorganisms; insect resistance; induced systemic resistance (ISR); systemic acquired resistance (SAR); decomposition of plant matter into the soil to increase usefulness Soil organic matter; and synthetic plant hormones such as guanidine-acetic acid (IAA), acetoin and 2,3-butanediol, which stimulate plant growth, development, and respond to environmental stresses such as drought. In addition, these microorganisms can be made by the precursor molecule 1-aminocyclopropane-1-carboxylate. (ACC) decomposes to interfere with the ethylene stress response of the plant, thereby stimulating plant growth and slowing fruit ripening. These beneficial microorganisms can improve soil quality, plant growth, yield and crop quality. Various microorganisms exhibit biological activity for use in controlling plant diseases. Such biocides (live organisms and naturally occurring compounds of such organisms) are safer and more biodegradable than synthetic fertilizers and insecticides.

Fungal plant pathogens, including but not limited to, Botrytis spp. (eg Botrytis cinerea ), Fusarium spp. (eg F. oxysporum ) F. graminearum , Rhizoctonia spp. (eg R. solani), Magnaporthe spp ., Mycosphaerella spp .), Puccinia spp . (eg P. recondita ), Phytopthora spp . and Phakopsora spp. (eg Phaeocysteen rust) ( P. pachyrhizi )) is a type of plant pest that can cause serious economic losses in the agricultural and horticultural industries. Chemical agents can be used to control fungal plant pathogens, but the use of chemical agents has the following disadvantages, including high cost, lack of efficacy, emergence of fungal resistant strains, and undesirable environmental effects. In addition, in addition to targeted plant pathogens, such chemical treatments are often indiscriminate and may have adverse effects on beneficial bacteria, fungi, and arthropods. The second type of plant pest is a bacterial pathogen including, but not limited to, Erwinia spp. (such as Erwinia chrysanthemi ), Pantoea spp. (such as green lemon) P. citrea , Xanthomonas (eg Xanthomonas campestris ), Pseudomonas spp. (such as Pseudomonas syringae ( P) .syringae )) and Ralstonia spp. (such as R. soleacearum ), causing severe economic losses in the agricultural and horticultural industries. Similar to pathogenic fungi, the use of chemical agents to treat such bacterial pathogens has disadvantages. Viruses and virus-like organisms comprise a third type of phytopathogenic agent that is difficult to control, but bacterial microorganisms can provide plant resistance thereto via induction systemic resistance (ISR). Therefore, microorganisms that can be used as biological fertilizers and/or biocides to control pathogenic fungi, viruses and bacteria are desirable and highly desirable for improving agricultural sustainability. The last type of plant pathogen includes phytopathogenic nematodes and insects that can cause serious damage and loss to plants.

Some members of the genus Bacillus have been reported to be biologically controlled strains, and some have been used in commercial products (Joseph W. Kloepper et al., 2004, Phytopathology Vol. 94, No. 11, 1259-1266). For example, strains currently used in commercial biological control products include: Bacillus pumilus strain QST2808, which is used as an active ingredient in SONATA and BALLAD-PLUS, produced by BAYER CROP SCIENCE; Bacillus licheniformis strain GB34, Used as an active ingredient in YIELDSHIELD, produced by BAYER CROP SCIENCE; Bacillus subtilis strain QST713, used as an active ingredient of SERENADE, produced by BAYER CROP SCIENCE; Bacillus subtilis strain GBO3, used as KODIAK and SYSTEM3 The active ingredient is produced by HELENA CHEMICAL COMPANY. Various strains of Bacillus thuringiensis and Bacillus firmus have been used as biological control agents against nematodes and vector insects, and these strains serve as the basis for many commercially available biological control products, including NORTICA and PONCHO-VOTIVO, produced by BAYER CROP SCIENCE. In addition, the current Bacillus strains used in commercial biostimulation products include: Bacillus amyloliquefaciens strain FZB42, which is used as an active ingredient in RHIZOVITAL 42, produced by ABiTEP GmbH; and various other Bacillus subtilis, including It acts as a whole cell, including its fermentation extract in biostimulating products, such as FULZYME manufactured by JH Biotech Inc.

The subject matter disclosed herein provides a microbial composition useful for plant growth and prevention and control of disease and methods of use thereof.

In one embodiment, there is provided a composition comprising a biologically pure culture of Bacillus amyloliquefaci RTI301 deposited with ATCC PTA-121165 or a mutant having all of its identifying characteristics, the composition being for application to a plant To achieve one or both of beneficial to plant growth or to confer protection to susceptible plants to avoid pathogenic infections.

In one embodiment, a plant seed coated with a composition comprising a spore of a biologically pure culture of a Bacillus amyloliquefaci RTI301 deposited with ATCC No. PTA-121165 or a mutant having all of its identifying characteristics is provided, The composition is present in an amount suitable to be beneficial for plant growth and/or to confer protection to susceptible plants to avoid pathogenic infections.

In one embodiment, a composition for achieving one or both of beneficial plant growth and/or protection of a susceptible plant to avoid a pathogenic infection is provided, the composition comprising: ATCC A biologically pure culture of Bacillus amyloliquefaciens RTI301 or a mutant having all of its identifying characteristics deposited under No. PTA-121165, which is suitable for plant growth and/or protection of susceptible plants to avoid pathogenic infections And one or a combination of microbial, biological or chemical insecticides, fungicides, nematicides, fungicides, herbicides, plant extracts, plant growth regulators or fertilizers, which are suitable for plant growth and / or protect the susceptible plants to avoid the amount of pathogenic infections.

In one embodiment, a method for achieving one or both of beneficial to plant growth or protection of a susceptible plant from pathogenic infection is provided, the method comprising: delivering a composition to a plant leaf, Plant bark, plant fruit, plant flower, plant seed, plant root, plant pruning, plant graft, plant healing tissue; soil or growth medium surrounding the plant; soil or growth medium prior to seeding the plant seed in soil or growth medium Or a soil or growth medium prior to planting a plant, plant pruning, plant graft or plant healing tissue in soil or growth medium, the composition comprising Bacillus amyloliquefaciens RTI301 deposited under ATCC PTA-121165 or having A biologically pure culture of all of its identified features, which are suitable for plant growth and/or susceptibility Diseased plants confer protection to avoid pathogenic infections.

In one embodiment, a method for achieving one or both of beneficial to plant growth or protection of a susceptible plant from pathogenic infection is provided, the method comprising: delivering a composition to a plant leaf, Plant bark, plant fruit, plant flower, plant seed, plant root, plant pruning, plant graft, plant healing tissue; soil or growth medium surrounding the plant; soil or growth medium prior to seeding the plant seed in soil or growth medium Or soil or growth medium prior to planting plants, plant pruning, plant transplants or plant healing tissues in soil or growth medium, the composition comprising the following: Bacillus amyloliquefaciens deposited under ATCC PTA-121165 RTI301 or a biologically pure culture of a mutant having all of its identifying characteristics, in an amount suitable for beneficial to plant growth and/or conferring protection against pathogenic infection; and microbial, biological or chemical insecticides, fungicides , one or a combination of nematicides, fungicides, herbicides, plant extracts, plant growth regulators or fertilizers Conducive to plant growth and / or confer protection against the amount of pathogenic infection.

In one embodiment, a method for achieving one or both of beneficial to plant growth or protection of a susceptible plant from pathogenic infection is provided, the method comprising: combining the first composition with the second The combination of substances is delivered to plant leaves, plant stems, plant fruits, plant flowers, plant seeds, plant roots, plant pruning, plant transplants, plant healing tissues; soil or growth medium around the plant; seeded in soil or growth medium Soil or growth medium prior to plant seeds; or soil or growth medium prior to planting plants, plant pruning, plant transplants or plant healing tissues in soil or growth medium, the first composition comprising ATCC PTA-121165 A biologically pure culture of the deposited Bacillus amyloliquefaciens RTI301 or a mutant thereof having all of its identifying characteristics, in an amount suitable for beneficial to plant growth and/or protection of susceptible plants to avoid pathogenic infection; The second composition comprises a microbial, biological or chemical insecticide, fungicide, nematicide, bactericide, herbicide, plant extract Plant growth regulators or fertilizers or a combination of one, was It is suitable for the benefit of plant growth and/or protection of susceptible plants to avoid pathogenic infections.

In one embodiment, a method for achieving one or both of beneficial to plant growth or protection of a susceptible plant to avoid pathogenic infection is provided, the method comprising: planting the plant seed in a suitable growth medium Or the nutrient pruning/tissue of the plant is regenerated in a suitable growth medium, wherein the seed has been coated with a composition or the nutrient pruning/tissue has been inoculated with the composition, the composition comprising the deposit of ATCC PTA-121165 A biologically pure culture of B. amyloliquesii RTI301 or a mutant thereof having all of its identifying characteristics, wherein the growth of the plant from the seed or nutrient pruning/tissue is beneficial and/or protection is provided to avoid pathogenic infection.

In one embodiment, there is provided a method of providing plant protection by protecting a susceptible plant from pathogenic infection or reducing pathogenic infection while minimizing the accumulation of resistance to treatment, the method comprising: The first composition and the second composition are delivered to the susceptible plant at separate intervals of application and alteration, wherein the first composition and the second composition are each adapted to impart protection to the plant to avoid or reduce Delivery of a pathogenic infection, wherein the first composition comprises a B. amyloliquefaci RTI301 deposited with ATCC No. PTA-121165 or a biologically pure culture thereof having all of its identifying characteristics, and wherein the second combination The composition comprises one or more chemical active agents having fungicidal or bactericidal properties, and wherein the first composition and the second composition are delivered to one or a combination of the following at varying time intervals: plant leaves, plant bark, plants Fruit, plant flower, plant seed, plant root, plant pruning, plant graft, plant healing tissue or soil or growth medium around the plant, Protection against and / or decrease the amount of chemical needed to pathogenic infections and to reduce the accumulation of active agents for the treatment of resistance to be minimized.

In one embodiment, a composition is provided, the composition comprising at least one of the following: an isolated abundance MA compound, an isolated abundance MB compound, an isolated abundance MC compound, and an isolated hydroxy abundance MA Compound, isolated hydroxy-hydroxytoxin MB compound, isolated hydroxy-hydroxytoxin MC compound, isolated abundance H compound, isolated hydroxy-hydroxytoxin H compound, isolated abundance I compound and isolated hydroxy-hydroxytoxin I A compound, in an amount suitable to achieve one or both of beneficial to plant growth or to confer protection to a susceptible plant to avoid a pathogenic infection having the following formula:

Wherein R is OH, n is in the range of 8 to 20, FA is linear, XOR or di-isomeric, and: for abundance MA, X ₁ is Ala, X ₂ is Thr, and X ₃ is Met; for abundance MB X ₁ is Val, X ₂ is Thr, and X ₃ is Met; for abundance MC, X ₁ is Aba, X ₂ is Thr, and X ₃ is Met; for abundance H, X ₁ is Val, X ₂ Is Thr and X ₃ is Hcy; and for abundance I, X ₁ is Ile, X ₂ is Thr, and X ₃ is Ile; or wherein R is H, n is in the range of 8 to 20, and FA is linear, XOR or singular, and: for dehydroxylectin MA, X ₁ is Ala, X ₂ is Thr, and X ₃ is Met; for dehydroxyl-rich MB, X ₁ is Val, X ₂ is Thr, and X ₃ Is Met; for dehydroxylectin MC, X ₁ is Aba, X ₂ is Thr, and X ₃ is Met; for dehydroxylectin H, X ₁ is Val, X ₂ is Thr, and X ₃ is Hcy; For dehydroxylectin I, X ₁ is Ile, X ₂ is Thr, and X ₃ is Ile.

In one embodiment, there is provided an extract of a biologically pure culture of a B. amyloliquefaciens strain, the extract comprising amylin-MA, amylin-MB, a protosan-MC, a protosan-H, and a protoxin-I compound And dehydroxy-hydroxytoxin-MA, hydroxy-hydroxytoxin-MB, hydroxy-hydroxytoxin-MC, hydroxy-hydroxytoxin-H and hydroxy-hydroxy-proto-I compounds, and other abundances and dehydroxylations listed in Table XIII One or a combination of a protoplast-like compounds.

In one embodiment, an extract of a biologically pure culture of Bacillus amyloliquefaciens RTI301 deposited under ATCC PTA-121165 is provided, the extract comprising amylin-MA, amylin-MB, abundance-MC, Fengyuan素-H and adiponectin-I compounds as well as hydroxy-hydroxytoxin-MA, hydroxy-hydroxytoxin-MB, hydroxy-hydroxytoxin-MC, hydroxy-hydroxytoxin-H and dehydroxy-hydroxy-proto-I compounds and Table XIII One or a combination of other abundances and dehydroxy-flavone-like compounds.

In one embodiment, a plant-promoting composition is provided, the composition comprising: Bacillus amyloliquefaciens RTI301 deposited with ATCC PTA-121165 or a biologically pure culture thereof having mutants of all of its identifying characteristics ; and bifenthrin insecticide.

In one embodiment, a plant-promoting composition is provided, the composition comprising: Bacillus amyloliquefaciens RTI301 deposited under ATCC PTA-121165 or a biologically pure culture thereof having all of its identifying characteristics And a fungicide comprising one or a combination of an extract from a white lupinus ( Lupinus albus ), a BLAD polypeptide or a BLAD polypeptide fragment.

In one embodiment, a product is provided comprising: a first composition comprising a biologically pure culture of Bacillus amyloliquefaci RTI301 deposited with ATCC PTA-121165 or a mutant having all of its identifying characteristics; a second composition of one or a combination of a microbial, biological or chemical insecticide, fungicide, nematicide, bactericide, herbicide, plant extract, plant growth regulator or fertilizer, wherein the first composition and The second composition is packaged separately and wherein the components are suitable for achieving an amount that is beneficial to plant growth and/or conferring protection to susceptible plants to avoid one or both of the pathogenic infections; and instructions as appropriate This specification is directed to the delivery of a combination of first and second compositions suitable for beneficial plant growth to: plant leaves, plant bark, plant fruits, plant flowers, plant seeds, plant roots, plant shears Branches, plant grafts, plant healing tissues; soil or growth medium around plants; seeding of plant seeds in soil or growth medium Pre-soil or growth medium; or soil or growth medium prior to planting plants, plant pruning, plant transplants, or plant healing tissue in soil or growth media.

1 shows, in accordance with one or more embodiments of the present invention, surrounded and included in B. amyloliquefaciens strain RTI301, as compared to the corresponding regions of two B. amyloliquefaciens reference strains B. amyloliquefaciens FZB42 and B. amyloliquefaciens TrigoCor1448 Schematic representation of the genomic structure of the unique lantibiotic biosynthesis operon found in the medium.

Figure 2A is a photograph showing a plant inoculated with the RTI301 strain. Figure 2B is a photograph showing a control plant. These photographs show the positive effects of B. amyloliquefaciens strain RTI301 on early plant growth in wheat according to one or more embodiments of the invention. The plants extracted after 13 days of growth are shown in the figure.

Figure 3A is a photograph showing a plant inoculated with RTI301. Figure 3B is a photograph showing a control plant. These photographs show the positive effect of B. amyloliquefaciens strain RTI301 on growth in wheat after 28 days according to one or more embodiments of the invention.

Figure 4 is a graph showing the % of disease control (average value) on the y-axis after 10 days of infection with soybean rust ( Uromyces appendiculatus ) after treatment with each of the following in accordance with one or more embodiments of the present invention. Plot: RTI301 stalk in waste broth (SFB) (administered at 1×10 ⁸ cfu/mL), SERENADE OPTIMUM (administered at 1×10 ⁸ cfu/ml), TACTIC (administered to all formulations at 0.1875%) Also used as a blank control), SERENADE OPTIMUM (administered at 4 x 10 ⁸ cfu/ml), Tebuconazole (administered at 50 g ai/ha) and tetrachloroisophthalonitrile (administered at 500 g ai/ha). Untreated controls produced 23% of the disease. The values followed by the same letter are not significantly different (p=0.10).

Figure 5 is a graph showing the increase in the amount of infection of soybean rust (Nicky rust) (50k to 300k conidia/ml) on the y-axis after 10 days of treatment according to one or more embodiments of the present invention. Histogram of % of disease control (average): decoction broth compared to TACTIC (administered to all formulations at 0.1875% and also used as a blank) and Kelly (HORIZON; applied at 50 g ai/ha) (SFB) RTI301 spores (administered at 1.0 x 10 ⁸ cfu/ml) and SERENADE OPTIMUM (administered at 1 x 10 ⁸ cfu/ml and 4 x 10 ⁸ cfu/ml). The % of diseases in the examination control were 50k=6%, 100k=6%, 150k=15%, 200k=15%, and 300k=7%, respectively. The values followed by the same letter are not significantly different (p=0.10).

Figure 6 shows an infection in an untreated control ("UT") in each of two independent tomato field trials determining the antagonism of the RTI301 strain against Botrytis cinerea pathogens in accordance with one or more embodiments of the present invention. A graph of the growth of the fruit of the pathogen over time.

Figure 7 shows infection in an untreated control ("UT") in each of four independent strawberry field trials determining antagonism of RTI301 strain against Botrytis cinerea pathogens in accordance with one or more embodiments of the present invention. A graph of the growth of the fruit of the pathogen over time.

Figure 8A shows the growth of Fusarium graminearum on a 869 agar plate. Figure 8B shows the growth of Fusarium graminearum on a 869 agar plate in the presence of 20 μl of RTI301 spore solution containing 1 x 10 ⁸ CFU/ml (left) or 1 x 10 ⁹ CFU/ml (right), respectively. Figure 8C shows the growth of 20 μl of a solution of Bacillus amyloliquefaciens RTI301 spores containing 1 x 10 ⁸ CFU/ml (left) or 1 x 10 ⁹ CFU/ml (right) on a 869 agar plate. Figure 8D shows the growth of Fusarium graminearum on a 869+1% FRACTURE agar plate. Figure 8E shows the growth of Fusarium graminearum on a 869+1% FRACTURE agar plate in the presence of 20 μl of RTI301 spore solution containing 1 x 10 ⁸ CFU/ml (left) or 1 x 10 ⁹ CFU/ml (right), respectively. Figure 8F shows the growth of 20 μl of a spore solution of Bacillus amyloliquefaciens RTI301 containing 1 x 10 ⁸ CFU/ml (left) or 1 x 10 ⁹ CFU/ml (right) on a 869 + 1% FRACTURE agar plate. These figures show images of disc analysis showing the control of Fusarium graminearum RTI301 in the presence and absence of FRACTURE in accordance with one or more embodiments of the invention.

Figure 9A shows the growth of Fusarium oxysporum on the 869 agar plate. Figure 9B shows the growth of Fusarium oxysporum on the 869 agar plate in the presence of 20 μl of RTI301 spore solution containing 1 x 10 ⁸ CFU/ml (left) or 1 x 10 ⁹ CFU/ml (right), respectively. Figure 9C shows the growth of 20 μl of a solution of spores of Bacillus amyloliquefaciens RTI301 containing 1 x 10 ⁸ CFU/ml (left) or 1 x 10 ⁹ CFU/ml (right) on a 869 agar plate. Figure 9D shows the growth of Fusarium oxysporum on a 869+1% FRACTURE agar plate. Figure 9E shows the growth of Fusarium oxysporum on a 869+1% FRACTURE agar plate in the presence of 20 μl of RTI301 spore solution containing 1 x 10 ⁸ CFU/ml (left) or 1 x ¹⁰⁹ CFU/ml (right), respectively. Figure 9F shows the growth of 20 μl of B. amyloliquefaciens RTI 301 solution containing 1 x 10 ⁸ CFU/ml (left) or 1 x 10 ⁹ CFU/ml (right) on a 869 + 1% FRACTURE agar plate. These figures show images of disc analysis showing the control of B. oxysporum RTI301 against Fusarium oxysporum in the presence and absence of FRACTURE in accordance with one or more embodiments of the present invention.

Figure 10 is a diagram showing a previously reported abundance of protosanoids and dehydroxyhydroxytoxin-type cyclic lipopeptides produced by a microbial species including Bacillus amyloliquefaciens, and by Bacillus amyloliquefaciens RTI301 according to one or more embodiments of the present invention Schematic representation of the new identification (shown in bold) of the isolates and the dehydroxyl-rich molecules.

Figure 11 is a graph showing the percentage of lipopeptides recovered from RTI301 spent broth (SFB) after acid precipitation, in accordance with one or more embodiments of the present invention. The terms "301-AP-spherulites" and "301-AP-supernatants" refer to resuspended pellets and supernatants obtained after acid precipitation and centrifugation of SFB, respectively. The percentage was calculated and based on the integrated ion abundance comparison of each lipopeptide from RTI301 waste broth (301-SFB).

The terms "a/an" and "the" are used in the context of the application (including the scope of the patent application) to mean "one or more." Thus, for example, reference to "a plant" includes a plurality of plants unless the context clearly dictates otherwise.

Throughout the specification and claims, the term "comprise/comprises/comprising" is used in a non-exclusive sense, unless the context requires otherwise. Similarly, the term "comprise" and its grammatical variations are intended to be non-limiting, such that the listing of items in the list does not exclude other similar items that can be substituted or added to the listed items.

For the purposes of this specification and the scope of the patent application, the term "about" is used with one or more The use of a range of numbers or ranges of values is understood to mean all such numbers, including all numbers in the range, and the extents are modified by extending the boundaries above and below the stated values. The recitation of numerical ranges by endpoints includes all numbers, including all integers, including the integers thereof, including the range of 1 to 5 including 1, 2, 3, 4, and 5, and Scores, such as 1.5, 2.25, 3.75, 4.1, and the like).

For the purposes of this specification and the scope of the patent application, the terms "metabolite" and "compound" are used interchangeably when used in combination with a compound having antimicrobial activity produced by a strain of RTI301 or other strains of Bacillus amyloliquefaciens.

For the purposes of this specification and the scope of the patent application, the phrase "biologically pure culture of bacterial strain" means one or a combination of the following: a spore of a biologically pure fermentation culture of a bacterial strain; a vegetative cell of a yeast culture; one or more products of a biologically pure fermentation culture of a bacterial strain; a culture solid of a biologically pure fermentation culture of a bacterial strain; a culture supernatant of a biologically pure fermentation culture of a bacterial strain a liquid; an extract of a biologically pure fermentation culture of a bacterial strain; and one or more metabolites of a biologically pure fermentation culture of the bacterial strain.

In certain embodiments of the invention, compositions and methods are provided comprising a biologically pure culture of a newly identified strain of Bacillus amyloliquefaciens for application to a plant to achieve a plant growth or to confer a susceptibility to a susceptible plant Protect from one or both of the pathogenic infections. In the compositions and methods of the present invention, plant growth benefits are enhanced by increased seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or Its combination is displayed.

Plant-associated bacteria identified as belonging to the species B. amyloliquefaciens were isolated from the vine roots of the vineyard in Long Island, NY, and the phytopathogenic antagonistic properties were subsequently tested. More specifically, the isolated bacterial strain was identified as a new strain of B. amyloliquefaciens strain by sequence analysis of the highly conserved 16S rRNA and rpoB genes (see Example 1). The 16S RNA sequence of the new bacterial isolate (referred to as "Bacillus amyloliquefaciens RTI301") and three other known strains of B. amyloliquefaciens, Bacillus amyloliquefaciens strain NS6 (KF177175), Bacillus amyloliquefaciens strain FZB42 ( NR_075005) and the 16S rRNA gene sequence of Bacillus subtilis subsp. subtilis strain DSM 10 (NR_027552) were identical. The rpoB gene sequence of RTI301 was also determined with the B. amyloliques plant subspecies TrigoCor1448 (CP007244) (99% sequence identity; 3 base pair differences), B. amyloliquefaciens plant subspecies AS43.3 (CP003838) (99% Sequence identity; 7 base pair differences), B. amyloliquefaciens CC178 (CP006845) (99% sequence identity; 8 base pair differences) and B. amyloliquefaciens FZB42 (CP000560) (99% sequence identity; The same genes in the 8 base pair differences have sequence similarity. The RTI301 strain was identified as B. amyloliquefaciens, however, the sequence difference of the rpoB gene at the DNA level indicated that RTI301 was a new strain of B. amyloliquefaciens. The strain of Bacillus amyloliquefaciens RTI301 was deposited with the American Type Culture Collection (ATCC) on April 17, 2014 in accordance with the terms of the Budapest Treaty on the International Recognition of Microorganisms for Patent Procedures (Manassas, Virginia, USA). And has the patent registration number PTA-121165.

Further sequence analysis of the genome of Bacillus amyloliquefaciens RTI301 strain revealed that the strain has a gene associated with lantibiotic biosynthesis that is homologous to other closely related B. amyloliquefaciens strains (see Example 2). This is illustrated in Figure 1, which shows the lantibiotic biosynthetic cluster found in B. amyloliquesii RTI301 and two known B. amyloliquefaciens reference strains FZB42 (middle) and TrigoCor1448 (bottom) (shown under RTI301 strain) Schematic diagram of the genomic structure of the corresponding region. It can be observed from Figure 1 that the FZB42 and TrigoCor1448 strains lack many of the genes present in this cluster, and many of the genes present have a low degree of sequence identity. The BLASTn analysis of this cluster for NCBI's non-redundant (nr) nucleotide library demonstrates high homology to the 5' and 3' flanking regions of B. amyloliquefaciens strains (similar to the high similarity in Figure 1). %). However, the lanthiopeptide biosynthetic cluster is unique to RTI301 and no previous sequencing with the NCBI nr library was observed. The DNA has significant homology. This data indicates that the newly identified RTI301 has a unique lantibiotic biosynthetic pathway.

Experiments were carried out to determine the growth-promoting and antagonistic activity of the B. amyloliquefaciens RTI301 strain in various plants and under varying conditions. The results of the experiments are provided in Figures 2 to 11 and Examples 3 to 16 herein, showing that Bacillus amyloliquefaciens RTI301 can be beneficial to plants compared to the commercially available SERENADE (BAYER CROP SCIENCE, INC) containing Bacillus subtilis strain QST713 as an active ingredient. Grow and protect to avoid or control plant pathogenic infections. The experimental results also provide a comparison of the benefits of the new RTI 301 strain with commercially available chemical fungicides/bactericides. In some cases, the efficacy of the application of the B. amyloliquefaciens RTI 301 strain alone is as good as the application of a chemical fungicide/bactericide. The experimental results also provide an example of the benefits of the new RTI301 strain enhancing the antagonistic properties of FRACTURE (CONSUMO EM VERDE (CEV), BIOTECNOLOGIA DAS PLANTAS S.A., PORTUGAL), a plant extract containing the BLAD polypeptide as an active ingredient.

In one embodiment of the present invention, there is provided a composition comprising a biologically pure culture of Bacillus amyloliquefaci RTI301 deposited with ATCC PTA-121165 or a mutant having all of its identifying characteristics, the composition being used for Application to plants to achieve one or both of beneficial to plant growth or to confer protection to susceptible plants to avoid pathogenic infections.

In another embodiment, a method for achieving one or both of beneficial to plant growth or protection of a susceptible plant from pathogenic infection is provided, the method comprising including ATCC No. PTA-121165 A composition of a deposited biologically pure culture of Bacillus amyloliquefaciens RTI301 or a mutant having all of its identifying characteristics is delivered to: plant leaves, plant bark, plant fruits, plant flowers, plant seeds, plant roots, plant pruning , plant grafts, plant healing tissue; soil or growth medium surrounding the plant; soil or growth medium prior to seeding the plant seed in soil or growth medium; or planting plants, plant pruning, plant transplants or in soil or growth medium The soil before the plant heals the tissue Or a growth medium, the composition being in an amount suitable to be beneficial for plant growth and/or to confer protection to susceptible plants to avoid pathogenic infections.

The growth promoting activity of the RTI301 isolate in wheat is described in Example 3. Germinated wheat seeds were inoculated in a suspension of about 2 x 10 ⁷ CFU/ml RTI 301 strain for 2 days and then planted in pots. A photograph of the plants extracted after 13 days of growth is shown in Figure 2. Figure 2A shows plants inoculated with RTI301 and Figure 2B shows control plants. The dry weight of wheat seedlings was determined. As a result, the dry weight of RTI301 treated plants increased by 8.1% compared with the uninoculated controls. In addition, the beneficial effects of B. amyloliquefaciens RTI301 strain on early growth in wheat are shown in the image in Figure 3. Figure 3A shows 28 day old wheat plants inoculated with RTI301 and Figure 3B shows control plants.

The antagonistic properties of B. amyloliquefaciens RTI301 against several major plant pathogens in the disc assay are described in Example 4 and the performance traits such as phytohormone production, acetoin and indole acetic acid (IAA) and nutrient cycling of the strain are described in Example 5. .

Beneficial plant-associated bacteria (root and endoparasite) are known to provide host plants with a variety of benefits ranging from disease and insect pests to environmental stresses including cold, salinity and drought stress. Since plants inoculated with plant growth-promoting bacteria obtain more water and nutrients from the soil, for example due to better developed root systems, plants grow healthier and less susceptible to biotic and abiotic stresses. Thus, the microbial compositions of the present invention can be applied alone or in combination with current crop management inputs, such as chemical fertilizers, herbicides, and insecticides, to maximize crop yield. Promoting the effects of plant growth translates into faster plant growth and increased above-ground biomass quality, which can be applied to improve early vigor. One benefit of improving early vigor is that plants are more competitive and outweigh weeds, directly reducing weed management costs by minimizing labor and herbicide application. The promotion of plant growth effects is also transformed into improved root development, including deeper and wider roots, with more fine roots involved in the absorption of water and nutrients. This feature allows for better use of agricultural resources and reduces water for irrigation needs and/or fertilization. Changes in root development and root structure affect the interaction of plants with other soil-borne micro-organisms, including Useful fungi and bacteria that absorb nutrients (including nitrogen fixation and phosphate dissolution) in plants. These beneficial microorganisms also compete with plant pathogens, improving overall plant health and reducing the need for chemical fungicides and pesticides.

In addition, various crops were studied in greenhouse and in field trials to determine the ability of B. amyloliquefaciens RTI301 to prevent and/or alleviate the natural or artificial infection effects of many common plant pathogens on plants. The results are described in Examples 6 to 13 and Figures 4 to 7. Other studies describe the antagonism of RTI301 in combination with a product containing an antifungal polypeptide against fungal pathogens (Example 14; Figures 8-9) and identify antimicrobial metabolites produced by the RTI 301 strain and in Examples 15 and 16 and Figure 10, respectively. And 11 in the separation.

Example 6 describes the ability of the B. amyloliquefaciens RTI 301 strain to alleviate the effects of plant pathogen bean rust (String rust) and the plant pathogen Botrytis blight (B. cinerea). In the first set of experiments, different formulations of B. amyloliquefaciens RTI301 strain were tested for the application of the RTI301 strain to the leaf surface control of R. solani and Botrytis cinerea. The experimental design was set up to assess the disease control % of the following individuals after nine days of infection with the pathogen: RTI301 spores ("RTI301+1% SFB") in the waste broth diluted with water alone, in the waste broth diluted with water RTI301 spore plus yeast extract ("RTI301+1% SFB+ yeast extract"), BRAVO WEATHER STIK (500g ai/ha tetrachloroisophthalonitrile), HORIZON (50g ai/ha derkeley) and the same spores as RTI301 strain Concentration of SERENADE OPTIMUM. An untreated control (water only) produced 28% of the disease. The results of the bean rust and Botrytis cinerea wilt disease experiments were similar. The results of the bean rust test are shown in Table III and indicate that the yeast extract was added to help the growth of the RTI301 strain on the plant leaf to increase disease control by about 40% compared to the RTI301 strain administered without the addition of the yeast extract. When administered at the same application rate (i.e., 1 x 10 ⁸ cfu/ml), the RTI301 + 1% SFB + yeast extract exhibited a disease control amount similar to that observed for SERENADE OPTIMUM, even though the amount of SFB in the RTI301 formulation Relatively low, it is 1%, and SFB is expected to contain secreted metabolites with antifungal activity. A similar experiment on disease control of RTI 301 strain against bean rust is described in Example 6, and the results are shown in Tables IV-V and Figures 4-5. The results show that the control of bean rust (leaf bean rust) with RTI301 spores is comparable to that treated with SERENADE OPTIMUM when treated at the same application rate.

Various crops were studied in field trials to determine the ability of B. amyloliquefaciens RTI301 to prevent and/or alleviate the natural or artificial infection effects of many common plant pathogens on plants. The results are described in Examples 7 to 9. Examples 7 and 8 describe the control of plant pathogen powdery mildew on cucumber and Xanthomonas in tomato, respectively. In the test, RTI 301 was applied to the crop at the same application rate as SERENADE OPTIMUM. The application is 1 to 6 times, wherein the time interval between administrations is 5 to 7 days, depending on the crop. The timing of the first application depends on the particular crop and is within the range of the crop, the weeks after the crop germination, the onset of flowering, the onset of the disease, or before the onset of the expected disease. The results in Examples 7 and VI show that RTI 301 has similar control for powdery mildew in cucumber as a single biofungicide when compared to SERENADE OPTIMUM when applied at the same application rate.

The results in Example 8 and Table VII show that RTI301 has similar control for bacterial spot disease (Homomonospora) in tomato as a single biofungicide when applied at the same application rate as compared to SERENADE OPTIMUM. RTI301, which is a separate biofungicide, demonstrates similar efficacy to the combination of copper hydroxide and tetrachloroisophthalonitrile.

The results shown in Example 9 and in Table VIII show that RTI301 acts as a single biofungicide, wheat gibberellosis, soybean rust, corn rust, cucumber white powder when applied at the same application rate compared to SERENADE OPTIMUM. The control of disease and tomato bacterial spot disease is improved or similar.

The experiments described in Example 10 and the results shown in Table IX are shown with only the chemical active agent CRUISERMAXX (SYNGENTA CROP PROTECTION, INC) plus methyl thiophanate fungicide (this combination is referred to herein as "chemical control") Compared with the treated seeds, the combination of RTI301 and chemical active agent improved the control of sudden death syndrome in soybean.

Example 11 describes a study conducted in a tomato field trial to determine the ability of the B. amyloliquefaciens RTI301 strain to prevent and/or alleviate the effects of the plant pathogen, Botrytis cinerea (B. cinerea). The RTI301 strain was compared with the application of a chemical active agent called "FARMER protocol" and the application of B. subtilis strain QST713 at a concentration 10 times higher than the REN301 strain of SERENADE MAX. The % of the fruit of the infected Botrytis cinerea pathogen in the untreated control ("UT") in each tomato test over time is shown in the graph of Figure 6. The results in Table X show that the B. amyloliquefaciens RTI301 and FARMER protocols have the best control of the brown mold on tomato and outperform the treatment with SERENADE MAX.

Example 12 describes a study conducted in a strawberry field trial to determine the ability of B. amyloliquefaciens RTI 301 strain to prevent and/or alleviate the effects of the plant pathogen, Botrytis cinerea (B. cinerea). The RTI301 strain was compared with the application of a chemical active agent called "FARMER protocol" and the application of B. subtilis strain QST713 at a concentration 10 times higher than the REN301 strain of SERENADE MAX. The % of the fruit of the infected Botrytis cinerea pathogen in the untreated control ("UT") in each strawberry test over time is shown in the graph of Figure 7. The results in Table XI show that all three treatments of B. amyloliquefaciens RTI301, SERENADE MAX and FARMER have improved control of B. cinerea on strawberries compared to untreated controls, with increased yields treated with RTI301 The value is slightly higher.

Example 13 describes a field trial in corn to investigate the effects on plant growth and development after treatment of plant seeds with B. amyloliquefaciens RTI301 strain. The experimental settings were as follows: 1) the seeds were untreated; 2) the seeds were treated with a combination of MAXIM, APRON XL and PONCHO (referred to as "chemical control"); and 3) the seeds were chemically treated plus plus 5.0 x 10 seeds per seed. ⁺⁵ cfu strain RTI301. Three field trials were conducted, one with natural disease stress, one with artificially inoculated Fusarium graminearum, and one with artificially inoculated Rhizoctonia. Notably, in the Rhizoctonia test, RTI301 plus chemical controls were observed to yield a significant yield benefit of 40.1 bushels per acre compared to the chemical control alone. In summary, treatment with the chemical control plus RTI 301 resulted in an increase in the yield of all three trials and a significant increase in the yield of the test for inoculation of Rhizoctonia (see Table XII).

Example 14 describes an in vitro disc assay demonstrating the ability of B. amyloliquefaciens RTI 301 strain to enhance the efficacy of a product sold by FRACTURE to control plant pathogens. The FRACTURE product is a plant extract containing a polypeptide (BLAD polypeptide) as an active ingredient which acts on susceptible fungal pathogens by causing damage to the fungal cell wall and destroying the inner cell membrane. For this analysis, RTI301 bacterial isolates and pathogenic fungi were grown side by side on agar plates in the presence and absence of 1% FRACTURE. The results of the analysis of the plant pathogens Fusarium graminearum and Fusarium oxysporum were shown in Figures 8A-8F and Figures 9A-9F, respectively. Although the addition of 1% FRACTURE to agar reduced the growth of both pathogens, no complete inhibition of fungal growth was achieved. The presence of 1% FRACTURE in the agar medium did not inhibit the growth of B. amyloliquefaciens RTI301. However, the presence of a combination of B. amyloliquefaciens RTI301 and FRACTURE caused additional inhibition of fungal growth of Fusarium graminearum and Fusarium oxysporum. Therefore, B. amyloliquefaciens RTI301 can be used to enhance FRACTURE potency.

Example 15 describes the study of the cyclic lipopeptides auretolide and dehydroxylectin produced by the B. amyloliquefaciens RTI 301 strain, and unexpectedly, the identification of several classes of molecules not previously reported. It was determined that B. amyloliquefaciens RTI301 produced previously reported protosulin A, abundance B and abundance C compounds as well as dehydroxylectin A, dehydroxylectin B and dehydroxylectin C compounds. Surprisingly, in addition to these known compounds, it was determined that the RTI 301 strain also produced derivatives of such compounds that were not previously identified, wherein L-isoleucine at position 8 of the cyclic peptide chain (referred to as X in Figure 10) ₃ ) Replacement with L-methionine. The new classes of protosan and dehydroxylectin are referred to herein as MA, MB and MC, and refer to class A, B and C derivatives, wherein L-isoleucine at X ₃ in Figure 10 is L- Methionine replacement. The newly identified molecular bold is shown in Figure 10 and in Table XIII. It was further determined that the RTI301 strain produced other classes of amylin and dehydroxylectin which have not been previously identified. In this category, fengycin B and activin B to the hydroxyl BBCA L- isoleucine (position of FIG. 10 X ₃₎ via L- homocysteine (of Hcy) replacement. These previously unidentified abundane and dehydroxylectin metabolites are referred to herein as protamine H and dehydroxylectin H and are shown in Figure 10 and Table XIII. It was further determined that the RTI 301 strain produced other classes of protosan and dehydroxylectin metabolites that were not previously identified. In this category, the position of the main chain structure of the cyclic peptide 4 (FIG. 10 in the position X ₁₎ is replaced by the amino acid L- isoleucine. These previously unidentified metabolites are referred to herein as protamine I and dehydroxylectin I and are shown in Figure 10 and Table XIII.

Example 16 describes the isolation of an antagonistic lipopeptide from B. amyloliquefaciens strain RTI301, and an in vitro disk assay showing that the isolated lipopeptide retains its activity against two common plant pathogens. RTI301 was cultured and the acid precipitate of the culture supernatant was analyzed by LCMS to compare the relative abundances of itallin, activin and amylin. Figure 11 is a graph showing the percentage of lipopeptide recovered from the RTI301 spent broth after acid precipitation. As a result, 80% of the total amount of lipopeptide was recovered by acid precipitation. Subsequently, for the Botrytis cinerea and Fusarium graminearum, disc bioassay was performed using the same sample analyzed by LCMS. The results showing that the acid precipitated sample had similar levels of resistance to the antagonism of Fusarium oxysporum and Fusarium graminearum to the starting waste mash.

In one embodiment, there is provided a composition comprising a biologically pure culture of B. amyloliquefaci RTI472 or a mutant having all of its identifying characteristics deposited under ATCC PTA-121166, for use in application to a plant To achieve one or both of beneficial to plant growth or to confer protection to susceptible plants to avoid pathogenic infections. Plant growth benefits and/or protection afforded by increased seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or combinations thereof Show.

In one embodiment, a composition is provided comprising a Bacillus amyloliquefaciens RTI301 deposited with ATCC No. PTA-121165 or a biologically pure culture thereof having mutants of all of its identifying characteristics for application to a plant To achieve one or both of beneficial to plant growth or to confer protection to susceptible plants to avoid pathogenic infections. plant growth Benefits and/or protection afforded may be manifested by increased seedling vigor, improved root development, improved plant growth, increased plant health, increased yield, improved appearance, increased resistance to plant pathogens, or reduced pathogenic infection or a combination thereof.

The compositions and methods of the present invention are beneficial to a variety of plants including, but not limited to, monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, feed corn, rice, wheat, barley , sorghum, asparagus, berries, blueberries, blackberries, raspberries, raspberries, cranberries, cranberries, gooseberries, elderberries, currants, cranberries, berry, cabbage, broccoli, kale Dish, broccoli, Brussels sprouts, broccoli, kale, mustard, broccoli, melon, cucumber, cantaloupe, melon, cantaloupe, squash, watermelon, pumpkin, eggplant, bulbous vegetables, onions, garlic, onions , tangerine peel, orange, grapefruit, lemon, red tangerine, tangerine, pomelo, fruit and vegetable, pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, green leafy vegetables, lettuce, celery, spinach, fragrant Parsley, chicory, legumes/vegetables (succulent and dry beans and peas), beans, green beans, food pods, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, peas Petals, lentils, oilseed crops, canola, ramie, coconut, cotton, flax, oil palm, olive, peanut, rapeseed, safflower, sesame, sunflower, soybean, pear, apple, jellyfish, pear, papaya, Summer flower hawthorn, roots/tubers and bulbous vegetables, carrots, potatoes, sweet potatoes, cassava, beets, ginger, horseradish, radish, ginseng, turnip, stone fruit, apricot, cherry, nectarine, peach, plum, plum, strawberry, nut , almonds, pistachios, pecans, walnuts, hazelnuts, chestnuts, cashews, beech nuts, walnuts, macadamia nuts, kiwis, bananas, (blue) agave, grass, turfgrass, ornamental plants, poinsettia, hydrangea, hard Branches, chestnuts, oak, maple, sugar cane and beets.

In one or more embodiments, the plant may include soybean, bean, pod bean, wheat, cotton, corn, pepper, tomato, potato, tapioca, grape, strawberry, banana, peanut, squash, pumpkin, eggplant, and cucumber. .

In the compositions and methods of the present invention, pathogenic infections can be caused by a variety of plant pathogens including, for example, but not limited to, plant fungal pathogens, plant bacterial pathogens, rust, Botrytis, Botrytis cinerea, Botrytis squamosa , Erwinia, Erwinia carotovora , Erwinia amylovora , Dickeya spp ., Dickeya Dadantii ), Dickeya solani , Agrobacterium spp ., Agrobacterium tumefaciens , Xanthomonas, Xanthomonas axonopodis , Xanthomonas campestris Xanthomonas campestris pv. carotae , Xanthomonas pruni , Xanthomonas arboricola , Xanthomonas oryzae pv. oryzae , Saccharomyces cerevisiae Genus ( Xylella spp .), Xylella fastidiosa , Candidatus spp., phloem ( Can Didatus liberibacter ), Fusarium colmorum , Fusarium oxysporum , Fusarium oxysporum , Fusarium oxysporum f.sp. Cubense , Fusarium oxysporum f .sp.Lycopersici ), Fusarium virguliforme , Sclerotinia spp ., Sclerotinia sclerotiorum , Sclerotinia minor , Sclerotinia Homeocarpa ), Cercospora/Cercosporidium spp., Uncinula spp., Uncinula necator (Powdery Mildew), Podosphaera spp () Powdery mildew), Podosphaera leucotricha , Poaosphaera clandestine , Phomopsis spp., Phomopsis Viticola ), Alternaria spp ., Alternaria tenuissima , Alternaria porri , Alternaria alternate , Alzheimer's Alternaria tenuis , Pseudomonas, Pseudomonas syringae pv. Tomato , Phytophthora spp., Phytophthora infestans ), Phytophthora parasitica , Phytophthora sojae , Phytophthora capsici , Phytophthora cinnamon , Phytophthora fragariae , Phytophthora, eucalyptus ( Phytophthora ramorum ), Phytophthora palmivara , Phytophthora nicotianae , Puccinia, Phakopsora pylori, Phakopsora meibomiae , Aspergillus spp .), Aspergillus flavus , Aspergillus niger , Uromyces spp ., Bean rust, Cladosporium spp ., Cladosporium Herbarum ), Rhizopus spp., Rhizopus arrhizus , Penicillium spp., Rhizoctonia Genus, Rhizoctonia solani , Rhizoctonia zeae, Rhizoctonia oryzae , Rhizoctonia caritae , Rhizoctonia cerealis Rhizoctonia crocorum , Rhizoctonia fragariae , Rhizoctonia ramicola , Rhizoctonia rubi , Rhizoctonia leguminicola , Rhizoctonia leguminicola , Rhizoctonia leguminicola Macrophomina phaseolina , Magnaorthe oryzae , Cochlear , Mycosphaerella graminocola , Mycosphaerella fijiensis (Black sigatoga) ), Mycosphaerella pomi , Mycosphaerella citri , Magnaporthe oryzae, Magnaporthe grisea , Monilinia spp., Peach brown rot Monilinia fruticola ), Monilinia vacciniicorymbosi , Monilinia laxa , Anthracnose ( Colletotrichum) Spp.), Colletotrichum gloeosporiodes , Colletotrichum acutatum , Colletotrichum Candidum , Diaporthe spp., Diaporthe citri , oxysporum genus rod (Corynespora spp.), multi-master rod cinerea (Corynespora Cassiicola), the genus gum rust (Gymnosporangium spp.), gum juniper plants by rust (Gymnosporangium juniperi-virginianae), Scarshield genus (Schizothyrium spp .), Schizothyrium pomi , Gloeodes spp., Gloeodes pomigena , Botryosphaeria spp., grape canker ( Botryosphaeria dothidea ), Neofabraea spp., Wilsonomyces spp., Wilsonomyces carpophilus , Sphaerotheca spp., spots Sphaerotheca macularis , Sphaerotheca pannosa , Erysiphe spp., Stagonospora spp ., S. serrata ( S) Tagonospora nodorum ), Pythium spp ., Pythium ultimum , Pythium aphanidermatum , Pythium irregularum , Pythium ulosum , aquatic rot Pythium lutriarium , Pythium sylvatium , Venturia spp , Venturia inaequalis , Verticillium spp., Ustilago spp .), Ustilago nuda , Ustilago maydis , Ustilago scitaminea , Claviceps spp ., Claviceps puprrea , alfalfa Tilletia spp ., Tilletia tritici , Tilletia laevis , Tilletia horrid , Tilletia controversa , Phoma spp ., Phoma glycinicola , Phoma exigua , Phoma lingam , Cocliobolus sativus , Wo Gaeumanomyces gaminis , Colleototricum spp., Rhychosporium spp ., Rhychosporium secalis , Biopolaris spp ., Helminthosporium ( Helminthosporium spp .), Helminthosporium secalis , Helminthosporium maydis , Helminthosporium solai , and Helminthosporium tritici-repentis or combinations thereof.

In some embodiments, the pathogenic infection can be caused by one or a combination of: soybean rust (Phaeophyllum serrata, Pseudomonas aeruginosa) and the plant comprises soybean; Botrytis cinerea (B. blight) and The plant comprises grapes; Botrytis cinerea (B. blight) and the plant comprises strawberries; Botrytis cinerea (B. blight) and the plant comprises tomato; Alternaria (such as Alternaria solani) and the plant comprises Tomato; Alternaria (such as Alternaria alternata) and the plant contains potato; bean rust (leaf bean rust) and the plant contains common beans; powdery mildew (soybean powdery mildew) and the plant contains soybean; (black banana leaf spot) or Fusarium oxysporum (Panama disease) and the plant contains bananas; Xanthomonas or Xanthomonas oryzae rice varieties and plants contain rice; Trichophyton rubrum and plants contain cassava Xanthomonas campestris and plant containing tomato; Botrytis cinerea (P. cinerea) and plant containing pepper; powdery mildew and plant containing gourd; S. cerevisiae (white mold) and plant containing pod bean; sunflower Sclerotinia White mold) and the plant contains potato; Sclerotinia sclerotiorum (coin spot disease) and the plant contains turfgrass; southern white mold and plants contain peanuts; leaf spot disease (C. cerevisiae / Cercospora) and plants contain peanuts; Fusarium oxysporum (French scab) and plants containing wheat; Phytophthora sinensis (S. cerevisiae) and plants containing wheat; Phytophthora capsici (Bacterial blight and Phytophthora capsici) and plants Wheat; starch-degrading Owen bacteria and plants containing apples, pears and other pear fruit; apple black scab and plants containing apples, pears and other pear fruit; or Rhizoctonia solani and plants containing wheat, rice, turf, Soybeans, corn, legumes and vegetable crops.

Compositions comprising the RTI 301 strain may be in the form of a liquid, an oily dispersion, a powder, a dry wettable powder, a spreadable granule or a dry wettable granule.

When the composition is applied to plant leaves, plant bark, plant fruits, plant flowers, plant seeds, plant roots, plant pruning, plant transplants, plant healing tissues; soil or growth medium surrounding the plant; seeding in soil or growth medium Soil or growth medium before plant seeds; or soil or growth medium prior to planting plants, plant pruning, plant transplants, or plant healing tissues in soil or growth medium, when suitable for plant growth and/or The composition is beneficial for plant growth when the susceptible plant is administered in an amount that provides protection against pathogenic infection.

The composition comprising the RTI 301 strain may further comprise one or a combination of a carrier, a surfactant, a dispersant or a yeast extract. For the purposes of this specification and the scope of the patent application, the terms "interacting agent" and "adjuvant" are used interchangeably. The yeast extract can be delivered at a rate suitable for plant growth in the range of about 0.01% to 0.2% w/w.

The composition can be in the form of a planting matrix. The planting substrate can be in the form of potting soil.

In one embodiment, a composition for achieving one or both of beneficial to plant growth or protection of a susceptible plant from pathogenic infection is provided, the composition comprising: being deposited under ATCC PTA-121165 A biologically pure culture of Bacillus amyloliquefaciens RTI301 or a mutant having all of its identifying characteristics, in an amount suitable for beneficial to plant growth and/or to confer protection to susceptible plants to avoid pathogenic infection; One or a combination of biological or chemical insecticides, fungicides, nematicides, bactericides, herbicides, plant extracts, plant growth regulators or fertilizers, suitable for plant growth and/or susceptible diseases Plants provide protection from the amount of pathogenic infections. In this embodiment, the biologically pure culture of Bacillus amyloliquefaciens RTI301 and one of microbial or chemical insecticides, fungicides, nematicides, bactericides, herbicides, plant extracts, plant growth regulators or fertilizers Or a combination of blends.

In one embodiment, the fungicide may comprise an extract from white lupin. In one embodiment, the fungicide may comprise a BLAD polypeptide. The BLAD polypeptide may be a fragment of a naturally occurring seed storage protein from sweet lupin (white lupin) by means of a fungus The cell wall causes damage and destroys the inner cell membrane and acts on susceptible fungal pathogens. The composition can include about 20% BLAD polypeptide.

In a composition comprising B. amyloliquesii RTI 301, the composition may be in liquid form and B. amyloliquefaciens RTI 301 may be present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. The composition may be in the form of a powder, a dry wettable powder, a spreadable granule or a dry wettable granule and the B. amyloliquefaciens RTI 301 may be in an amount from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. presence. The composition may be in the form of an oily dispersion, and the B. amyloliquefaciens RTI301 may be present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. Bacillus amyloliquefaciens RTI301 can be in the form of spores or vegetative cells.

In one embodiment, a method for achieving one or both of beneficial to plant growth or protection of a susceptible plant to avoid pathogenic infection is provided, the method comprising: comprising ATCC No. PTA-121165 A composition of a deposited biologically pure culture of Bacillus amyloliquefaciens RTI301 or a mutant having all of its identifying characteristics is delivered to: plant leaves, plant bark, plant fruits, plant flowers, plant seeds, plant roots, plant pruning , plant grafts, plant healing tissue; soil or growth medium surrounding the plant; soil or growth medium prior to seeding the plant seed in soil or growth medium; or planting plants, plant pruning, plant transplants or in soil or growth medium The soil or growth medium prior to the healing of the tissue, the composition being present in an amount suitable to benefit the growth of the plant and/or to confer protection to the susceptible plant to avoid pathogenic infection. The composition can be delivered to plant leaves.

In this method, the composition comprising B. amyloliquefaciens RTI 301 may further comprise one or a combination of a carrier, a surfactant, a dispersing agent or a yeast extract. The yeast extract can be delivered at a rate suitable for plant growth in the range of about 0.01% to 0.2% w/w.

In another embodiment of the invention, a method for achieving one or both of beneficial to plant growth or protection of a susceptible plant to avoid pathogenic infection is provided, the method comprising delivering a composition to Plant leaves, plant bark, plant fruits, plants Flowers, plant seeds, plant roots, plant pruning, plant grafts, plant healing tissues; soil or growth medium surrounding the plant; soil or growth medium prior to seeding the plant seeds in soil or growth medium; or in soil or growth medium A soil or growth medium prior to planting a plant, plant pruning, plant graft or plant healing tissue, the composition comprising the following: A. faecalis RTI301 deposited under ATCC PTA-121165 or having all of its identification a biologically pure culture of a mutant of a characteristic that is suitable for beneficial to plant growth and/or to confer protection to a susceptible plant to avoid pathogenic infection; and microbial, biological or chemical insecticides, fungicides, kills One or a combination of nematodes, bactericides, herbicides, plant extracts, plant growth regulators or fertilizers is in an amount suitable to be beneficial for plant growth and/or to confer protection against pathogenic infections.

In one embodiment, a method for achieving one or both of beneficial to plant growth or protection of a susceptible plant to avoid pathogenic infection is provided, the method comprising: combining the first composition with the second The combination of substances is delivered to plant leaves, plant stems, plant fruits, plant flowers, plant seeds, plant roots, plant pruning, plant transplants, plant healing tissues; soil or growth medium around the plant; seeded in soil or growth medium Soil or growth medium prior to plant seeds; or soil or growth medium prior to planting plants, plant pruning, plant transplants or plant healing tissues in soil or growth medium, the first composition comprising ATCC PTA-121165 A biologically pure culture of the deposited Bacillus amyloliquefaciens RTI301 or a mutant thereof having all of its identifying characteristics, in an amount suitable for beneficial to plant growth and/or protection of susceptible plants to avoid pathogenic infection; The second composition comprises a microbial, biological or chemical insecticide, fungicide, nematicide, bactericide, herbicide, plant extract Plant growth regulators or fertilizers or one combination, was adapted to benefit plant growth and / or to impart protection to the susceptible plants in order to avoid an amount of pathogenic infections. In one embodiment, the first and second compositions can be delivered to the foliage of the plant.

The first composition may further comprise one or a combination of a carrier, a surfactant, a dispersant, or a yeast extract. Yeast extract can be in the range of about 0.01% to 0.2% w/w Conducive to the application rate of plant growth.

The fungicide of the second composition may comprise an extract from white lupin. The fungicide of the second composition can include a BLAD polypeptide. The BLAD polypeptide may be a fragment of a naturally occurring seed storage protein from sweet lupin (white lupin) that acts on a susceptible fungal pathogen by causing damage to the fungal cell wall and disrupting the inner cell membrane. The fungicide of the second composition can include about 20% BLAD polypeptide.

In the compositions and methods of the present invention for delivering RTI 301 in combination with a microbial, biological or chemical insecticide, fungicide, nematicide, bactericide, herbicide, plant extract, plant growth regulator or fertilizer, Plant growth benefits and/or protection afforded by increased seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or combinations thereof Show.

In one embodiment, the method may further comprise applying a liquid fertilizer to: soil or growth medium surrounding the plant; soil or growth medium prior to seeding the plant seed in the soil or growth medium; or planting in soil or growth medium Soil or growth medium prior to plant, plant pruning, plant graft or plant healing tissue.

In a method for transporting RTI 301 in combination with a microbial, biological or chemical insecticide, fungicide, nematicide, bactericide, herbicide, plant extract, plant growth regulator or fertilizer, the composition may be in the form of a liquid, Oily dispersions, powders, dry wettable powders, spreadable granules or dry wettable granules. Bacillus amyloliquefaciens RTI301 can be in the form of spores or vegetative cells. Bacillus amyloliquefaciens RTI301 be about 1.0 × ¹⁰ 10 CFU / ha to about 1.0 × 10 ¹⁴ CFU / benefit ha application rate of delivery of plant growth. The yeast extract can be delivered at a rate suitable for plant growth in the range of about 0.01% to 0.2% w/w.

In the compositions and methods of the present invention for delivering RTI 301 with microbial, biological or chemical insecticides, fungicides, nematicides, bactericides, herbicides, plant extracts, plant growth regulators or fertilizers, insecticides It can contain Bifenin. In one or more embodiments, The nematicide may comprise cadusafos. In one or more embodiments, the pesticide may comprise bifenin and clothianidin. In one or more embodiments, the pesticide can comprise diphennin and the composition can be formulated as a liquid. In one or more embodiments, the pesticide can comprise diphennin and cotinine. In one or more embodiments, the pesticide can comprise diphennin and cotinine and the composition can be formulated as a liquid. In one or more embodiments, the pesticide may comprise diphennin or acetonin. In one or more embodiments, the composition can be formulated as a liquid and the pesticide can comprise diphennin or acetonin.

The insecticide may be bifenin, and the composition formulation may further comprise hydrated aluminum-magnesium citrate and at least one dispersant selected from the group consisting of sucrose esters, lignosulfonates, alkyl polyglycosides, naphthalenes. Sulfonic acid formaldehyde condensate and phosphate ester. Bifenin insecticides can be present in concentrations ranging from 0.1 g/ml to 0.2 g/ml. The bifenin insecticide can be present at a concentration of about 0.1715 g/ml. The application rate of the Bifenin insecticide may range from about 0.1 gram per gram of bifenine (g ai/ha) to about 1000 g ai/ha, more preferably from about 1 g ai/ha to about 100 g ai/ha.

In one embodiment, the Phenfenin composition may comprise: phenfenamicin; hydrated aluminum-magnesium citrate; and at least one dispersant selected from the group consisting of sucrose esters, lignosulfonates, alkyl polyglycosides, naphthalenesulfonic acid formaldehyde Condensate and phosphate.

Bifenin may preferably be present at a concentration of from 1.0% to 35% by weight, more particularly from 15% to 25% by weight, based on the total weight of all components in the composition. The Bifenin insecticide composition can be formulated to be mixed with a fertilizer to form a liquid. The Phenfenin insecticidal composition can be present in the liquid formulation at a concentration ranging from 0.1 g/ml to 0.2 g/ml. The bifenin insecticide can be present in the liquid formulation at a concentration of about 0.1715 g/ml. The terms "may be suitable for mixing with fertilizers as a liquid" and "formulations compatible with liquid fertilizers" are used interchangeably throughout the specification and patent application and are intended to mean that the formulation is capable of dissolving or dispersing. Emulsified in an aqueous solution to allow mixing with the fertilizer for delivery to the plant in a liquid formulation.

The dispersing agent may preferably be present in a total concentration of from about 0.02% by weight to about 20% by weight based on the total weight of all components of the composition.

In some embodiments, the hydrated aluminum-magnesium silicate may be selected from the group consisting of montmorillonite and attapulgite.

In some embodiments, the phosphate ester can be selected from the group consisting of nonylphenol phosphate and tridecyl alcohol ethoxylated phosphate potassium salt.

Other embodiments may further include at least one of an antifreeze, an antifoaming agent, and a biocide.

In one embodiment, a plant-promoting composition is provided, the composition comprising: Bacillus amyloliquefaciens RTI301 deposited with ATCC PTA-121165 or a biologically pure culture thereof having mutants of all of its identifying characteristics ; and pesticides. Insecticides may be pyrethroid, bifennin, tefluthrin, guanidine, organophosphate, chlorethoxyfos, chlorpyrifos, butyl pyrimidine One or a combination of (tebupirimfos), cyfluthrin, fiproles, fipronil, nicotinoids, or cotinine. The insecticide can include diphennin. The composition may be in a formulation compatible with liquid fertilizers. The composition comprising bifennin may further comprise hydrated aluminum-magnesium citrate and at least one dispersant. The bifenin insecticide can be present in a concentration ranging from 0.1 g/ml to 0.2 g/ml. The bifenin insecticide can be present at a concentration of about 0.1715 g/ml.

Additionally, in one or more embodiments, suitable insecticides, herbicides, fungicides, and nematicides of the compositions and methods of the present invention can include the following: insecticides: A0) various insecticides, including Agricultural insecticide (agrigata), aluminum phosphide, blunt cockroach, cockroach bee, cockroach bee, eater bee, artemisinin, N. glabrata NPV, azocyclotin, Bacillus subtilis, Bacillus thuringiensis species Asha China and Asia (Bacillus thuringiensis-spp.aizawai), Bacillus thuringiensis库斯塔克亚species (Bacillus thuringiensis spp.kurstaki), Bacillus thuringiensis, Beauveria (BEAUVERIA), Beauveria Beauveria bassiana , betacyfluthrin, biologics, bisultap, brofluthrinate, bromophos-e, bromopropylate, Bt-corn-GM, Bt-soybean-GM, capsaicin, cartap, southern snake vine extract, chlorantraniliprole, chlorbenzuron, chlorophosphonate (chlorethoxyfos), chlorfluazuron, taosson-e Osthole, cryolite, cyanophos, cyantraniliprole, cyhalothrin, cyhexatin, cytosine, geranium, DCIP, dichloropropene , dicofol, scorpion, scorpion fly + scorpion bee, dimethacarb, disulfide, dodecyl acetate, emamectin, striata, EPN, pulp Hornbill bees, dibromoethylene, eucalyptol, fatty acids, fatty acids/salts, fentanyl, zhongdingwei (BPMC), fenfluramine, brommethrin, fluoxazine, chlorpyrifos, ango , furosemide, γ-cylonine, garlic juice, granulosis virus, heterochromatic ladybug, cotton bollworm NPV, inactivated bacteria, indole-3-butyric acid, methyl iodide, iron, hirudin, different Amphetamine, isopropylamine-m, isoprocarb, isophosphorus, kaolin, lingdan, liuyangmycin, matrine, diammine, polyacetal, Metarhizium, damascus, acesulfame MTMC), mineral oil, mirex, isothiocyanate, insecticide, Fusarium oxysporum, dibromophosphorus, Fuxinji wasp, nicotine, nicotine insecticide, oil, oleic acid Omethoate Small calyx, oxymatrine, Paecilomyces, paraffin oil, parathion-e, pasteurella, petroleum, pheromone, phosphoric acid, light bacillus, phoxim, small plant绥螨, ultimisol-e (pirimiphos-e), vegetable oil, Plutella xylostella GV, polyhedrosis virus, polyphenol extract, potassium oleate, profenofos, psorale, prothiofos , pyrazophos, pyrethrin, pyridaphenthione, pyrimidine, pyriproxyfen, soap tree extract, chlorpyrifos, canola oil, rotenone, saponin, saponozit, sodium compound, Sodium fluoroantimonate, starch, nematode, Streptomyces, sulfluramid, sulfur, butyl pyrimidine, tefluthrin, dithiphos, tetrachloromethane, chlorpyrifos, methyl ethion Transgenic (eg Cry3Bb1), zodiazepam, Trichoderma, Trichogramma, Sanfulong, Verticillium, Veratrine, Isomerized Insecticides (eg κ-biphennin, κ-hetamethrin), mesophore Insecticides, biguanide insecticides, pyrazilflumid; A1) urethanes, including aldicarb, alanycarb, benfurac Arb), carbaryl, carbofuran, carbosulfan, metiocarb, methodyl, oxamyl, bicap Pirimicarb), propoxur and thiodicarb; A2) organophosphates, including acephate, azinphos-ethyl, tamarind-methyl Azinphos-methyl), chlorfenvinphos, chlorpyrifos, chlorpyrifos-methyl, demeton-S-methyl, diazinon, two Dichlorvos/DDVP, dicrotophos, dimethoate, disulfoton, ethion, fenitrothion, fenthion, plus Isoxathion, malathion, methamidaphos, methidathion, mevinphos, monocrotophos, oxymethoate, oxydemeton- Methyl), parathion, parathion-methyl, phenthoate, phorate, phosalon e), phosmet, phosphamidon, pirimiphos-methyl, quinalphos, terfufos, tetrachlorvinphos, three falling Triazophos and trichlorfon; A3) cyclopentadiene organochlorine compounds, such as endosulfan; A4) non-poor, including ethiprole, fepni, Pyrafluprole and pyriprole; A5) neonicotinoid analogues, including acetamiprid, cotinine, dinotefuran, imidacloprid, acetamiprid Nitenpyram, thiacloprid and thiamethoxam; A6) spinosad, such as spinosad and spinetoram; A7) chloride channel activator from statins , including abaftin (abamectin), benzoic acid, statin, rividine, citidine and methiophene; A8) juvenile hormone mimics, such as methionine, enediyl ester, methine Methoprene), fenoxycarb and pyriproxyfen; A9) selective homopteran feeding blockers, such as pymetrozine, Flonicamid and pyrifluquinazon; A10) sputum growth inhibitors, such as ketopurine, dexamethasone and acetonide; A11) mitochondrial ATP synthase inhibitors, such as Nock, fenbuxil and hydrazine; oxidative phosphorylation decoupling agents, such as chlorfenapyr; A12) nicotinic acetylcholine receptor channel blockers, such as benzultap, hydrochloric acid Pedan, thiocyclam and insecticidal disodium; A13) Chitin biosynthesis type 0 inhibitors from benzamidine, including ditrifluorobacillus, diflubenzuron, fluorofenon, six volts Long, Lucendon, Norvaron and Defolon; A14) Chitin biosynthesis type 1 inhibitors, such as buffingin; A15) quercerin, such as racemic cleansing; A16) quercetin receptor agonist , such as fentanyl, defenofol, phenymidine and cyclazone; A17) octopamine receptor agonist, such as triterpenoids; A18) mitochondrial complex electron transport inhibitor darridaben , tebufenpyrad, tolfenpyrad, flufenine, nitrile pyridin, butaflurane, amemusone, azine or pyrimidin A19) voltage-dependent sodium channel blockers, such as indoxacarb and cyprofenil; A20) lipid synthesis inhibitors, such as chlorfenapyr, spironolactone, and spirotetramat; A21) from A linoredine receptor modulator of guanamines, including flubendiamide, phthalimin compound (R)-3-chloro-N1-{2-methyl-4-[1,2, 2,2-tetrafluoro-1-(trifluoromethyl)ethyl]phenyl}-N2-(1-methyl-2-methylsulfonylethyl)phthalic acid and (S) -3-chloro-N1-{2-methyl-4-[1,2,2,2-tetrafluoro-1-(trifluoromethyl)ethyl]phenyl}-N2-(1-methyl- 2-methylsulfonylethyl) phthalimide, chloranthraniliprole, and cyanarylpron; A22) Compounds of unknown or indeterminate mode of action, such as azadirachtin, amphetamine (amidoflumet), bifenazate, fluthiazide, synergistic sperm, acetamiprid, fluramide; or A23) sodium channel regulators from pyrethroids, including ananin, Pyrethroid, Bifenin, Saifu Ning, λ-Xelonine, Sai Ningning, α-赛灭宁, β-赛灭宁, ξ-赛灭宁, 灭宁宁, Yihuali, Yifen Ning, Fenpning, Fenhua Li, Husaining, τ-Fuhuali, Baishenning, Qifufen and Taininging.

Fungicide: B0) Benzoene fluconazole, downy mildew, fluzinamide, carbazolam, copper salt (such as copper hydroxide, copper oxychloride, copper sulfate, copper peroxosulfate), Boscalid, thiflumazide, flutianil, furalaxyl, thiabendazole, benodanil, mepronil, Aesop Isofetamid, fenfuram, bixafen, fluxapyroxad, penflufen, sedaxane, syringolide (isofetamid) Coumoxystrobin), enoxastrobin, flufenoxystrobin, pyraoxystrobin, pyramatetostrobin, triclopyricarb, fenaminstrobin , metominostrobin, pyribencarb, meptyldinocap, fentin acetate, fentin chloride, triphenyltin hydroxide (fentin) Hydroxide), oxytetracycline, chlozolinate, chloroneb, tetrachloronitrobenzene (tecnaze) Ne), etridiazole, iodocarb, prothiocarb, Bacillus subtilis synthesis, Bacillus amyloliquefaciens (eg strain QST 713, FZB24, MBI600, D747), from alternate leaf white Extract of Melaleuca alternifolia , extract from white lupinus ( Lupinus albus doce ), BLAD polypeptide, pyrisoxazole, oxpoconazole, etaconazole, albendazole Fenpyrazamine, naftifine, terbinafine, validamycin, pyrimorph, valifenalate, philistine (fthalide), probenazole, isotianil, laminarin, extract from Polygonum cuspidatum, phosphorous acid and salt, teclofthalam, triazoxide, nail Pyrofenone, organic oil, potassium bicarbonate, tetrachloroisophthalonitrile, fluoroimide; B1) azoles, including more than peas, carbendazole, cyclazone, holly, dar Klee, Enconazole, Epto, Fluoroquinazole, Fenke, Guardian, and Protection , Fickley, imidazole, inocastazole, chlorpyrifos, micney, pingke, pugliine, prothioconazole, flurazol, tripafin, three tailong, dexclily, tikili, sterilized azole, Imiamine, pedicure, idylindole, safty, squadron, smectite, befenbut, rot, wheat ear, ethaboxam, edulis and chlorpyrifos , Dakli-M, Imidazole, Barkerol, Uniconazole, 1-(4-Chloro-phenyl)-2-([1,2,4]triazol-1-yl)-cycloheptanol and sulfuric acid B2) opiate, including atorenosin, noxylamine, enestrobin, fluoxastrobin, kresoxim-methyl, phenoxybenzamine, oxime, Pyridoxine, hexagram, trifluoro-sensitive, enestrobin, methyl 2-(2-chloro-5-[1-(3-methylbenzyloxyimino)ethyl]benzyl)carbamate, Methyl 2-chloro-5-[1-(6-methylpyridin-2-ylmethoxyimino)ethyl]benzyl)carbamate and 2-(o-(2,5-dimethyl) Methyl phenoxymethylene)-phenyl)-3-methoxy acrylate, 2-(2-(6-(3-chloro-2-methyl-phenoxy)-5-fluoro-pyrimidine- 4-yloxy)-phenyl)-2-methoxyimino-N-methyl-acetamide and 3-methoxy-2- (2-(N-(4-methoxy-phenyl)-cyclopropane)-imidylthiomethyl)-phenyl)-methyl acrylate; B3) Carboxamide, including wilting, present Benalaxyl, Bendula-M, cycloheximide, flutanil, furametpyr, mepronil, metalaxyl, mefenoxam ), furosemide, oxadixyl, oxycarboxin, penthiopyre, pyrazolamide, thifluzamide, tiadinil, 3,4-dichloro -N-(2-cyanophenyl)isothiazol-5-formamide, dimethomorph, flumorph, flufenamide, fluoprazamide (picobenzamid), Amine, gupamine, diclosan, dipropionamide, N-(2-(4-[3-(4-chlorophenyl)prop-2-ynyloxy)-3-methoxy Phenyl)ethyl)-2-methanesulfonyl-amino-3-methylbutanamine, N-(2-(4-[3-(4-chloro-phenyl)prop-2-ynyloxy) 3-yloxy-phenyl)ethyl)-2-ethanesulfonylamino-3-methylbutanamine, 3-(4-chlorophenyl)-3-(2-isopropoxy Carbonyl-amino-3-methyl-butylammonium)propionic acid methyl ester, N-(4'-bromobiphenyl-2-yl)-4-difluoromethyl-methylthiazole-δ-formamidine amine N-(4'-trifluoromethyl-biphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-formamide, N-(4'-chloro-3'-fluoro Biphenyl-2-yl)-4-difluoromethyl-2-methyl-thiazole-5-carboxamide, N-(3\4'-dichloro-4-fluorobiphenyl-2-yl)- 3-Difluoro-methyl-1-methyl-pyrazole-4-carboxamide, N-(3',4'-dichloro-5-fluorobiphenyl-2-yl)-3-difluoromethyl 1-methylpyrazole-4-carboxamide, N-(2-cyano-phenyl)-3,4-dichloroisothiazol-5-carboxamide, 2-amino-4-methyl Base-thiazole-5-methylanilide, 2-chloro-N-(1,1,3-trimethyl-indan-4-yl)-nicotinium amide, N-(2-(1,3- Dimethylbutyl)-phenyl)-1,3-dimethyl-5-fluoro-1H-pyrazole-4-carboxamide, N-(4'-chloro-3',5-difluoro- Biphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N-(4'-chloro-3',5-difluoro-biphenyl- 2-yl)-3-trifluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N-(3',4'-dichloro-5-fluoro-biphenyl-2-yl )-3-trifluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N-(3',5-difluoro-4'-methyl-biphenyl-2-yl)- 3-Difluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N-(3',5-difluoro-4'-methyl-biphenyl-2-yl)-3- Trifluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N-(cis-2-bicyclopropyl-2-yl-benzene )-3-Difluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N-(trans-2-dicyclopropyl-2-yl-phenyl)-3-difluoro-A -1-methyl-1H-pyrazole-4-carboxamide, flupirtine, N-(3-ethyl-3,5-5-trimethyl-cyclohexyl)-3-carboxamidine Amino-2-hydroxy-benzamide, oxytetracycline, thiophanate, N-(6-methoxy-pyridin-3-yl)cyclopropanecarbamide, 2-iodo-N-phenyl- Benzylamine, N-(2-bicyclo-propyl-2-yl-phenyl)-3-difluoromethyl-1-methylpyrazol-4-ylcarboxamide, N-(3', 4',5'-Trifluorobiphenyl-2-yl)-1,3-dimethylpyrazole-4-ylcarboxamide, N-(3',4',5'-trifluorobiphenyl- 2-yl)-1,3-dimethyl-5-fluoropyrazole-4-yl-carbenamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-5 -Chloro-1,3-dimethyl-pyrazol-4-ylcarboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-3-fluoromethyl-1 -methylpyrazol-4-ylformamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-3-(chlorofluoromethyl)-1-methylpyrazole 4-ylformamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methylpyrazole-4-ylcarboxamide , N-(3',4',5'-trifluorobiphenyl-2-yl)-3-difluoromethyl-5-fluoro-1-methylpyrazole-4-ylcarboxamide, N- (3',4',5'-trifluorobiphenyl-2-yl)-5-chloro-3- Fluoromethyl-1-methylpyrazole-4-ylcarboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-3-(chlorodifluoromethyl)- 1-methylpyrazol-4-ylcarboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-1-methyl-3-trifluoromethylpyrazole- 4-ylformamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-5-fluoro-1-methyl-3-trifluoromethylpyrazol-4-yl Formamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-5-chloro-1-methyl-3-trifluoromethylpyrazole-4-ylcarboxamide , N-(2',4',5'-trifluorobiphenyl-2-yl)-1,3-dimethylpyrazol-4-ylcarboxamide, N-(2',4',5 '-Trifluorobiphenyl-2-yl)-1,3-dimethyl-5-fluoropyrazole-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl- 2-yl)-5-chloro-1,3-dimethylpyrazole-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-3- Fluoromethyl-1-methylpyrazole-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-3-(chlorofluoromethyl)-1 -methylpyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methylpyrazole-4 -carbamamine, N-(2',4',5'-trifluorobiphenyl-2-yl)-3-difluoromethyl-5-fluoro-1-methylpyrazole-4-yl Indoleamine, N-(2',4',5'-trifluorobiphenyl-2-yl)-5-chloro-3-difluoromethyl-1-methylpyrazole-4-ylcarboxamide N-(2',4',5'-trifluorobiphenyl-2-yl)-3-(chlorodifluoromethyl)-1-methylpyrazol-4-ylcarboxamide, N-(2 ',4',5'-Trifluorobiphenyl-2-yl)-1-methyl-3-trifluoromethylpyrazole-4-ylcarboxamide, N-(2',4',5' -Trifluorobiphenyl-2-yl)-5-fluoro-1-methyl-3-trifluoromethylpyrazole-4-ylcarboxamide, N-(2',4',5'-trifluoro Biphenyl-2-yl)-5-chloro-1-methyl-3-trifluoromethylpyrazole-4-ylcarboxamide, N-(3',4'-dichloro-3-fluorobiphenyl -2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-dichloro-3-fluorobiphenyl-2-yl )-1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-difluoro-3-fluorobiphenyl-2-yl)-1- Methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-difluoro-3-fluorobiphenyl-2-yl)-1-methyl-S -difluoromethyl-1H-pyrazole-4-carboxamide, N-(3'-chloro-4'-fluoro-3-fluorobiphenyl-2-yl)-1-methyl-3-difluoro Methyl-1H-pyrazole-4-carboxamide, N-(3',4'-dichloro-4-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H -pyrazole-4-carbamide, N-(3',4'-difluoro-4-fluorobiphenyl-2-yl)-1-methyl-S-trifluoromethyl-1H-pyrazole- 4-Protonamine, N-(3',4'-dichloro-4-fluorobiphenyl-2-yl)-1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxamidine Amine, N -(3',4'-difluoro-4-fluorobiphenyl-2-yl)-1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxamide, N-(3'-Chloro-4'-fluoro-4-fluorobiphenyl-2-yl)-1-methyl-S-difluoromethyl-1H-pyrazole-4-carboxamide,N-(3',4' -Dichloro-5-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-difluoro- 5-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-dichloro-5-fluoro Benz-2-yl)-1-methyl-S-difluoromethyl-1H-pyrazole-formamide, N-(3',4'-difluoro-5-fluorobiphenyl-2-yl) -1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-dichloro-5-fluorobiphenyl-2-yl)-1,3 -Dimethyl-1H-pyrazole-4-carboxamide, N-(3'-chloro-4'-fluoro-5-fluorobiphenyl-2-yl)-1-methyl-3-difluoromethyl -1H-pyrazole-4-carboxamide, N-(4'-fluoro-4-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4 -Procarbamide, N-(4'-fluoro-5-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-( 4'-Chloro-5-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(4'-methyl-5- Fluorbiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(4'-fluoro-5-fluorobiphenyl-2-yl) -1,3- Methyl-1H-pyrazole-4-carboxamide, N-(4'-chloro-5-fluorobiphenyl-2-yl)-1,3-dimethyl-1H-pyrazole-4-carboxamidine Amine, N-(4'-methyl-5-fluorobiphenyl-2-yl)-1,3-dimethyl-1H-pyrazole-4-carboxamide, N-(4'-fluoro-6 -fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(4'-chloro-6-fluorobiphenyl-2-yl )-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-[2-(1,1,2,3,3,3-hexafluoropropoxy)- Phenyl]-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N-[4'-(trifluoromethylthio)-biphenyl-2-yl]-3 -Difluoromethyl-1-methyl-1 H-pyrazole-4-carboxamide and N-[4'-(trifluoromethylthio)-biphenyl-2-yl]-1-methyl- 3-trifluoromethyl-1-methyl-1H-pyrazole-4-carboxamide; B4) heterocyclic compounds including chlorinamine, pyrifenox, bupirimate, azoxystrobin Cyclic amine, fenarimol, ferimzone, azoxystrobin, nuarimol, pyrimethanil, triforine, seed dressing, protection, Aldimorph, carbendazim, fenfluramine, darfusin, fenpropidin, ipproxil, chlorhexidine, chlorhexidine, chlorhexidine, imidacloprid, octyl ketone, Pyrogen, 5-chloro-7-(4-methyl-piperidin-1-yl)-6-(2,4,6-trifluorophenyl)-[1,2,4]triazolo[1 , 5-a] pyrimidine, carbendazim, chlorhexidine, baiweilong, propoxyquinoline, trioxazole, 2-butoxy-6-iodo-3-propyl Ace-4-one, acidified benzothiadiazole-S-methyl ester, tetrachlorodan, captan, melon, folfet, cyanamide, quinoxyfen, N , N-dimethyl-3-(3-bromo-6-fluoro-2-methylindole-1-sulfonyl)-[1,2,4]triazole-1-sulfonamide, 5- Ethyl-6-octyl-[1,2,4]triazolo[1,5-a]pyrimidine-2,7-diamine, 2,3,5,6-tetrachloro-4-methylsulfonate Base-pyridine, 3,4,5-trichloro-pyridine-2,6-dicarbonitrile, N-(1-(5-bromo-3-chloro-pyridin-2-yl)-ethyl)-2, 4-Dichloro-nicotine decylamine, N-((5-bromo-3-chloropyridin-2-yl)-methyl)-2,4-dichloro-nicotinium decylamine, difluoroline, chloridine , carbendazim acetate, oxazolin, blasticidin-S, saponin, imiprozil, wild swallow, wild swallow-methyl sulfate, oxolinic acid and powdery disease; B5) urethanes, including zinc-manganese, manganese-niobium, Weibaimu, sulfavir, elixirs, fumei, methyl zinc, Pu'er, zinc, Pu, Yisui, and B. Granules, carbendazim, phenothiamine, propamocarb, chlorfenapyr hydrochloride, N-(1-(1-(4-cyanophenyl)-ethanesulfonyl)butan-2-yl) 4-fluorophenyl carbamic acid, 3-(4-chloro-phenyl)-3-(2-isopropoxycarbonylamino) Methyl 3-methyl-butyl-decylamino)propionate; or B6) other fungicides, including guanidine, tannin, toxin free base, ke shui, biguanide; antibiotics: kasugamycin, oxytetracycline And its salt, streptomycin, granulin, vilimycin A; nitrophenyl derivatives: Baiqi grams, white powder grams, large aphid; sulfur-containing heterocyclic compounds: nitrile sulfonium, rice blast Ling; organometallic compounds: dip and tin salts; organophosphorus compounds: granules pine, propyl chelsson, fosetyl, foss-aluminum, phosphorous acid and its salts, white pine, methyl gram Pine; organochlorine compounds: Yifaling, fluorosulfuron, hexachlorobenzene, benzoquinone, berberine, pentachloronitrobenzene, chlorhexidine, methyl chlorhexidine, methyl valproate; others: cycloflufenamide , ketone, xylenol, irilimole, furosemide, fenfenin and spirocycline, bisindole acetate, gramacetal acetate, gram-net-paraben (alkylbenzene sulfonate), Kasugamycin hydrochloride hydrate, dichlorophenol, pentachlorophenol and its salt, N-(4-chloro-2-nitro-phenyl)-N-ethyl-4-methyl-benzenesulfonamide, Chloramine, isopropyl rustamine, tetrachloronitrate Benzene, biphenyl, bromide, diphenylamine, midodinomycin, copper porphyrin, calcium cyclate, N-(cyclopropylmethoxyimino-(6-difluoromethoxy-2,3) -difluoro-phenyl)-methyl)-2-phenylacetamide, N'-(4-(4-chloro-3-trifluoromethyl-phenoxy)-2,5-dimethyl -phenyl)-N-ethyl-N-methylformamidine, N'-(4-(4-fluoro-3-trifluoromethyl-phenoxy)-2,5-dimethyl-phenyl )-N-ethyl-N-methylformamidine, N'-(2-methyl-5-trifluoromethyl-4-(3-trimethyldecyl-propoxy)-phenyl)- N-ethyl-N-methylformamidine and N'-(5-difluoromethyl-2-methyl-4-(3-trimethyldecyl-propoxy)-phenyl)-N- Ethyl-N-methylformamidine.

Herbicide: C1) Acetyl-CoA carboxylase inhibitor (ACC), such as cyclohexenone oxime ether, such as grass, ketone, chlorpropoxy, cyclohexane, chlorpyrifos, valerian Ketone, butyl ketone, benzophenone or pyridone; phenoxyphenoxypropionate, such as clodinafop, cyhalofop, oxacillin, oxazolyl, oxazol Cao Ling, thiazol-grass, flurazepam, flupirtine, flupirtine, flupirtine, flupirtine, clomazone, valerate, quin Orphan, quizalofop-p-ethyl or quinoxalate; or arylaminopropionic acid, such as wheat fluoromethyl ester or wheat straw fluoroisopropyl ester; C2) acetamidine lactate synthase inhibitor (ALS), such as imidazolinone, Such as acesulfame, imazaquin, imidate (imidate), imazamox, methyl imazethapyr or imazethapyr; pyrimidinyl ether, such as pyruvic acid, sodium thiosulfate, double grass Ether sodium, KIH-6127 or pyribenzoxym; sulfonamide, such as diflufenacil, flufensulfuron or sulfazinamide; or sulfonylurea, such as sulfometuron, tetrazole Sulfoxsulfuron, bensulfuron-methyl, chlorsulfuron-methyl, chlorsulfuron Ethersulfuron, cyclosulfuron, ethametsulfuron, ethoxysulfuron, pyrimidine, chlorpyrifos, oxazolidine, metsulfuron, nicosulfuron, flusulfuron, fluoride Sulfonamide, pyrazosulfuron, sulfometuron, sulfometuron, thifensulfuron, ether benzosulfuron, bensulfuron, flufensulfuron, trifluoromethanesulfuron, sulfonium sulfonate, formazan Sulfon or iodosulfuron; C3) guanamine, such as chlorpyrifos (CDAA), neoyanling, brombutachlor, chiorthiamid, grass, and ethidium (etobenzanid (benzchlomet) )), thiazolinamide, phosphine (fosamin or heptylamine; C4) auxin herbicide, such as picolinic acid, such as chlorpyrifos or gramin; or 2,4-D or herbicide; C5) auxin transport inhibitors, such as naproxil or diflupiron; C6) carotenoid biosynthesis inhibitors, such as piracetone, clomazone/dimethazone, diflufenic acid, fluroxypyr Ketone, fluroxypyr, pyrazol, benzoxazole, isoxaflutole, isoxafluzate, mesotrione, sulcotrione/chlormesulone, ketospiradox, fur Grass ketone Or killing grass; C7) enol acetone oxalic acid-3-phosphate synthase inhibitor (EPSPS), such as glyphosate or phosphinothricin; C8) glutamate synthetase inhibitor, such as bilberry ( Bilanafos/bialaphos) or glufosinate; C9) lipid biosynthesis inhibitors, such as indoline, such as samarium or mefenacet; chloroacetanilide, such as methotrexate, S-methicillin Amine, acetochlor, alachlor, butachlor, butachlor, acetochlor, dimethylamine, metazachlor, metolachlor, S-metolachlor, propylidene Amine, acetochlor, acetochlor, chlorpyrifos, methoxymethoxamine or xanthin; thiourea, such as sulbactam, chlorhexidine, oatmeal, piperidan, EPTC, pentosan, Grass chlorhexidine, chlorpyrifos, benzobendan, thiobencarb/benthiocarb, wild wheat or chlorpyrifos; or furosemide or flufensulfate; C10) mitotic inhibitors, such as urethane , such as Huangcao Ling, long-killing grass, chlorpheniramine, pingcao Dan, pronamid/propyzamid, aniline or ketone; dinitroaniline, such as fluramide, daramine, dexamethasine, Fluoride, flufenazone, huangcaoxiao, pendimethalin, alanine or trifluralin; pyridine, such as flurazepam or thiacildine; or oxacillin, diquat (DCPA) or Calyxate; C11) protoporphyrinogen IX oxidase inhibitor, such as diphenyl ether, such as acifluorfen, acifluorfen sodium, dipivoxil, bifenox, chlorhexidine (CNP) ), flufenacet, herbicide ether, flufenacetate, flufenazone, flufenacetate, flufenacil, herbicide ether, nitrofurazone or oxyfluorophene; oxadiazole, Such as propyne oxalofenone or oxacillin; cyclic quinone imine, such as oxafluridine, buffing grass, oxafluridone, oxadiazon, flumethacic acid, propargyl flufenate, flumicil , flupropacil, flumethoate, mesoxalam or thiadiazole; or pyrazole, such as ET-751.JV 485 or flufenacet; C12) photosynthesis inhibitors, such as enemies Anthraquinone, valerate or pyridinol; benzothiadiazinone, such as bentazone; dinitrophenol, such as chlorfenapyr, dinose, dilofen, tetral or DNOC; Pyridine, such as sedge fast, benzene fast, enemy grass fast or Herbaceous; urea, such as chlorbromide, chloromeron, acetonide, oxazolone, diuron, thiadiazol, oxadiazepine, oxaflurazine, isoproturon, isoxron, ligulon , thiazolyl, chlorpyrifos, pyrrolidine, methoxylated, green valley, grass, Cycloheximide or decapone; phenol, such as bromoxynil or iodobenzonitrile; chloramphenicol; triazines such as chlorpyrifos, atrazine, cyanazine, desmein, dimethamethryn , hexazinone, chlorpyrifos, chlorpyrifos, chlorpyrifos, simazine, cisplatin, methoxy oxacillin, chlorpyrifos, terbutin or chlorpyrifos; triazinone, such as oxazinone Or selezine; uracil, such as herbicidal, cyclohexidine or terpene; or bis-carbamate, such as beet or beetin; C13) synergist, such as ethylene oxide, such as herbicide Ring; C14) CIS cell wall synthesis inhibitors, such as isoxachlor or dichlorfen; C15) various other herbicides, such as dichloropropionic acid, such as thatch; dihydrobenzofuran, such as ethigen yellow; benzene Acetic acid, such as chlorfenac/fenac; or azidazine, oatmeal, disperse phosphorus, benzothiazide, flufenazone, buminafos, terazosole, cloacazone, oxazinamide , chlorsulfurin, oatmeal ester, haysulfuron, cycloheptyl ether, benzalkon, guanylon, oxadiazepine, tricyclic thiamatine, dibenzyluron, kill Net, vanilla, grass, chlorpyrifos, oxazinone, flufensulfuron, fluorbentranil, fluoxazole, chlorfluralin, lycopene, carbendazim, fluorosulfonate Valeramine, chlorpyrifos, chlorpyrifos, dichlorfen, chloramphenicol, oxaciclomefone, ketamine, piperacite, cyprodinil, ciprofloxacin, valerian, dense Caotong, gram death (CDEC), bud root, triazine fenacetamide, triazofenamid or trimethyl ketone; or an environmentally compatible salt thereof.

Nematicides or bio-nematicides: free of lysine, chlorpyrifos, annihilation, sputum, diamine, fensin, sulphur, phosphorus, pusson, phosphos, Thiazophos, acesulfame, isamimidofof, chlorfenapyr, chlorpyrifos, nicotinic thiophosphorus, methyl phosphazone, acetaminophen, phenylchlorothiophene, chlorination Bitter, Mellon, fluthiazem, 1,3-dichloropropene (dichloropropene), dithiodimethane, Weibaimu sodium, Weibaimu potassium, Weibaimu salt (all MITC generators), bromine Methane, biological soil improver (such as mustard seed, mustard seed extract), soil steam fumigation, allyl isothiocyanate (AITC), dimethyl sulfate, furfural (aldehyde).

Suitable plant growth regulators of the invention include the following: plant growth regulator: D1) anti-auxin, such as treasic acid, 2,3,5-triiodobenzoic acid; D2) auxin, such as 4-CPA , 2,4-D, 2,4-DB, 2,4-DEP, 2,4-dipropionic acid, dipropionic acid, IAA, IBA, naphthylamine, α-naphthylacetic acid, 1- Naphthol, naphthyloxyacetic acid, potassium naphthenate, sodium naphthenate, 2,4,5-T; D3) cytokinins, such as 2iP, benzyl adenine, 4-hydroxyphenylethanol, activin, corn D4) defoliant, such as calcium cyanamide, thiazide, sedum, auxin, defoliated phosphorus, methoxylated, pentachlorophenol, thiabendazole, defoliated phosphorus; D5) ethylene inhibition Agents, such as aviglycine, 1-methylcyclopropene; D6) ethylene release agents, such as ACC, vinyl hydrazine, auxin, acetonide; D7) gametocides, such as potassium oxazinone , sulphate; D8) gibberellin, such as gibberellic acid, gibberellic acid; D9) growth inhibitors, such as abscisic acid, pyrimidine alcohol, butralin, plus poly, tributyl chlorophosphonazo, chlorine Aniline, diquat, fluorohexamine, glycosaminoglycan Killing phosphine, glyphosate, pyripramine, jasmonic acid, sulphate, strong cotton, piperazine, jasmonone, aniline, tiaojiean, 2,3,5-triiodobenzene Formic acid; D10) morphines, such as chlorofluoric acid, chloromethyl sulphate, dichloromethyl sulphate, sulphate; D11) growth retardants, such as chlormequat, butyl hydrazine, chlorfluazurol, flufenacetamide , bucksin, tetracyclazole, uniconazole; D12) growth stimulators, such as brassinolide, brassinolide-ethyl, DCPTA, chlorpyrifos, carbendazim, psorale, triacontane Alcohol; D13) Plant growth regulators of unknown class, such as methamine, sulfamethoxazole, tek grass, carvone, choline chloride, benzalkonium, benzoxazinic acid, cyanamide, ring Indole propionate, cycloheximide, ciprofloxacin, propican lactone, benzoic acid ester, ethylene, furose thiourea, ethylene glycol acetal, sputum, chloroethane sulfonate Acid (holosulf), enaxin, lead arsenate, chlorhexidine, cyclohexanoic acid, pydanon, chlordamine, oxazole, anti-pivoxil.

The fertilizer can be a liquid fertilizer. The term "liquid fertilizer" means a fluid or liquid form of fertilizer containing nitrogen, phosphorus and potassium in various ratios (such as, but not limited to, 10% nitrogen, 34% phosphorus and 0% potassium) and micronutrients, commonly referred to as High phosphorus and promotes rapid and strong root growth fat.

The chemical formulations of the present invention may be in any suitable conventional form, such as emulsion concentrates (EC), suspension concentrates (SC), suspoemulsion emulsions (SE), capsule suspensions (CS), water dispersible granules (WG). ), emulsifiable particles (EG), water-in-oil emulsion (EO), oil-in-water emulsion (EW), microemulsion (ME), oily dispersion (OD), oily miscible flowable agent (OF), Oily miscible solution (OL), soluble concentrate (SL), ultra low volume suspension (SU), ultra low volume liquid (UL), dispersible concentrate (DC), wettable powder (WP) or Any technically feasible formulation in combination with an agriculturally acceptable adjuvant.

In another embodiment of the present invention, there is provided a plant seed coated with a composition comprising Bacillus amyloliquefaciens RTI301 deposited under ATCC PTA-121165 or a mutant thereof having all of its identifying characteristics Spores of pure cultures are present in an amount suitable to be beneficial for plant growth and/or to confer protection to susceptible plants to avoid pathogenic infections. Plant growth benefits and/or protection afforded by increased seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or combinations thereof Show.

The composition coated on the plant seed may comprise from about 1.0 x 10 ² CFU per seed to about 1.0 x ¹⁰⁹ CFU per seed.

Plant seeds may include, but are not limited to, monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, feed corn, rice, wheat, barley, sorghum, kale, broccoli , kale, broccoli, Brussels sprouts, broccoli, kale, mustard, broccoli, bulbous vegetables, onions, garlic, onions, fruit vegetables, pepper, tomato, eggplant, ground cherry, tree tomatoes, okra, grapes, Vanilla/spice, melon, cucumber, cantaloupe, melon, cantaloupe, squash, watermelon, pumpkin, eggplant, green leafy vegetables, lettuce, celery, spinach, parsley, chicory, legumes/vegetables (succulent and Dried beans and peas), beans, green beans, pod beans, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, peas, lentils, oil Seed crops, mustard, ramie, cotton, flax, peanuts, rapeseed, safflower, sesame, sunflower, soybean, root/tuber and bulbous vegetables, carrots, potatoes, sweet potatoes, beets, ginger, horseradish, radish, ginseng, Seeds of turnip, sugar cane, sugar beet, grass or turf grass.

In one or more embodiments, the plant seeds can include seeds of dried beans, corn, wheat, soybeans, rapeseed, rice, cucumber, pepper, tomato, squash, cotton, grass, and turfgrass.

Pathogenic infections treated by coated plant seeds may be caused by plant pathogens including, for example, but not limited to, plant fungal pathogens, plant bacterial pathogens, Botrytis, Botrytis cinerea, Botrytis cinerea Bacteria, Owens, Carrot soft rot, Owen bacteria, Fusarium oxysporum, Fusarium, Fusarium oxysporum, Fusarium graminearum, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum , Xanthomonas, Rhizoctonia solani, Xanthomonas camphora, carcinogenic lesions, P. oleracea, P. oleracea, P. xylori, rice varieties, Pseudomonas, Pseudomonas syringae tomato pathogen, Phytophthora, Phytophthora infestans, Phytophthora parasitica, Phytophthora sojae, Phytophthora capsici, Phytophthora capsici, Phytophthora strawberry, Phytophthora, eucalyptus, palm plague Mold, Phytophthora nicotiana, Rhizoctonia, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Strawberry silk nucleus Bacteria Phytophthora, Rhizoctonia solani, Rhizoctonia solani, Coccidioides, Magnaporthe oryzae, Pythium, Pythium, Pythium, Pythium, Pythium, Pythium , forest Pythium, black powder fungus, naked black powder fungus, maize black powder fungus, sugar cane whip black powder fungus, ergot fungus, ergot fungus, black fungus genus, wheat black powder fungus, smooth black Powder fungus, rice black powder fungus, wheat dwarf smut, stalk genus, soybean stalk, mildew, sphaerophyll, sphaerophyll, Helminthosporium, sclerotium, anthrax Genus or a combination thereof.

The composition applied to the plant seeds may further comprise microorganisms, organisms or chemistry One or a combination of an insecticide, fungicide, nematicide, bactericide or plant growth regulator present in an amount suitable to be beneficial for plant growth and/or to confer protection to susceptible plants to avoid pathogenic infections. . The insecticide can include diphennin. Nematicides may include sulphur phosphorus. Insecticides may include bifenin and clothianidin.

In another embodiment of the present invention, there is provided a method for achieving one or both of beneficial to plant growth or to confer protection to susceptible plants to avoid pathogenic infections, the method comprising: planting plant seeds in Suitable for growing in a medium or for regenerating a plant's nutrient pruning/tissue in a suitable growth medium, wherein the seed has been coated with a composition or nutrient pruning/tissue has been inoculated with the composition comprising ATCC PTA-121165 A biologically pure culture of the deposited B. amyloliquefaciens RTI 301 or a mutant thereof having all of its identifying characteristics, wherein the growth of the plant from the seed or nutrient pruning/tissue is beneficial and/or protection is provided to avoid pathogenic infection. Plant growth benefits and/or protection afforded by increased seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or combinations thereof Show.

In one embodiment, the method may further comprise applying a liquid fertilizer to: soil or growth medium surrounding the plant; soil or growth medium prior to seeding the plant seed in the soil or growth medium; or planting in soil or growth medium Soil or growth medium prior to plant, plant pruning, plant graft or plant healing tissue.

In one embodiment, a method of protecting plant growth by avoiding or reducing pathogenic infections while imparting protection to susceptible plants while minimizing the accumulation of resistance to treatment is provided. The method comprises delivering the first composition and the second composition to a susceptible plant at separate application and varying time intervals, wherein each of the first and second compositions is adapted to confer protection to the plant to avoid or reduce pathogenicity The amount of infection is delivered. The first composition comprises a biologically pure culture of Bacillus amyloliquefaciens RTI301 deposited with ATCC PTA-121165 or a mutant having all of its identifying characteristics. The second composition includes one or more A chemical active agent that kills fungi or bactericidal properties. In this method, the first and second compositions are delivered to plant leaves, plant stems, plant fruits, plant flowers, plant seeds, plant roots, plant pruning, plant transplants, plant healing tissues or plants at varying time intervals. One or a combination of surrounding soil or growth medium. In this method, the total amount of chemical active agent required to impart protection to avoid and/or reduce pathogenic infection is reduced and the accumulation of resistance to treatment is minimized. Plant growth benefits and/or protection afforded by increased seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or combinations thereof Show.

The first composition may further comprise one or a combination of a carrier, a surfactant, a dispersant, or a yeast extract. The yeast extract can be delivered at a rate suitable for plant growth in the range of about 0.01% to 0.2% w/w.

In a method of protecting a susceptible plant to avoid or reduce pathogenic infections that are beneficial to plant growth while minimizing the accumulation of resistance to treatment, the time interval of change can be in the range of 1 day to 10 days apart and It can be 5 to 7 days apart. The timing of the first application may depend on the particular crop and may be within the range of planting, weeks after germination of the crop, flowering, disease, or pre-exposure. Each of the first and second compositions can be delivered to a plant leaf, plant fruit or plant flower. Suitable for plants to confer protection to avoid or reduce the amount of pathogenic infections delivery may be about 1.0 × ¹⁰ 10 CFU / ha to about 1.0 × 10 ¹⁴ CFU ha Solution / Bacillus amyloliquefaciens RTI301. Delivery rates suitable for conferring protection to plants to avoid or reduce pathogenic infections may range from about 1.0 x 10 ¹⁰ CFU/ha to about 1.0 x 10 ¹⁴ CFU/ha B. amyloliquefaciens RTI 301 and about 0.01% to 0.2% w/w. Yeast extract.

One or more chemically active agents delivered to the susceptible plant at separate administrations and varying time intervals may include, for example, but not limited to, one or a combination of the following: strobilurine, triazole, dimethoate ( Flutriafol), dextran, prothiaconazole, expoxyconazole, fluopyram, tetrachloroisophthalonitrile, methyl thiophanate, copper hydroxide fungicide, EDBC-based fungicide Agent, zinc manganese Mancozeb, succinase dehydrogenase (SDHI) fungicide, chlorpyrifos, iprodione, dafoxdine or velifenate.

One or more chemically active agents delivered to the susceptible plant at separate administrations and varying time intervals may include flupirtine plus dekli and a first composition comprising RTI 301 for delivery of a replaceable tetrachloroisophthalonitrile fungicide Delivery of the agent. The plant can be a gourd and the pathogenic infection can be caused by powdery mildew.

One or more chemically active agents delivered to the susceptible plant at separate application and varying time intervals may comprise a methyl thiophanate fungicide and the delivery of the first composition comprising RTI 301 may be substituted for the delivery of the prothioconazole fungicide .

One or more chemically active agents delivered to the susceptible plant at separate application and varying time intervals may comprise a copper hydroxide fungicide and the delivery of the replaceable tetrachloroisophthalonitrile fungicide comprising the first composition of RTI301 .

In one embodiment, a product is provided comprising: a first composition comprising a biologically pure culture of Bacillus amyloliquefaci RTI301 deposited with ATCC PTA-121165 or a mutant having all of its identifying characteristics; a second composition of one or a combination of a microbial, biological or chemical insecticide, fungicide, nematicide, bactericide, herbicide, plant extract, plant growth regulator or fertilizer, wherein the first and second The compositions are packaged separately and wherein each composition is in an amount suitable to achieve one or both of beneficial to plant growth and/or to confer protection to susceptible plants to avoid pathogenic infection; and for use in a plant that would be suitable for benefiting The combined amount of the first and second compositions is delivered to the following instructions: plant leaves, plant bark, plant fruits, plant flowers, plant seeds, plant roots, plant pruning, plant grafts, plant healing tissues; Soil or growth medium around the plant; soil or growth medium before planting the seed in the soil or growth medium; or planting the plant in soil or growth medium, Was pruning the plant or plant healing of graft tissue prior to soil or growth medium.

The insecticide in this product can be pyrethroid, bifennin, tefluthrin, ξ-赛 One or a combination of Ning, organophosphate, oxyphosphonate, taosson, butyl pyrimidine, cyprofen, non-Piro, fepnini, nicotine or cotinine.

The first composition in the product may further comprise one or a combination of a carrier, a surfactant, a dispersant or a yeast extract.

In this product, the first and second compositions may be in the form of a liquid, a powder, a spreadable granule, a dry wettable powder or a dry wettable granule. In one embodiment, the first composition is in liquid form and the B. amyloliquefaciens RTI301 can be present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. In one embodiment, the first composition is in the form of a powder, a dry wettable powder, a spreadable granule or a dry wettable granule, and the B. amyloliquefaciens RTI 301 can be from about 1.0 x 10 ⁸ CFU/g to about 1.0 x. A concentration of 10 ¹² CFU/g is present. In one embodiment, the first composition is in the form of an oily dispersion and B. amyloliquefaciens RTI301 is present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml.

In one embodiment of the present invention, there is provided a composition comprising at least one of the following: isolated abundance-MA compound, isolated abundance MB compound, isolated abundance MC compound, isolated Hydroxy-prolamin MA compound, isolated dehydroxy-hydroxytoxin MB compound, isolated dehydroxy-hydroxytoxin MC compound, isolated abundance H compound, isolated dehydroxy-hydroxy-prohalogen H compound, isolated protamine I compound and isolated A hydroxytolanin I compound, which is suitable for achieving one or both of beneficial plant growth or protection of a susceptible plant to avoid a pathogenic infection having the following formula:

Wherein R is OH, n is in the range of 8 to 20, FA is linear, XOR or di-isomeric, and: for abundance MA, X ₁ is Ala, X ₂ is Thr, and X ₃ is Met; for abundance MB X ₁ is Val, X ₂ is Thr, and X ₃ is Met; for abundance MC, X ₁ is Aba, X ₂ is Thr, and X ₃ is Met; for abundance H, X ₁ is Val, X ₂ Is Thr and X ₃ is Hcy; and for alanogen I, X ₁ is Ile, X ₂ is Thr, and X ₃ is Ile; and wherein R is H, n is in the range of 8 to 20, and FA is linear, XOR or singular, and: for dehydroxylectin MA, X ₁ is Ala, X ₂ is Thr, and X ₃ is Met; for dehydroxyl-rich MB, X ₁ is Val, X ₂ is Thr, and X ₃ Is Met; for dehydroxylectin MC, X ₁ is Aba, X ₂ is Thr, and X ₃ is Met; for dehydroxylectin H, X ₁ is Val, X ₂ is Thr, and X ₃ is Hcy; For dehydroxylectin I, X ₁ is Ile, X ₂ is Thr, and X ₃ is Ile.

In another embodiment, the composition further comprises one or a combination of other isolated abundance-like and dehydroxy-norformin-like compounds listed in Table XIII, suitable for imparting benefits to plants for growth or protection of susceptible plants To avoid the amount of one or both of the pathogenic infections.

Plant growth benefits and/or protection afforded by increased seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or combinations thereof Show.

Fengyuansu-MA, Fengyuansu-MB, Fengyuansu-MC, Fengyuansu-H and Fengyuansu-I compounds and hydroxy-hydroxytoxin-MA, hydroxy-hydroxytoxin-MB, hydroxy-hydroxytoxin-MC, dehydroxy-fuyuan One or a combination of a pro-H and a dehydroxyl-richin-I compound, as well as other abundance-like and dehydroxy-norformin-like compounds, may be by suitable conditions well known to those skilled in the art, such as those described in the examples herein. Under the conditions, firstly culture RTI301 strain of Bacillus amyloliquefaciens or produce abundance-MA, amylin-MB, amylin-MC, amyloin-H and amylin-I compound, and dehydroxyl-producin-MA, dehydroxy-hydroxytoxin- Another strain of Bacillus amyloliquefaciens of MB, dehydroxyhydroxytoxin-MC, dehydroxylungin-H and dehydroxylungin-I compounds, including but not limited to cultured strains in 869 or M2 medium for 3 to 6 days To separate. The adiponectin-like and dehydroxyl-rich-like cyclic lipopeptides present in the supernatant of B. amyloliquefaciens culture can then be further isolated using methods well known to those skilled in the art. For example, the B. amyloliquefaciens culture supernatant can be acidified to pH 2 as described herein in Example 16, or with CaCl ₂ (Ajesh, K et al., 2013, "Purification and characterization of antifungal lipopeptide from a soil isolated Strain of Bacillus cereus.", Worldwide research efforts in the fighting against microbial pathogens: from basic research to technological developments. A. Mendez-Vilas (eds.) pp. 227-231) or NH ₄ SO ₄ (Kim, SH et al. , 2000, Biotechnol Appl Biochem . 31 (Pt 3): 249-253), with or without organic extraction steps (Kim, PI et al, 2004, J Appl Microbiol. 97(5): 942-949), Combinations such as various phase separation formats include, but are not limited to, direct liquid dispensing, membrane ultrafiltration, and foam fractionation (Baker, SC et al, 2010, Adv Exp Med Biol. 672:281-288).

In one embodiment, amylin-MA, amylin-MB, amylin-MC, amylin-H, and a protoxin-I compound and a dehydroxyl-producin-MA, a dehydroxyl-producin-MB, a dehydroxy-hydroxytoxin -MC, dehydroxyhydroxytoxin-H and dehydroxylungin-I compounds, and one or a combination of other abundance-like and dehydroxy-norformin-like compounds listed in Table XIII can produce self-producing starch spores of such compounds The biologically pure culture of the Bacillus strain is isolated.

In one embodiment, amylin-MA, amylin-MB, amylin-MC, amylin-H, and a protoxin-I compound and a dehydroxyl-producin-MA, a dehydroxyl-producin-MB, a dehydroxy-hydroxytoxin -MC, dehydroxylectin-H and dehydroxylectin-I compounds, and one or a combination of other abundance-like and dehydroxy-norformin-like compounds listed in Table XIII can be deposited with ATCC PTA-121165 The biologically pure culture of Bacillus amyloliquefaciens RTI301 was isolated.

In one embodiment, an extract of a biologically pure culture of a B. amyloliquefaciens strain is provided, the extract comprising amylin-MA, amylin-MB, abundance-MC, Fengyuan 素-H and abundance-I compounds and hydroxy-hydroxytoxin-MA, hydroxy-hydroxytoxin-MB, hydroxy-hydroxytoxin-MC, hydroxy-hydroxytoxin-H and hydroxy-hydroxytoxin-I compounds and Table XIII One or a combination of other abundances and dehydroxy-flavone-like compounds.

In one embodiment, an extract of a biologically pure culture of Bacillus amyloliquefaciens RTI301 deposited under ATCC PTA-121165 is provided, the extract comprising amylin-MA, amylin-MB, abundance-MC, Fengyuan素-H and abundance-I compounds and hydroxy-hydroxytoxin-MA, hydroxy-hydroxytoxin-MB, hydroxy-hydroxytoxin-MC, hydroxy-hydroxytoxin-H and hydroxy-hydroxytoxin-I compounds and Table XIII One or a combination of other abundances and dehydroxy-flavone-like compounds.

Including abundance-MA, amylin-MB, abundance-MC, abundance-H and abundance-I compounds and dehydroxy-hydroxytoxin-MA, dehydroxy-prolamin-MB, dehydroxy-hydroxytoxin-MC, dehydroxyl The composition of at least one of the abundance-H and the dehydroxyl-richin-I compound and optionally one or a combination of other isolated abundance-like and dehydroxy-norformin-like compounds may further comprise microbial, biological or chemical insecticidal One or a combination of agents, fungicides, nematicides, bactericides, herbicides, plant extracts, plant growth regulators or fertilizers, suitable for beneficial to plant growth and/or protection of susceptible plants to avoid The amount of pathogenic infection exists.

The fungicide may include an extract from white lupin. The fungicide can include a BLAD polypeptide. The BLAD polypeptide may be a fragment of a naturally occurring seed storage protein from sweet lupin (white lupin) that acts on a susceptible fungal pathogen by causing damage to the fungal cell wall and disrupting the inner cell membrane. The fungicide may comprise about 20% BLAD polypeptide.

Including abundance-MA, amylin-MB, abundance-MC, abundance-H and abundance-I compounds and dehydroxy-hydroxytoxin-MA, dehydroxy-prolamin-MB, dehydroxy-hydroxytoxin-MC, dehydroxyl The composition of at least one of the abundance-H and the dehydroxyl-prohalin-I compound may be in the form of a liquid, an oily dispersion, a powder, a dispersible granule or a dry wettable granule.

In one embodiment, a method is provided for benefiting plant growth and/or conferring protection against phytopathogenic infections, comprising: comprising an effective amount comprising isolated proemin-MA, amylin-MB, amylin-MC, Fengyuansu-H and adiponectin-I compounds and dehydroxyl-precipitin-MA, hydroxy-hydroxytoxin-MB, hydroxy-hydroxytoxin-MC, hydroxy-hydroxytoxin-H and dehydroxy-hydroxy-proto-I compounds and other separations An extract or composition of one or a combination of aureus-like and dehydroxy-norformin-like compounds is applied to the soil surrounding the roots or roots of the plant or fruit or plant to benefit plant growth and/or to confer protection against phytopathogenic infections. . Plant growth benefits and/or protection afforded by increased seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or combinations thereof Show.

In the effective amount, the isolated abundance of protoxin-MA, abundance-MB, abundance-MC, abundance-H and amylin-I compound and dehydroxyl-proto- s-MA, hydroxy-hydroxytoxin-MB, dehydroxyl In the method of extracting or extracting a prostaglandin-MC, a dehydroxyl-prohalin-H and a dehydroxyl-prohalin-I compound and one of the other isolated abundance-like or dehydroxy-norformin-like compounds, the plant may comprise For example, monocots, dicots, grains, corn, sweet corn, popcorn, seed corn, silage corn, feed corn, rice, wheat, barley, sorghum, asparagus, berries, blueberries, blackberries, raspberries, roganberries , cranberry, cranberry, gooseberry, elderberry, black currant, cranberry, raspberry, kale, broccoli, kale, broccoli, Brussels sprouts, broccoli, kale, mustard, bulb Cabbage, melon, cucumber, cantaloupe, melon, cantaloupe, squash, watermelon, pumpkin, eggplant, bulbous vegetables, onions, garlic, onions, citrus, oranges, grapefruit, lemon, red orange, orange grapefruit, Grapefruit, fruits and vegetables , pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, leafy vegetables, lettuce, celery, spinach, parsley, chicory, legumes/vegetables (succulent and dry beans and peas) , beans, green beans, pod beans, shell beans, soybeans, dried beans, back beans, lima beans, peas, chickpeas, pea petals, lentils, oilseed crops, canola, nettle, coconut, cotton, Asian Hemp, oil palm, olive, peanut, rapeseed, safflower, sesame, sunflower, soybean, pear fruit, apple, jellyfish, pear, alfalfa, summer flower hawthorn, root/tuber and bulbous vegetables, carrot, potato, sweet potato, Cassava, beet, ginger, horseradish, radish, ginseng, turnip, stone fruit, apricot, cherry, nectarine, peach, plum, plum, strawberry, nut, almond, pistachio, pecan, walnut, hazelnut, chestnut, cashew, Beech nuts, walnuts, macadamia nuts, kiwi, bananas, (blue) agave, grass, turf grass, ornamental plants, poinsettia, hardwood pruning, chestnut, oak, maple, sugar cane or beet.

In the effective amount, the isolated abundance of protoxin-MA, abundance-MB, abundance-MC, abundance-H and amylin-I compound and dehydroxyl-proto- s-MA, hydroxy-hydroxytoxin-MB, dehydroxyl Pathogenic infection in a method of extracting or extracting a protosulin-MC, a dehydroxyl-prohalin-H and a dehydroxyl-proto-I compound and other isolated abundance-like or dehydroxy-norformin-like compounds May be caused by plant pathogens including, for example, plant fungal pathogens, plant bacterial pathogens, rust fungi, Botrytis, Botrytis cinerea, Botrytis cinerea, Owenium, Carrot soft rot, Erwinia , Dick's genus, D. dandelion, black smut, Agrobacterium, Agrobacterium tumefaciens, Xanthomonas, P. sphaeroides, Xanthomonas campestris, carrot-induced disease, peach and plum Xanthomonas, Monochamus humilis, Rice Variety of Xanthomonas oryzae, Saccharomyces, Saccharomyces cerevisiae, Carterella, phloem, Fusarium, Fusarium oxysporum, Fusarium graminearum Fusarium, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Sclerotinia, S. sclerotiorum, Sclerotinia sclerotiorum, Sclerotinia sclerotiorum, Cercospora, H. genus, Grape white powder Pathogens (powder disease), sclerotium sclerotium (powder disease), white stalk single sac, clams, single sac, stalk, sphaeroides, Phytophthora Genus, fine-stranded chain bacteria, onion chain bacteria, Alternaria alternata, Alternaria alternata, Alternaria, Pseudomonas, Pseudomonas syringae, tomato Phytophthora Phytophthora infestans, Phytophthora infestans, Phytophthora sojae, Phytophthora sojae, Phytophthora capsici, Phytophthora capsici, Phytophthora infestans, Phytophthora, eucalyptus, Phytophthora palmatum, Phytophthora nicotiana Phytophthora, Puccinia genus, Phakopsora pachyrhizi, P. sylvestris, Aspergillus, Scutellaria, Aspergillus niger, Puccinia rot, Bean rust, Cladosporium, Multi-spore Mildew, Rhizopus, Rhizopus, Penicillium, Rhizoctonia, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia cerealum, Violet Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia sclerophylla, Rhizoctonia solani, Coccidioides, Magnaporthe oryzae, Glomus, Trichoderma, Fiji ball Cavity (black banana leaf spot), apple spotted bacterium, lemon bacterium, rice blast, rice blast, chain genus, brown rot, blueberry fruit pathogen, stone fruit chain Phytophthora, anthrax, anthrax, anthrax, anthracnose, anthrax, genus, citrus, genus, genus, genus, genus, rust, plant Phytophthora rust, Shield genus, Phytophthora, Helminthosporium, Phytophthora, Phytophthora, Grape canker, Clostridium, Phytophthora, Phytophthora velutipes, Phytophthora, Phytophthora, Phytophthora capsici, Polygonum genus, Polyporus genus, Phytophthora, Pythium Genus, ultimate Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Pseudomonas, Escherichia coli, Verticillium, Sphaerotheca, Naked black powder fungus, maize black powder fungus, sugarcane whip black powder fungus, ergot fungus, ergot fungus, sphaerotheca fuliginea, wheat black powder fungus, smooth black powder fungus, rice black powder fungus, wheat Dwarf sphaeroides, Pleurotus ostreatus, soybean stems, variegated shoots, cabbage stems, Helminthosporium, sclerotium, anthracnose, genus, genus, barley Phytophthora, genus Helminthosporium, Helminthosporium, Helminthosporium longum, Helminthosporium fulvus, Helminthosporium serrata and Helminthosporium hirsutum or a combination thereof.

Instance

The following examples are included to provide guidance for the general practitioner to implement representative embodiments of the subject matter disclosed herein. In view of the present invention and the general technical level of the present invention, it is to be understood by those skilled in the art that the following examples are intended to be illustrative only and that many variations, modifications and changes may be employed without departing from the scope of the presently disclosed subject matter.

Example 1 Bacillus amyloliquefaciens

Plant-associated strains referred to herein as RTI301 were isolated from vine roots soil grown in Wynyard, Long Island, New York. The 16S rRNA and rpoB genes of the RTI301 strain were sequenced and subsequently compared to other known bacterial strains in the NCBI and RDP databases using BLAST. To determine the 16S RNA partial sequence of RTI301 (SEQ ID NO: 1) and the 16S rRNA of B. amyloliquefaciens strain NS6 (KF177175), B. amyloliquefaciens strain FZB42 (NR_075005) and Bacillus subtilis subsp. subtilis strain DSM 10 (NR_027552) The gene sequence is identical. The rpoB gene sequence of RTI301 (SEQ ID NO: 2) was also determined with B. amyloliques plant subspecies TrigoCor1448 (CP007244) (99% sequence identity; 3 base pair difference), B. amyloliquefaciens plant subspecies AS43. 3 (CP003838) (99% sequence identity; 7 base pair differences), B. amyloliquefaciens CC178 (CP006845) (99% sequence identity; 8 base pair differences) and B. amyloliquefaciens FZB42 (CP000560) The same genes in the (99% sequence identity; 8 base pair differences) have sequence similarity. The RTI301 strain was identified as B. amyloliquefaciens. The sequence difference of the rpoB gene at the DNA level indicates that RTI301 is a new strain of B. amyloliquefaciens. Bacillus amyloliquefaciens RTI301 was deposited with the American Type Culture Collection (ATCC) (Manassas, Virginia, USA) on April 17, 2014 under the terms of the Budapest Treaty on the International Recognition of Microorganisms for the Purposes of Patent Procedures and has the patent registration number PTA- 121165.

Example 2 Genes related to lantibiotic biosynthesis in Bacillus amyloliquefaciens RTI301 strain

Further sequence analysis of the genome of B. amyloliquefaciens RTI 301 strain revealed that this strain has genes associated with lantibiotic biosynthesis in other closely related B. amyloliquefaciens strains without homologs. This display is shown in Figure 1 and shows the genes surrounding the lantibiotic biosynthesis operon found in B. amyloliquesii RTI301 Schematic diagram of the group structure. In Figure 1, the top arrow group indicates the protein coding region of the RTI301 strain, indicating relative transcriptional orientation. For comparison, the corresponding regions of the two B. amyloliquefaciens reference strains FZB42 and TrigoCor1448 are shown below the RTI301 strain. The genes in the lantibiotic synthesis operon in the RTI 301 strain were initially identified using RAST and their identity was then refined using BLASTp. The degree of amino acid identity of the protein encoded by the gene of the RTI 301 strain compared to the two reference strains is indicated by both the shaded degree of the representative arrow and the percent identity indicated within the arrow. It can be observed from Figure 1 that there is a high degree of sequence identity in the region around the lantibiotic synthesis operon in genes from three different strains, and only the degree of sequence identity in the lantibiotic synthesis operon is low (ie, lantibiotic synthesis) Less than 40% in the operon but greater than 99% in the surrounding area). BLASTn analysis of this cluster was performed against NCBI's non-redundant (nr) nucleotide library, and the analysis showed high homology to the 5' and 3' flanking regions of B. amyloliquefaciens strains (similar to Figure 1). Medium height similarity %). However, the lanthiopeptide biosynthetic cluster was unique to RTI301 and no significant homology to any previously sequenced DNA in the NCBI nr library was observed. Therefore, this lantibiotic synthesis operon is a unique feature of B. amyloliquefaciens strain RTI301.

Example 3 Growth of Bacillus amyloliquefaciens RTI301 isolate in wheat

The effect of application of bacterial isolates on early plant growth and vigor of wheat was determined. The experiment was carried out by inoculating surface-sterilized germinated wheat seeds in a suspension of about 2 x 10 ⁷ CFU/ml bacteria in the dark at room temperature for 2 days (also in the absence of bacteria). Subsequently, the inoculated and control seeds were planted in 6" pots filled with sand. 10 seeds were planted per pot and 1 pot was planted per treatment, and alternately watered and modified Hoagland's solution as needed. The pots were incubated for 13 days under a natural light/dark cycle at approximately 21 ° C in a laboratory window sill, at which time the plants were recovered and the parameters were measured. The dry weight was determined as the total weight of each 9 plants, and the results were determined by Bacillus amyloliquefaciens. The total average plant dry weight of the plants inoculated with the RTI 301 strain was equal to 36.1 mg, compared to the weight of the uninoculated control equal to 33.38 mg and the dry weight increased by 8.1% compared to the uninoculated control. Photographs of the plants extracted after 13 days of growth Shown in Figure 2. Figure 2A shows plants inoculated with RTI301 and Figure 2B shows control plants.

In addition, another experiment was conducted to demonstrate the beneficial effects of the B. amyloliquefaciens RTI301 strain on early growth in wheat. The experiments were carried out by inoculating surface-sterilized germinated wheat seeds under shaking for 10 days at room temperature in a suspension of 10 ⁺⁸ CFU/ml bacteria. Subsequently, the inoculated seeds were planted in a 1 gallon filled pot of PROMIX BX treated with lime to pH 6.5. For each treatment, 12 seeds were sprinkled with 9 pots, and the seeds were planted at a depth of 2.5 cm. The pots were incubated in a greenhouse at 22 ° C under 14/10 hr light and dark cycles and watered twice as needed. A photograph of the plants after 28 days of growth is shown in Figure 3. Figure 3A shows plants inoculated with RTI301 and Figure 3B shows control plants.

Example 4 Antimicrobial properties of Bacillus amyloliquefaciens RTI301 isolates

The antagonistic ability of the RTI301 isolate against major plant pathogens was measured in disk assays. Disc analysis against antagonism of plant fungal pathogens was evaluated by growing bacterial isolates and pathogenic fungi side by side on a 869 agar plate at 4 cm distance. The plates were incubated at room temperature and examined for growth behavior for up to two weeks, such as growth inhibition, status occupancy or no effect. In the case of screening for antagonistic properties against bacterial pathogens, the pathogen first spreads to the lawn on the 869 agar plate. Subsequently, a 20 μl aliquot of the culture of each isolate was spotted on a culture plate. The plate was incubated at room temperature and the zone of inhibition in the lawn around the site where RTI301 had been applied was periodically checked for up to two weeks. Table I below shows an overview of antagonistic activity. The RTI 301 strain exhibits antagonistic properties against a wide range of phytopathogenic microorganisms.

+++ very strong activity, strong ++ activity, + activity, weak activity -- no activity observed

Example 5 Performance traits of Bacillus amyloliquefaciens RTI301 isolates

In addition to the positive effects on plant growth and antagonistic properties, various performance traits of Bacillus amyloliquefaciens RTI301 strains were also measured and the data are shown in Table II below. Analyze according to the procedure described in the text below Table II.

+++ is very strong, ++ strong, + some, +-weak, - not observed

Acid and acetoin test. 20 μl of 869 medium containing bacterial culture was transferred to 1 ml of methyl red-Voges Proskauer medium (Sigma Aldrich 39484). The culture was incubated at 30 ° C, 200 rpm for 2 days. 0.5 ml of the culture was transferred, and 50 μl of 0.2 g/l methyl red was added. Red indicates acid production. The remaining 0.5 ml of the culture was mixed with 0.3 ml of 5% alpha-naphthol (Sigma Aldrich N1000) followed by 0.1 ml of 40% KOH. Samples were interpreted after 30 minutes of incubation. Red development indicates the occurrence of acetoin. For the acid and acetoin tests, the uninoculated medium was used as a negative control (Sokol et al., 1979, Journal of Clinical Microbiology. 9: 538-540).

Indole-3-acetic acid . 20 μl of the rich 869 medium containing the bacterial culture was transferred to 1 ml of 1/10 869 medium (Sigma Aldrich T0254) supplemented with 0.5 g/l of tryptophan. Cultures were incubated for 4-5 days at 30 ° C, 200 RPM in the dark. The sample was centrifuged and 0.1 ml of the supernatant was mixed with 0.2 ml of Salkowski's Reagent (35% perchloric acid, 10 mM FeCl3). After incubation for 30 minutes in the dark, the pink-producing sample was recorded as positive for IAA synthesis. IAA dilution (Sigma Aldrich I5148) was used as a positive comparison; uninoculated medium was used as a negative control (Taghavi et al, 2009, Applied and Environmental Microbiology 75: 748-757).

Phosphate Dissolution Test . Bacteria were inoculated on Picokovskaya (PVK) agar medium consisting of 10 g of glucose per liter, 5 g of calcium triphosphate, 0.2 g of potassium chloride, 0.5 g of ammonium sulfate, 0.2 g of chlorination. Sodium, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of yeast extract, 2 mg of manganese sulfate, 2 mg of iron sulfate and 15 g of agar, pH 7, and autoclaved. The clearance zone indicates phosphate lytic bacteria (Sharma et al, 2011, Journal of Microbiology and Biotechnology Research 1: 90-95).

Chitinase activity . Add 10% wet weight gelatinous shell to the modified PVK agar medium (10g glucose, 0.2g potassium chloride, 0.5g ammonium sulfate, 0.2g sodium chloride, 0.1g magnesium sulfate per liter) Heptahydrate, 0.5 g yeast extract, 2 mg manganese sulfate, 2 mg iron sulfate and 15 g agar, pH 7, autoclaved). Bacteria are plated on these hull disks; the clearance zone indicates chitinase activity (NKSMurthy and Bleakley., 2012. "Simplified Method of Preparing Colloidal Chitin Used for Screening of Chitinase Producing Microorganisms". The Internet Journal of Microbiology .10 ( 2)).

Protease activity . Bacteria were spread on 869 agar medium supplemented with 10% milk. The clearance zone indicates the ability to decompose proteins, indicating protease activity (Sokol et al., 1979, Journal of Clinical Microbiology. 9: 538-540).

Example 6 Antagonistic effects of Brassica glabrata RTI301 and Botrytis cinerea RTI301

Soybeans were studied in a greenhouse to determine the ability of the B. amyloliquefaciens RTI301 strain to prevent and/or alleviate the effects of the plant pathogen bean rust (Nicky rust fungus) and the plant pathogen Botrytis cinerea.

In the first set of experiments, different formulations of Bacillus amyloliquefaciens RTI301 strain were tested for foliar application to control the plant pathogen Phytophthora rust and Botrytis cinerea. The experimental design and formulation are as follows:

Formulation :

The spores of Bacillus amyloliquefaciens RTI301 in waste broth (SFB) were diluted about 100 times in water and applied to the leaves at an application rate of 1 x 10 ⁸ cfu/ml.

The spores of Bacillus amyloliquefaciens RTI301 in waste broth (SFB) were diluted about 100-fold in water and yeast extract was added and applied to the leaves at an application rate of 1 x 10 ⁸ cfu/ml and about 0.2% yeast extract.

The SERENADE OPTIMUM (BAYER CROP SCIENCE, INC) product was applied at a rate of application of 1 x 10 ⁸ cfu/ml of B. subtilis strain QST713 spores.

HORIZON (HORIZON AG-PRODUCTS) product is applied at 50g a.i./ha Rate (Dickley) applied.

The BRAVO WEATHER STIK (SYNGENTA CROP PROTECTION, INC) product was applied at an application rate of 500 g a.i./ha (tetrachloroisophthalonitrile).

TACTIC (LOVELAND PRODUCTS, INC) products are included in all of the above formulations at a concentration of 0.1875% v/v.

Treatment application method :

A track sprayer was used to inoculate 21 day old common legumes (with two trifolials) with various top treatments with a single top nozzle (TeeJet SS8001E flat fan) and pressure = 276 kPa (40 psi). The nozzle height was 36 cm (14") above the leaves of the legumes. The application volume was 200 L/ha and the number of replicates in the experiment was equal to six times. The single treatment of the treated plants and the control plants did not receive any treatment.

Infection rate :

One day after the treatment, the test plants were infected with bean rust (Nicky rust) at a vaccination rate of 200 k conidia/ml.

Nine days after infection with bean rust (Nicky rust), the disease control % of each of the following was evaluated: RTI301 spores ("RTI301+1% SFB") in 100% of the waste broth diluted with water, diluted 100 times with water. In the yeast solution, RTI301 spore plus yeast extract ("RTI301+1% SFB+ yeast extract"), BRAVO WEATHER STIK, HORIZON and SERENADE OPTIMUM according to the above application rates. An untreated control (water only) produced 28% of the disease.

The experimental results are shown in Table III. The results indicate that for the RTI301 strain, the addition of the yeast extract resulted in an increase in disease control of about 40% compared to the RTI301 strain administered without the addition of the yeast extract. When administered at the same application rate (i.e., 1 x 10 ⁸ cfu/ml), the RTI301 + 1% SFB + yeast extract exhibited a disease control amount similar to that observed for SERENADE OPTIMUM, even though the amount of SFB in the RTI301 formulation Relatively low, it is 1%, and SFB is expected to contain secretory compounds with antifungal activity.

A similar result was observed using the same formulation and experimental design listed above in pepper plants, which measured the disease control of bacillus blight spores caused by the pathogen Botrytis cinerea (data not shown) .

The following experiment describes the disease control of RTI301 against bean rust caused by the phytopathogen Phytophthora rust. The experimental design and formulation are as follows:

Formulation :

The B. amyloliquefaciens RTI301 spores were diluted about 100-fold in water in spent broth (SFB) and yeast extract was added and applied to the leaves at an application rate of 1 x 10 ⁸ cfu/ml and about 0.2% yeast extract.

The SERENADE OPTIMUM (BAYER CROP SCIENCE, INC) product was applied at an application rate of 1 x 10 ⁸ cfu/ml and 4 x 10 ⁸ cfu/ml.

TACTIC (LOVELAND PRODUCTS, INC) was applied to all formulations at a concentration of 0.1875% v/v.

HORIZON (HORIZON AG-PRODUCTS) was applied at an application rate of 50 g a.i./ha (Dickley).

Tetrachloroisophthalonitrile was applied at an application rate of 500 g a.i./ha.

Treatment application method :

A track sprayer was used to inoculate 21 day old legumes (with two trifolials) with various top treatments with a single top nozzle (TeeJet SS8001E flat fan) and pressure = 276 kPa (40 psi). The nozzle height was 36 cm (14") above the leaves of the soybean plant. The application volume was 200 L/ha and the number of replicates in the experiment was equal to six times. The single treatment of the treated plants and the control plants did not receive any treatment.

Infection rate:

One day after the treatment, the test plants were infected with bean rust (Nicky rust) at a vaccination rate of 200 k conidia/ml. Ten days after infection with soybean rust (Rh. solani), the disease control % of each of the following were evaluated: RTI301 spores (administered at 1 × 10 ⁸ cfu/ml) and SERENADE OPTIMUM (1 × 10) in waste broth (SFB) ⁸ cfu/ml application), SERENADE OPTIMUM (administered at 4 x 10 ⁸ cfu/ml), Dekli (administered at 50 g ai/ha) and tetrachloroisophthalonitrile (administered at 50 g ai/ha). TACTIC (administered at 0.1875%) was also included as a control for all formulations. The check control produced 23% of the disease. The experimental results are shown in Table IV below and Figure 4.

In another similar experiment, soybean plants were studied in a greenhouse to determine the ability of the B. amyloliquefaciens RTI301 strain to prevent and/or alleviate the effects of plant pathogen bean rust (Nicky rust fungus) under varying pathogen inoculation rates. Formulations and treatment methods used The same was stated in previous experiments.

Infection rate:

One day after the treatment, the test plants were infected with soybean rust (Nicky rust) at an inoculation rate ranging from 50 k to 300 k conidia/ml. Ten days after infection with soybean rust (Nicky rust), the disease control % of each of the following: RTI301 spores (administered at 1 × 10 ⁸ cfu/ml) and SERENADE OPTIMUM (with 1 ×) in waste broth (SFB) were evaluated. 10 ⁸ cfu/ml and 4×10 ⁸ cfu/ml application) and HORIZON (dickley; administered at 50 g ai/ha). TACTIC (administered at 0.1875%) was also included as a control for all formulations. The % of the disease in the control was 50k=6%, 100k=6%, 150k=15%, 200k=15%, and 300k=7%. The experimental results are shown in Table V below and Figure 5.

Example 7 Antagonistic effect of Bacillus amyloliquefaciens RTI301 on cucumber powdery mildew in cucumber in a field trial in Florida

The study was conducted in a cucumber field trial to determine the ability of B. amyloliquefaciens RTI301 to prevent and/or alleviate the powdery mildew effect of the plant pathogen cucurbit disease.

Overall, each product was applied 6 times with a time interval between 5 and 7 between applications. day. The timing of the first application depends on the particular crop and is within the range of the crop, the weeks after the crop is germinated, the flowering, the disease, or the expected disease.

Formulation:

SERENADE OPTIMUM (BAYER CROP SCIENCE, INC ) to 1400g / ha, corresponding to 1.8 × 10 ⁺¹³ CFU / ha, of administration.

The Bacillus amyloliquefaciens RTI301 spore is in the waste mash (SFB) and the yeast extract is added, and the application rate to the leaves corresponds to the yeast extract based on the application rate of about 1400 g/ha and about 0.01% to 0.2%, The same colony forming units/ha (CFU/ha) are recommended by SERENADE OPTIMUM.

TACTIC (LOVELAND PRODUCTS, INC) was applied at a concentration of 0.1875% v/v and was included in all treatments.

LUNA EXPERIENCE (BAYER CROP SCIENCE, INC) was applied at an application rate of 500 g a.i./ha (fluopyram and dextrobacter fungicide).

BRAVO WEATHER STIK (SYNGENTA CROP PROTECTION, INC) was applied at an application rate of 2240 g a.i./ha (tetrachloroisophthalonitrile).

The experimental design is as follows: untreated control, RTI301+TACTIC, SERENADE OPTIMUM+TACTIC, and BRAVO WEATHER STIK+LUNA EXPERIENCE+TACTIC.

Treatment application method:

Six applications were performed as described above. The sprayer was set to deliver 30 gallons per acre. Individual plots were sprayed at a ground speed of 4 mph using a CO ₂ backpack sprayer with a flat fan nozzle (type 8004) and each nozzle was 18 inches apart.

Disease score: For each treatment, the average degree of disease severity of the 5 leaves of the plants was evaluated. Evaluation of crop response and disease control of powdery mildew from four plots of each treatment. After each application, the test will be rated just prior to the next application. Powdery mildew comes from natural infections.

The results of the experiment are shown in Table VI below and indicate that the control of powdery mildew in cucumber is similar when applied at the same application rate as a separate biofungicide compared to SERENADE OPTIMUM.

Example 8 Antagonistic effect of Bacillus amyloliquefaciens RTI301 on tomato bacterial spot disease (Homomonospora) in a tomato field trial

Studies were conducted in a tomato field trial to determine the ability of B. amyloliquefaciens RTI301 to prevent and/or alleviate the effects of the plant pathogen tomato bacterial spot disease (Homomonospora).

A total of 4 applications were applied to the crop with a time interval between administrations of 5 to 7 days. In the case of Scheme 4 in which the bioactive component is combined with the administration of the chemically active ingredient, the first and third administrations are carried out with the biologically active ingredient, and the second and fourth administrations are carried out with the chemically active ingredient.

Formulation:

SERENADE OPTIMUM an application rate of 1400g / ha of, corresponding to 1.8 × 10 ⁺¹³ CFU / ha administration.

Bacillus amyloliquefaciens RTI301 spores are in waste mash (SFB) and yeast extract is added, and the application rate to the leaves corresponds to an application rate based on 1400 g/ha and about 0.01% to 0.2% yeast extract for SERENADE The same colony forming units/ha are recommended by OPTIMUM.

TACTIC (LOVELAND PRODUCTS, INC) is applied at a concentration of 0.1875% v/v use.

KOCIDE 3000 (DUPONT USA) was applied at an application rate of 1850 g a.i./ha (copper hydroxide fungicide).

BRAVO WEATHER STIK (SYNGENTA CROP PROTECTION, INC) was applied at an application rate of 2240 g a.i./ha (tetrachloroisophthalonitrile).

The experimental design is as follows: untreated control, RTI301+TACTIC, SERENADE OPTIMUM+TACTIC, SERENADE OPTIMUM+KOCIDE 3000+TACTIC and BRAVO WEATHER STIK+KOCIDE 3000+TACTIC.

Treatment application method:

Four separate treatment applications were delivered to the crop with a time interval between 5 and 7 for each application. The sprayer was set to deliver 40 gallons per acre. Individual plots were sprayed at 3 mph using a CO ₂ backpack sprayer with a conical nozzle and each nozzle was 12 相 apart. The carrier that transports the chemical is water mixed in a 2.5 liter bottle.

The severity of the disease was measured by assessing the canopy. The average % of disease severity was assessed in the middle of each treated plant. The percentage of disease control was based on 100% of diseased, untreated control plants. The data is shown in Table VII below. Treatments include: untreated control, RTI301+TACTIC, SERENADE OPTIMUM+TACTIC, SERENADE OPTIMUM+KOCIDE 3000+TACTIC, and BRAVO WEATHER STIK+KOCIDE 3000+TACTIC.

The experimental results are shown in Table VII below. The RTI301+TACTIC, SERENADE OPTIMUM+TACTIC and BRAVO WEATHER STIK+KOCIDE 3000+TACTIC treatments were used to control tomato bacterial spot disease (Homomonospora) in tomatoes equally well.

Table VII. Control Results of Bacillus amyloliquefaciens RTI301 on Tomato Bacterial Spot Disease (Homomonospora) in Tomatoes Compared with SERENADE OPTIMUM and Other Chemical Active Agents

Example 9 Antagonistic effects of Bacillus amyloliquefaciens RTI301 on plant pathogens in field trials

Studies in wheat, soybean, corn, cucumber, and tomato field trials in Georgia to determine that B. amyloliquefaciens RTI301 strain prevents and/or alleviates plant pathogens caused by Pseudomonas aeruginosa, wheat scab, soybean rust , the ability of corn rust, cucumber powdery mildew and tomato bacterial spot disease.

For each of RTI 301 and SERENADE OPTIMUM, SERENADE OPTIMUM used an application rate of 1400 g/ha, and RTI 301 was applied using an application rate corresponding to the same colony forming unit/ha for SERENADE OPTIMUM based on an application rate of 1400 g/ha.

One or more treatments are administered to the crop with a time interval between administrations of 5 to 7 days. The number of applications and the timing of the first application are dependent on the particular crop and are within the range of the time of planting, weeks after the germination of the crop, the onset of flowering, the onset of disease, or the onset of the expected disease. The application sprayer was set to deliver 20-30 gallons per acre (1891/ha). Individual plots were sprayed at 3-4 mph using a CO ₂ backpack sprayer (Model 8003 or 8004) with double flat fan nozzles.

For wheat scab, the plants are applied in a single treatment when the crop begins to flower. After three days of treatment, the plants were artificially infected with the pathogen of the scab, Zea mays (also known as Fusarium graminearum). Disease severity was measured by determining the percentage of wheat head affected by scab (white spikes). The percent control of disease severity is based on considering that the % disease severity in untreated control plants is 100%. The data is shown in Table VIII below.

For soybean rust, the plants were applied 6 times. Initial application in growth R1 Segment delivery. This test has a natural infection. The severity of the disease was measured by assessing the stem and canopy. In particular, six 2呎 segments per plot were evaluated as subsamples and the severity was determined by assessing the percentage of leaves showing rust symptoms from the entire canopy. The percent control of disease severity is based on considering that the % disease severity in untreated control plants is 100%. The data is shown in Table VIII below.

For corn rust, plants were treated 6 times. This test has a natural infection. The severity of the disease was measured by assessing the canopy. Six 2 呎 sections of each plot were evaluated as subsamples and the severity was determined by assessing the percentage of leaves showing rust symptoms from the entire canopy. The severity of the disease was scored as the number of hits per 1 m column. The crop response from the four plots of each treatment and the disease control of corn rust were assessed. The percent control of disease severity is based on considering that the % disease severity in untreated control plants is 100%. The data is shown in Table VIII below.

For cucumber powdery mildew, the plants were applied 6 times after the crop was germinated for about 3 weeks. The mean % disease severity was assessed in 5 leaves of each treated plant. Evaluation of crop response and disease control of powdery mildew from four plots of each treatment. After each application, the test was rated just prior to the next application. Powdery mildew comes from natural infections. The percent control of disease severity is based on considering that the % disease severity in untreated control plants is 100%. The data is shown in Table VIII below.

For tomato bacterial spot disease, the plants were applied 6 times. Disease severity was measured by observing in the canopy and assessing the percentage of affected leaves. The percent control of disease severity is based on considering that the % disease severity in untreated control plants is 100%. The data is shown in Table VIII below.

Table VIII. Results of disease control of B. amyloliquefaciens RTI301 against wheat scab, soybean rust, corn rust, cucumber powdery mildew and tomato bacterial spot disease compared to SERENADE OPTIMUM

* The percentage control of disease severity is based on considering that the % disease severity in untreated control plants is 100%.

⁺ PMD: Powdery mildew

RTI301 better controls wheat scab and soybean rust than SERENADE OPTIMUM, as measured by untreated control %. RTI301 is comparable to SERENADE OPTIMUM in controlling cucumber powdery mildew, corn rust, and tomato bacterial spot, as measured by untreated control %. In the treatment administration protocol, RTI301 was not indicated to have a negative crop response.

Example 10 Antagonistic effect of Bacillus amyloliquefaciens RTI301 on sudden death syndrome in soybean

Experiments were conducted in soybean to determine the ability of B. amyloliquefaciens RTI301 strain to prevent and/or alleviate the effects of sudden death syndrome in soybean. Experiments were performed using RTI301 spores as described below. For the experiment, the strain formed spores in 2XSG in a 14L fermentation tank. Spores were collected, washed with H ₂ O and concentrated in a concentration of 1.0 × ¹⁰ 10 CFU / mL.

The experimental settings in soybean were as follows: 1) the seed was untreated; 2) the seed was passed through CRUISERMAXX (insecticide plus fungicide, containing thiamethoxam, shi ning plus statin-M; SYNGENTA CROP PROTECTION , INC) combined with methyl thiophanate fungicide (which is typical soybean seed treatment) (combined with CRUISERMAXX and methyl thiophanate as "chemical control"); and 3) seed treated with chemical control plus each seed 5.0 x 10 ^{+ 5} cfu strain RTI301 was inoculated.

Field trials were conducted on soils inoculated with Fusarium oxysporum (a causative agent of soy dying syndrome) in Ames, Iowa. Fusarium oxysporum grows on wet high pressure treated granule seeds. After the granule seeds are covered by mycelial growth, the seeds are air dried and then grown. The prepared ground inoculum was planted with soybean seeds at a specified application rate to ensure a higher and more uniform infection rate. Although the symptoms themselves are not obvious, the disease is infected early in the season until the end of the season. After 119 days, the disease incidence, disease severity and disease index of the Sudden Death Syndrome were determined. In addition, the soybean yield of each treatment was determined.

The results in Table IX show that soybean seed vaccination with B. amyloliquefaciens RTI301 has a positive effect on disease control, as measured by various parameters; and has a positive effect on the overall yield of soybean compared to seeds treated with chemical control alone. And resulted in a 7.8% increase in soybean yield (55.2 to 59.5 bushels per acre) compared to the separate chemical controls.

Soybean disease incidence, disease severity, disease index, and yield were determined after 119 days of planting under conditions of soil inoculation with Fusarium oxysporum (a causative agent of soybean sudden death syndrome).

Example 11 Antagonistic effect of Bacillus amyloliquefaciens RTI301 on the gray mold of Botrytis cinerea in Italy in field trials in Italy

The study was conducted in a tomato field trial to determine the ability of the B. amyloliquefaciens RTI301 strain to prevent and/or alleviate the effects of the plant pathogen, Botrytis cinerea (B. cinerea).

A total of 4 applications were applied to the crop with a time interval between administrations of 7 days.

Formulation:

SERENADE MAX was applied at an application rate of 4000 g/ha corresponding to 2.0 x 10 ^{+ 14} CFU/ha of Bacillus subtilis strain QST713.

Spores of Bacillus amyloliquefaciens RTI301 the spent fermentation broth Po (SFB) and the yeast extract, and the application rate is from about 0.01 to 0.2% yeast extract was under 2.0 × 10 ⁺¹³ CFU / ha.

The first two applications of the crop were carried out with SWITCH (cytosine 375 g/kg plus shining 250 g/kg; SYNGENTA CROP PROTECTION, INC) at an application rate of 0.8 kg/ha, followed by an application rate of 1.8 kg/ha. SIGNIUM was administered twice (white 267 g/kg plus 100 g/67 g/kg; BAYER CROP SCIENCE, INC). This is referred to herein as the "FARMER program."

SILWET L77 (HELENA CHEMICAL) is a nonionic organopolyfluorene surfactant used in all treatments at an application rate of 0.15 liters per 100 liters of spray solution, except for SERENADE MAX.

The experimental design was as follows: untreated control (UTC), FARMER protocol + SILWET L77, RTI301 + SILWET L77 and SERENADE MAX.

Treatment application method:

Two independent field trials were performed, each with 4 replicates, and the two trial results were combined and presented as average results. Four separate treatment applications were delivered to the crop with a time interval between each application of 7 days. Three days before the start of treatment, all small plots, including untreated controls, were treated with SWITCH to contain the initial disease progression. The sprayer was set to deliver 53 gallons per acre. Individual plots were sprayed at 1.7 mph (0.8 m/s or 2.9 km/h) using a CO ₂ backpack sprayer with conical nozzles and each nozzle was 2 吋 (5.5 cm) apart.

All tomatoes were harvested, counted, weighed, and divided into marketable or diseased to determine yield. Incidence of disease (% of fruit affected by brown mold) by grading on 5 separate dates Estimate the fruit of each treatment and measure it as "area under the disease stress curve" (AUDPC). Disease incidence and cumulative yield data are shown in Table X below. The change in disease incidence over time in UTC is shown in the following figure (Figure 6), and shows that disease stress increases during the two trials, ending with extremely high disease stress, ie, the % of each infected fruit in each of the two trials It is 51.5% and 44.5%.

The results showed that it was observed that B. amyloliquefaciens RTI301 and FARMER based on chemical active agents had the best control of B. cinerea (Botrytis cinerea), and it was better than using Bacillus subtilis strain QST713 which was 10 times higher than RTI301. Processing of SERENADE MAX.

Example 12 Antagonistic effects of Bacillus amyloliquefaciens RTI301 on the gray mold of Botrytis cinerea in Italy (Italy) and Spain (Spain)

A study was conducted in a strawberry field trial to determine the ability of B. amyloliquefaciens RTI301 to prevent and/or alleviate the effects of the plant pathogen, Botrytis cinerea (B. cinerea).

A total of 4 applications were applied to the crop with a time interval between administrations of 7 days.

Formulation :

SERENADE MAX was applied at an application rate of 4000 g/ha corresponding to 2.0 x 10 ^{+ 14} CFU/ha of Bacillus subtilis strain QST713.

Spores of Bacillus amyloliquefaciens RTI301 the spent fermentation broth Po (SFB) and the yeast extract and the application rate is from about .01 to 0.2% yeast extract was under 2.0 × 10 ⁺¹³ CFU / ha. Further, a nonionic organopolyfluorene surfactant SILWET L77 (HELENA CHEMICAL) was added at an application rate of 0.15 liter per 100 liters of the spray solution.

The first two applications of the crop were carried out with SWITCH (cytosine 375 g/kg plus shining 250 g/kg; SYNGENTA CROP PROTECTION, INC) at an application rate of 0.8 kg/ha, followed by an application rate of 1.8 kg/ha. SIGNIUM was administered twice (white 267 g/kg plus 100 g/67 g/kg; BAYER CROP SCIENCE, INC). This is referred to herein as the "FARMER program."

The experimental design is as follows: untreated control (UTC), FARMER protocol, RTI301+SILWET L77 and SERENADE MAX.

Treatment application method :

Four independent field trials were performed, each with 4 replicates, and the test results were combined and presented as average results. Four separate treatment applications were delivered to the crop with a time interval between each application of 7 days. Three days before the start of treatment, all small plots, including untreated controls, were treated with SWITCH to contain the initial disease progression. The application sprayer is set to deliver between 53 and 107 gallons per acre depending on the crop density at the time of application. Individual plots were sprayed at a ground speed of 0.56 mph (0.25 m/s or 0.9 km/h) using a CO ₂ backpack sprayer with a conical nozzle and each nozzle was 2 吋 (5.5 cm) apart.

All strawberries were harvested, counted, weighed, and divided into marketable or diseased to determine yield. The incidence of disease (% of fruit affected by B. cinerea) was measured by assessing the fruits of each treatment at the time of harvest on 6 separate dates, and was expressed as "area under the disease pressure curve" (AUDPC). The % increase in disease incidence and yield relative to the untreated control is shown in Table XI below. The change in UTC disease incidence over time is shown in the following figure (Figure 7) and shows that the disease pressure progressing during the trial reaches the highest disease stress of 20% to 45% of the infected fruit.

The results in Table XI below show that for all three treatments, B. amyloliquefaciens RTI301, SERENADE MAX and FARMER protocols, all observed on the strawberry (G. cinerea) was better controlled than the untreated control, where the yield increase with RTI301 treatment was slightly higher.

Example 13 Treatment of corn seeds with Bacillus amyloliquefaciens RTI301

Experiments were conducted to investigate the effects on plant growth and development in maize after treatment of plant seeds with B. amyloliquefaciens RTI301 strain.

Specifically, the experimental settings in corn are as follows: 1) the seeds are not treated; 2) the seeds are subjected to MAXIM (broad-spectrum seed treatment fungicides as the active ingredient, 0.0625 mg per seed; SYNGENTA CROP PROTECTION, INC) , combination of APRON XL (active ingredient, halal-M, 0.0625 mg per seed; SYNGENTA CROP PROTECTION, INC) and PONCHO (canidin insecticide, 0.25 mg per seed; BAYER CROPSCIENCE, INC) A typical corn seed treatment, a combination of MAXIM, APRON XL, and PONCHO is referred to as a "chemical control"treatment; and 3) a seed is chemically treated plus a seed of 5.0 x 10 ^{+ 5} cfu strain RTI301 per seed. Three trials were conducted at the field location in Shawneetown, Ill., with 5 replicates per treatment. The conditions of the three trials were natural disease stress or soil inoculation with one of Fusarium graminearum or Rhizoctonia. Fusarium graminearum and Rhizoctonia solani are separately grown on wet high pressure treated granule seeds and then air dried. The dried inoculum used in the selected assay is mixed with the seed at a specified application rate to provide infection as the seed begins to grow.

The average corn yield results for the field trials (bushel/acre) are presented in Table XII below. The results in Table XII show that inoculation of the chemical control plus B. amyloliquefaciens RTI301 had an effect on the overall yield of corn compared to the seeds treated with the chemical control alone. Statistical relevance (in letters) is based on P = 0.1. Notably, a significant yield benefit of RTI301 plus 40.1 bushels per acre under the chemical control was observed for the test for inoculation of Rhizoctonia compared to the chemical control alone. In addition, the test for the artificial inoculation of Fusarium graminearum and the natural disease stress increased by 3.3 bushels per acre and 8.4 bushels per acre, respectively. In summary, treatment with the chemical control plus RTI 301 resulted in an increase in yield in all three trials and a significant increase in the yield of the maize plant inoculated with Rhizoctonia.

Example 14 Antagonistic effect of combination of Bacillus amyloliquefaciens RTI301 and FRACTURE on fungal pathogens

Studies were performed using in vitro disc assay to determine the antagonistic ability of B. amyloliquefaciens RTI301 to enhance FRACTURE (CONSUMO EM VERDE (CEV), BIOTECNOLOGIA DAS PLANTAS S.A., PORTUGAL) to control the efficacy of fungal plant pathogens. FRACTURE is a plant extract based formulation containing 20% BLAD polypeptide as the active ingredient. The BLAD polypeptide is a fragment of a naturally occurring seed storage protein in sweet lupin (white lupin) that acts on a susceptible fungal pathogen by causing damage to the fungal cell wall and disrupting the inner cell membrane.

Disk assays against antagonism of plant fungal pathogens were evaluated by growing bacterial isolates and pathogenic fungi side by side on 869 agar plates or on 869 + 1% FRACTURE agar plates. On the opposite side of each disk, 20 μl of RTI301 spore solution containing 1 × 10 ⁸ CFU/ml or 1 × 10 ⁹ CFU/ml was spotted at a distance of 4 cm from the center of the disk. Subsequently, 20 μl of the fungal spore solution or the agar plug inoculated with the fungal mycelium was placed in the center of the disk. The plates were incubated at 25 ° C for 7 days and the growth behavior was checked regularly, such as growth inhibition, ecological habitat occupancy or no effect.

The results of the antagonistic activity against Fusarium graminearum and Fusarium oxysporum provided by the combination of Bacillus amyloliquefaciens RTI301 and FRACTURE are shown in Figures 8A-8F (Fusarium graminearum) and Figures 9A-9F (Fusarium oxysporum) .

Figures 8A-8F are images showing the analysis of B. amyloliquefaciens RTI 301 to control Fusarium graminearum in the presence and absence of FRACTURE. A) Growth of Fusarium graminearum on 869 agar plate; B) 869 agar plate in the presence of 20 μl of RTI301 spore solution containing 1×10 ⁸ CFU/ml (left) or 1×10 ⁹ CFU/ml (right) Growth of Fusarium graminearum; C) 20 μl of 869 agar plate containing 1×10 ⁸ CFU/ml (left) or 1×10 ⁹ CFU/ml (right) growth of Bacillus amyloliquefaciens RTI301 spore solution; D) Growth of Fusarium graminearum on 869+1% FRACTURE agar plate; E) 869+ in the presence of 20 μl of RTI301 spore solution containing 1×10 ⁸ CFU/ml (left) or 1×10 ⁹ CFU/ml (right) Growth of Fusarium graminearum on 1% FRACTURE agar plate; F) 20 μl on 869+1% FRACTURE agar plate containing 1×10 ⁸ CFU/ml (left) or 1×10 ⁹ CFU/ml (right) starch Growth of the spore solution of Bacillus RTI301.

Figures 9A-9F are images showing the analysis of Bacillus cerevisiae RTI301 controlling Fusarium oxysporum in the presence and absence of FRACTURE. A) Growth of Fusarium oxysporum on the 869 agar plate; B) 869 agar plate in the presence of 20 μl of RTI301 spore solution containing 1 × 10 ⁸ CFU/ml (left) or 1 × 10 ⁹ CFU/ml (right) Growth of Fusarium oxysporum on C; 869; 20 μl of a solution on the 869 agar plate containing 1×10 ⁸ CFU/ml (left) or 1×10 ⁹ CFU/ml (right) of Bacillus amyloliquefaciens RTI301 spore solution; D) 869+1% FRACTURE agar plate on the growth of Fusarium oxysporum; E) in 20μl containing 1 × 10 ⁸ CFU / ml (left) or 1 × 10 ⁹ CFU / ml (right) of the RTI301 spore solution Growth of Fusarium oxysporum on the 869+1% FRACTURE agar plate; F) 869+1% FRACTURE agar plate containing 20μl of 1×10 ⁸ CFU/ml (left) or 1×10 ⁹ CFU/ml (right) The growth of the spore solution of Bacillus amyloliquefaciens RTI301.

Results The presence of 1% FRACTURE in 869 agar medium did not inhibit the growth of B. amyloliquefaciens RTI301. The lack of inhibition of RI301 was contrasted with that observed for other strains of B. amyloliquefaciens, where the presence of 1% FRACTURE inhibited strain growth. In addition, although the addition of 1% FRACTURE to 869 medium reduced the growth of Fusarium graminearum and Fusarium oxysporum, no complete inhibition of fungal growth was achieved. However, the presence of B. amyloliquefaciens RTI301 cumulatively inhibits fungal growth of Fusarium graminearum and Fusarium oxysporum. Therefore, B. amyloliquefaciens RTI301 can be used to enhance FRACTURE potency. Similar results were observed for the control of Astragalus membranaceus (data not shown).

Example 15 Identification of new metabolites produced by B. amyloliquefaciens RTI301 isolates

It has previously been reported that five classes of abundane-type metabolites and dehydroxy-hydroxytoxin-type metabolites are produced by microbial species including Bacillus amyloliquefaciens (see, for example, Li, Xing-Yu et al., 2013, J. Microbiol . Biotechnol. 23 (3), 313-321; Pecci Y et al, 2010 , Mass Spectrom ., 45(7): 772-77). These metabolite cyclic lipopeptides are cyclic peptide molecules which also contain fatty acid groups. The five classes of abundance and dehydroxyl-producing metabolites are called A, B, C, D and S. The backbone structure of these metabolites and the specific amino acid sequence for each of the five classes are shown in Figure 10.

The abundance of the amylobacterin and the dehydroxyl-precipitate metabolite produced by B. amyloliquesii RTI301 were analyzed using UHPLC-TOF MS. The molecular weight of the abundane-type metabolite produced by the RTI 301 strain after growth for 6 days at 30 ° C in M 2 medium was compared to the theoretical molecular weight expected for the abundance of the abundance and the dehydroxy-hydroxytoxin-type metabolite. In addition, to determine The amino acid composition of various abundane-type metabolites produced by the RTI 301 strain was subjected to peptide sequencing using LC-MS-MS for each of the abundane-type metabolites previously identified by UHPLC-TOF MS. In this manner, it was determined that B. amyloliquefaciens RTI301 produced abundance of A, abundance of B, and abundance of C, and a dehydroxyl-producin A, a dehydroxyl-producin B, and a dehydroxy-hydroxytoxin C. Surprisingly, in addition to these known compounds, it was determined that the RTI 301 strain also produced previously unidentified compound derivatives.

For example, it is determined that the B. amyloliquefaciens RTI301 strain produces a protosanoid-like and dehydroxy-norformin-like compound in which the L-isoleucine acid at position 8 (referred to as X _{3 in} FIG. 10) of the cyclic peptide chain is L- Methionine replacement. The new classes of protosan and dehydroxylectin are referred to herein as MA, MB and MC, and refer to class A, B and C derivatives, wherein L-isoleucine at X ₃ in Figure 10 is L- Methionine replacement. The newly identified molecules are shown in bold in Figure 10 and in Table XIII below.

It was further determined that the RTI301 strain produced other classes of previously unidentified abundance and dehydroxylectin. In this category, fengycin B and activin B to the hydroxyl BBCA L- isoleucine (position of FIG. 10 X ₃₎ via L- homocysteine (of Hcy) replacement. These previously unidentified abundane and dehydroxylectin metabolites are referred to herein as proemin H and dehydroxylectin H and are shown in Figure 6 and Table XIII.

It was further determined that the RTI301 strain produced other classes of previously unidentified abundance and dehydroxylectin metabolites. In this category, the position of the main chain structure of the cyclic peptide 4 (FIG. 10 in the position X ₁₎ is replaced by the amino acid L- isoleucine. These previously unidentified metabolites are referred to herein as aureoline I and dehydroxylectin I and are shown in Figure 10 and Table XIII.

An overview of the amino acid sequences of the previously reported proepine-type and dehydroxy-hydroxy-protopin-type lipopeptides and newly identified metabolites is provided in Table XIII below.

Table XIII. Overview of UHPLC-TOF MS identification of abundance of lipopeptides in Bacillus amyloliquefaciens RTI301

Example 16 Antimicrobial activity of isolated lipopeptide metabolites of RTI301

The antagonist lipopeptide from B. amyloliquefaciens strain RTI301 was isolated from the RTI301 waste mash and was shown to retain its activity.

In this experiment, the procedure described in Smyth, TJP et al., 2010, "Isolation and Analysis of Lipopeptides and High Molecular Weight Biosurfactants.": Handbook of Hydrocarbon and Lipid Microbiology, KNTimmis (eds.), pp. 3687-3704. The B. amyloliquefaciens RTI301 culture supernatant was acidified to pH 2. The recovery of lipopeptides was analyzed by UHPLC-TOF MS and tested for antagonistic activity against Botrytis cinerea and Fusarium graminearum.

RTI301 was cultured in M2 sporulation medium at 30 ° C for six days, and spent broth (301-SFB) was centrifuged at 18,514 g for 20 minutes to remove spores. The supernatant was then acidified to pH 2.0 by the addition of concentrated HCl and precipitated overnight at 4 °C. The sample was then centrifuged at 18,514 g for 20 minutes to obtain a solid crude lipopeptide. The pellets were lyophilized overnight, dissolved in the original volume of M2 medium and analyzed by LCMS. Extract the quality of italin (C14, C15, C16), surfactant (C12, C13, C14, C15, C16, C17) and agrosperm (A, B, C, D, S), integrate and calculate the sum To compare the relative abundance of lipopeptides from each sample. Figure 11 is a graph showing the percentage of lipopeptides recovered from RTI301 spent broth (SFB) after acid precipitation. The terms "301-AP-spherule" and "301-AP-supernatant" refer to resuspended pellets and supernatant obtained after acid precipitation of SFB in centrifugation, respectively. The results in the graph of Figure 11 show that 80% of the total amount of lipopeptide is recovered by acid precipitation. For the imipenin, 35% was precipitated, and by the acid precipitation method, 59% of the avermectin was not recovered. Surfactant and abundance are 100% recovered using acid precipitation.

To confirm that the LCMS results correlate with the antagonistic activity, bioassays were performed using the same samples analyzed by LCMS. For this bioassay, 20 [mu]l of Botrytis cinerea or Fusarium graminearum inoculum was spotted in the middle of the plates, with 301-AP-spherule samples spotted in 10 [mu]l, 20 [mu]l and 40 [mu]l aliquots. For the Botrytis cinerea and Fusarium graminearum disks, the antifungal activity was examined after incubation at 30 ° C for 5 or 7 days, respectively. Results show samples of acid precipitation (301- AP-spherulites have similar degrees of antagonistic activity against Botrytis cinerea and Fusarium graminearum in the same degree as the starting waste mash. Bioanalytical results are well correlated with LCMS data.

references

All publications, patent applications, patents, and other references cited herein are hereby incorporated by reference in their entirety.

Although the above-described subject matter has been described in considerable detail for the purpose of illustration and example, it will be understood by those skilled in the art that certain changes and modifications can be made within the scope of the claimed invention.

【Biomaterial Storage】 Domestic registration information [please note according to the registration authority, date, number order]

1. Center for Biological Resource Conservation and Research, Institute of Food Industry Development; April 26, 105; BCRC 910728

Foreign deposit information [please note according to the country, organization, date, number order]

1. United States: American Type Culture Collection (ATCC); April 17, 2014; PTA-121165

<110> American Fumanxi Company

<120> Bacillus amyloliquefaciens RTI301 composition and method for using the same for plant growth and treatment of plant diseases

<130> 61083-PCT

<160> 2

<170> PatentIn version 3.5

<210> 1

<211> 1554

<212> DNA

<213> Bacillus amyloliquefaciens

<400> 1

<210> 2

<211> 3246

<212> DNA

<213> Bacillus amyloliquefaciens

<400> 2

Claims (95)

A composition comprising a Bacillus amyloliquefaciens RTI301 deposited with ATCC PTA-121165 or a biologically pure culture of a mutant having all of its identifying characteristics for application to a plant to achieve a plant growth or Protect susceptible plants from one or both of the pathogenic infections. The composition of claim 1, wherein the plant growth benefit and/or the conferred protection is manifested by improved seedling vigor, improved root development, improved plant growth, improved plant health, Increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or a combination thereof. The composition of claim 1 wherein the composition is in the form of a liquid, a powder, a dry wettable powder, a spreadable granule or a dry wettable granule. The composition of claim 1, wherein the composition is in a liquid form, and the B. amyloliquefaciens RTI301 is present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. The composition of claim 1 wherein the composition is in the form of a powder, a dry wettable powder, a spreadable granule or a dry wettable granule, and the B. amyloliquefaciens RTI 301 is at about 1.0 x 10 ⁸ CFU/g. It is present in an amount of about 1.0 x 10 ¹² CFU/g. The composition of claim 1, wherein the composition is in the form of an oily dispersion, and the B. amyloliquefaciens RTI301 is present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. The composition of claim 1, wherein the B. amyloliquefaciens RTI301 is in the form of spores or vegetative cells. The composition of claim 1, which further comprises one of the following amounts present in an amount suitable for beneficial to plant growth and/or to confer protection to susceptible plants to avoid pathogenic infections. Or combination: microbial, biological or chemical insecticides, fungicides, nematicides, fungicides, herbicides, plant extracts, plant growth regulators or fertilizers. The composition of claim 1, which further comprises one or a combination of: a carrier, a surfactant, a dispersant, or a yeast extract. The composition of claim 1 wherein the composition is in the form of a planting matrix. The composition of claim 10, wherein the planting substrate is in the form of potting soil. A plant seed coated with a composition comprising the presence of an amount suitable for beneficial to plant growth and/or protection of a susceptible plant to avoid pathogenic infection: deposited under ATCC PTA-121165 A spore of a biologically pure culture of B. amyloliquefaciens RTI301 or a mutant thereof having all of its identifying characteristics. The plant seed of claim 12, wherein the plant growth benefit and/or the conferred protection is manifested by improved seedling vigor, improved root development, improved plant growth, enhanced plant health, Increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or a combination thereof. The plant seed of claim 12, wherein the composition comprises from about 1.0 x 10 ² CFU per seed to about 1.0 x 10 ⁹ CFU per seed of B. amyloliquefaciens spores. The plant seed of claim 12, wherein the seed comprises the following seeds: monocot, dicot, grain, corn, sweet corn, popcorn, corn, silage corn, feed corn, rice, wheat, barley, sorghum , kale-type vegetables, broccoli, kale, broccoli, Brussels sprouts, broccoli, kale, mustard, broccoli, bulbous vegetables, onions, garlic, onions, fruit vegetables, pepper, tomato, eggplant, ground cherry, tree Tomato, okra, grape, vanilla/spice, melon, cucumber, cantaloupe, melon, cantaloupe, squash, watermelon, pumpkin, eggplant, green leafy vegetables, lettuce, celery, spinach, parsley, chrysanthemum Borage, legumes/vegetables (succulent and dry beans and peas), beans, green beans, pod beans, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, pea petals, Lentils, oilseed crops, canola, ramie, cotton, flax, peanuts, rapeseed, safflower, sesame, sunflower, soybean, roots/tubers and bulbous vegetables, carrots, potatoes, sweet potatoes, beets, ginger, horseradish, radish , ginseng, turnip, sugar cane, sugar beet, grass or turf grass. A plant seed according to claim 15 wherein the plant comprises the following seeds: dried beans, corn, wheat, soybean, canola, rice, cucumber, pepper, tomato, squash, cotton, grass or turf grass. The plant seed of claim 12, wherein the composition further comprises one or a combination of the presence of an amount suitable for beneficial to plant growth and/or protection of the susceptible plant to avoid pathogenic infection: microbial, biological or chemical Insecticides, fungicides, nematicides, fungicides or plant growth regulators. A composition for achieving one or both of beneficial to plant growth or protection of a susceptible plant to avoid pathogenic infection, the composition comprising: Bacillus amyloliquefaciens RTI301 deposited under ATCC PTA-121165 Or a biologically pure culture of a mutant having all of its identifying characteristics in an amount suitable for beneficial to plant growth and/or conferring protection to the susceptible plant to avoid pathogenic infection; and microbial, biological or chemical insecticidal One or a combination of a fungicide, a fungicide, a nematicide, a bactericide, a herbicide, a plant extract, a plant growth regulator or a fertilizer, which is suitable for beneficial to plant growth and/or to confer protection to the susceptible plant Avoid the amount of pathogenic infections. The composition of claim 18, wherein the plant growth benefit and/or the conferred protection is manifested by enhanced seedling vigor, improved root development, improved plant growth, enhanced plant health, Increased production, improved Appearance, increased resistance to plant pathogens, reduced pathogenic infection, or a combination thereof. The composition of claim 18, wherein the composition is in liquid form, and the B. amyloliquefaciens RTI301 is present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. The composition of claim 18, wherein the composition is in the form of a powder, a dry wettable powder, a spreadable granule or a dry wettable granule, and the Bacillus amyloliquefaciens RTI 301 is about 1.0 x 10 ⁸ CFU/g. It is present in an amount of about 1.0 x 10 ¹² CFU/g. The composition of claim 18, wherein the composition is in the form of an oily dispersion, and the B. amyloliquefaciens RTI301 is present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. The composition of claim 18, wherein the B. amyloliquefaciens RTI301 is in the form of spores or vegetative cells. The composition of claim 18, wherein the plant comprises monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, feed corn, rice, wheat, barley, sorghum, asparagus, berries , blueberry, blackberry, raspberry, raspberry, cowberry, cranberry, gooseberry, elderberry, black currant, cranberry, raspberry, kale, broccoli, kale, broccoli, Brussels sprouts , broccoli, kale, mustard, stalks, melons, cucumbers, cantaloupe, melon, cantaloupe, squash, watermelon, pumpkin, eggplant, bulbous vegetables, onions, garlic, onions, tangerine peel, oranges, Grapefruit, lemon, red orange, tangerine, grapefruit, fruit vegetable, pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, green leafy vegetables, lettuce, celery, spinach, parsley, chicory, beans Plants/vegetables (succulent and dry beans and peas), beans, green beans, pod beans, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, pea petals, lentils, oil Seed crop, canola, castor, coconut, Cotton, flax, oil palm, olive, peanut, rapeseed, safflower, sesame, sunflower, soybean, pear, apple, jellyfish, pear, alfalfa, summer flower hawthorn, roots/tubers and bulbous vegetables, carrots, potatoes, Sweet potato, cassava, beet, ginger, horseradish, radish, ginseng, turnip, stone fruit, apricot, cherry, nectarine, peach, plum, plum, strawberry, nut, almond, pistachio, pecan, walnut, hazelnut, chestnut, Cashew nuts, beech nuts, walnuts, macadamia nuts, kiwis, bananas, (blue) agave, grass, turfgrass, ornamental plants, poinsettia, hardwood pruning, chestnut, oak, maple, sugar cane or beet. A method for achieving one or both of beneficial to plant growth or to confer protection to a susceptible plant to avoid a pathogenic infection, the method comprising: delivering a composition to: a leaf of the plant, a stem of the plant Skin, the fruit of the plant, the flower of the plant, the seed of the plant, the root of the plant, the pruning of the plant, the graft of the plant, the healing tissue of the plant; the soil or growth medium surrounding the plant; Or the soil or growth medium prior to seeding the seed of the plant in the growth medium; or planting the plant, the plant pruning, the plant graft or the soil or growth medium prior to healing the tissue in the soil or growth medium, The composition comprises a biologically pure culture of Bacillus amyloliquefaciens RTI301 deposited with ATCC No. PTA-121165 or a mutant having all of its identifying characteristics, which is suitable for benefiting the growth of the plant and/or the susceptible disease Plants provide protection from the amount of pathogenic infections. The method of claim 25, wherein the composition is delivered to the leaves of the plant. The method of claim 25, wherein the growth benefit of the plant and/or the protection conferred by the plant is manifested by enhanced seedling vigor, improved root development, improved plant growth, improved plant health, Increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or a combination thereof. The method of claim 25, wherein the B. amyloliquefaciens RTI301 is delivered at a rate of from about 1.0 x 10 ¹⁰ CFU/ha to about 1.0 x 10 ¹⁴ CFU/ha. The method of claim 25, wherein the B. amyloliquefaciens RTI301 is in the form of spores or vegetative cells. The method of claim 25, wherein the composition is in the form of a liquid, a powder, a dry wettable powder, a spreadable granule, a dry wettable granule or an oily dispersion. The method of claim 25, wherein the composition further comprises one or a combination of the presence of an amount suitable for beneficial to plant growth and/or protection of the susceptible plant to avoid pathogenic infection: microbial, biological or chemical Insecticides, fungicides, nematicides, fungicides, herbicides, plant extracts, plant growth regulators or fertilizers. The method of claim 25, wherein the composition further comprises one or a combination of: a carrier, a surfactant, a dispersant, or a yeast extract. The method of claim 25, wherein the plant comprises monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, forage corn, rice, wheat, barley, sorghum, asparagus, berries, Blueberries, blackberries, raspberries, raspberries, cranberries, cranberries, gooseberries, elderberries, currants, cranberries, berry, cabbage, broccoli, kale, broccoli, Brussels sprouts, Cabbage, kale, mustard, stalk, melon, cucumber, cantaloupe, melon, sweet melon, squash, watermelon, pumpkin, eggplant, bulbous vegetables, onions, garlic, onions, tangerine peel, oranges, grapes Pomelo, lemon, red orange, tangerine, grapefruit, fruit vegetable, pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, green leafy vegetables, lettuce, celery, spinach, parsley, chicory, legume Plants/vegetables (succulent and dry beans and peas), beans, green beans, pod beans, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, peas, lentils, oil Sub-crop, canola, castor, coconut, Cotton, flax, oil palm, olive, peanut, rapeseed, safflower, sesame, sunflower, soybean, pear, apple, jellyfish, pear, alfalfa, summer flower hawthorn, roots/tubers and bulbous vegetables, carrots, potatoes, Sweet potato, cassava, beet, ginger, horseradish, radish, ginseng, turnip, stone fruit, apricot, cherry, nectarine, peach, plum, plum, strawberry, nut, almond, pistachio, pecan, walnut, hazelnut, chestnut, Cashew nuts, beech nuts, walnuts, macadamia nuts, kiwis, bananas, (blue) agave, grass, turfgrass, ornamental plants, poinsettia, hardwood pruning, chestnut, oak, maple, sugar cane or beet. The method of claim 33, wherein the plant comprises soybean, bean, pod bean, wheat, cotton, corn, pepper, tomato, potato, tapioca, grape, strawberry, banana, peanut, squash, pumpkin, eggplant or cucumber. The method of claim 25, wherein the pathogenic infection is caused by a plant fungal pathogen, a plant bacterial pathogen, a rust fungus, Botrytis spp ., Botrytis cinerea , and Botrytis cinerea ( Botrytis squamosa ), Erwinia spp ., Erwinia carotovora , Erwinia amylovora , Dickeya spp ., Dickeya Dadantii ), Dickeya solani , Agrobacterium spp ., Agrobacterium tumefaciens , Xanthomonas spp ., Xanthomonas axonopodis Xanthomonas campestris pv. carotae , Xanthomonas pruni , Xanthomonas arboricola , Xanthomonas oryzae pv. Oryzae ), Xylella spp ., Xylella fastidiosa , Candidatus spp., phloem ( C) Andidatus liberibacter ), Fusarium spp ., Fusarium colmorum , Fusarium graminearum , Fusarium oxysporum , Fusarium oxysporum f.sp .Cubense ), Fusarium oxysporum f.sp. Lycopersici , Fusarium virguliforme , Sclerotinia spp ., Sclerotinia sclerotiorum , small nucleus Sclerotinia minor , Sclerotinia homeocarpa , Cercospora/Cercosporidium spp., Uncinula spp., Uncinula necator (Powdery) Mildew)), Podosphaera spp. ( powder disease ), white stalk shell ( Podosphaera leucotricha ), Claude stalked shell ( Podosphaera clandestine ), Pseudomonas sp. Phomopsis spp.), Phomopsis viticola , Alternaria spp ., Alternaria tenuissima , Alterna solani ( Alterna) Ria porri ), Alternaria alternate , Alternaria solani , Alternaria tenuis , Pseudomonas spp., Pseudomonas syringae Tomato cultivar ( Pseudomonas syringae pv. Tomato ), Phytophthora spp., Phytophthora infestans , Phytophthora parasitica , Phytophthora sojae , Phytophthora capsici ), Phytophthora cinnamon , Phytophthora fragariae , Phytophthora spp ., Phytophthora ramorum , Phytophthora palmivara , Phytophthora nicotianae ), Phakopsora spp., Phakopsora pachyrhizi , Phakopsora meibomiae , Aspergillus spp ., Aspergillus flavus , Aspergillus niger , Uromyces spp ., Uromyces appendiculatus ), Cladosporium spp., Cladosporium herbarum , Rhizopus spp., Rhizopus arrhizus , Penicillium spp., silk Rhizoctonia spp ., Rhizoctonia solani , Rhizoctonia zeae , Rhizoctonia oryzae , Rhizoctonia caritae , cereal Rhizoctonia cerealis , Rhizoctonia crocorum , Rhizoctonia fragariae , Rhizoctonia ramicola , Rhizoctonia rubi , Beanlike silk solani (Rhizoctonia leguminicola), bean shell fungus Beauveria (Macrophomina phaseolina), Pyricularia oryzae (Magnaorthe oryzae), Mycosphaerella (Mycosphaerella spp.), Mycosphaerella graminicola (Mycosphaerella graminocola), Fiji Leptosphaeria (Mycosphaerella fijiensis) (black Sigatoka (black sigatoga)), apple scab Leptosphaeria (Mycosphaerella pomi), lemon Leptosphaeria (Mycosphaerella citri), rice blast species ( Magnaporthe spp .), Magnaporthe grisea , Monilinia spp., Monilinia fruticola , Monilinia vacciniicorymbosi , Monilinia Laxa ), Colletotrichum spp., Colletotrichum gloeosporiodes , Colletotrichum acutatum , Colletotrichum Candidum , Diaporthe spp., Citrus Block globosum (Diaporthe citri), the genus Micromonospora rod (Corynespora spp.), multi-master rod cinerea (Corynespora Cassiicola), the genus gum rust (Gymnosporangium spp.), gum juniper plants by rust (Gymnosporangium juniperi-virginianae) , Schizothyrium spp., Schizothyrium pomi , Gloeodes spp., Gloeodes pomigena , Botryosphaeria spp.), grape canker pathogen (Botryosphaeria dothidea), Ming Sclerotinia cinerea (Neofabraea spp.), Wilson genus Fusarium shell (Wilsonomyces spp.), Weir fruit addicted Spore globosum (Wilsonomyces carpophilus), Sphaerotheca spp (Sphaerotheca spp.), Sphaerotheca spot fungus (Sphaerotheca macularis), fungus Sphaerotheca rose (Sphaerotheca pannosa), Erysiphe (Erysiphe spp.), Chitin Stagonospora spp ., Stagonospora nodorum , Pythium spp ., Pythium ultimum , Pythium aphanidermatum, Pythium pyogenes Pythium irregularum ), Pythium ulosum , Pythium lutriarium , Pythium sylvatium , Venturia spp , Venturia inaequalis , Verticillium Verticillium spp., Ustilago spp ., Ustilago nuda , Ustilago maydis , Ustilago scitaminea , ergots ( Claviceps spp .), Claviceps puprrea , Tilletia spp ., Tilletia tritici , Tilletia laevis , rice black Tilletia horrid , Tilletia controversa , Phoma spp ., Phoma glycinicola , Phoma exigua , Cabbage stalk Phoma lingam , Cocliobolus sativus , Gaeumanomyces gaminis , Colleototricum spp., Rhychosporium spp ., Barley grisea Rhychosporium secalis ), Biopolaris spp ., Helminthosporium spp ., Helminthosporium secalis , Helminthosporium maydis , Helminthosporium solai ), Helminthosporium tritici-repentis or a combination thereof. The method of claim 35, wherein the pathogenic infection is caused by one or a combination of: soybean rust (Pythium humilis, Pseudomonas aeruginosa) and the plant comprises soybean; Botrytis cinerea (grape) Spore plague) and the plant comprises a grape; Botrytis cinerea (B. blight) and the plant comprises a strawberry; Botrytis cinerea (B. blight) and the plant comprises a tomato; the genus Alternaria (eg, the genus And the plant comprises tomato; Alternaria (such as Alternaria alternata) and the plant comprises potato; bean rust (String rust) and the plant contains common beans; powdery mildew (soybean powdery mildew) And the plant comprises soybean; Ficus faecium (black banana leaf spot) or Fusarium oxysporum (Panama disease) and the plant comprises a banana; a yellow variant of Xanthomonas or Xanthomonas oryzae and the plant comprises Rice; Phytophthora sojae and the plant comprising cassava; Xanthomonas campestris and the plant comprising tomato; Botrytis cinerea (P. cinerea) and the plant comprising pepper; powdery mildew and the plant comprising a gourd; Sclerotinia sclerotiorum The plant comprises a pod bean; a sunflower sclerotiorum (white mold) and the plant comprises a potato; a sclerotinia (coin spot disease) and the plant comprises a turf grass; a southern white mold and the plant comprises peanut; leaf spot (C. cerevisiae / Cercospora) and the plant comprises peanuts; Fusarium graminearum (branches of wheat) and the plant comprises wheat; Phytophthora sinensis (S. cerevisiae) and the plant comprises wheat; Phytophthora (Bacterial blight and Phytophthora capsici) and the plant comprises wheat; Starch auxin and the plant comprises apple, pear and other pear fruit; apple black spot disease and the plant comprises apple, pear and other pear fruit; or Rhizoctonia solani and the plant comprises wheat, rice, turf grass , soybeans, corn, legumes and vegetable crops. A method for achieving one or both of beneficial to plant growth or to confer protection to a susceptible plant to avoid a pathogenic infection, the method comprising: planting the seed of the plant in a suitable growth medium or causing the plant to The nutrient pruning/tissue is regenerated in a suitable growth medium, wherein the seed has been coated with a composition or the nutritional pruning/tissue has been inoculated with the composition comprising Bacillus amyloliquefaciens deposited with ATCC PTA-121165 A biologically pure culture of RTI 301 or a mutant thereof having all of its identifying characteristics, wherein the plant is beneficial for growth and/or protection from the seed or the nutritional pruning/tissue to avoid pathogenic infection. The method of claim 37, wherein the plant growth benefit is manifested by improved seedling vigor, improved root development, improved plant growth, improved plant health, increased yield, improved Appearance, increased resistance to plant pathogens, reduced pathogenic infection, or a combination thereof. The method of claim 37, wherein the B. amyloliquefaciens RTI301 is present in the form of spores in an amount of from about 1.0 x 10 ² CFU per seed to about 1.0 x 10 ⁹ CFU per seed. The method of claim 37, wherein the plant comprises monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, forage corn, rice, wheat, barley, sorghum, asparagus, berries, Blueberries, blackberries, raspberries, raspberries, cranberries, cranberries, gooseberries, elderberries, currants, cranberries, berry, cabbage, broccoli, kale, broccoli, Brussels sprouts, Cabbage, kale, mustard, stalk, melon, cucumber, cantaloupe, melon, sweet melon, squash, watermelon, pumpkin, eggplant, bulbous vegetables, onions, garlic, onions, tangerine peel, oranges, grapes Pomelo, lemon, red orange, orange Pomelo, grapefruit, fruit vegetables, pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, green leafy vegetables, lettuce, celery, spinach, parsley, chicory, legumes/vegetables (succulent and dry) Beans and peas), beans, green beans, pod beans, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, peas, lentils, oilseed crops, canola, ramie, Coconut, cotton, flax, oil palm, olive, peanut, rapeseed, safflower, sesame, sunflower, soybean, pear, apple, jellyfish, pear, alfalfa, summer flower hawthorn, roots/tubers and bulbous vegetables, carrots, Potato, sweet potato, cassava, beet, ginger, horseradish, radish, ginseng, turnip, stone fruit, apricot, cherry, nectarine, peach, plum, plum, strawberry, nut, almond, pistachio, pecan, walnut, hazelnut, Chestnut, cashew, beech nuts, walnut, macadamia, kiwi, banana, (blue) agave, grass, turf, ornamental, poinsettia, hardwood pruning, chestnut, oak, maple, sweet Cane or beet. The method of claim 40, wherein the plant comprises soybean, bean, pod bean, wheat, cotton, corn, pepper, tomato, potato, tapioca, grape, strawberry, banana, peanut, squash, pumpkin, eggplant or cucumber. The method of claim 37, wherein the pathogenic infection is caused by a plant fungal pathogen, a plant bacterial pathogen, a rust fungus, a Botrytis, a Botrytis cinerea, a Botrytis cinerea, an Erwinia, and a carrot soft rot , starch-degrading Owen bacteria, Dick's genus, D. dandelion, black smut, Agrobacterium, Agrobacterium tumefaciens, Xanthomonas, Phytophthora sojae, Xanthomonas campestris Pathogenic species, Rhizoctonia solani, Rhizoctonia solani, Rice variety of Xanthomonas oryzae, Saccharomyces, Saccharomyces cerevisiae, Cartilian, Phloem phlei, Fusarium, Fusarium oxysporum, Wo Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium solani, Sclerotinia, S. sclerotiorum, Sclerotinia sclerotiorum, Sclerotinia sclerotiorum, Cercospora Genus, genus, genus, grape white powder Pathogens (powder disease), sclerotium sclerotium (powder disease), white stalk single sac, clams, single sac, stalk, sphaeroides, Phytophthora Genus, Alternaria alternata, Alternaria alternata, Alternaria alternata, Alternaria alternata, Alternaria, Pseudomonas, Pseudomonas syringae, tomato Phytophthora , Phytophthora infestans, Phytophthora infestans, Phytophthora sojae, Phytophthora capsici, Phytophthora capsici, Phytophthora infestans, Phytophthora infestans, Phytophthora, eucalyptus, Phytophthora palm, Phytophthora nicotianae, Puccinia genus, Bean Rust fungus, Pseudomonas aeruginosa, Aspergillus, Trichoderma spp., Aspergillus niger, Puccinia rot, Bean rust, Cladosporium, Cladosporium, Rhizopus, Less root Rhizopus, Penicillium, Rhizoctonia, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Strawberry silk Nuclear bacterium, Rhizoctonia solani, Rhizoctonia platensis, Rhizoctonia solani, Coccidioides, Magnaporthe oryzae, Glomus, G. sphaeroides, G. globosa (black banana leaf spot), apple spotted bacillus, lemon bacterium, rice blast, rice blast, chain genus, brown rot, blueberry fruit pathogen, nucleus , anthrax, anthrax, anthrax, anthracnose, anthrax, genus, citrus, genus, genus, genus, genus, rust, plant Rust fungus, Shield genus, Rhizoctonia solani, Myxosporium, Phaeocystis, Plasmodium, Grape canker, Clostridium, Wilson genus, Phytophthora sclerophylla, Phytophthora, Phytophthora, Phytophthora, Rhizoctonia solani, Powdery mildew, genus Polyporus, Needle, Pythium, Pythium, Papaya Pythium, Pythium, Pythium, Pythium, Pythium, Pseudomonas, Arthropod, Verticillium, Black Powder, Black Powder, Jade Black Powder fungus, sugarcane whistle, ergot, ergot, smut, wheat smut, smooth smut, rice black Bacteria, Tilletia Tilletia, the genus Phoma, Phoma raw soybeans, Phoma varied, cabbage Phoma, Helminthosporium, Hessian sac, Anthracnose, genus, genus, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium , Wheat Helminthosporium or a combination thereof. The method of claim 37, wherein the composition further comprises one or a combination of the presence of an amount suitable for beneficial to plant growth and/or protection of the susceptible plant to avoid pathogenic infection: microbial, biological or chemical killing. Insecticides, fungicides, nematicides, fungicides or plant growth regulators. A method for achieving one or both of beneficial to plant growth or protection of a susceptible plant to avoid pathogenic infection, the method comprising: delivering a combination of the first composition and the second composition to: the plant Leaf, the bark of the plant, the fruit of the plant, the flower of the plant, the seed of the plant, the root of the plant, the pruning of the plant, the graft of the plant, the healing tissue of the plant; Soil or growth medium; the soil or growth medium prior to sowing the seed of the plant in soil or growth medium; or planting the plant, the plant pruning, the plant graft or the plant healing tissue in the soil or growth medium The soil or growth medium, the first composition comprising a B. amyloliquefaci RTI301 deposited with ATCC No. PTA-121165 or a biologically pure culture thereof having all of its identifying characteristics, is suitable for benefiting the plant Growing and/or imparting protection to the susceptible plant to avoid pathogenic infection; and the second composition comprises a microbial, biological or chemical insecticide, fungicide One or a combination of a nematicide, a bactericide, a herbicide, a plant extract, a plant growth regulator, or a fertilizer, which is suitable for benefiting the growth of the plant and/or conferring protection to the susceptible plant to avoid pathogenic infection. the amount. The method of claim 44, wherein the combination is delivered to the foliage of the plant. The method of claim 45, wherein the first composition comprises one or a combination of: a carrier, a surfactant, a dispersant, or a yeast extract. The method of claim 44, wherein the plant growth benefit and/or the conferred protection is manifested by improved seedling vigor, improved root development, improved plant health, enhanced plant quality, Increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infections, or a combination thereof. The method of claim 44, wherein the amount suitable for plant growth and/or protection is from about 1.0 x 10 ¹⁰ CFU/ha to about 1.0 x 10 ¹⁴ CFU/ha B. amyloliquefasic RTI301. The method of claim 44, wherein the B. amyloliquefaciens RTI301 is in the form of spores or vegetative cells. The method of claim 44, wherein the plant comprises monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, forage corn, rice, wheat, barley, sorghum, asparagus, berries, Blueberries, blackberries, raspberries, raspberries, cranberries, cranberries, gooseberries, elderberries, currants, cranberries, berry, cabbage, broccoli, kale, broccoli, Brussels sprouts, Cabbage, kale, mustard, stalk, melon, cucumber, cantaloupe, melon, sweet melon, squash, watermelon, pumpkin, eggplant, bulbous vegetables, onions, garlic, onions, tangerine peel, oranges, grapes Pomelo, lemon, red orange, tangerine, grapefruit, fruit vegetable, pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, green leafy vegetables, lettuce, celery, spinach, parsley, chicory, legume Plants/vegetables (succulent and dry beans and peas), beans, green beans, pod beans, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, peas, lentils, oil Sub-crop, canola, castor, coconut, cotton, flax, oil palm, olive, peanut, rapeseed, safflower, sesame, to Japanese sunflower, soybean, pear fruit, apple, jellyfish, pear, alfalfa, summer flower hawthorn, root/tuber and bulbous vegetables, carrot, potato, sweet potato, cassava, beet, ginger, horseradish, radish, ginseng, turnip, Drupe, apricot, cherry, nectarine, peach, plum, plum, strawberry, nut, almond, pistachio, pecan, walnut, hazelnut, chestnut, cashew, beech nut, walnut, macadamia, kiwi, banana, (blue) Agave, grass, turf grass, ornamental plants, poinsettia, hardwood pruning, chestnut, oak, maple, sugar cane or beet. The method of claim 50, wherein the plant comprises soybean, bean, pod bean, wheat, cotton, corn, pepper, tomato, potato, tapioca, grape, strawberry, banana, peanut, squash, pumpkin, eggplant or cucumber. The method of claim 50, wherein the pathogenic infection is caused by a plant fungal pathogen, a plant bacterial pathogen, a rust fungus, a Botrytis, a Botrytis cinerea, a Botrytis cinerea, an Owenium genus, and a carrot soft rot Starch Amoeba, Diracium, D. dandelion, black smut, Agrobacterium, Agrobacterium tumefaciens, Xanthomonas, P. sphaeroides, Xanthomonas campestris Pathogenic species, Rhizoctonia solani, Rhizoctonia solani, Rice variety of Xanthomonas oryzae, Saccharomyces, Saccharomyces cerevisiae, Cartilian, Phloem phlei, Fusarium, Fusarium oxysporum, Wo Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium solani, Sclerotinia, S. sclerotiorum, Sclerotinia sclerotiorum, Sclerotinia sclerotiorum, Cercospora Genus, genus, genus, powdery mildew (white powder), genus of the genus of the genus of the genus (white powder), white stalk, single sac, clad, single sac, stalk, stalk Phomopsis solani, Alternaria, Alternaria alternata, onion chain Alternaria, Alternaria alternata, Alternaria alternata, Alternaria, Pseudomonas, Pseudomonas syringae, tomato Phytophthora, Phytophthora infestans, Phytophthora infestans, Phytophthora sojae Phytophthora capsici, Phytophthora capsici, Phytophthora strawberry, Phytophthora, eucalyptus, Phytophthora palmatum, tobacco Phytophthora fungi, rust fungus, rust fungus, rust layer pseudomonas, Aspergillus, Astragalus, Aspergillus niger, Puccinia rot, Bean rust, Cladosporium, Cladosporium, Rhizopus, Rhizopus, Penicillium, Rhizoctonia, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Cedar nucleus Fungi, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia platensis, Rhizoctonia solani, Coccidioides, Magnaporthe oryzae, Glomus , Fiji Glomus (Black Banana Leaf Spot), Apple Spotted Glomus, Lemon Glomus, Magnaporthe oryzae, Magnaporthe oryzae, Pseudomonas, Peach Brown Rot Pathogen, Blueberry Fruit Pathogen Spirulina platensis, anthracis, anthrax, anthrax, anthracnose, anthrax, genus, citrus, genus, genus, genus, genus Phytophthora, Phytophthora, Schizophyllum, Rhizoctonia solani, Myxosporium, Phaeocystis, Plasmodium, Grape canker, Camphor Genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Escherichia, Verticillium, Verticillium, Black Powder Genus, naked black powder fungus, maize black powder fungus, sugarcane whip black powder fungus, ergot genus, ergot fungus, sphaerotheca fuliginea, wheat black powder fungus, smooth black powder fungus, rice black powder fungus , wheat dwarf smut, stalk genus, soybean stalk, stalk, stalk, mildew, sphaerophyll, Helminthosporium, sclerotium, anthracnose, genus Barley genus, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium hirsutum or a combination thereof. The method of claim 52, wherein the pathogenic infection is caused by one or a combination of: soybean rust (Pythium humilis, Pseudomonas aeruginosa) and the plant comprises soybean; Botrytis cinerea (B. blight) and the plant comprises grapes; Botrytis cinerea (B. blight) and the plant comprises a strawberry; Botrytis cinerea (B. blight) and the plant comprises tomato; Alternaria (eg, Alternaria solani) and the plant comprises tomato; Alternaria (such as Alternaria solani) and the plant comprises potato; soybean rust (String rust) and the plant contains common beans; Phytophthora (soybean powdery mildew) and the plant comprises soybean; Fibroin bacterium (black banana leaf spot) or Fusarium oxysporum (Panama disease) and the plant contains banana; Xanthomonas or Xanthomonas oryzae Rice variety and the plant comprises rice; Rhizoctonia solani and the plant comprises cassava; Xanthomonas campestris and the plant comprises tomato; Botrytis cinerea (P. cinerea) and the plant comprises pepper; powdery mildew And the plant comprises a gourd; S. cerevisiae (White mold) and the plant comprises a pod bean; a S. cerevisiae (White mold) and the plant comprises a potato; Sclerotinia sclerotiorum (coin spot disease) and the plant comprises Turf grass; southern white mold and the Containing peanuts; leaf spot (C. cerevisiae / Cercospora) and the plant comprises peanuts; Fusarium graminearum (branches of wheat) and the plant comprises wheat; Phytophthora sinensis (S. cerevisiae) The plant comprises wheat; A. sinensis (Bacterial blight and Phytophthora capsici) and the plant comprises wheat; the starch is worm-infected and the plant comprises apple, pear and other pear fruit; apple black scab The plant comprises apple, pear and other pear fruit; or Rhizoctonia solani and the plant comprises wheat, rice, turf, soybean, corn, legume and vegetable crops. A method for achieving one or both of beneficial plant growth or protection of a susceptible plant to avoid a pathogenic infection, the method comprising: delivering a composition to the leaf of the plant, the bark of the plant , the fruit of the plant, the flower of the plant, the seed of the plant, the root of the plant, the pruning of the plant, the graft of the plant, the healing tissue of the plant; the soil or growth medium surrounding the plant; The soil or growth medium prior to seeding the seed of the plant in the growth medium; or planting the plant, the plant pruning, the plant graft or the soil or growth medium prior to healing the tissue in the soil or growth medium, The composition comprises the following: a biologically pure culture of Bacillus amyloliquefaciens RTI301 deposited with ATCC PTA-121165 or a mutant having all of its identifying characteristics, which is suitable for plant growth and/or protection Avoid the amount of the pathogenic infection; and microbial, biological or chemical insecticides, fungicides, nematicides, fungicides, herbicides, plant extracts, plants Growth regulators or fertilizers or one combination, which was the amount of plant growth and / or confer protection against infection disease immunogenic adapted to benefit. The method of claim 54, wherein the composition is delivered to the leaves of the plant. The method of claim 54, wherein the plant growth benefit and/or the protection conferred by the avoidance of the pathogenic infection is manifested by improved seedling vigor, improved root development, improved plant health, and improved Plant quality, increased yield, improved appearance, increased resistance to plant pathogens, reduced pathogenic infection, or a combination thereof. The method of claim 54, wherein the B. amyloliquefaciens RTI301 is in the form of spores or vegetative cells. The method of claim 54, wherein the plant comprises monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, feed Corn, rice, wheat, barley, sorghum, asparagus, berries, blueberries, blackberries, raspberries, raspberries, cranberries, cranberries, gooseberries, elderberries, currants, cranberries, berries, kale Vegetables, broccoli, kale, broccoli, Brussels sprouts, loose cabbage, kale, mustard, broccoli, melon, cucumber, cantaloupe, melon, sweet melon, pumpkin, watermelon, pumpkin, eggplant, bulb Vegetables, onions, garlic, onions, tangerine peel, oranges, grapefruit, lemon, red tangerine, tangerine, grapefruit, fruit and vegetable, pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, green leafy vegetables , lettuce, celery, spinach, parsley, chicory, legumes/vegetables (succulent and dry beans and peas), beans, green beans, food pods, shell beans, soybeans, dried beans, back beans, Lima Beans, peas, chickpeas, pea petals, lentils, oilseed crops, canola, ramie, coconut, cotton, flax, oil palm, olive, peanut, rapeseed, safflower, sesame, sunflower, soybean, pear, apple , Jelly, pear, alfalfa, summer flower hawthorn, root/tuber and bulbous vegetables, carrots, potatoes, sweet potatoes, cassava, beets, ginger, horseradish, radish, ginseng, turnip, stone fruit, apricots, cherries, nectarines, peaches , plums, plums, strawberries, nuts, almonds, pistachios, pecans, walnuts, hazelnuts, chestnuts, cashews, beech nuts, walnuts, macadamia, kiwi, bananas, (blue) agave, grass, turf grass, ornamental Plant, poinsettia, hardwood pruning, chestnut, oak, maple, sugar cane or beet. The method of claim 58, wherein the plant comprises soybean, bean, pod bean, wheat, cotton, corn, pepper, tomato, potato, tapioca, grape, strawberry, banana, peanut, squash, pumpkin, eggplant or cucumber. The method of claim 54, wherein the pathogenic infection is caused by a plant fungal pathogen, a plant bacterial pathogen, a rust fungus, a Botrytis, a Botrytis cinerea, a Botrytis cinerea, an Erwinia, and a carrot soft rot , starch-degrading Owen bacteria, Dick's genus, D. dandelion, black smut, Agrobacterium, root cancer Alcaligenes, Xanthomonas, Rhizoctonia solani, Xanthomonas camphora, carcinogenic species, P. oleracea, P. oleracea, rice xanthophyll rice variety, Saccharomyces, Leaf sclerotium, Carterella, phloem, Fusarium, Fusarium graminearum, Fusarium graminearum, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, nuclear disk Phytophthora, S. cerevisiae, Sclerotinia sclerotiorum, Sclerotinia sclerotiorum, Cercospora, H. genus, Powdery mildew (powdery), Trichophyton (powdery), White fork Silk single-shell shell, Crande fork silk single-shell shell, Pseudomonas spp., Pseudomonas cepacii, Alternaria, Alternaria alternata, Alternaria alternata, Alternaria alternata Bacillus, Alternaria alternata, Alternaria, Pseudomonas, Pseudomonas syringae, tomato Phytophthora, Phytophthora infestans, Phytophthora parasitica, Phytophthora sojae, Phytophthora capsici, 樟Phytophthora, Phytophthora strawberry, Phytophthora, eucalyptus, Phytophthora palm, Phytophthora nicotianae, rust fungus, beans Potato leaf rust, layer rust layer Pseudomonas, Aspergillus, Astragalus, Aspergillus niger, Puccinia genus, Bean rust, Cladosporium, Cladosporium, Rhizopus, less Rhizopus, Penicillium, Rhizoctonia, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Strawberry Rhizoctonia, Rhizoctonia solani, Rhizoctonia platensis, Rhizoctonia solani, Coccidioides, Magnaporthe oryzae, Glomus, G. sphaeroides, Ficus sclerotium (black banana leaf spot) Disease), apple spotted bacterium, lemon bacterium, rice blast fungus, rice blast fungus, genus Streptomyces, brown rot fungus, blueberry fruit pathogen, nucleus of the genus Serum, anthracnose, Anthrax solani, anthracnose, anthracnose, anthrax, genus, citrus, genus, genus, genus, rust, rust, rust Genus, Phytophthora, Phytophthora, Phytophthora, Phytophthora, Phytophthora, Phytophthora, Phytophthora If the spore globosum L. Wilson, Sphaerotheca spp., Sphaerotheca fungus spots, rose Sphaerotheca fungus, Erysiphe, Phytophthora, Phytophthora, Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Pseudostellaria, Apple Black Star disease, Verticillium, sphaerotheca, naked black powder, black smut, sugarcane, black bacterium, ergot, ergot, sphaerotheca, wheat smut , smooth black powder fungus, rice black powder fungus, wheat dwarf smut, stalk genus, soybean stalk mold, variegated shoot mold, cabbage stem mold, Helminthosporium, Woding Capsule, anthracis, genus, genus, genus, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium var. Its combination. The method of claim 60, wherein the pathogenic infection is caused by one or a combination of: soybean rust (Phaeophyllum serrata, Pseudomonas sphaeroides) and the plant comprises soybean; Botrytis cinerea (grape Spore plague) and the plant comprises a grape; Botrytis cinerea (B. blight) and the plant comprises a strawberry; Botrytis cinerea (B. blight) and the plant comprises a tomato; the genus Alternaria (eg, the genus And the plant comprises tomato; Alternaria (such as Alternaria alternata) and the plant comprises potato; bean rust (String rust) and the plant contains common beans; powdery mildew (soybean powdery mildew) And the plant comprises soybean; Ficus faecium (black banana leaf spot) or Fusarium oxysporum (Panama disease) and the plant comprises a banana; a yellow variant of Xanthomonas or Xanthomonas oryzae and the plant comprises Rice; Phytophthora sojae and the plant comprising cassava; Xanthomonas campestris and the plant comprising tomato; Botrytis cinerea (P. cinerea) and the plant comprising pepper; powdery mildew and the plant comprising a gourd; S. cerevisiae (White mold) and the plant comprises a pod bean; S. cerevisiae (White mold) and the plant comprises potato; Sclerotinia sclerotiorum (coin spot disease) and the plant comprises turfgrass; Southern white mold And the plant comprises peanut; leaf spot disease (S. cerevisiae / Cercospora) and the plant comprises peanut; Fusarium graminearum (Schistosomiasis) and the plant comprises wheat; and the bacterium of the genus Corydalis And the plant comprises wheat; Phytophthora sinensis (Bacterial blight and Phytophthora capsici) and the plant comprises wheat; the starch is worm-infected and the plant comprises apple, pear and other pear fruit; apple black star A pathogen and the plant comprises apple, pear, and other pear fruit; or Rhizoctonia solani and the plant comprises wheat, rice, turf, soybean, corn, legume, and vegetable crops. The method of claim 54, wherein the amount suitable for plant growth and/or protection from pathogenic infection is from about 1.0 x 10 ¹⁰ CFU/ha to about 1.0 x 10 ¹⁴ CFU/ha Bacillus amyloliques RTI301 . A method for benefiting plant growth by imparting protection to susceptible plants to avoid pathogenic infections or reducing pathogenic infections while minimizing the accumulation of resistance to treatment, the method comprising: treating the first composition with The second composition is delivered to the susceptible plant at separate application and varying time intervals, wherein the first composition and the second composition are each adapted to confer protection to the plant to avoid pathogenic infection or reduce pathogens Delivery of a sexual infection, wherein the first composition comprises a B. amyloliquefaci RTI301 deposited with ATCC No. PTA-121165 or a biologically pure culture thereof having a mutant having all of its identifying characteristics; and wherein the second composition Containing one or more compounds with fungicidal or bactericidal properties An active agent, and wherein the first composition and the second composition are delivered to the leaves of the plant, the bark of the plant, the fruit of the plant, the flower of the plant, the seed of the plant at the time interval of the change One or a combination of the root of the plant, the pruning of the plant, the graft of the plant, the healing tissue of the plant, or the soil or growth medium surrounding the plant, wherein protection is provided to avoid the pathogenic infection and/or to reduce The total amount of the (s) chemically active agent required for pathogenic infection is reduced and the accumulation of resistance to treatment is minimized. The method of claim 63, wherein the plant comprises monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, forage corn, rice, wheat, barley, sorghum, asparagus, berries, Blueberries, blackberries, raspberries, raspberries, cranberries, cranberries, gooseberries, elderberries, currants, cranberries, berry, cabbage, broccoli, kale, broccoli, Brussels sprouts, Amaranth, kale, mustard, stalk, pumpkin, eggplant, melon, cucumber, cantaloupe, melon, sweet melon, squash, watermelon, bulbous vegetables, onions, garlic, onions, tangerine peel, oranges, grapes Pomelo, lemon, red orange, tangerine, grapefruit, fruit vegetable, pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, green leafy vegetables, lettuce, celery, spinach, parsley, chicory, legume Plants/vegetables (succulent and dry beans and peas), beans, green beans, pod beans, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, peas, lentils, oil Sub-crops, canola, ramie, coconut, cotton, flax, oil palm, olive, peanut, rapeseed, safflower, sesame, sunflower, soybean, pear, apple, jellyfish, pear, alfalfa, summer flower haw, Roots/tubers and bulbs vegetables, carrots, potatoes, sweet potatoes, cassava, beets, ginger, horseradish, radish, ginseng, turnip, stone fruit, apricots, cherries, nectarines, peaches, Plum, plum, strawberry, nut, almond, pistachio, pecan, walnut, hazelnut, chestnut, cashew, beech nut, walnut, macadamia, kiwi, banana, (blue) agave, grass, turf, ornamental Poinsettia, hardwood pruning, chestnut, oak, maple, sugar cane or beet. The method of claim 63, wherein the pathogenic infection is caused by a plant fungal pathogen, a plant bacterial pathogen, a rust fungus, a Botrytis, a Botrytis cinerea, a Botrytis cinerea, an Erwinia, and a carrot soft rot Starch Amoeba, Diracium, D. dandelion, black smut, Agrobacterium, Agrobacterium tumefaciens, Xanthomonas, P. sphaeroides, Xanthomonas campestris Pathogenic species, Rhizoctonia solani, Rhizoctonia solani, Rice variety of Xanthomonas oryzae, Saccharomyces, Saccharomyces cerevisiae, Cartilian, Phloem phlei, Fusarium, Fusarium oxysporum, Wo Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium solani, Sclerotinia, S. sclerotiorum, Sclerotinia sclerotiorum, Sclerotinia sclerotiorum, Cercospora Genus, genus, genus, powdery mildew (white powder), genus of the genus of the genus of the genus (white powder), white stalk, single sac, clad, single sac, stalk, stalk Phomopsis solani, Alternaria, Alternaria alternata, onion chain Alternaria, Alternaria alternata, Alternaria alternata, Alternaria, Pseudomonas, Pseudomonas syringae, tomato Phytophthora, Phytophthora infestans, Phytophthora infestans, Phytophthora sojae Phytophthora capsici, Phytophthora capsici, Phytophthora infestans, Phytophthora, eucalyptus, Phytophthora palmatum, Phytophthora nicotianae, rust fungus, rust fungus, rust layer pseudomonas, koji Fungi, Astragalus, Aspergillus niger, Puccinia, Phytophthora rust, Cladosporium, Cladosporium, Rhizopus, Rhizopus, Penicillium, Rhizoctonia Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia cerealis, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia granulosus , Rhizoctonia solani, Coccidioides, Magnaporthe oryzae, Glomus Glomus bacillus, Filipino sclerotium (black banana leaf spot), apple spotted bacterium, lemon bacterium, rice blast, rice blast, chain genus, brown rot, blueberry Fruit pathogens, Sclerotinia sclerotiorum, Anthrax, Anthrax anthracis, Capsicum anthracis, Anthracnose, Aphis citrinum, Citrus genus, Corynebacterium, Multi-C. Genus, Phytophthora, Phytophthora, Schizophyllum, Rhizoctonia solani, Myxosporium, Phaeocystis, Plasmodium, Grape canker, Brucella Genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus, genus Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Pythium, Escherichia, Verticillium, Verticillium, Black Powder Genus, naked black powder fungus, maize black powder fungus, sugarcane whip black powder fungus, ergot genus, ergot fungus, sphaerotheca fuliginea, wheat smut, Phytophthora black fungus, rice black powder fungus, wheat dwarf smut, stalk genus, soybean stalk, mildew, sphaerophyll, cabbage stem, Helminthosporium, sulcus Shell, anthracnose, genus of genus, genus, genus, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium, Helminthosporium ssp. combination. The method of claim 63, wherein the change intervals are 5 to 7 days apart. The method of claim 63, wherein the first composition and the second composition are each delivered to a leaf of the plant, a fruit of the plant, or a flower of the plant. The method of claim 63, wherein the amount suitable for imparting a protective system to the plant to avoid pathogenic infection or reducing pathogenic infection is from about 1.0 x 10 ¹⁰ CFU/ha to about 1.0 x 10 ¹⁴ CFU/ha of amylopectin spores. Bacillus RTI301. The method of claim 63, wherein the chemically active agent comprises one or a combination of: strobilurine, triazole, flutriafol, dextran, prothiaconazole, fluoro Expoxazole (expoxyconazole) Bacteramide, tetrachloroisophthalonitrile, methyl thiophanate, copper hydroxide fungicide, EDBC-based fungicide, zinc-manganese (mancozeb), succinase dehydrogenase (SDHI) fungicide Agent, bixafen, iprodione, dimethomorph or valifenalate. The method of claim 69, wherein the chemically active agent comprises flupirtine plus dextran, and delivery of the first composition comprising the RTI 301 replaces delivery of the tetrachloroisophthalonitrile fungicide. The method of claim 70, wherein the plant comprises a gourd and the pathogenic infection is caused by powdery mildew. The method of claim 69, wherein the chemically active agent comprises a methyl thiophanate fungicide, and the delivery of the first composition comprising the RTI 301 replaces the delivery of the prothioconazole fungicide. The method of claim 69, wherein the chemically active agent comprises a copper hydroxide fungicide, and the delivery of the first composition comprising the RTI 301 replaces the delivery of the tetrachloroisophthalonitrile fungicide. The method of claim 63, wherein the first composition further comprises one or a combination of: a carrier, a surfactant, a dispersant, or a yeast extract. A composition comprising at least one of the following: an isolated abundance-MA compound, an isolated abundance MB compound, an isolated abundance MC compound, an isolated dehydroxyl-protonin MA compound, an isolated dehydroxyeductin The MB compound, the isolated hydroxytoxin MC compound, the isolated abundance H compound, the isolated hydroxy abbreviated H compound, the isolated abundance I compound, and the isolated hydroxy-free prostaglandin I compound are suitable for the susceptibility The disease plant confers protection against pathogenic infections and the compound has the formula: Wherein R is OH, n is in the range of 8 to 20, FA is linear, XOR or di-isomeric, and: for abundance MA, X ₁ is Ala, X ₂ is Thr, and X ₃ is Met; for abundance MB X ₁ is Val, X ₂ is Thr, and X ₃ is Met; for abundance MC, X ₁ is Aba, X ₂ is Thr, and X ₃ is Met; for abundance H, X ₁ is Val, X ₂ Is Thr and X ₃ is Hcy; and for abundance I, X ₁ is Ile, X ₂ is Thr, and X ₃ is Ile; and wherein R is H, n is in the range of 8 to 20, and FA is linear, XOR or singular, and: for dehydroxylectin MA, X ₁ is Ala, X ₂ is Thr, and X ₃ is Met; for dehydroxyl-rich MB, X ₁ is Val, X ₂ is Thr, and X ₃ Is Met; for dehydroxylectin MC, X ₁ is Aba, X ₂ is Thr, and X ₃ is Met; for dehydroxylectin H, X ₁ is Val, X ₂ is Thr, and X ₃ is Hcy; For dehydroxylectin I, X ₁ is Ile, X ₂ is Thr, and X ₃ is Ile. The composition of claim 75, further comprising one or a combination of the presence of an amount suitable for beneficial to plant growth and/or protection of the susceptible plant to avoid pathogenic infection: microbial, biological or chemical insecticidal Agents, fungicides, nematicides, fungicides, herbicides, plant extracts, white lupin extracts, BLAD polypeptides, fragments of BLAD polypeptides, plant growth regulators or fertilizers. The composition of claim 75, wherein the composition is in the form of a liquid, dust, sprinkable particles or dry wettable particles. A kind of Bacillus amyloliquefaciens RTI301 deposited under ATCC No. PTA-121165 An extract of a pure culture, the extract comprising at least one of the following: an isolated abundance-MA compound, an isolated abundance MB compound, an isolated abundance MC compound, an isolated dehydroxyl-protonin MA compound, The isolated hydroxytobarenin MB compound, the isolated hydroxy-hydroxytoxin MC compound, the isolated abundance H compound, the isolated hydroxy-hydroxytoxin H compound, the isolated abundance I compound, and the isolated hydroxy-hydroxytoxin I compound. A method of protecting a plant or fruit from a pathogenic infection or treating a pathogenic infection of a plant or fruit, comprising administering an effective amount of a composition or extract of any one of claims 75, 76, 77 or 78 to The plant or fruit, or the soil around the roots or roots of the plants. The method of claim 79, wherein the plant comprises monocots, dicots, grains, corn, sweet corn, popcorn, corn, silage corn, forage corn, rice, wheat, barley, sorghum, asparagus, berries, Blueberries, blackberries, raspberries, raspberries, cranberries, cranberries, gooseberries, elderberries, currants, cranberries, berry, cabbage, broccoli, kale, broccoli, Brussels sprouts, Cabbage, kale, mustard, stalk, melon, cucumber, cantaloupe, melon, sweet melon, squash, watermelon, pumpkin, eggplant, bulbous vegetables, onions, garlic, onions, tangerine peel, oranges, grapes Pomelo, lemon, red orange, tangerine, grapefruit, fruit vegetable, pepper, tomato, ground cherry, tree tomato, okra, grape, vanilla/spice, green leafy vegetables, lettuce, celery, spinach, parsley, chicory, legume Plants/vegetables (succulent and dry beans and peas), beans, green beans, pod beans, shell beans, soybeans, dried beans, peas, lima beans, peas, chickpeas, peas, lentils, oil Sub-crops, canola, ramie, coconut, cotton, flax, oil palm, olive, peanut, rapeseed, safflower, sesame, sunflower, soybean, pear, apple, jellyfish, pear, alfalfa, summer flower haw, Roots/tubers and bulbs vegetables, carrots, potatoes, sweet potatoes, cassava, beets, ginger, Horseradish, radish, ginseng, turnip, stone fruit, apricot, cherry, nectarine, peach, plum, plum, strawberry, nut, almond, pistachio, pecan, walnut, hazelnut, chestnut, cashew, beech nut, walnut, hawaii Fruit, kiwi, banana, (blue) agave, grass, turf grass, ornamental plant, poinsettia, hardwood pruning, chestnut, oak, maple, sugar cane or beet. The method of claim 79, wherein the pathogenic infection is caused by: a plant fungal pathogen, a plant bacterial pathogen, a rust fungus, a Botrytis, a gray grape, a Bacillus licheniformis, an Owenium, a carrot, a soft rot Starch Amoeba, Diracium, D. dandelion, black smut, Agrobacterium, Agrobacterium tumefaciens, Xanthomonas, P. sphaeroides, Xanthomonas campestris Pathogenic species, Rhizoctonia solani, Rhizoctonia solani, Rice variety of Xanthomonas oryzae, Saccharomyces, Saccharomyces cerevisiae, Cartilian, Phloem phlei, Fusarium, Fusarium oxysporum, Wo Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium oxysporum, Fusarium solani, Sclerotinia, S. sclerotiorum, Sclerotinia sclerotiorum, Sclerotinia sclerotiorum, Cercospora Genus, genus, genus, powdery mildew (white powder), genus of the genus of the genus of the genus (white powder), white stalk, single sac, clad, single sac, stalk, stalk Phomopsis solani, Alternaria, Alternaria alternata, onion chain Alternaria, Alternaria alternata, Alternaria alternata, Alternaria, Pseudomonas, Pseudomonas syringae, tomato Phytophthora, Phytophthora infestans, Phytophthora infestans, Phytophthora sojae Phytophthora capsici, Phytophthora capsici, Phytophthora infestans, Phytophthora, eucalyptus, Phytophthora palmatum, Phytophthora nicotianae, rust fungus, rust fungus, rust layer pseudomonas, koji Fungi, Astragalus, Aspergillus niger, Puccinia, Phytophthora rust, Cladosporium, Cladosporium, Rhizopus, Rhizopus, Penicillium, Rhizoctonia Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia cerealis, Rhizoctonia solani, Rhizoctonia solani, Rhizoctonia solani, Raspberry Rhizoctonia, Rhizoctonia solani, Coccidioides, Magnaporthe oryzae, Glomus, G. sphaeroides, Ficus sclerotium (black banana leaf spot), apple spotted bacterium, lemon ball Phytophthora, Magnaporthe oryzae, Magnaporthe oryzae, Sclerotinia, Peach Brown Rot Pathogen, Blueberry Fruit Pathogen, Sclerotinia sclerotiorum, Anthrax, Anthrax anthracis, Capsicum anthracis, Anthracnose Bacteria, genus, citrus, genus, genus, genus, genus, genus, rust, rust, genus, genus, genus, genus Helicobacter, Phytophthora, Phytophthora, Phytophthora, Phytophthora, genus, genus, genus, genus, genus, genus, singular, single sac Phytophthora, Rhizoctonia solani, Powdery mildew, genus Polyporus, Phytophthora, Pythium, Pythium, Pythium, Pythium, Pythium, Pythium Mold, forest Pythium, Black genus, Apple black spot, Verticillium, Black powder, Naked black powder, Jade black powder, sugar cane Black powder fungus, ergot fungus, ergot fungus, sphaerotheca fuliginea, wheat black powder fungus, smooth black powder fungus, rice black powder fungus, wheat dwarf black powder fungus, stem mold, soybean Spirulina platensis, variegated shoot mold, cabbage stem mold, Helminthosporium, Hessian shell, Anthrax, Rhizoctonia, Barley grisea, Helminthosporium, Helminthosporium , Barley Helminthosporium, Zea mays, Helminthosporium, Wheat Helminthosporium or a combination thereof. A composition for plant growth comprising: a Bacillus amyloliquefaciens RTI301 deposited with ATCC No. PTA-121165 or a biologically pure culture thereof having mutants having all of its identifying characteristics; and bifenthrin killing Insecticide. The composition of claim 82, wherein the composition is in a formulation compatible with a liquid fertilizer. The composition of claim 83, wherein the composition further comprises hydrated aluminum-magnesium silicate and at least one dispersant. The composition of claim 83, wherein the Phenfenin insecticide is present in a concentration ranging from 0.1 g/ml to 0.2 g/ml. The composition of claim 83, wherein the Phenfenin insecticide is present at a concentration of about 0.1715 g/ml. A composition useful for plant growth, comprising: a Bacillus amyloliquefaciens RTI301 deposited with ATCC No. PTA-121165 or a biologically pure culture thereof having mutants having all of its identifying characteristics; and comprising from a white lupin One or a combination of fungicides of extracts of beans, BLAD polypeptides or fragments of BLAD polypeptides. The composition of claim 87, wherein the composition comprises about 20% of the BLAD polypeptide or a fragment of the BLAD polypeptide. A product comprising: a first composition comprising a Bacillus amyloliquefaciens RTI301 deposited with ATCC PTA-121165 or a biologically pure culture thereof having mutants having all of its identifying characteristics; a second combination The second composition comprises one or a combination of a microorganism, a biological or chemical insecticide, a fungicide, a nematicide, a bactericide, a herbicide, a plant extract, a plant growth regulator or a fertilizer, wherein the first The composition and the second composition are packaged separately, and wherein each composition is an amount suitable for beneficial to plant growth and/or to confer protection to susceptible plants to avoid pathogenic infection; and instructions for Delivering a combination of the first composition and the second composition in an amount suitable for plant growth to: a leaf of the plant, a bark of the plant, a fruit of the plant, a flower of the plant, a seed of the plant , the root of the plant, the pruning of the plant, the graft of the plant, the healing tissue of the plant; the soil or growth medium surrounding the plant; sowing the plant in soil or growth medium The growing medium or soil before the seeds; or soil or growth medium The plant, the plant pruning, the plant graft or the soil or growth medium prior to healing the tissue of the plant is planted in the plastid. The product of claim 89, wherein the insecticide is one or a combination of the following: pyrethroid, bifenin, tefluthrin, anthraquinone, organophosphate, oxychloride Chlorethoxyphos, chlorpyrifos, tebupirimphos, cyfluthrin, fiproles, fipronil, nicotinoids or cotinine Clothianidin). The product of claim 89, wherein the first composition further comprises one or a combination of: a carrier, a surfactant, a dispersant, or a yeast extract. The product of claim 89, wherein the first composition and the second composition are each in the form of a liquid, a powder, a spreadable granule, a dry wettable powder or a dry wettable granule. The product of claim 89, wherein the first composition is in liquid form, and the B. amyloliquefaciens RTI301 is present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml. The product of claim 89, wherein the first composition is in the form of a powder, a dry wettable powder, a dispersible granule or a dry wettable granule and the B. amyloliquefaciens RTI 301 is at about 1.0 x 10 ⁸ CFU/g to It is present in an amount of about 1.0 x 10 ¹² CFU/g. The product of claim 89, wherein the first composition is in the form of an oily dispersion, and the B. amyloliquefaciens RTI301 is present at a concentration of from about 1.0 x 10 ⁸ CFU/ml to about 1.0 x 10 ¹² CFU/ml.