Biological pesticides

Abstract

A strain of Bacillus licheniformis (named "K-357") acted as an insecticide. Surprisingly, application of a single strain of Bacillus licheniformis K-357 reduced nematode hatching and increased nematode mortality in vitro, and reduced nematode numbers while increasing plant vigor and yield in cotton field trials. The nematicidal activity and cotton yield of seeds treated with Bacillus licheniformis K-357, seeds treated with aldicarb and fluopyram in furrows, and seeds treated with abamectin as a nematicide were compared. The nematicidal activity of Bacillus licheniformis K-357 was comparable to, and in some cases superior to, chemical treatment. Cotton yield was increased by seeds treated with K-357 as a nematicide compared to those treated with furrow application and chemical nematicides. Maize seeds treated with Bacillus licheniformis K-357 in combination with chemical insecticides and fungicides reduced root node damage in field trials of maize severely infested with maize rootworms.

Description

[0001] Cross-reference of related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 061,906, filed August 6, 2020; U.S. Provisional Patent Application No. 63 / 210,086, filed June 14, 2021; and U.S. Provisional Patent Application No. 63 / 215,239, filed June 25, 2021, all of which are incorporated herein by reference in their entirety. Technical Field

[0003] This invention relates to the use of microbial compositions for controlling plant pathogenic insects and increasing crop yields. More specifically, this invention relates to applying compositions containing microbial strains to plants, seeds from which plants grow, or the environment in which plants grow. Background Technology

[0004] Crops' responses to global warming indicate that yields will decline as growing season temperatures rise. Deutsche et al. point out that pests may exacerbate this impact (C. Deutsch, et al. Dryad (2018); https: / / dx.doi.org / 10.5061 / dryad.b7q3g2q). Insects already consume 5% to 20% of major cereal crops. The authors' model shows that for the three most important cereal crops—wheat, rice, and maize—insect-induced yield losses will increase by 10% to 25% for every 1 degree Celsius increase in temperature, with the greatest impact on temperate regions. These findings provide estimates of further potential climate impacts on global food supply and set a benchmark for future regional and sector-specific studies on the interactions of crops, pests, and climate.

[0005] In the American Corn Belt, corn rootworms (Diabrotica spp.) are the number one insect threatening corn yields for farmers. Studies over many years estimate that these pests cause nearly $1 billion in economic losses annually. There are three species of corn rootworms: the western corn rootworm, the northern corn rootworm, and the southern corn rootworm. All species share a similar life cycle, consisting of four stages: egg, larva, pupa, and adult. They each lay eggs, which hatch into larvae (worms) that feed on the roots of corn plants.

[0006] Furthermore, due to its worldwide distribution and economic impact, Potato Virus Y (PVY) is one of the ten most important plant viruses affecting Solanaceae crops. The most effective way to control PVY is to prevent the virus from entering farmland. However, once a plant is infected, it remains infected throughout its life and becomes an inoculum for aphids to spread the PVY virus. Approximately 65 aphid species have been documented to spread the PVY virus to varying degrees (Pelletier, Y et al. J. Econ. Entomol. 105, 1909-1914, doi:10.1603 / ec12085 (2012)). Using insecticides and horticultural mineral oils to control aphids can reduce the spread of PVY in the field. Nevertheless, most aphid vectors of PVY do not reside on potatoes and are therefore unaffected by systemic insecticides. Commercially available virus-resistant varieties only confer moderate resistance to PVY. Therefore, it is necessary to develop new strategies to control the seasonal spread of PVY.

[0007] It is estimated that plant parasitic nematodes cause far greater losses annually than pests (Singh et al., Procedia Environmental Sciences 29(2015)215-216). Crop yield losses in many countries are enormous due to these tiny, invisible pests. They contribute to an estimated 12.3% ($157 billion) in global yield losses. Growers consider pests and other limiting factors as production problems, but overlook plant parasitic nematodes. Nematode diseases are difficult to control because of their insidious nature, making them easier to ignore. Plant parasitic nematodes not only cause damage individually but also form disease complexes with other microorganisms, increasing crop losses.

[0008] The chemical standard for effectively protecting crops from nematode infestation is aldicarb, a carbamate insecticide and the active ingredient in pesticides formerly sold under the trade name TEMIK. Aldicarb is banned in Europe, and in 2010, the U.S. Environmental Protection Agency (EPA) banned it due to its high toxicity. A new aldicarb pesticide, AGLOGIC 15G, was approved by the EPA in December 2011 and entered the market in 2015. Aldicarb is one of the most widely used pesticides internationally and also one of the most environmentally toxic. Aldicarb poisoning in agricultural runoff leads to the destruction of healthy ecosystems and irreversible damage to fertile farmland.

[0009] Much of the recent interest in bioproducts for controlling agricultural pests stems from the desire to replace synthetic fertilizers and chemical pesticides to improve crop health and limit negative environmental impacts. Research has demonstrated the economic benefits of bioproducts and the reliable performance of integrated crop management systems. Microbial products are a class of bioproducts derived from naturally occurring microorganisms, such as bacteria and fungi. These products are live strains of microorganisms that can be used in seeding, in-furrow application, or sprayed onto crops to grow alongside them, preventing pests and diseases, or enhancing plant productivity and fertility. Microbial products have the potential to provide sustainable, cost-effective solutions that help increase yields while using fewer inputs. However, much work remains to be done in utilizing microorganisms to control plant pests, particularly nematodes.

[0010] A range of bacterial and fungal bionematicides already used in integrated nematode management strategies include Aspergillus niger, Paecilomyces lilacinus, Trichoderma harzianum, Trichoderma viride, Pochonia chlamydosporia, Pasteuria penetrans, Pseudomonas flurorescens, Bacillus firmus, B. thuringiensis, B. velzensis, B. mojavensis, and B. subtilis (Xiang et al., Plant Disease (2017) 101:774; Abd-Elgawad and Askary, Egyptian Journal of Biological Pest Management). Control (2018) 28:74). In one instance, European Patent No. EP2603086B1 describes the use of Bacillus subtilis strain DSM17231 in combination with Bacillus licheniformis strain DSM17236 (marketed under the trade name NEMIX) for the control of plant nematodes. This combination of strains showed satisfactory results in inhibiting nematode movement, but its effect on hatching was not significant; only the chemical product TEMIK could inhibit hatching. Another example is the product MELOCON WG (CERTIS USA) for the control of soil nematodes. MELOCON WG is based on the fungus Paecilomyces lilacinus, which is a parasite of common plant nematodes at all developmental stages, especially eggs and infective larvae.

[0011] As mentioned above, many microbial strains have been tested as nematicides, and several bionematicide products are commercially available. However, existing bionematicides and biopesticides are insufficient to address the serious global threat posed to plants by plant-parasitic nematodes and pathogenic insects.

[0012] Therefore, there is a need for improved microbial insecticides to reduce insect damage to crops, including nematodes, corn root borers, and aphids, some of the most troublesome insects, without the toxicity of chemical insecticides. This disclosure provides such improved microorganisms and methods of application to reduce pests and improve plant health and crop yield. Summary of the Invention

[0013] In one embodiment, a method for controlling plant pests is provided, the method comprising: planting a plant or plant seed in a suitable growing environment, the plant or seed having a coating or partial coating of a composition comprising: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients or microbial crop protectants, bio-crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0014] In one embodiment, a method for controlling plant pests is provided, the method comprising: delivering a composition to the seeds, foliage, roots, or soil or growing environment surrounding a plant, the composition comprising: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients, or microbial crop protectants, bio-crop protectants, or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0015] In one embodiment, a method for controlling plant pests is provided, the method comprising: planting a plant or plant seed in a suitable growing environment, the plant or seed having a coating or partial coating of a composition substantially consisting of: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and one or more carriers, excipients or nutrients, wherein the composition increases plant yield or reduces plant pests, or both, in the presence of plant pathogenic insects.

[0016] In one embodiment, a method for controlling plant pests is provided, the method comprising: delivering a composition to the seeds, leaves, roots, or soil or growing environment surrounding a plant, the composition consisting essentially of: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics, and one or more carriers, excipients, or nutrients, wherein the composition increases plant yield or reduces plant pests, or both, in the presence of plant pathogenic insects.

[0017] In one embodiment, a method for treating plants or plant seeds is provided, comprising applying a composition coating to the plants or plant seeds, said composition coating comprising a biologically pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics, said composition optionally comprising one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, said composition not comprising Bacillus subtilis, and said Bacillus licheniformis K-357 being present in the coating in an amount suitable for increasing plant yield or reducing plant pests or both in the presence of plant pathogenic insects.

[0018] In one embodiment, a plant or plant seed is provided having a composition coating comprising: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in the coating in an amount suitable for increasing plant yield or reducing plant pests or both in the presence of plant pathogenic insects.

[0019] In one embodiment, a method for treating plants or plant seeds is provided, comprising applying a composition coating to the plants or plant seeds, said composition coating comprising a biologically pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics, said composition optionally comprising one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, said composition not comprising Bacillus subtilis, and said Bacillus licheniformis K-357 being present in the coating in an amount suitable for increasing plant yield or reducing plant pests or both in the presence of plant pathogenic insects.

[0020] In one embodiment, a plant or plant seed is provided having a composition coating comprising: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in the coating in an amount suitable for increasing plant yield or reducing plant pests or both in the presence of plant pathogenic insects.

[0021] In one embodiment, a method for controlling plant pathogenic nematodes is provided, the method comprising planting plant seeds in a suitable growing environment, the seeds having a coating comprising: spores of a biopure culture of a single microbial strain, said single microbial strain being *Bacillus licheniformis* K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients, crop protectants, fungicides, insecticides, or nematicides. *Bacillus licheniformis* K-357 is present in the seeds in amounts suitable for increasing plant yield, increasing plant resistance to plant pathogenic nematodes, or reducing plant infection by plant pathogenic nematodes, and combinations thereof, in the presence of plant pathogenic nematodes.

[0022] In one embodiment, a method for controlling plant pathogenic nematodes is provided, the method comprising delivering a composition to the seeds, roots, or growing environment surrounding a plant, the composition comprising: a biopure culture of a single microbial strain, said microbial strain being Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more vectors, excipients, nutrients, crop protectants, fungicides, insecticides, or nematicides. In the presence of plant pathogenic nematodes, delivery of said composition increases plant yield, enhances plant resistance to plant pathogenic nematodes, or reduces plant infection by plant pathogenic nematodes, and combinations thereof.

[0023] In one embodiment, a method for controlling plant pathogenic nematodes is provided, the method comprising delivering a composition to the seeds, roots, or soil or growing environment surrounding a plant, the composition consisting essentially of: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and one or more carriers, excipients, or nutrients. In the presence of plant pathogenic nematodes, the delivery of the composition increases plant yield, enhances plant resistance to plant pathogenic nematodes, or reduces plant infection by plant pathogenic nematodes, or combinations thereof.

[0024] In one embodiment, a method for treating seeds is provided, the method comprising applying a coating of the seeds with a biopure culture of a single microbial strain, said microbial strain being Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics, said coating optionally comprising one or more carriers, excipients, nutrients, crop protectants, fungicides, insecticides, or nematicides. Bacillus licheniformis K-357 is present on the treated seeds in amounts suitable for increasing plant yield, increasing plant resistance to plant pathogenic nematodes, or reducing plant infection by plant pathogenic nematodes, and combinations thereof, in the presence of plant pathogenic nematodes.

[0025] In one embodiment, a coated plant seed is provided, the coating comprising: a biopure culture of a single microbial strain, said single microbial strain being *Bacillus licheniformis* K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients, crop protectants, fungicides, insecticides, or nematicides. *Bacillus licheniformis* K-357 is present in the seed in an amount suitable for increasing plant yield, enhancing plant resistance to plant nematodes, or reducing plant infection by plant nematodes, and combinations thereof, in the presence of plant pathogenic nematodes.

[0026] In one embodiment, a composition for improving plant health and / or yield is provided, the composition comprising: i) a biopure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients or microbial crop protectants, bio-crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0027] In one embodiment, a plant or plant seed is provided with a composition coating comprising: i) a biologically pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients, or microbial crop protectants, biological crop protectants, or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in the coating in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0028] In one embodiment, a method of treating plants or plant seeds is provided, the method comprising: applying a composition coating to the plants or plant seeds, the composition coating comprising: i) a biologically pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in the coating in an amount suitable for increasing plant yield or reducing plant pests in the presence of plant pathogenic insects, or both.

[0029] In one embodiment, a method is provided for formulating a microbial crop protectant to improve plant health and / or yield, comprising: adding together a bioculture of a soybean protein hydrolysate, chelated ferrous sulfate, and one or more microbial crop protectants, wherein the one or more microbial crop protectants are present in the formulation in an amount suitable for increasing plant yield or reducing plant pests in the presence of plant pathogenic insects, or both.

[0030] In one embodiment, a formulation of a microbial crop protectant is provided to improve plant health and / or yield, comprising: i) a bioculture of one or more microbial crop protectants; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients, or nonmicrobial crop protectants, wherein the one or more microbial crop protectants are present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects. Attached Figure Description

[0031] Figure 1 This is demonstrated by the in vitro application of different doses of Bacillus licheniformis K-357 spores (0.02561 L / ha - equivalent to 5.5 × 10⁻⁶ spores per 50 mL) to soybean cyst nematode (Heterodera glycines), southern root-knot nematode (Meloidogyne incognita), and kidney nematode (Rotylenchulus reniformis). 6 CFU; 0.1428 5 L / ha - equivalent to 2.75 × 10⁵ CFU per 50 mL 7 CFU; 0.333310 L / ha - equivalent to 5.5 × 10⁻⁶ CFU per 50 mL. 7 A graph showing the percentage of nematode hatching counts 10 days after CFU (Centralized Fuel Cell).

[0032] Figure 2Image A is an image of nematodes without the addition of Bacillus licheniformis K-357, showing nematode eggs, larvae, and adults.

[0033] Figure 2 Image B is a nematode image without Bacillus licheniformis K-357, showing unhatched larvae in eggs.

[0034] Figure 2 Image C is an image of nematodes without Bacillus licheniformis K-357 added, showing hatched larvae.

[0035] Figure 2 Image D is an image of nematodes without the addition of Bacillus licheniformis K-357, showing adult nematodes.

[0036] Figure 2 E is an image of nematodes without the addition of Bacillus licheniformis K-357, showing adult nematodes.

[0037] Figure 2 F is an image of a nematode without the addition of Bacillus licheniformis K-357, showing the mouth of an adult nematode.

[0038] Figure 3 This is an image magnified 1000 times showing an adult nematode inhabited by Bacillus licheniformis K-357.

[0039] Figure 4 This is a 1000x magnified image of Bacillus licheniformis K-357, stained red, surrounding fixed kidney-shaped nematode eggs.

[0040] Figure 5 This is an image magnified 1000 times of a kidney-shaped egg surrounded by purple-stained Bacillus licheniformis K-357. The larval worm is visible in the center of the egg and is fixed under the microscope.

[0041] Figure 6This is a graph showing the percentage of kidney nematode counts 55 days after emergence in cotton seeds treated with chemical nematicides such as aldicarb or fluopyram in furrows, compared to seeds treated with K-357, in a field trial in Arkansas: 1) with fungicides (Azoxystrobin, Fludioxinol, Mefenoxam, Sedaxane), insecticides (Thiamethoxam), and furrow nematicides aldicarb (1) Seeds treated with 5 lbs / acre, 2) seeds treated with fungicides (pyraclostrobin, fludioxonil, metalaxyl, fluoxastrobin), insecticides (thiamethoxam, imidacloprid) and furrow nematicide fluopyram (16 oz / acre), and 3) seeds treated with fungicides (metalaxyl, fludioxonil, myclobutanil), insecticides (imidacloprid) and K-357 as a nematicide at 2 fl oz per 100 lbs of cotton seed (“2,000 Fl.Oz. / CWT”).

[0042] Figure 7 It is a display Figure 6 A graph showing the percentage of cotton yield at harvest time.

[0043] Figure 8 This is a graph showing root-knot nematode larval populations at sowing, 60 days later, and harvest in a cotton field trial conducted in Texas. It compares seeds treated with abamectin with seeds treated with K-357 as a nematicide. Specifically, the seeds were treated with: 1) fungicides (pyraclostrobin, fludioxonil, metalaxyl, fluoxastrobin), insecticides (imidacloprid, thiamethoxam), and AVICTA ELITEPLUS containing VIBRANCE (abamectin) as a nematicide; or 2) fungicides (metalaxyl, fludioxonil, cyazofamid), insecticides (imidacloprid), and 2 fl oz (“2,000 Fl.Oz. / CWT”) of K-357 per 100 lbs of cotton seed as a nematicide.

[0044] Figure 9 This represents the percentage of nematode counts 25 days after planting cotton seeds treated with fungicides (metalaxyl, fludioxonil, cyproconazole), insecticides (imidacloprid), and 2 (“2,000 Fl.Oz. / CWT”) or 3 (“3,000 Fl.Oz. / CWT”) fluid ounces of K-357 as a nematicide, compared to seeds treated with fungicides (pyraclostrobin, fludioxonil, metalaxyl, fluoxastrobin), insecticides (imidacloprid, thiamethoxam), and abamectin as a nematicide.

[0045] Figure 10 It is a display Figure 9 A graph showing cotton yield at harvest time.

[0046] Figure 11 It is displayed Figure 9 A graph showing plant vigor calculated 1–9 days, 12 days, and 25 days after planting cotton seeds in a field trial.

[0047] Figure 12 This is a graph showing the percentage mortality of each of the four plant pathogenic nematodes treated in vitro with Avicta or K-357, with water used as a control.

[0048] Figure 13 This is a graph showing the average stand count of maize seeds treated with K-357 and chemical pesticides / fungicides at marked ratios 37 days after planting at the University of Illinois at Monmouth in fields severely infected with CRW, compared to maize seeds treated with chemical pesticides / fungicides alone.

[0049] Figure 14 This chart shows the average score of maize root node damage assessment in a field with severe CRW infection, 64 days after maize seeds treated with K-357 and chemical pesticides / fungicides at marked ratios at the University of Illinois at Monmouth, compared to maize seeds treated with chemical pesticides / fungicides alone.

[0050] Figure 15 This is a schematic diagram of the sequential steps of an experimental setup used to study the effects of Bacillus licheniformis K-357 on the transmission of potato virus Y (PVY) by the green peach aphid (GPA). Detailed Implementation

[0051] To facilitate an understanding of the principles of this disclosure, reference will now be made to preferred embodiments, and these embodiments will be described using specific language. However, it should be understood that this is not intended to limit the scope of this disclosure, and such changes and further modifications to this disclosure as illustrated herein are considered to be commonly thought of by those skilled in the art to which this disclosure pertains.

[0052] As used herein, the phrase “biopure culture” includes one or a combination of spores and vegetative cells of a biopure fermentation culture of a microbial strain. “Biopure” means substantially biopure as understood in the art. Furthermore, “biopure culture” includes mutants of microbial strains that possess all of their identifying characteristics.

[0053] Following long-standing patent law practice, the terms “a,” “an,” and “the” are used in this application (including the claims) to mean “one or more.” Thus, for example, reference to “plant or seed of a plant” includes multiple plants or seeds unless the context clearly indicates otherwise, and so on.

[0054] Throughout the specification and claims, the terms “comprise,” “comprises,” and “comprising” are used in a non-exclusive sense unless the context requires otherwise. Similarly, the terms “having” and “including,” and their grammatical variations, are intended to be non-restrictive, such that the listing of items does not exclude other similar items that may substitute for or be added to the listed items.

[0055] For the purposes of this specification and claims, when used in conjunction with one or more numbers or numerical ranges, the term "about" should be understood to refer to all such numbers, including all numbers within the range, and to modify the range by extending the boundaries above and below the value. A numerical range expressed in terms of endpoints includes all numbers contained within the range, such as integers, including their decimals (e.g., the expression for 1 to 5 includes 1, 2, 3, 4, and 5, and their decimals, such as 1.5, 2.25, 3.75, 4.1, etc.), and any range within the range. Furthermore, as used herein, the term "about," when referring to a value, may include a variation from a specified amount, in some embodiments + / - 20%, in some embodiments + / - 10%, in some embodiments + / - 5%, in some embodiments + / - 1%, in some embodiments + / - 0.5%, in some embodiments + / - 0.1%, variations suitable in the disclosed compositions and methods. Alternatively, particularly in relation to biological systems or processes, the term may indicate a range within an order of magnitude, preferably within 5 times a value, more preferably within 2 times. When a particular value is described in this application and claims, unless otherwise stated, the term “about” should be considered to mean that the particular value is within an acceptable range of error.

[0056] Throughout the specification and claims, the phrase "microbial crop protectant, biological crop protectant, or chemical crop protectant" refers to any microbial, biological, or chemical agent used to protect crops from plant pests and / or plant pathogens, including agents used as one or more of biocides, insecticides, fungicides, or nematicides.

[0057] For the purposes of this specification and claims, the term "insect" includes nematodes.

[0058] For the purposes of this specification and claims, the phrase "the branches and leaves of a plant" refers to any part of a plant other than the seeds and roots.

[0059] For the purposes of this specification and claims, the terms "pesticide" and "insecticide" are used interchangeably.

[0060] In order to replace or reduce synthetic fertilizers and chemical pesticides to improve crop health and limit negative environmental impacts, this paper provides live microbial strains, compositions containing said strains, and seeds and plants coated with said strains to control plant-parasitic nematodes and other pathogenic insects, including but not limited to aphids and corn rootworms. Studies have demonstrated the economic benefits and reliable performance of such live microbial products in integrated crop management systems. The live microbial strains provided herein can be used on seeds, in furrows, or applied to the top or roots of plants to grow alongside them, thereby protecting plants from nematodes and insects and / or increasing plant productivity and reproductive capacity. The live microbial products and related methods provided herein have the potential to offer a sustainable, cost-effective solution that can increase yields with less input while avoiding the toxic effects on users and the environment associated with chemical alternatives.

[0061] Bacillus licheniformis strain NRRL B-23318, originally isolated from the Sonoran Desert, was obtained from the NRRL Culture Collection. Surprisingly, the inventors discovered that this Bacillus licheniformis strain (hereinafter referred to as "K-357") can be used as a biological nematicide. More specifically, application of Bacillus licheniformis K-357 reduced in vitro nematode hatching, reduced nematode soil counts in field trials, and increased cotton yield. The nematicidal activity of Bacillus licheniformis K-357 is equivalent to or significantly superior to that of standard chemical nematicide treatments. The increase in cotton yield due to Bacillus licheniformis K-357 is superior to that of standard chemical treatments. Exemplary experiments and data are described herein.

[0062] Example 1 describes an in vitro experiment that tested the effectiveness of Bacillus licheniformis K-357 in controlling three different types of plant nematodes: soybean cyst nematode (Heterodera glycines), southern root-knot nematode (Meloidogyneincognita), and kidney nematode (Rotylenchulus reniformis). Figure 1 The results shown indicate that K-357 can reduce or prevent nematode hatching in a dose-dependent manner, including an 87% reduction in kidney-shaped nematodes and complete inhibition of southern root-knot nematode hatching. Figure 2-5K-357 was shown to reside on nematodes and nematode eggs. K-357 showed a significant inhibitory / preventive effect on the hatching of *Bacillus reniformis* / southern root-knot nematode, superior to the effects observed in in vitro application of *Bacillus subtilis* strain DSM17231 in combination with *Bacillus licheniformis* strain DSM17236, where only one of the three tested root-knot nematode species showed a minor effect (see EP2603086B1). What makes the K-357 discovery described herein even more surprising is that it is generally accepted by those skilled in the art that any bionematicidal effect produced by the strain combination described in EP2603086B1 (i.e., Bacillus subtilis strain DSM 17231 + Bacillus licheniformis strain DSM17236) is derived from the Bacillus subtilis strain, rather than from the Bacillus licheniformis strain, since there are no known reports of Bacillus licheniformis strains possessing nematicidal properties (see, for example, Xiang et al. (2017); Abd-Elgawad and Askary (2018)).

[0063] Example 2 describes a field trial of cotton to evaluate the effectiveness of chemical nematicides, including aldicarb (which is the standard of effectiveness), and Bacillus licheniformis K-357. Each treatment was applied to the furrows at sowing. Results are as follows: Figure 6 and 7 As shown in the figure. The results showed that 55 days after emergence, K-357 reduced the number of nematodes in the soil to about 70% of the chemical standard aldicarb, and was superior to fluopyram. Figure 7 This is a chart showing the percentage of cotton yield at harvest. Surprisingly, cotton treated with Bacillus licheniformis K-357 yielded higher yields than cotton treated with fluopyram (approximately 4.5% higher) and aldicarb (approximately 2.5% higher).

[0064] Example 3 describes a cotton field trial to evaluate the efficacy of Bacillus licheniformis K-357 applied to seeds in reducing nematode populations and increasing yield. In the trial, seeds treated with two different proportions of Bacillus licheniformis K-357 were compared with seeds treated with avermectin. Results are as follows: Figures 8 to 11 As shown. Figure 8 K-357 showed a more sustained effect than the chemical nematicide abamectin in suppressing juvenile root-knot nematode populations. Specifically, although the number of nematodes treated with abamectin was lower than that treated with K-357 after 60 days, this relationship was reversed at harvest. At harvest, the number of nematodes treated with K-357 decreased to less than 15, while the number of nematodes treated with abamectin was approximately 40. Furthermore, the nematode population in the K-357 group was initially higher than that in the abamectin group and steadily declined as the experiment progressed. In contrast, the nematode population in the abamectin group reversed and rapidly increased after 60 days.

[0065] Figure 9 The results showed that, compared with seeds treated with abamectin, seeds treated with K-357 at 2-oz / cwt and 3-oz / cwt had 88% and 91% control effects on nematode numbers, respectively, 25 days after planting. Figure 10 The study demonstrated that treatment with 2 Fl.Oz. / CWT and 3,000 Fl.Oz. / CWT of Bacillus licheniformis K-357 resulted in unexpectedly significant increases in cotton yield of 54 lb / acre and 102 lb / acre, respectively. Furthermore, Figure 11 K-357 showed that it improved plant vigor between 12 and 25 days after planting, while no improvement in vigor was observed with abamectin.

[0066] Example 4 describes an in vitro experiment that tested the effectiveness of Bacillus licheniformis K-357 against four different types of plant nematodes compared to the chemical nematicide Avicta: soybean cyst nematode, southern root-knot nematode, kidney nematode, and Lesion nematode. The nematodes were placed in petri dishes and treated with Avicta, water, or K-357 spores for 48 hours. Data are as follows: Figure 12 As shown in the figure, the mortality rate of each of the four nematode species is illustrated. Surprisingly, the K-357 treatment was as effective as or significantly more effective than Avicta in killing nematodes. Specifically, K-357 was equally effective in killing southern root-knot nematodes, soybean cyst nematodes, and disease nematodes, and significantly superior to Avicta in killing kidney-shaped nematodes (84.6% and 35.4%, respectively).

[0067] In addition to nematodes, Bacillus licheniformis K-357 also exhibits insecticidal activity against other plant pathogenic insects. Example 5 describes field trials assessing the ability of seeds treated with Bacillus licheniformis K-357 alone and in combination with chemical insecticides and fungicides to reduce maize root-knot weevils (CRW) damage and increase yield. These trials were conducted at the University of Illinois at Monmouth in fields with high CRW virus infestations. Stand counts and root node damage analyses at 37 and 64 days after planting were shown in… Figure 13 and 14 In the middle, the mean stand count for both seed treatment group #1 (insecticide / fungicide) and seed treatment group #2 (K-357 + insecticide / fungicide) was 33.8. Figure 13 ).However, Figure 14The root node damage plots show that the addition of K-357 to the insecticide / fungicide mixture reduced root node damage. Specifically, the root node damage score was 1.12 for seed treatment group #2 (K-357 + insecticide / fungicide) and 1.47 for seed treatment group #1 (insecticide / fungicide).

[0068] Example 6 describes another example of Bacillus licheniformis K-357 with insecticidal activity. In this experiment, the ability of Bacillus licheniformis K-357 to reduce or prevent the spread of Potato Virus Y (PVY) was evaluated using a virus transmission test on detached leaves of Nicotiana benthamiana. The green peach aphid (GPA, Myzus persicae) is the most effective vector of PVY, and this experiment aimed to assess whether Bacillus licheniformis K-357 could weaken hempeptipteran pests (e.g., aphids), thereby completely inhibiting and / or reducing the spread of PVY.

[0069] In one embodiment, a method for controlling plant pests is provided, the method comprising: planting a plant or plant seed in a suitable growing environment, the plant or seed having a coating or partial coating of a composition comprising: a pure culture of Bacillus licheniformis K-357 or a mutant having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0070] In one embodiment, a method for controlling plant pests is provided, the method comprising: delivering a composition to the seeds, foliage, roots, or soil or growing environment surrounding a plant, the composition comprising: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients, or microbial crop protectants, bio-crop protectants, or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0071] In one embodiment, a method for controlling plant pests is provided, the method comprising: planting a plant or plant seed in a suitable growing environment, the plant or seed having a coating or partial coating of a composition substantially consisting of: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and one or more carriers, excipients or nutrients, wherein the composition increases plant yield or reduces plant pests, or both, in the presence of plant pathogenic insects.

[0072] In one embodiment, a method for controlling plant pests is provided, the method comprising: delivering a composition to the seeds, leaves, roots, or soil or growing environment surrounding a plant, the composition consisting essentially of: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics, and one or more carriers, excipients, or nutrients, wherein the composition increases plant yield or reduces plant pests, or both, in the presence of plant pathogenic insects.

[0073] In one embodiment, a method for treating plants or plant seeds is provided, comprising applying a composition coating to the plants or plant seeds, said composition coating comprising a biologically pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics, said composition optionally comprising one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, said composition not comprising Bacillus subtilis, and said Bacillus licheniformis K-357 being present in the coating in an amount suitable for increasing plant yield or reducing plant pests or both in the presence of plant pathogenic insects.

[0074] In one embodiment, a plant or plant seed is provided having a composition coating comprising: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in the coating in an amount suitable for increasing plant yield or reducing plant pests or both in the presence of plant pathogenic insects.

[0075] In one embodiment, a method for treating plants or plant seeds is provided, comprising applying a composition coating to the plants or plant seeds, said composition coating comprising a biologically pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics, said composition optionally comprising one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, said composition not comprising Bacillus subtilis, and said Bacillus licheniformis K-357 being present in the coating in an amount suitable for increasing plant yield or reducing plant pests or both in the presence of plant pathogenic insects.

[0076] In one embodiment, a plant or plant seed is provided having a composition coating comprising: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in the coating in an amount suitable for increasing plant yield or reducing plant pests or both in the presence of plant pathogenic insects.

[0077] In one embodiment of this disclosure, a method for controlling plant pathogenic nematodes is provided. The method includes delivering a composition to plant seeds, plant roots, or the soil or growing environment surrounding the plant, said composition consisting essentially of: a pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; and one or more carriers, excipients, or nutrients. Delivering the composition to plant seeds, roots, or the growing environment can increase plant yield and plant resistance to plant pathogenic nematodes in the presence of plant pathogenic nematodes, or both. In some cases, delivery of said composition can also reduce plant infection by plant pathogenic nematodes.

[0078] The phrase "composition consisting essentially of a biopure culture of Bacillus licheniformis K-357" means that the composition comprises only a biopure culture of a single microbial strain (i.e., Bacillus licheniformis K-357). Therefore, the phrase "consistently of" refers only to the microbial strain, meaning that the microbial strain contained in the composition is limited to Bacillus licheniformis K-357. While the composition contains only the single microbial strain Bacillus licheniformis K-357, the composition may contain any number of other non-microbial strain components, including but not limited to one or more carriers, excipients, nutrients, crop protectants, insecticides, nematicides, or fungicides.

[0079] In one embodiment of this disclosure, a method for controlling plant pathogenic nematodes is provided, the method comprising delivering a composition of a pure culture of a single microbial strain, namely Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics, to plant seeds, plant roots, or the growing environment surrounding the plant. Delivering the composition to plant seeds, roots, or the growing environment can increase plant yield and plant resistance to plant pathogenic nematodes in the presence of plant pathogenic nematodes, or both. In some cases, delivery of the composition can also reduce plant infection by plant pathogenic nematodes.

[0080] Similar to the description above, "a composition comprising a biopure culture of a single microbial strain, wherein the single microbial strain is Bacillus licheniformis K-357" is intended to indicate that while the composition may contain an unlimited number of components, the microbial strain component is limited to a biopure culture of a single strain of Bacillus licheniformis K-357. The composition may contain any number of other non-microbial components, including but not limited to one or more carriers, excipients, nutrients, crop protectants, insecticides, nematicides, or fungicides.

[0081] In one embodiment of this disclosure, a method for controlling plant pathogenic nematodes is provided, the method comprising planting coated seeds in a suitable growing environment. In this embodiment, the seeds have a coating comprising: spores of a biopure culture of a single microbial strain, said single microbial strain being *Bacillus licheniformis* K-357 or a mutant thereof having all of its identifying characteristics; and optionally one or more carriers, excipients, nutrients, crop protectants, fungicides, insecticides, or nematicides. *Bacillus licheniformis* K-357 is present in one or more amounts suitable for increasing plant yield, increasing plant resistance to plant pathogenic nematodes, or reducing plant infection by plant pathogenic nematodes in the presence of plant pathogenic nematodes. The composition comprising coated seeds may include one or more carriers, excipients, or nutrients, and one or more crop protectants, insecticides, nematicides, or fungicides.

[0082] Bacillus licheniformis K-357 can be produced at a rate of 1.0 × 10⁻⁶. 12 CFU / g up to 1.0×10 1 CFU / g, 9.0×10 7 CFU / g up to 2.0×10 4 CFU / g or 4.0×10 8 CFU / g up to 1.0×10 4 The amount of CFU / g seed present in the seed.

[0083] The compositions of this disclosure containing Bacillus licheniformis K-357 can be in liquid form. The concentration of Bacillus licheniformis K-357 present in the liquid composition ranges from about 1.0 × 10⁻⁶. 12 Up to 1.0×10 1 CFU / ml, 1.0×10 11 CFU / ml up to 1.0×10 2 CFU / ml, or 1.0×10 10 CFU / ml up to 1.0×10 4 CFU / ml.

[0084] The compositions of this disclosure containing Bacillus licheniformis K-357 can also be in the form of powder, dry wettable powder, spreadable granules, or dry wettable granules. The content of Bacillus licheniformis K-357 present in these non-liquid forms of the compositions ranges from 1.0 × 10⁻⁶. 12 CFU / g up to 1.0×10 10 CFU / g, 5.0×10 11 CFU / g up to 1.0×10 3 CFU / g, or 1.25×10 11 CFU / g up to 1.0×10 8 CFU / g.

[0085] In one embodiment of this disclosure, a method for treating seeds is provided, the method comprising applying a coating of the seeds with a biopure culture of a single microbial strain, said single microbial strain being *Bacillus licheniformis* K-357 or a mutant thereof having all of its identifying characteristics. While the coating is limited to a biopure culture of a single microbial strain (*Bacillus licheniformis* K-357), the coating may optionally include one or more carriers, excipients, nutrients, crop protectants, fungicides, insecticides, or nematicides. *Bacillus licheniformis* K-357 is present in the coating in a manner suitable for increasing plant yield, enhancing plant resistance to plant nematodes, and / or reducing the amount of plant nematode infection in the presence of plant pathogenic nematodes.

[0086] Embodiments of this disclosure also include coated plant seeds, said coating comprising a biopure culture of a single microbial strain, said single microbial strain being *Bacillus licheniformis* K-357 or a mutant thereof having all of its identifying characteristics. Optionally, the seed coating may also comprise one or more crop protectants, including but not limited to insecticides, nematicides, fungicides, or nutrients, or other carriers or excipients. *Bacillus licheniformis* K-357 is present in the seed coating in a manner suitable for increasing plant yield, enhancing plant resistance to plant nematodes, and / or reducing the amount of plant nematode infection in the presence of plant pathogenic nematodes.

[0087] In the methods and compositions disclosed herein, the plant may be a monocotyledonous plant or a dicotyledonous plant, or any of them, but not limited to cotton, rice, soybean, tomato, cereals, root / tuberous and corn vegetables, brassica vegetables, cucurbit vegetables, bulb vegetables, citrus, fruit vegetables, herbs / spices, leafy vegetables, legumes / vegetables (fleshly and dried legumes and peas), oil crops, pome fruits, stone fruits, strawberries, sugarcane, beets, nuts, kiwifruit, bananas, grasses, ornamental plants or hardwood cuttings.

[0088] In one embodiment, the plant or plant seed having a coating containing Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics is corn, cotton, peanut, soybean, sorghum, wheat, rice, potato, sunflower, or onion.

[0089] In one embodiment of the methods and compositions disclosed herein, plant root tips or transplanted roots may be coated with a composition comprising Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics.

[0090] In one embodiment of this disclosure, the composition may be applied to the seeds of a plant, the leaves and branches of a plant, the roots of a plant, or the growing environment surrounding the plant.

[0091] In the methods and compositions disclosed herein, the plant pathogenic nematodes may include, but are not limited to, southern root-knot nematodes, M. paranaensis, Javan root-knot nematodes (M. javonica), soybean cyst nematodes (SCN), soybean cyst nematodes (SCN), kidney-shaped nematodes (Rotylenchuitis reniformis), or disease-causing nematodes.

[0092] In one embodiment of the methods and compositions disclosed herein, the pathogenic insect may include, but is not limited to, maize rootworm, western maize rootworm (WCR), northern maize rootworm (NCR), southern maize rootworm (SCR), aphid, green peach aphid (peach aphid) or cotton aphid (Aphis gossypii).

[0093] In some implementations, Bacillus licheniformis K-357 is formulated with soybean protein hydrolysate and chelated ferrous sulfate.

[0094] For example, in one embodiment, a composition for improving plant health and / or yield is provided, comprising: i) a biopure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients or microbial crop protectants, bio-crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0095] In one embodiment, a composition for improving plant health and / or yield is provided, comprising: i) a biopure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients or microbial crop protectants, bio-crop protectants or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0096] In one embodiment, a plant or plant seed is provided with a composition coating comprising: i) a biologically pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients, or microbial crop protectants, biological crop protectants, or chemical crop protectants, wherein the composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in the coating in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0097] In one embodiment, a method of treating plants or plant seeds is provided, comprising: applying a composition coating to the plants or plant seeds, said composition coating comprising: i) a biologically pure culture of Bacillus licheniformis K-357 or a mutant thereof having all of its identifying characteristics; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients or microbial crop protectants, biological crop protectants or chemical crop protectants, wherein said composition does not include Bacillus subtilis, and wherein Bacillus licheniformis K-357 is present in the coating in an amount suitable for increasing plant yield or reducing plant pests in the presence of plant pathogenic insects, or both.

[0098] In one embodiment, a method is provided for formulating a microbial crop protectant to improve plant health and / or yield, comprising: adding together a bioculture of a soybean protein hydrolysate, chelated ferrous sulfate, and one or more microbial crop protectants, wherein the one or more crop protectants are present in the formulation in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0099] In one embodiment, a formulation of a microbial crop protectant is provided to improve plant health and / or yield, comprising: i) a bioculture of one or more microbial crop protectants; ii) a soybean protein hydrolysate; iii) chelated ferrous sulfate; and iv) optionally, one or more carriers, excipients, nutrients, or nonmicrobial crop protectants, wherein the one or more microbial crop protectants are present in an amount suitable for increasing plant yield or reducing plant pests, or both, in the presence of plant pathogenic insects.

[0100] In the compositions disclosed herein, the soybean protein hydrolysate may contain about 4% water-soluble nitrogen.

[0101] In the compositions disclosed herein, the content of the soy protein hydrolysate present in the composition or formulation may be from about 60% by weight to about 99% by weight, and the content of the chelated ferrous sulfate present may be from about 0.05% by weight to about 1.5% by weight.

[0102] In the compositions disclosed herein, the content of the soy protein hydrolysate present in the composition or formulation may be from about 85% by weight to about 95% by weight, and the content of the chelated ferrous sulfate present may be from about 0.05% by weight to about 0.3% by weight.

[0103] The compositions and formulations disclosed herein can be in liquid form, and Bacillus licheniformis K-357 can be in a concentration of 1.0 × 10⁻⁶. 12 Up to 1.0×10 1 CFU / ml, 1.0×10 11 CFU / ml up to 1.0×10 2 CFU / ml or 1.0×10 10 CFU / ml up to 1.0×10 4 A concentration of CFU / ml is present.

[0104] The compositions and formulations disclosed herein may be in the form of powder, dry wettable powder, spreadable granules, or dry wettable granules, and Bacillus licheniformis K-357 may be in the form of 1.0 × 10⁻⁶ granules. 12 CFU / g up to 1.0×10 10 CFU / g, 5.0×10 11 CFU / g up to 1.0×10 3CFU / g or 1.25×10 11 CFU / g up to 1.0×10 8 The content of CFU / g is present.

[0105] In the method disclosed herein, Bacillus licheniformis K-357 can be used at a concentration of 1.0 × 10⁻⁶. 12 CFU / g up to 1.0×10 1 CFU / g, 9.0×10 7 CFU / g up to 2.0×10 4 CFU / g, or 4.0×10 8 CFU / g up to 1.0×10 4 The amount of CFU / g seed applied to or present on the seed.

[0106] Furthermore, in one or more embodiments, suitable insecticides, fungicides, and nematicides for the compositions and methods of the present invention may include: Insecticides: Al-agrigata, aluminum phosphide, amblyseius, apherinus, aphidinus, aphidletes, artemisinin, autographa californica NPV, azocyclotin, Bacillus subtilis, Bacillus thuringiensis subsp. aizawai, Bacillus thuringiensis subsp. kurstaki, Bacillus thuringiensis thuringiernsis), Beauveria, Beauveria bassiana, Beauveria bassiana, β-cyfluthrin, biologics, bislutap, broflutrinate, bromophos-e, bromopropylate, Bt transgenic corn, Bt transgenic soybean, capsaicin, fenitrothion, celastru s) extract, chloranthraniprole, chlorfenapyrone, chlorethoxyphos, chlorfluazuron, chiorpyrifos-e, cinidiadin, cryolite, cyanophos, cyantraniliprole, cyhalothrin, cihexatin, cypermethrin, Dakunusa, DCIP, dichloropropene, dicophor, Digliphas, Digliphas + Dakunusa, Dimetacarb, Dithioether, dodecyl acetateAcetate, abamectin, Encarsia, EPN, Eretmocerus, ethylene dibromide, eucalyptol, fatty acids, salts, fenazacquin, fenobucarb (BPMC), fenpyroximate, flubrocythrinate, flufensin, formethanate, formeotethione, furthiocarb, gamma cyhalothrin, garlic juice, granulosis virus, harmonia, heliothis armigera NPV, inactive bacteria, indol-3-ylbutyric acid acid), iodomethane, iron, isocarbofos, isofenphos, isofenphos-M, isoprocarb, isothioate, kaolin, lindane, liuyangmycin, matrine, mephospholane, metaaldehyde, metalicdium-anisoprie, methamicdophos, metorcarb (MTMC), mineral oil, mirex, m-isothiocyanate, monosultap, myrothecium verrucaria, dibromophos, bees (neochrysocharis)Formosan formos, nicotine, nicotinoids, oils, oleic acid, ometoate, *Orius*, oxymatrine, *Pesilomyces*, paraffin oil, parathion-e, *Pasteuria*, petroleum, pheromones, phosphates, *Photolabdas*, foxime, *Phytoseiulus*, pyrimimiphos-e, vegetable oils, *GV* of *Gastropoda*, polyhedrosis virus. Virus), polyphenol extract, potassium oleate, profenofos, prosuler, prothiophos, pyracrofos, pyrethrin, pyridafenthione, pyrimidifene, pyriproxyfen, quillay extract, quinomethionate, rapeseed oil, rotenone, saponin, saponozit, sodium compoundsCompound), sodium fluorosilicate, starch, nematodes, streptomyces, sulfuramide, sulfur, tebupyrimphos, tefluthrin, temefos, tetradiphone, thiophanox, thiometone, genetically modified organisms (e.g., Cry38b1), triazamate, Trichoderma, Trichogramma, triflumuron, vericilliim, beltrin, isomeric insecticides (e.g., ... A1) Carbamates, including aldicarb, aranicarb, benfturacarb, carbaryi, carbofuran, carbosulfan, methiocarb, mesomil, oxamyl, pirimicarb, propoxur, and thiocdicarb; A2) Acephate, azine phos-ethyl, azine phos-methyl, chlorfenvin phos, chiorpyrifos, chlorpyrifos-methyl, and methomyl emulsion.Meton-S-methyl, diazinon, dichlorvos / DDVP, didrotophos, dimethoate, disulfotone, ethion, fenitrothion, fenthion, isoxathion, malathion, metamidaphos, methidathion, mevinphos, monocrotophos, oxymethoate, oxyciemethone-methyl, parathion, organophosphates including phenol salts, folic acid, hosalon, phosmet, phosphamidone, pyrimiphos-methyl, quinalphos, terbufos, tetrachlortbinphos, triazole A3) Cyclopentadiene organochlorides, such as endosulfan; A4) Ethylene, fipronil, and fiprolols, including pyrafluprole and pyriprole; A5) Acetamiprid and neonicotinoids, including rotianidin, dinotefuran, imidacloprid, acetamiprid, thiamethoxam, and thiamethoxam; A6) Spinosyns, such as spinosad and ethyl spinosad; A7) Chlorides derived from mectins, including abamectin and methylamino... A8) Avermectin benzoate, ivermectin, lepimectin, and milkemectin; A9) Juvenile hormone mimics, such as acetamiprid, quinolones, metoprene, phenoxycarb, and difenoconazole; A0) Selective homologous feeding blockers, such as pymetrozine, flonicamid, and pyrifluquinazone; A11) Tetramethrin, hexamethonium, and etoxazole; A12) Propramide, fenbutatin, and propargite mitochondrial ATP synthase inhibitors; Oxidative phosphorylation uncoupling agents, such as bromfenoxam; A13) Nicotinic acetylcholine receptor channel blockers, such as bensultap and cartap.A13) Hydrochloride, thiosultap sodium; A14) Bistriflulone, a type 0 inhibitor of chitin biosynthesis derived from benzoylurea, including diflubenzuron, flufenoxuron, flufenoxuron, lufenuron, nobarulone, and teflubenzuron; A15) Type 1 inhibitors of chitin biosynthesis, such as thiamethoxam; A16) Cypermethrin; A17) Ecdysone receptor agonists, such as methoxyphenozide, tebufenozide, chlorfenapyr, and cyclotebufenozide; A18) Octopus amine receptors, such as diflubenzuron, thiosultap sodium, and thiosultap sodium. A18) Mitochondrial complex electron transport inhibitors such as pyridaben, pymetrozine, azoxystrobin, flufenelim, sienopyrafen, cyflumetofene, hydramethylnon, acequinosyl, or fluacrylpyrim; A19) Voltage-dependent sodium channel blockers, such as indoxacarb and cyflumetrozine; A20) Lipid synthesis inhibitors, such as spirodiclofen, spirodiclofen, and spirotetramat; A21) Flufenoxuron (f... ulvendiamide), phthalamide compounds (R)-3-chloro-N1-{2-methyl-4-[1,2,2,2-tetrafluoro-1-(trifluoromethyl)ethyl]phenyl}-N2-(1-methyl-2-methylsulfonylethyl)phthalamide and (S)-3-chloro-N1-{2-methyl-4-[1,2,2,2-tetrafluoro-1-(trifluoromethyl)ethyl]phenyl}-N2-(1-methyl-2-methanesulfonylethyl)phthalamide derived from diamides, chlorantraniliprole and cyanantraniliprole ryanodine receptor modulators, including: A22) having unknown or Compounds with uncertain mechanisms of action, such as azadirachtin, amidoflumet, biphenazate, fluenesulfone, piperonyl butyl ether, trifluralin, fludioxonil; or (A23) acrinathrin, allethrin, bifenthrin, cypermethrin, lambda-cyhalothrin, cypermethrin, α-cypermethrin, β-cypermethrin, ζ-cypermethrin, deltamethrin, fenvalerate, phenropatorin, fenvalerate, fenvalerate, tau-fluvalinate, permethrin, sodium channel modulator-derived fluorosilylpyrethrin, and tetrabromopyrethrin.

[0107] Fungicides: BO) benzobindiflupyr, antiperonosporic, amethoxazole, amisulbrom, copper salts (e.g., copper hydroxide, copper chloride, copper sulfate, copper persulfate), boscalid, thiflumazide, fluthianyl, flaxilyl, thiabendazole, benodanyl, mepronyl, isofetamide, fenflam, bixafen, fluapiroxad, penflufen, sedaxane, succinoxyxine, enoxastrobin, fluphenoxystrobin, pyroxystrobin, pyramidazole Tostrobin), Triclopyricarb, Phenamine Strobin, Metominostrobin, Pyribencarb, Meptyldinocap, fentin acetate, Fentin chloride, fentin hydroxide, oxytate Cyclin, Chlozolinate, chloronebu, Technazen, etridiazole, iodocarb, prothiocarb, Bacillus subtilis synonym, Bacillus amyloliquefaciens (e.g., QST) 713, FZB24, MBI600, D747 strains), Melaleuca alternifolia extract, Lupinus officinalis extract, BLAD peptide, pyrisoxazole, oxpoconazole, etaconazole, fenpyrazamine, naphthifine, terbinafine hydrochloride, validamycin, pyrimorph, varifenalate, phthalide.Probenazole, isotianil, kelp polysaccharide, lisianthus extract, phosphorous acid and its salts, teclofthalam, triazine, pyriophenone, organic oil, potassium bicarbonate, chlorothalonil, fluoroimide, bifenthrinol, bromconazole, cyproconazole, difenoconazole, diniconazole, enilconazole, epoxiconazole, fluquinazole, cyproconazole, flusilazole, flutriazole, hexaconazole, imibenconazole, ipconazole zole), metconazole, microbutanyl, penconazole, propiconazole, prothioconazole, econazole, triadimethone, triadimenol, tebuconazole, flufenoxuron, methamidophos, imazalil, pefazoate, imazalyl, triflumizole, cyazofamid, benomyl Carbendazim, thiabendazole, fuberidazole, ethaboxam, etridiazole, hymexazole, azaconazole, diniconazole-M, oxpoconazole, paplobutrazole, uniconazole, 1-(4-chlorophenyl)-2-([1,2,4]triazol-1-yl)-azole containing cycloheptamol and thiamethoxam sulfate; B2) azoxystrobin, dimoxystrobin, enestrobrin, fluoxastrobin, cresoxime-methyl, methinostrobin, orisatrobin.Picoxystrobin, Pyraclostrobin, Trifloxystrobin, Enstrobrin, Methyl (2-chloro-5-[1-(3-methylbenzyloxyimino)ethyl]benzyl)carbamate, (2-c Ro-5-[1-(6-methylpyridin-2-ylmethoxyimino)ethyl]benzyl)carbamate, 2-(o-(2,5-dimethylphenoxymethylene)-phenyl)-3-methoxyacrylate, 2-(2-(6-(3-chloro-2-methylphenoxy)-5-fluoro-pyrimidin-4-yloxy)-phenyl)-2-methoxyimino-N-methylacetamide and 3-methoxy-2-(2-(N-(4-methoxy-phenyl)-cyclopropaneformimidesulfonylmethyl)-phenyl)-acrylate, carboxin; B3) carboxin, benzalkonium chloride, benzalkonium chloride. xyl-M), phenhexamide, flutolanil, furametopil, mepronil, metalaxyl, mefenoxam, Off-race, oxadixyl, oxycarboxyl, penthiopyrad, isopyrazam, tifluzamide, thiazinyl ), 3,4-dichloro-N-(2-cyanophenyl)isothiazolyl-5-carboxamide, dimethomorph, flumorph, flumetober, picobenzamide, zoxamide, carpropamide, diclocimet, mandipropamide, N-(2-(4-[3-(4-chlorophenyl)prop-2-alkynoxy]-3-methoxyphenyl)ethyl)-2-methanesulfonyl- Amino-3-methylbutyramide, N-(2-(4-[3-(4-chlorophenyl)prop-2-alkynoxy]-3-methoxy-phenyl)ethyl)-2-ethanesulfonylamino-3-methylbutyramide, methyl 3-(4-chlorophenyl)-3-(2-isopropoxycarbonyl-amino-3-methyl-butyrylamino)propionate, N-(4'-bromobiphenyl-2-yl)-4-difluoromethyl-methylthiazole-δ-carboxamide, N-(4'-trifluoromethylbiphenyl-2-yl)-4-difluoromethyl-2-methylthiazole-5-carboxamide, N-(4'-chloro-3'-fluorobiphenyl-2-yl)-4-difluoromethyl-2-methyl-thiazole-5-carboxamide,N-(3,4'-dichloro-4-fluorobiphenyl-2-yl)-3-difluoro-methyl-1-methylpyrazole-4-carboxamide, N-(3',4'-dichloro-5-fluorobiphenyl-2-yl)-3-difluoromethyl-1-methylpyrazoleRu-4-carboxamide, N-(2-cyanophenyl)-3,4-dichloroisothiazol-5-carboxamide, 2-amino-4-methyl-thiazol-5-carboxyaniline, 2-chloro-N-(1,1,3-trimethyl-indan-4-yl)-nicotinamide, N-(2-(1,3-dimethylbutyl)-phenyl)-1,3-dimethyl-5-fluoro-1H-pyrazole-4-carboxamide, N-(4'-chloro-3',5-difluorobiphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole N-(4'-chloro-3',5-difluorobiphenyl-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-phenyl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N- (trans-2-bicyclopropyl-2-yl-phenyl)-3-difluoro-methyl-1-methyl-1H-pyrazole-4-carboxamide, fluopyram, N-(3-ethyl-3,5-5-trimethyl-cyclohexyl)-3-carboxylammonamide-2-hydroxybenzamide, oxytetracycline, silthiophane, N-(6-methoxy-pyridin-3-yl)cyclopropanecarboxamide, 2-iodo-N-phenylbenzamide, N-(2-bicyclopropyl-2-yl-phenyl)-3-difluoromethyl-1-methylpyrazole-4-ylcarboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-1,3-dimethylpyrazole-4-ylcarboxamide, N-(3',4',5'-trifluorobiphenyl) N-(3',4',5'-trifluorobiphenyl-2-yl)-5-chloro-1,3-dimethyl-pyrazole-4-ylcarboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-3-fluoromethyl-1-methylpyrazole-4-ylcarboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-3-(chlorofluoromethyl)-1-methylpyrazole-4-ylcarboxamide, 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)-3-difluoromethyl-5-fluoro-1-methylpyrazole-4-ylcarboxamideN-(3',4',5'-trifluorobiphenyl-2-yl)-5-chloro-3-difluoromethyl-1-methylpyrazole-4-yl carboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-3-(chlorodifluoromethyl)-1-methylpyrazole-4-yl carboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-1-methyl-3-trifluoromethylpyrazole-4-yl carboxamide, N-(3',4',5'-trifluorobiphenyl-2-yl)-5-fluoro-1-methyl-3-trifluoromethylpyrazole-4-yl carboxyl N-(2',4',5'-trifluorobiphenyl-2-yl) -1,3-Dimethylpyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl)-2-yl)-1,3-dimethyl-5-fluoropyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-5-chloro-1,3-dimethylpyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-3-fluoromethyl-1-methylpyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-3-(chlorofluoromethyl)-1-methylpyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-3-difluoromethyl ... Benzene-2-yl)-3-difluoromethyl-5-fluoro-1-methylpyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-5-chloro-3-difluoromethyl-1-methylpyrazol-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-trifluoromethylpyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-5-furan-1-methyl-3-trifluoromethylpyrazol-4-ylcarboxamide, N-(2',4',5'-trifluorobiphenyl-2-yl)-5-chloro-1-methyl-3-trifluoromethylpyrazol-4-ylcarboxamide Fluoromethylpyrazol-4-ylcarboxamide, N-(3,4'-dichloro-3-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazol-4-carboxamide, N-(3',4'-dichloro-3-fluorobiphenyl-2-yl)-1-methyl-3-difluoromethyl-1H-pyrazol-4-carboxamide, N-(3',4'-difluoro-3-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazol-4-carboxamide, N-(3',4'-difluoro-3-fluorobiphenylRu-2-yl)-1-methyl-S-difluoromethyl-1H-pyrazol-4-carboxamide, N-(3'-chloro-4'-fluoro-3-fluorobiphenyl-2-yl)-1-methyl-3-difluoromethyl-1H-pyrazol-4-carboxamide,N-(3',4'-dichloro-4-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-difluoro-4-fluorobiphenyl-2-yl)-1-methyl-S-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-dichloro-4-fluorobiphenyl-2-yl)-1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxamide, 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 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-fluorobiphenyl-2-yl)-1-methyl-S-difluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-dichloro-5-fluorobiphenyl-2-yl)-1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxamide, N-(3',4'-dichloro-5-fluorobiphenyl-2-yl)-1,3-dimethyl-1 H-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-carboxamide, N-(4'-fluoro-5-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(4'-chloro-5-fluoroo-biphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(4'-methyl-5-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole-4-carboxamide, N-(4'-methyl-5-fluorobiphenyl-2-yl)-1-methyl-3-trifluoromethyl-1H-pyrazole N-(4'-fluoro-5-fluorobiphenyl-2-yl)-1,3-dimethyl-1H-pyrazole-4-carboxamide, 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-πsol-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-1H-pyrazole-4-carboxamide and N-[4'-(trifluoromethylthio)-biphenyl-2-yl]-1-methyl-3-trifluoromethylcarboxamide include 1-methyl-1H-pyrazole-4-carboxamide; B4) fluazinam, pyrifenox, bupyrimetromethorphan sulfonate Prodinil, Phenarimol, Phenarimol, Phenarimol, Nuarimol, Pyrimethanyl, Trifolin, Fenpicuronyl, Fluopyram, Ardimorph, Cyclomorpholine, Butylmorpholine, Tridemorpholine, Benzoyl, Iprodione, Procimidone, Imidacloprid, Oxadiazon, Fenamiden, Octopronone-Azole ), 5-chloro-7-(4-methylpiperidin-1-yl)-6-(2,4,6-trifluorophenyl)-[1,2,4]triazolo[1,5-a]pyrimidine, dichromedin, pyroxylone, propoxyquinoline, tricyclazole, 2-butoxy-6-iodo-3-propylchromene-4-one, acibenzoral-S-methyl, captahol, dazomet, holpet, phenoxanyl, benzene Quinoxyphene, N,N-dimethyl-3-(3-bromo-6-fluoro-2-methylindole-1-sulfonyl)-[1,2,4]triazole-1-sulfonamide, ethyl-6-octyl-[1,2,4]triazolo[1,5-a]pyrimidine-2,7-diamine, 2,3,5,6-tetrachloro-4-methanesulfonyl-pyridine, 3,4,5-trichloropyridin-2,6-dionitrile, N-(1-(5-bromo-3-chloro-pyridin-2-yl)-ethyl)-2,4-dichloronicotinamide, N-((5-bromo-3-chloropyridin-2-yl)-methyl)-2,4-dichloronicotinamide, difluorophenamine (diflumetrim), nitrapirine, dimethomorph, fluoroimide heterocyclic compounds, including blast fungicide S, acaricide, imidacloprid, difenoquinone, diphenzoquat-methylsulfate, oxophosphate, and piperalin; B5) mancozeb, mancozeb, methylam, ferrous sulfate, felvam, propineb, tyram.Zinc bismuth subtilis, zinc thiram, ethoxycarb, iprovaricarb, benchavaricarb, propamocarb propionamide, propamocarb hydrochloride, 4-fluorophenyl-N-(1-(1-(4-cyanophenyl)-ethanesulfonyl)butyr-2-yl)carbamate, carbamates including methyl 3-(4-chlorophenyl)-3-(2-isopropoxycarbonylamino-3-methyl-butyrylamino)propionate, or B6) guanidine, dodecyl guanidine monoacetate, dodine free base Base), iminotadine, biguanide salts, antibiotics: kasugamycin, streptomycin, polyoxin, effectivemycin A, nitrobenzene derivatives: chlorfenapyr, diclofenac, chlorfenapyr, sulfur-containing heterocyclic compounds: dithionine, isoprothiolane, organometallic compounds: fenbutatin salts, organophosphorus compounds: dimethoate, iprobenphos, fosetyl-aluminum, phosphorous acid and its salts, pyraclostrobin, tolcrofos-methyl, organochlorine compounds: dichlorofluanid, fursulfamide, hexachlorobenzene, phthalide, pencyclon, pentachloronitrobenzene, thiophanate-methyl, tolylfluani, and others: cycloflufenicol, simoxanyl, dimethylylmol, ethirimol, flamel, methamidophos. Metraphenone and Spiroxa, biguanide octanoic acid salt, biguanide octane acetate hydrochloride, biguanide trioctylbenzene sulfonate (iminoctadine-tris (albesylate)), kasugamycin hydrochloride monohydrate, diclofenac, pentachlorophenol and its salts, N-(4-chloro-2-nitrophenyl)-N-ethyl-4-methylbenzenesulfonamide, chloronitrobenzene, nitrotar-isopropyl, and tetraoxonyl nitrobenzene (techna) Zen), biphenyl, bromonitol, diphenylamine, myrdiomycin, copper hydroxyquinoline, calcium cyclohexanoate, 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-methylformamidin, N'-(4-(4-fluoro-3-trifluoromethyl-phenoxy)-2,5-dimethyl-phenyl)-N-ethyl-N-methylformamidinN'-(2-methyl-5-trifluoromethyl-4-(3-trimethylsilyl-propoxy)-phenyl)-N-ethyl-N-methylformamidin and N'-(5-difluoromethyl-2-methyl-; other fungicides, including 4-(3-trimethylsilyl-propoxy)-phenyl)-N-ethyl-N-methylformamidin.

[0108] Nematicides or biological nematicides: Benomyl, Cloetocarb, Aldimethoate, Tiluperamide, Fenamifos, Kazusafos, Diclofenthion, Etoprophos, Fensophos, Phostiazete, Isamidophos, Chlorpyrifos, Phosphocarbimisiafos Mecalphone, acetaminophen, benclothiazide, chloropicrin, dazomet, fluenesulfone, 1,3-dichloropropene, dimethyl disulfide, methyl dithiomethanine potassium, methyl dithiomethanine salt (all produced by MITC), methyl bromide, bio-soil amendments (e.g., mustard seed, mustard extract), soil steam fumigation, allyl isothiocyanate (AITC), dimethyl sulfate, furfural (acetaldehyde).

[0109] Suitable plant growth regulators of the present invention include: plant growth regulators: D1) antitoxins, such as clofibrate, 2,3,5-triiodobenzoic acid; D2) 4-CPA, 2,4-D, 2,4-DB, 2,4-DEP, diChlorpropionic acid, 2,4,5-tetrapropionic acid, IAA, IBA, naphthylacetamide, α-naphthylacetic acid, 1-naphthol, naphthoxyacetic acid, potassium naphthenate, sodium naphthenate, auxins such as 2,4,5-T; D3) 2iP, benzyladenine. Cytokinins, such as 4-hydroxyphenylethanol, kinetin, and zeatin; D4) defoliants, such as calcium cyanamide, dimethipine, endal, ethephon, melphos, methoxuron, pentachlorophenol, thiazulone, and defoliant; D5) abiglycine and 1-methylethylene, such as cyclopropene inhibitors; D6) ethylene release agents, such as ACC, etaceracyl, ethephon, and dioxime; D7) gametosides, such as fenridazone and maleic acid hydrazide; D8) gibberellins, such as gibberellin and gibberellic acid; D9) abscisic acid, ansimidol, diphenhydramine, carbaryl, chlorophonium, and chloroprofen. FAM), dikeglac, flubendiamide, fluoridamide, glufosinate, glyphosdine, isopyrimole, jasmonic acid, maleic hydrazine, phytohexidine, phenoxybenzidine, propyl jasmonic acid, profam, thiaojian, 2,3,5-growth inhibitors such as triiodobenzoic acid; D10) morphactins such as chlorflurane. D11) Chlormecote, growth inhibitors such as nozide, furazolidone, flulenol, paclobutrazol, tetracycline, uniconazole; D12) Growth stimulants such as brassinolide, high-brassinolide, DCPTA, chlorpyrifos, tebuconazole, prosuler, triacontanol, etc.D13) Bacmedesh, benzofluor, buminafos, carvone, choline chloride, siobide, clofenset, cyanamide, cyclanilide, cyclanilide, cyprosulfamide, epoccholeon, etizlozate, ethylene, fufenthiourea, fullerane, heptopargyl, holosulfuric acid, inabenfide, caletazan, lead arsenate, metasulfocarb, cyclohexane, pida Down, shintofen, styrazolium, unclassified plant growth regulators, such as trinexapac.

[0110] In one embodiment, the crop protectant disclosed herein may comprise one or a combination of imidacloprid, bifenthrin, metalaxyl, tebuconazole, azoxystrobin, carbendazim, thiram, fludioxonil, cyazofamid, metalaxyl-M, fluoxastrobin, thiamethoxam, lpconazole, or abamectin.

[0111] The compositions, formulations, and methods disclosed herein can be used to control pests from one or more of the following plant pathogenic insects, including but not limited to: plant pathogenic nematodes, southern root-knot nematodes, soybean cyst nematode (SCN), kidney nematode, corn rootworm, western corn rootworm (WCR), northern corn rootworm (NCR), southern corn rootworm (SCR), aphids, peach aphid, cotton aphid, diamondback moth, beet armyworm, red flour beetle, fall armyworm, brown planthopper, bean weevil, European corn borer (Ostinia nubilalis), tobacco hawk moth, rice stem borer, rice brown planthopper, rice green leafhopper (Nephotettix cinciteps), and potato leafhopper. hopper (bean leafhopper (Empoascafabae)), peach aphid (Myzus persicae)), pea aphid (Acyrthosiphon pisum)), wheat stem fly (Chlorop pumilionis), cricket, locust, termite, thrips, ant, mite (mites), lepidopteran insects (cabbage moth), wheat seed fly (Delia coarctata)), wheat aphid (wheat long-tubed aphid), and Colorado potato beetle (potato beetle).

[0112] In one implementation, the pathogenic insect includes one or a combination of plant pathogenic nematodes, southern root-knot nematodes, cyst nematodes (SCN), kidney-shaped nematodes, disease nematodes, maize rootworms, western maize rootworms (WCR), northern maize rootworms (NCR), southern maize rootworms (SCR), aphids, green peach aphids (peach aphids), or cotton aphids (cotton aphids).

[0113] Example

[0114] Example 1

[0115] K-357 inhibits the in vitro hatching of nematodes.

[0116] In vitro experiments were conducted to evaluate the effectiveness of Bacillus licheniformis K-357 in controlling three different types of plant nematodes: soybean cyst nematode, southern root-knot nematode, and kidney-shaped nematode. Nematode eggs were grown in a hatchery and treated with different doses of K-357 spores for 10 days.

[0117] The experiment involved four treatments:

[0118] 1. Control: Water

[0119] 2. K-357@0.0256 1L / ha (equivalent to 5.5×10 in 50ml) 6 CFU)

[0120] 3. K-357@0.1428 5L / ha (equivalent to 2.75×10 in 50ml) 7 CFU)

[0121] 4. K-357@0.3333 10L / ha (equivalent to 5.5×10 in 50ml) 7 CFU)

[0122] The lyophilized K-357 was dispersed in water to provide a concentrated solution (10). 8 The concentrate (CFU / ml) was diluted to a total of 50 ml, resulting in the three different concentrations shown above. Each 50 ml solution was applied to three different species of nematode eggs present in a culture dish and left for 10 days. The eggs were then evaluated under a microscope to obtain hatch counts.

[0123] Data such as Figure 1-5 As shown. Figure 1 The graph shows the percentage of nematode hatching counts after applying different doses of K-357 to soybean cyst nematode, southern root-knot nematode, and kidney nematode, indicating that K-357 can reduce nematode hatching in a dose-dependent manner, including an 87% reduction in kidney nematode hatching and complete inhibition of southern root-knot nematode hatching.

[0124] Figure 2 A-2F is an image of nematodes without the addition of Bacillus licheniformis K-357, showing: A) nematode eggs, larvae, and adults; B) unhatched larvae in eggs; C) hatched larvae; D) adult nematodes; E) adult nematodes; and F) the mouth of an adult nematode.

[0125] Figure 3 This is an image magnified 1000 times showing an adult nematode inhabited by Bacillus licheniformis K-357.

[0126] Figure 4 This is a 1000x magnified image of Bacillus licheniformis K-357, stained red, surrounding fixed kidney-shaped nematode eggs.

[0127] Figure 5 This is an image magnified 1000 times of a kidney-shaped egg surrounded by purple-stained Bacillus licheniformis K-357. The larval worm is visible in the center of the egg and is fixed under the microscope.

[0128] Example 2

[0129] K-357-treated seeds outperformed aldicarb-treated seeds in cotton yield.

[0130] A cotton field trial was conducted at five different locations in the United States to evaluate the effectiveness of seed treatment with Bacillus licheniformis K-357 as a nematicide compared to furrow application of the chemical nematicides aldicarb (15 lb / acre) and fluopyram (16 oz / acre). At sowing, aldicarb and fluopyram were applied in the furrows. Nematode numbers and cotton yield were measured.

[0131] The treatment groups in the field trial are as follows:

[0132] 1. Fungicide (pyraclostrobin, fludioxonil, metalaxyl, fluoxastrobin) + insecticide (thiamethoxam) + nematicide (aldicarb 15 lb / acre)

[0133] 2. Fungicides (pyraclostrobin, fludioxonil, metalaxyl, fluoxastrobin) + insecticides (thiamethoxam, imidacloprid) + nematicides (fluopyram (BAYER) 16 oz / acre)

[0134] 3. Fungicide (Methoxyfenozide, Fludioxonil, Cyazofamid) + Insecticide (Imidacloprid) + Nematicide K-357 @ 2,000Fl.Oz. / CWT

[0135] Field trials were conducted in a randomized complete block design, with each treatment replicated five or six times (depending on collaborator settings). The planting density was 2.5 seeds per row (ft), and the plot size was 4 rows × 50 ft.

[0136] Method for treating seeds with K-357. Cotton seeds treated with Bacillus licheniformis K-357 were prepared as follows: Bacillus licheniformis spores were dispersed in a liquid fertilizer solution containing 89.9% by weight of soybean protein hydrolysate (containing 4% soluble nitrogen) and 0.15% by weight of chelated ferrous sulfate, at a concentration of 7.0 × 10⁻⁶ ppm. 9 CFU (designated "K-357 Liquid"). Apply K-357 Liquid to cottonseed at a ratio of 2 fluid ounces / 100 pounds of cottonseed (designated "2,000 Fl.Oz. / CWT").

[0137] Figure 6-7 An example of field trial data is shown.

[0138] Figure 6 This graph shows the percentage of kidney-shaped nematode counts in cotton seeds treated with the chemical nematicides aldicarb or fluopyram in furrows 55 days after emergence, compared to seeds treated with K-357, in a field trial in Arkansas. The results showed that Bacillus licheniformis K-357 reduced nematode numbers to approximately 70% of the chemical standard aldicarb and was superior to fluopyram.

[0139] Figure 7 This is a graph showing the percentage of cotton yield at harvest in this experiment. Surprisingly, seeds treated with K-357 yielded higher yields than cotton treated with fluopyram (~4.5%) and aldicarb (~2.5%) in the furrows.

[0140] Example 3

[0141] K-357-treated seeds outperformed AVICTA ELITE containing VIBRANCE in cotton yield. PLUS

[0142] A cotton field trial was conducted at five different locations in the United States to evaluate its effectiveness and efficacy. contain The efficacy of seeds treated with Bacillus licheniformis K-357 as a nematicide in reducing nematode numbers and increasing yield was compared to that of seeds treated with AVICTA ELITE PLUS nematicide (referred to as avermectin in this study). In the experiment, seeds treated with two different proportions of Bacillus licheniformis K-357 were compared with seeds treated with avermectin applied at the indicated proportion.

[0143] In the field trial, the cotton seed treatment groups were as follows:

[0144] 1. Fungicide (pyraclostrobin, fludioxonil, metalaxyl, fluoxastrobin) + Insecticide (imidacloprid, thiamethoxam) + Nematicide (avermectin, AVICTA ELITE PLUS from Syngenta containing VIBRANCE, applied according to the indicated ratio)

[0145] 2. Fungicide (Methoxyfenozide, Fludioxonil, Cyazofamid) + Insecticide (Imidacloprid) + Nematicide (K-357@2,000Fl.Oz. / CWT)

[0146] 3. Fungicide (Methoxyfenozide, Fludioxonil, Cyazofamid) + Insecticide (Imidacloprid) + Nematicide (K-357@3.000Fl.Oz. / CWT)

[0147] Field trials were conducted in a randomized complete block design, with each treatment replicated five or six times (depending on collaborator settings). The planting density was 2.5 seeds per row (ft), and the plot size was 4 rows × 50 ft.

[0148] K-357 seed treatment method: Bacillus licheniformis spores are dispersed in a liquid fertilizer solution containing 89.9% by weight of soybean protein hydrolysate (containing 4% soluble nitrogen) and 0.15% by weight of chelated ferrous sulfate, at a concentration of 7.0 × 10⁻⁶ ppm. 9CFU (designated "K-357 Liquid"). K-357 Liquid is applied to cottonseed at a ratio of 2 or 3 fluid ounces per 100 pounds of cottonseed (designated "2,000 Fl.Oz. / CWT" and "3,000 Fl.Oz. / CWT" respectively).

[0149] Figure 8-11 Examples of field trial results are shown.

[0150] Figure 8 This study presents juvenile root-knot nematode populations at sowing, 60 days later, and harvest in a cotton field trial conducted in Texas, comparing seeds treated with abamectin with those treated with K-357 as a nematicide. While the number of nematodes treated with abamectin was lower than that treated with K-357 at 60 days, this relationship reversed at harvest. At harvest, the number of nematodes treated with K-357 decreased to below 15%, while the number of nematodes treated with abamectin was 40%. In fact, the nematode population in the K-357 group started higher and steadily declined as the trial progressed. In contrast, the nematode population in the abamectin group reversed and rapidly increased after 60 days. These data suggest that K-357 has a more durable inhibitory effect compared to abamectin.

[0151] Figure 9 This graph shows the percentage of nematode counts 25 days after planting cotton seeds treated with avermectin or K-357 at 2 Fl.Oz. / CWT or 3,000 Fl.Oz. / CWT as a nematicide in a field trial in Mississippi. Surprisingly, the nematode counts in seeds treated with 2,000 Fl.Oz. / CWT K-357 and 3,000 Fl.Oz. / CWT K-357 were approximately 8% and 12% lower, respectively, than those in seeds treated with avermectin.

[0152] Figure 10 This is a graph showing cotton yield at harvest for three different seed treatments. The effects of K-357 treatments at 2,000 Fl.Oz. / CWT and 3,000 Fl.Oz. / CWT had a more significant impact on yield than on nematode counts, increasing yield by 54 lb / acre and 102 lb / acre, respectively.

[0153] Figure 11 It is displayed Figure 9 The graph shows plant vigor calculated 1–9 days, 12 days, and 25 days after cotton seed planting in a field trial. The data show that both dosages of K-357 improved plant vigor between 12 and 25 days post-planting, while no improvement was observed with abamectin.

[0154] Example 4

[0155] K-357 is more effective than Avicta in killing nematodes in vitro.

[0156] In vitro experiments were conducted to evaluate the efficacy of Bacillus licheniformis K-357 against four different types of plant nematodes: soybean cyst nematode, southern root-knot nematode, kidney-shaped nematode, and disease-causing nematode, and compared with the chemical nematicide Avicta. Nematodes were placed in petri dishes and treated with Avicta, water, or K-357 spores for 48 hours as described below.

[0157] The experiment involved three treatments:

[0158] 1. Avicta (Syngenta, 0.031%)

[0159] 2. Control: Water

[0160] 3. K-357@0.3333 10L / ha (equivalent to 50ml containing 5.5×10) 7 CFU)

[0161] The lyophilized K-357 was dispersed in water to provide a concentrated solution (10). 8 The concentrated solution (CFU / ml) was diluted to a total of 50 ml. Similarly, Avicta was diluted to 50 ml, and each of the above treatments (1)-(3) was applied to four different species of nematode eggs present in a petri dish and left to stand for 48 hours. Mortality was assessed by stereomicroscopy. Nematodes exhibiting any mobility or appearing in a curved shape were considered alive, while those that did not exhibit any mobility and whose body shape was straight were considered dead.

[0162] Data shows that Figure 12 The figure shows the percentage mortality rate for each of the four nematode types. Surprisingly, the K-357 treatment was as effective as or significantly better than Avicta in killing nematodes. Specifically, K-357 was equally effective in killing southern root-knot nematodes, soybean cyst nematodes, and disease nematodes, and significantly superior to Avicta in killing kidney-shaped nematodes (84.6% and 35.4%, respectively).

[0163] Example 5

[0164] Field trials of maize seeds treated with K-357 and chemical insecticides / fungicides.

[0165] Corn field trials were conducted in the United States to evaluate the ability of seeds treated with Bacillus licheniformis K-357 alone, as well as seeds treated with Bacillus licheniformis K-357 in combination with chemical insecticides and fungicides, to reduce corn root-worm (CRW) damage and increase yield. These trials were conducted at the University of Illinois at Monmouth, in fields with high CRW virus infestations. In the trials, chemical insecticides and fungicides were applied at labeled dosages. Corn plant number, mean root damage score, and yield were determined.

[0166] In the field trial, the treated maize seed groups were as follows:

[0167] 1. Fungicides (pyraclostrobin, metalaxyl, tebuconazole, vitavax (37.5% carboxin + 37.5% DS thiram)) + insecticides (bifenthrin @ 3.6 FL.Oz. / CWT, imidacloprid @ 4.8 FL.Oz. / CWT)

[0168] 2. K-357 (2.00 FL.Oz. / CWT) + fungicide (azoxystrobin, metalaxyl, tebuconazole, vitavax (37.5% carboxin + 37.5% DS thiram)) + insecticide (bifenthrin @ 3.6 FL.Oz. / CWT, imidacloprid @ 4.8 FL.Oz. / CWT)

[0169] 3. K-357 (2.00 FL.Oz. / CWT) + fungicide (pyraclostrobin, metalaxyl, tebuconazole, vitavax (37.5% carboxin + 37.5% DS thiram))

[0170] 4. K-357 (2.00 FL.Oz. / CWT)

[0171] The field trial was conducted in a randomized complete block design, with each treatment replicated four times. The planting density was 4 rows × 30 feet (30-inch row spacing), with 5-foot unplanted paths between plots. Stand counts were recorded 37 days after planting, and root node damage scores were recorded 64 days after planting.

[0172] K-357 seed treatment method: Bacillus licheniformis spores were dispersed in a liquid fertilizer containing 89.9% by weight of soybean protein hydrolysate (containing 4% soluble nitrogen) and 0.15% by weight of chelated ferrous sulfate, at a concentration of 7.0 × 10⁻⁶. 9 CFU / mL (named "K-357 Liquid"). Apply K-357 Liquid to corn seeds at a ratio of 2 or 3 fluid ounces / 100 lbs of corn seeds (named "2,000 Fl.Oz. / CWT" and "3,000 Fl.Oz. / CWT" respectively).

[0173] During the early vegetative stage, record stand counts and plant height at least once. Monitor plots for early seasonal pests; if natural pests occur, collect data from all plots. Collect and wash maize roots (5 per plot) and rate maize rootworm damage using a 0-3 node damage scale; this rating is based on peak loss estimates determined by degreedrays and preliminary assessments of untreated plots (typically mid-to-late July). Record yields from row 2 using plot harvest. Perform ANOVA and post-mean separation on the data.

[0174] The results of stand counts and root node damage analysis at 37 and 64 days after planting showed that... Figure 13 and 14 In the middle, the mean stand count for both seed treatment group #1 (insecticide / fungicide) and seed treatment group #2 (K-357 + insecticide / fungicide) was 33.8. Figure 13 ).However, Figure 14 The root node damage plots show that the addition of K-357 to the insecticide / fungicide mixture reduced root node damage. Specifically, the root node damage score was 1.12 for seed treatment group #2 (K-357 + insecticide / fungicide) and 1.47 for seed treatment group #1 (insecticide / fungicide).

[0175] Example 6

[0176] K-357 inhibits potato virus Y transmitted by the green peach aphid.

[0177] The purpose of the experiments outlined below is to determine, using virus transmission assays on detached leaves of *Tobacco Benzoenta*, whether *Bacillus licheniformis* K-357 can reduce or prevent the spread of Potato Virus Y (PVY). The green peach aphid (GPA, peach aphid) is the most effective vector of PVY. It is hypothesized that *Bacillus licheniformis* K-357 can weaken hemiptera pests (such as aphids), thereby completely inhibiting and / or reducing PVY transmission.

[0178] Aphid and virus isolates: A cohort of GPAs was cultured and maintained on a mixture of radish and mustard seedlings in a climate-controlled indoor environment (14h light; 20±3℃; 70% relative humidity). The PVY strain used in this study was a recombinant strain NTN, previously collected from infected potato plants in the St. Louis Valley, Colorado.

[0179] Analysis of artificial feed: A rearing chamber was constructed by stretching a sheet of plastic wrap across a petri dish (60 mm in diameter). Then, 1.2 ml of artificial feed (Dadd, R. & Mittler, T. Cellular and Molecular Life Sciences 22, 832-833 (1966)) was added, and a second layer of stretched plastic wrap was placed to form a feeding bag through which the aphids could feed. An O-ring was placed on the feeding bag, and five aphids were placed on the bag. The entire apparatus was covered by the lid of the 60 mm petri dish.

[0180] Experiment #1: To determine whether K-357 acts as an aphid killer, the mortality rate of GPAs fed with K-357 was first determined. In short, age-synchronized GPAs were fed artificial diets containing three different concentrations (low, medium, and high) of K-357. After 24 and 48 hours, the aphids were transferred to detached leaves of *N. benthamiana*. Aphid growth and mortality were monitored over two weeks. Artificial diets without K-357 served as a control. The concentration of K-357 used in all future experiments will result in a mortality rate of less than 15%, similar to the control diet level.

[0181] Objective 1: Determination of virus transmission efficiency: To standardize the artificial feed experiments, first-instar nymphs (age 0-24h) were selected at the beginning of each experiment. The experiments for Objective 1 were conducted in a climate-controlled indoor environment. A complete experimental procedure was used (see [link to experimental procedure]). Figure 15 ). Figure 15 The following describes the sequential steps of the experimental setup used to study the effect of K-357 on the spread of PVY by the green peach aphid.

[0182] Step 1: By placing first-instar nymphs on a mixture of radish and mustard seedlings, aphids of synchronized age are produced.

[0183] Step 2: Once each aphid reaches the third instar (after 24-48 hours), the aphids are transferred to a rearing room containing a concentration of K-357 that results in a mortality rate of less than 15%. A diet without K-357 is used as a control. The aphids are allowed to feed for 24 and 48 hours to ensure K-357 is absorbed by the aphids. For each treatment (4 treatments x 2 feeding time points), five age-synchronized GPAs are used.

[0184] Step 3: After the appropriate feeding times (24 and 48 hours), carefully transfer the GPA to the PVY. NTN Positive or infected tobacco plants are allowed to be fed for 48 hours. This feeding period is the viral acquisition and entry period, or AAP.

[0185] Step 4: After a 48-hour PVY acquisition period, transfer individual GPAs (one GPA per healthy leaf) to a single healthy detached Nicotiana leaf and incubate for 30 minutes to begin the feeding period. Under laboratory and growth chamber conditions, a single detached Nicotiana leaf placed in a petri dish with moist filter paper can survive for up to one month without any harmful effects. Remove, collect, and store the individual GPAs for further analysis (if needed).

[0186] Step 5: Monitor the development of symptoms on isolated tobacco leaves daily over three weeks. At the onset of symptom development, collect a small sample of leaves and test for the presence of PVY using PVY immunostrips (AGDIA, Inc., Indiana).

[0187] All experiments were repeated 5 times, with 10 replicates containing one aphid (total: 400 GPA individuals and detached tobacco leaves). ANOVA analysis was used to determine the effect of K-357 on virus transmission (number of infected plants per batch).

[0188] Therefore, although the methods and systems have been described with reference to specific implementations, features and illustrative implementations, it should be understood that the utility of this subject matter is not limited thereto, but extends to and includes many other variations, modifications and alternative implementations, as would be conceived by one of ordinary skill in the art based on the disclosure herein.

[0189] Various combinations and sub-combinations of the compositions, methods, and features described herein are contemplated and will be apparent to those skilled in the art upon which this disclosure is known. Unless otherwise indicated herein, any of the various features and elements disclosed herein may be combined with one or more other disclosed features and elements. Accordingly, the subject matter claimed below is intended to be broadly understood and interpreted to include all such variations, modifications, and alternative embodiments within its scope, and to include equivalents of the claims.

Claims

1. A method for controlling plant pests, the method comprising: Plants or plant seeds are planted in a suitable growing environment, the plants or seeds having a coating or partial coating of a composition comprising: A pure biological culture of Bacillus licheniformis K-357, with accession number NRRL B-23318; and One or more excipients, nutrients, biological crop protectants, or chemical crop protectants. The composition does not include Bacillus subtilis, and the pests are soybean cyst nematode, southern root-knot nematode, kidney nematode, and diseased nematode.

2. A method for controlling plant pests, the method comprising: Delivering a composition to the seeds, leaves, roots, or soil or growing environment of a plant, said composition comprising: A pure biological culture of Bacillus licheniformis K-357, with accession number NRRL B-23318; and One or more excipients, nutrients, biological crop protectants, or chemical crop protectants. The composition does not include Bacillus subtilis, and the pests are soybean cyst nematode, southern root-knot nematode, kidney nematode, and diseased nematode.

3. The method according to claim 1, wherein the Bacillus licheniformis K-357 is at a concentration of 1.0 × 10⁻⁶. 12 CFU / g up to 1.0×10 1 The amount of CFU / g seed present in the seed.

4. The method according to claim 1 or 2, wherein the chemical crop protectant comprises one or a combination of imidacloprid, bifenthrin, metalaxyl, tebuconazole, azoxystrobin, carbendazim, thiram, fludioxonil, cyazofamid, metalaxyl-M, fluoxastrobin, thiamethoxam, styraclostrobin, or abamectin.

5. The method of claim 2, wherein the composition is in liquid form, and the Bacillus licheniformis K-357 is expressed at a concentration of 1.0 × 10⁻⁶. 12 Up to 1.0×10 1 A concentration of CFU / ml is present.

6. The method of claim 2, wherein the composition is in the form of a powder, a dry wettable powder, spreadable granules, or dry wettable granules, and the Bacillus licheniformis K-357 is in the form of 5.0 × 10⁻⁶. 11 CFU / g up to 1.0×10 3 A CFU / g level is present.

7. The method according to claim 1 or 2, wherein the plant comprises cereals, brassica vegetables, cucurbitaceous vegetables, bulbous vegetables, citrus fruits, leafy vegetables, legumes, pome fruits, or grasses.

8. The method according to claim 1 or 2, wherein the plant is selected from cotton, rice, soybean, tomato, sunflower, sugarcane, sugar beet, strawberry, kiwi, banana or ornamental plants.

9. A method for controlling plant pests, the method comprising: Applying a composition coating to plants or plant seeds, said composition coating comprising: i. A pure biological culture of Bacillus licheniformis K-357, the accession number of Bacillus licheniformis K-357 being NRRL B-23318; ii. Soybean protein hydrolysates; iii. Chelated ferrous sulfate; iv. One or more excipients, nutrients, biological crop protectants, or chemical crop protectants, The composition does not include Bacillus subtilis, and the pests are soybean cyst nematode, southern root-knot nematode, kidney nematode, and diseased nematode.

10. The method of claim 9, wherein the soybean protein hydrolysate contains 4% water-soluble nitrogen.

11. The method of claim 9, wherein the soybean protein hydrolysate is present in the composition at 60% to 99% by weight, and the chelated ferrous sulfate is present in the composition at 0.05% to 1.5% by weight.

12. The method of claim 9, wherein the soybean protein hydrolysate is present in the composition at 85% to 95% by weight, and the chelated ferrous sulfate is present in the composition at 0.05% to 0.3% by weight.

13. The method of claim 9, wherein the composition is in liquid form, and Bacillus licheniformis K-357 is present at 1.0 × 10⁻⁶. 12 Up to 1.0×10 1 A concentration of CFU / ml is present.

14. The method of claim 9, wherein the Bacillus licheniformis K-357 is at a concentration of 1.0 × 10⁻⁶. 12 CFU / g up to 1.0×10 1 The amount of CFU / g is present in seeds or plants.

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