Serratia marcescens, microbial agent, pesticide and application thereof

Abstract

The present application provides a Serratia marcescens, an Enterococcus sp., and a Morganella morganii. The present application also provides a microbial agent, which includes one or more of Serratia marcescens, Enterococcus sp., and Morganella morganii. The microbial agent can be applied to the prevention and direct killing of fall armyworms in the corn seedling stage.

Description

Technical Field

[0001] The present application belongs to the field of microbial technology, and specifically relates to a kind of Serratia marcescens, a microbial agent, a pesticide and their application. Background Art

[0002] Fall armyworm (Spodoptera frugiperda), belonging to the Lepidoptera, Noctuidae, Spodoptera genus, also known as fall armyworm, is a major agricultural pest newly invaded my country and is rated as one of the world's top ten plant pests by CABI International. The pest is native to tropical and subtropical regions of America. After it was first discovered in Yunnan, my country on January 11, 2019, it quickly spread to 1,366 counties (cities and districts) in 24 provinces in my country.

[0003] The fall armyworm has a strong ability to adapt to its host, and its larvae can feed on more than 80 kinds of crop plants such as corn, rice, and peanuts, causing serious damage. In the existing technology, the prevention and control of the fall armyworm mainly adopts chemical control methods, which has risks such as pesticide residues and the development of drug resistance. Screening more new strain resources that inhibit the hatching of fall armyworm eggs and the death of larvae can provide an effective way to achieve efficient, green, and sustainable prevention and control of the fall armyworm. The bacterial microbial agents currently screened are mainly Bacillus thuringiensis and Cryptobacterium brevis, which are far from enough for the needs of production prevention and control applications.

[0004] In view of this, this application is hereby filed. Summary of the invention

[0005] The purpose of the present application is to solve the above problems, and the first purpose is to provide Serratia marcescens.

[0006] The second purpose of the present application is to provide a microbial agent.

[0007] The third object of the present application is to provide a pesticide or a plant growth promoter.

[0008] The fourth object of the present application is to provide an application of a pesticide or a plant growth promoter.

[0009] To achieve the above objectives, this application adopts the following technical solutions:

[0010] 1. A Serratia marcescens, whose deposit number is CGMCC24567.

[0011] 2. The Serratia marcescens according to item 1, wherein the 16S rDNA gene sequence is shown in SEQ ID NO: 1.

[0012] 3. An Enterococcus sp., whose accession number is CGMCC24566.

[0013] 4. The enterococcus according to item 3, whose 16S rDNA gene sequence is shown in SEQ ID NO: 2.

[0014] 5. A Morganella morganii, whose deposit number is CGMCC24593.

[0015] 6. The Morganella morganii according to item 5, wherein the 16S rDNA gene sequence is shown in SEQ ID NO: 3.

[0016] 7. A microbial agent, comprising one or more of Serratia marcescens, Enterococcus and Morganella;

[0017] Preferably, the bacterial agent includes Serratia marcescens, Enterococcus and Morganella;

[0018] Further preferably, the bacterial agent consists of Serratia marcescens, Enterococcus and Morganella;

[0019] More preferably, calculated by weight, the amount of Serratia marcescens relative to the amount of Enterococcus is 1-10 parts, and the amount of Morganella relative to the amount of Enterococcus is 1-10 parts;

[0020] More preferably, the bacterial agent is composed of Serratia marcescens with a preservation number of CGMCC24567, Enterococcus with a preservation number of CGMCC24566, and Morganella morganii with a preservation number of CGMCC24593.

[0021] 8. A pesticide or a plant growth promoter, which comprises the microbial agent described in item 7.

[0022] 9. The pesticide or plant growth promoter according to item 8,

[0023] The pesticide or plant growth promoter also includes an adjuvant;

[0024] Preferably, the auxiliary agent is selected from one or more of methyl methacrylate and methacrylic acid copolymer nanoparticles, nanogold particles, urea, wettable powder cuprous oxide, ferrous sulfate, sodium alkyl sulfonate, and soapberry extract, preferably methyl methacrylate and methacrylic acid copolymer nanoparticles;

[0025] Further preferably, the adjuvant is 0.1-10wt% based on the mass percentage of the total weight of the pesticide or plant growth promoter.

[0026] 10. The pesticide or plant growth promoter according to item 8 or 9, wherein the plant growth promoter comprises Serratia marcescens.

[0027] 11. A method for promoting plant growth or controlling fall armyworm or promoting cannibalism among fall armyworm larvae, comprising applying the microbial agent described in item 7 or the pesticide or plant growth promoter described in item 8 or 9 to plants;

[0028] Preferably, the plant is selected from one or more of corn, rice, sorghum, barley, cotton, rapeseed, and legumes, preferably corn;

[0029] The application method is root dredging and / or spraying.

[0030] Effects of the Invention

[0031] 1. This application isolates a strain of Serratia marcescens, and studies have found that Serratia marcescens has preventive and insecticidal effects on the fall armyworm; on this basis, the effect of directly killing the fall armyworm was tested and improved, and ultimately the insecticidal effects of Enterococci and Morganella, their synergistic effects on Serratia marcescens, and the optimal preparation method and application method of microbial agents were determined.

[0032] 2. Serratia marcescens also has a certain effect of promoting plant growth. The microbial insecticide in this application is non-toxic and pollution-free, not easy to produce drug resistance, and has high biological safety, and is suitable for the prevention and direct killing of fall armyworms in the seedling stage of crops.

[0033] 3. This application confirms that a strain of Serratia marcescens extracted from the intestines of insects can improve the growth of corn and its preventive effect on the fall armyworm; on this basis, the effect of directly killing the fall armyworm was tested and improved, and finally it was determined that the microbial agent was configured with methyl methacrylate and methacrylic acid copolymer nanoparticles.

[0034] 4. The present application provides a microbial agent or pesticide with significant insecticidal effect. The present application is applicable to the prevention and direct killing of fall armyworms in the corn seedling stage.

[0035] Collection Information

[0036] The Serratia marcescens AHPC29 described in this application was deposited on March 22, 2022 at the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. The strain name is Serratia marcescens AHPC29 and the deposit number is CGMCC24567.

[0037] The Enterococcus sp. 10A described in this application was deposited on March 22, 2022 at the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. The strain name is Enterococcus sp. 10A and the deposit number is CGMCC24566.

[0038] The Morganella morganii CDHu described in this application was deposited on March 25, 2022 at the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. The strain name is Morganella morganii CDHu and the deposit number is CGMCC24593. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 The effect of different temperatures on the growth of Serratia marcescens strains in Example 2;

[0040] Figure 2 The effect of different culture time on the growth of Serratia marcescens strain in Example 2;

[0041] Figure 3 The effect of Serratia marcescens on the height of corn seedlings in Example 5;

[0042] Figure 4 The effect of Serratia marcescens on the number of leaves of corn seedlings in Example 5;

[0043] Figure 5 The effect of Serratia marcescens on the leaf width of corn seedlings in Example 5;

[0044] Figure 6 is the ratio of the number of leaves of corn seedlings treated with Serratia marcescens in Example 6 that were eaten by the fall armyworm;

[0045] Figure 7 is the proportion of the leaf surface of the corn seedlings treated with Serratia marcescens in Example 6 that was eaten by the fall armyworm;

[0046] Figure 8 The Serratia marcescens in Example 6 reduces the mortality rate of fall armyworm to corn seedlings;

[0047] Fig. 9 The synergistic effect of different adjuvants on promoting crop growth by Serratia marcescens in Example 7;

[0048] Fig.10 The synergistic effect of different concentrations of methyl methacrylate and methacrylic acid copolymer nanoparticles on the bacterial agent in Example 8;

[0049] Fig.11 The direct insecticidal effect of Serratia marcescens, Enterococcus and Morganella on Spodoptera frugiperda in Example 9;

[0050] Fig.12 The effect of different preparation ratios of the microbial agent in Example 10 on the mortality rate of fall armyworm;

[0051] Fig.13 The figure shows the effect of different preparation ratios of the microbial agent in Example 11 on the cannibalism rate of the fall armyworm. DETAILED DESCRIPTION

[0052] The present application is further described below in conjunction with examples. It should be understood that the examples are only used to further describe and illustrate the present application and are not used to limit the present application.

[0053] Unless otherwise defined, the technical and scientific terms in this specification have the same meaning as those generally understood by those skilled in the art. Although similar or identical methods and materials as described herein may be used in experiments or practical applications, materials and methods are described herein below. In the event of a conflict, the present specification including the definitions therein shall prevail. In addition, materials, methods and examples are provided for illustration only and are not restrictive. The present application is further described below in conjunction with specific embodiments, but is not intended to limit the scope of the present application.

[0054] The application method of the pesticide of the present application can be appropriately selected according to the type of applied plant, the type of pest, the application site, the application period, the dosage form, etc. The application method in the present application is preferably root irrigation and / or spraying.

[0055] The pesticide of the present application can be directly applied as it is, or applied after dilution with water or a carrier. The application method can be listed as follows: spreading on plant stems and leaves, spreading on the base of the plant (plant element), spreading on the soil surface, soil mixing, soil irrigation, water surface application, seed powder coating (seed dressing), coating, dipping, root irrigation, spraying and other methods, but are not limited to these methods. Moreover, the pesticide formulation composition of the present application can also be mixed with other fungicides, insecticides, nematicides, herbicides, plant growth regulators, fertilizers, soil improvement materials, etc., applied alternately, or applied simultaneously, in which case it sometimes shows a more excellent effect.

[0056] The application site of the pesticide of the present application can be applied to nursery beds, farmlands, paddy fields, orchards, hydroponic culture facilities, etc. for cultivating agricultural and horticultural plants, but is not limited to these.

[0057] The application period of the pesticide of the present application is not limited to the planting period. In the case of a nursery, it can be applied at any time before, during, or after planting. In the case of the seedling raising period, it can be applied at any time before, during, or after sowing.

[0058] The application amount of the pesticide in this application varies according to the type of plant, the type of pest or weed, the state of the soil, the application period, the planting density, the dosage form, etc., so it cannot be generally specified. For example, in the case of the seedling stage, the application amount of the pesticide per 1m 2 You can use about 100 to 1000g of soil. In the case of a nursery, use about 100 to 1000g of soil per 1m 2 For soil of about 5 to 1000 g, about 1 to 1000 g can be used. In addition, when powder coating is formed on seeds (including seed potatoes, tubers, bulbs, bulbs, etc.), the crushed material can be used directly or after diluting with water, etc., at about 1 to 100 g per 1 kg of seeds.

[0059] Specific examples of plants to which the pesticide of the present application is applied include: cereals (e.g., rice, wheat, barley, rye, oats, corn, sorghum, millet, sorghum, millet, foxtail millet, millet, buckwheat), tubers (e.g., potatoes, sweet potatoes, taro, yam, konjac), legumes (e.g., soybeans, red beans, kidney beans, peas, broad beans, peanuts, cowpeas, chickpeas, pigeon peas), vegetables (e.g., eggplant, tomato, green pepper, hot pepper, cucumber, melon, watermelon, pumpkin, zucchini, blueberry, gourd, winter melon, bitter melon, cabbage, Chinese cabbage, broccoli, cauliflower, white radish, turnip, pakchoi, komatsuna, mizuna, green onions, onions, leeks, garlic, wild garlic, etc.) chive), asparagus, lettuce, burdock, chrysanthemum, butterbur, carrot, celery, celery, parsley, strawberry, spinach, okra, basil, basil, mint, ginger, cumin), fruit trees (e.g., apple, pear, pear, quince, papaya, sweet cherry, peach, plum, apricot, chestnut, walnut, almond, pecan, grape, kiwi, akebia, persimmon, fig, pomegranate, raspberry, blackberry, blueberry, cranberry, citrus, loquat, olive, bayberry, mango, guava, avocado, date palm, coconut, banana, pineapple, papaya, passion fruit, acerola), specialty crops (e.g., cotton, , flax, rush, rapeseed, sunflower, sesame, palm oil, beet, sugar cane, tea, coffee, cocoa, hops, tobacco), flowers (e.g., cosmos, morning glory, marigold, impatiens, baby's breath, sweet pea, chrysanthemum, carnation, tulip, lily, narcissus, gladiolus, cyclamen, begonia, water lily, dahlia, rose, cymbidium, Cattleya), lawn grass (e.g., zoysia, korean grass, zoysia, Bermuda grass, chaff grass, fescue, ryegrass, bluegrass), trees (e.g., cherry, azalea, oak, beech, cypress, beech), etc., but are not limited to these examples. In addition, it can also be applied to plants that are endowed with resistance to pests and diseases, resistance to herbicides, and resistance to environmental stresses such as dryness by traditional breeding methods, genetic recombination technology, etc.

[0060] In the present application, the plant is preferably one or more of corn, rice, sorghum, barley, cotton, rapeseed, and legumes, and most preferably corn.

[0061] In the present application, "control" means to avoid plant diseases and the like by preventing the infection of bacterial plant disease bacteria and the like that target crops (plants).

[0062] The present application provides a Serratia marcescens, which is isolated from the intestinal microorganisms of insects, and the Serratia marcescens is named Serratia marcescens AHPC29. The strain was deposited on March 22, 2022 at the General Microbiological Center of the China Microbiological Culture Collection Administration, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the strain name is Serratia marcescens AHPC29, and the deposit number is CGMCC24567.

[0063] The "Serratia marcescens" in this article, also known as Bacillus thuringiensis, belongs to the genus Serratia of the family Enterobacteriaceae and is a Gram-negative facultative anaerobic bacillus. Serratia marcescens is a non-demanding microorganism that can utilize a variety of carbon sources, such as chitosan, glucose, sucrose, mannose, cellobiose, citric acid glycerol, etc. Such a wide spectrum of carbon sources provides favorable conditions for its fermentation production. The nuclease from Serratia marcescens consists of 266 amino acids. Unlike other Gram-negative bacteria, the signal peptide composed of 1-21 amino acids helps the nuclease to be secreted into the culture medium. The proteins of other Gram-negative bacteria are secreted into the plasma membrane space rather than the surrounding culture medium. During the secretion process, the signal peptide (1-21 amino acids) is removed; the 22-266 amino acid part with hydrolytic nuclease activity is secreted into the culture medium, with a molecular weight of about 26.7kDa.

[0064] According to the present application, the method for isolating the Serratia marcescens may be a conventional method for screening new strains in the art. For example, the separation method may include: Serratia marcescens AHPC29 may be screened and obtained by the following method:

[0065] In a clean bench, rinse the larvae from which intestinal bacteria are to be removed in 75% alcohol and sterile water for about 3 seconds respectively; use sterile dissecting scissors to slowly cut open the larval body wall, remove the intestine, and place it in a sterile centrifuge tube; add 1 mL of sterile water to grind; dilute the grinding solution by 10 6 Then take 10 μL of the dilution and spread it on the plate, and place the plate in a 28°C incubator in the dark for 2 to 3 days.

[0066] The 16SrDNA gene sequence of Serratia marcescens AHPC29 is shown in SEQ ID NO: 1:

[0067] SEQ ID NO: 1

[0068]

[0069] The DNA template for full-length identification of bacterial strain 16S involved in this embodiment is prepared quickly and with high quality by Mighty Prep reagent for DNA (Takara). The PCR amplification primers for the full length of 16S rDNA use 8F 5'-GCGGATCCGCGGCCGCTGCAGAGTTTGATCCTGGCTCAG) (recorded as SEQ ID NO: 4) and 1492R 5'-GGCTCGAGCGGCCGCCCGGGTTACCTTGTTACGACTT) (recorded as SEQ ID NO: 5) (Weisburg et al., 1991), and the primers are synthesized by Beijing Qingke Biotechnology Co., Ltd. The PCR product is detected using 1% agarose gel, and the fragment size is 1500bp.

[0070] The present application provides an enterococcus (Enterococcus sp.), which is isolated from the intestine of an insect, and the enterococcus (Enterococcus sp.) is named Enterococcus (Enterococcus sp.) 10A. The strain was deposited on March 22, 2022 at the General Microbiological Center of the China Microbiological Culture Collection Administration, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the strain name is Enterococcus sp.10A, and the deposit number is CGMCC24566.

[0071] According to the present application, the method for isolating the enterococci may be a conventional method for screening new strains in the art. For example, the isolation method may include: Enterococcus sp. 10A may be screened by the following method:

[0072] In a clean bench, rinse the larvae from which intestinal bacteria are to be removed in 75% alcohol and sterile water for about 3 seconds respectively; use sterile dissecting scissors to slowly cut open the larval body wall, remove the intestine, and place it in a sterile centrifuge tube; add 1 mL of sterile water to grind; dilute the grinding solution by 10 6 Then take 10 μL of the dilution and spread it on the plate, and place the plate in a 28°C incubator in the dark for 2 to 3 days.

[0073] The 16S rDNA gene sequence of the Enterococcus sp. 10A is shown in SEQ ID NO: 2:

[0074] SEQ ID NO: 2

[0075]

[0076] The DNA template for full-length identification of bacterial strain 16S involved in this embodiment is prepared quickly and with high quality by Mighty Prep reagent for DNA (Takara). The PCR amplification primers for the full length of 16S rDNA use 8F 5'-GCGGATCCGCGGCCGCTGCAGAGTTTGATCCTGGCTCAG) (recorded as SEQ ID NO: 4) and 1492R 5'-GGCTCGAGCGGCCGCCCGGGTTACCTTGTTACGACTT) (recorded as SEQ ID NO: 5) (Weisburg et al., 1991), and the primers are synthesized by Beijing Qingke Biotechnology Co., Ltd. The PCR product is detected using 1% agarose gel, and the fragment size is 1500bp.

[0077] The present application provides a Morganella morganii, which is isolated from the intestine of an insect, and the Morganella morganii is named Morganella morganii CDHu. The strain was deposited on March 25, 2022 at the General Microbiological Center of the China Microbiological Culture Collection Administration, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the strain name is Morganella morganii CDHu, and the preservation number is CGMCC24593.

[0078] Morganella morganii (abbreviated as Morganella or Morganella) is a facultative anaerobic bacterium, oxidase negative. Its colonies are off-white and opaque. When grown on agar plates, Morganella is a straight rod with a diameter of about 0.6-0.7 microns and a length of about 10-17 microns. It moves by means of pericarial flagella, but some strains do not form flagella at 30°C.

[0079] According to the present application, the method for isolating the Morganella morganii may be a conventional method for screening new strains in the art. For example, the isolation method may include: Morganella morganii CDHu may be screened and obtained by the following method:

[0080] In a clean bench, rinse the larvae from which intestinal bacteria are to be removed in 75% alcohol and sterile water for about 3 seconds respectively; use sterile dissecting scissors to slowly cut open the larval body wall, remove the intestine, and place it in a sterile centrifuge tube; add 1 mL of sterile water to grind; dilute the grinding solution by 10 6 Then take 10 μL of the dilution and spread it on the plate, and place the plate in a 28°C incubator in the dark for 2 to 3 days.

[0081] The 16S rDNA gene sequence of the Morganella morganii CDHu is shown in SEQ ID NO: 3:

[0082] SEQ ID NO: 3

[0083]

[0084] The DNA template for the full-length identification of the bacterial strain 16S involved in this example was prepared quickly and with high quality by MightyPrep reagent for DNA (Takara). The PCR amplification primers for the full-length 16S rDNA used 8F 5'-GCGGATCCGCGGCCGCTGCAGAGTTTGATCCTGGCTCAG) (denoted as SEQ ID NO: 4) and 1492R 5'-GGCTCGAGCGGCCGCCCGGGTTACCTTGTTACGACTT) (denoted as SEQ ID NO: 5) (Weisburg et al., 1991), and the primers were synthesized by Beijing Qingke Biotechnology Co., Ltd. The PCR product was detected using 1% agarose gel, and the fragment size was 1500bp.

[0085] The present application provides a microbial agent, which includes one or more of Serratia marcescens, Enterococcus and Morganella.

[0086] In some specific embodiments of the present application, the bacterial agent includes Serratia marcescens, Enterococcus and Morganella.

[0087] In some specific embodiments of the present application, the bacterial agent consists of Serratia marcescens, Enterococcus and Morganella.

[0088] In some specific embodiments of the present application, the weight parts are calculated based on the same OD 600 Calculated based on the volume ratio under the premise of , the Serratia marcescens is 1-10 parts relative to the Enterococcus, and the Morganella is 1-10 parts relative to the Enterococcus;

[0089] For example, the amount of the Serratia marcescens relative to the Enterococcus can be 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts or any range therebetween;

[0090] The Morganella may be present in an amount of 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts or any range therebetween relative to the Enterococcus.

[0091] The weight of different strains can be calculated based on methods known to those skilled in the art, such as based on the same OD 600 It is calculated based on the volume ratio.

[0092] In some specific embodiments of the present application, the bacterial agent is composed of Serratia marcescens with a preservation number of CGMCC24567, Enterococcus with a preservation number of CGMCC24566, and Morganella morganii with a preservation number of CGMCC24593.

[0093] The present application also provides a pesticide, which includes the above-mentioned microbial agent. In addition, the pesticide of the present application may also include other suitable pesticides.

[0094] Other suitable pesticides are biologically active compounds used to control agricultural pests and include, for example, herbicides, plant growth regulators, crop desiccants, fungicides, bactericides, bacteriostats, insecticides, and repellents, as well as their water-soluble salts and esters. Suitable pesticides include, for example, aryloxyphenoxy-propionate herbicides such as haloxyfop-butyl, cyhalofop-butyl, and quizalofop-butyl, triazine herbicides such as metribuzin, hexaxinone, or atrazine; sulfonylurea herbicides such as chlorsulfuron; uracils such as cypermethrin, cypermethrin, or terbacil; urea herbicides such as linuron, diuron, cypermethrin, or chloranil; acetanilide herbicides such as alachlor or isopropyl metolachlor; thiocarbamate herbicides such as cypermethrin and cypermethrin; oxadiazolone herbicides such as oxadiazolone; isoxathiocarb herbicides, phenoxycarboxylic acid herbicides such as dichlorophenoxyacetic acid ("2,4-D"), dichlorophenoxybutyric acid, ("2,4-DB"), 2-methyl-4-chlorophenoxyacetic acid ("MCPA"), 4-(4-chloro-2-methylphenoxy)butanoic acid ("MCPB"), 2,4-dichlorprop, and 2-methyl-4-chlorophenoxypropionic acid, diphenyl ether herbicides such as fluazifop, acifluorfen, chlorfenapyr, or oxyfluorfen; dinitroaniline herbicides such as trifluralin; organophosphonate herbicides such as glufosinate salts and esters and glyphosate salts and esters; dihalobenzonitrile herbicides such as bromoxynil or ioxynil, benzoic acid herbicides such as dicamba, bipyridinium herbicides such as paraquat, and pyridine and pyridyloxycarboxylic acid herbicides such as clopyralid, fluazifop, piclopram, triclopyr, and aminopyralid. Suitable fungicides include, for example, nitrilo-oxime fungicides such as cymoxanil; imidazole fungicides such as benomyl, carbendazim, or thiophanate-methyl; triazole fungicides such as triadimefon; sulfenamide fungicides such as captan; dithiocarbamate fungicides such as maneb, mancozeb, or thiram; chlorinated aromatic fungicides such as chlorpyrifos; dichloroaniline fungicides such as isoprodinil, strobilurin fungicides such as kresoxim-methyl, trifloxystrobin, or azoxystrobin; thiophanate-methyl; copper salt fungicides such as copper oxychloride; sulfur; aniline; and amide fungicides such as metalaxyl or metalaxyl-M. Suitable insecticides include, for example, carbamate insecticides such as methomyl, carbaryl, furadan, or aldicarb; organothiophosphate insecticides such as EPN, isopropylphos, isoxathiophos, chlorpyrifos, or chlormethoate; organophosphate insecticides such as terbufos, monocrotophos, or stirofos; fully chlorinated organic insecticides such as methoxychlor; synthetic pyrethroid insecticides such as fenvalerate, avermectin or emamectin benzoate, neonicotinoid insecticides such as thiamethoxam or imidacloprid; pyrethroid insecticides such as lambda-cyhalothrin, cypermethrin or bifenthrin, and oxadiazine insecticides such as indoxacarb, imidacloprid, or fipronil.Suitable miticides include, for example, propargyl sulfite miticides, such as propargyl; triazapentadiene miticides, such as amitraz; chlorinated aromatic hydrocarbon miticides, such as ethyl chlorfenapyr, or tetrachlorofenapyr; and dinitrophenol miticides, such as binachlor. Suitable nematicides include carbamate nematicides, such as oxamyl.

[0095] Pesticide compounds are generally referred to herein by the names assigned by the International Organization for Standardization (ISO). ISO common names can be cross-referenced with the International Union of Pure and Applied Chemistry ("IUPAC") and Chemical Abstracts Service ("CAS") names through a number of sources.

[0096] In some specific embodiments of the present application, the pesticide further includes an adjuvant.

[0097] In some specific embodiments of the present application, the auxiliary agent is selected from one or more of methyl methacrylate and methacrylic acid copolymer nanoparticles, nanogold particles, urea, wettable powder cuprous oxide, ferrous sulfate, sodium alkyl sulfonate, and soapberry extract, preferably methyl methacrylate and methacrylic acid copolymer nanoparticles.

[0098] In some specific embodiments of the present application, the adjuvant is 0.1-10wt% in terms of mass percentage of the total weight of the pesticide;

[0099] For example, based on the mass percentage of the total weight of the pesticide, the adjuvant can be 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 wt % or any range therebetween.

[0100] The present application also provides a plant growth promoting agent, which includes the above-mentioned microbial agent.

[0101] In some specific embodiments of the present application, the plant growth promoting agent also includes an adjuvant.

[0102] In some specific embodiments of the present application, the auxiliary agent is selected from one or more of methyl methacrylate and methacrylic acid copolymer nanoparticles, nanogold particles, urea, wettable powder cuprous oxide, ferrous sulfate, sodium alkyl sulfonate, and soapberry extract, preferably methyl methacrylate and methacrylic acid copolymer nanoparticles.

[0103] In some specific embodiments of the present application, the auxiliary agent is 0.1-10wt% in terms of mass percentage of the total weight of the plant growth promoting agent;

[0104] For example, based on the mass percentage of the total weight of the plant growth promoter, the auxiliary agent can be 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 wt % or any range therebetween.

[0105] In some specific embodiments of the present application, the plant growth promoting agent includes Serratia marcescens.

[0106] The present application also provides a method for preparing the above-mentioned Serratia marcescens, which comprises the following steps:

[0107] The liquid fermentation method is used to ferment the Serratia marcescens strain in a culture medium to obtain a fermentation liquid; during the fermentation, the temperature is controlled at 28-30°C, the fermentation time is 36-48 hours, and then the fermentation liquid is prepared into an aqueous solution.

[0108] The present application also provides a method for preparing the above-mentioned enterococci, which comprises the following steps:

[0109] The enterococcus strain is fermented in a culture medium by a liquid fermentation method to obtain a fermentation liquid; the temperature is controlled at 28°C and the fermentation time is 48 hours during the fermentation, and then the fermentation liquid is prepared into an aqueous solution.

[0110] The present application also provides a method for preparing the above-mentioned Morganella, which comprises the following steps:

[0111] The Morganella strain is fermented in a culture medium by a liquid fermentation method to obtain a fermentation liquid; the temperature is controlled at 28°C and the fermentation time is 48 hours during the fermentation, and then the fermentation liquid is prepared into an aqueous solution.

[0112] The present application provides a method for promoting plant growth, comprising applying the above-mentioned microbial agent or the above-mentioned pesticide on the plants.

[0113] In some specific embodiments of the present application, the plant is selected from one or more of corn, rice, sorghum, barley, cotton, rapeseed, and legumes, preferably corn.

[0114] In some specific embodiments of the present application, the application method is root irrigation and / or spraying.

[0115] The present application provides a method for controlling fall armyworm, comprising applying the above-mentioned microbial agent or the above-mentioned pesticide on plants.

[0116] In some specific embodiments of the present application, the plant is selected from one or more of corn, rice, sorghum, barley, cotton, rapeseed, and legumes, preferably corn.

[0117] In some specific embodiments of the present application, the application method is root irrigation and / or spraying.

[0118] The present application provides a method for promoting cannibalism among fall armyworm larvae, comprising applying the above-mentioned microbial agent or the above-mentioned pesticide on plants.

[0119] In some specific embodiments of the present application, the plant is selected from one or more of corn, rice, sorghum, barley, cotton, rapeseed, and legumes, preferably corn.

[0120] In some specific embodiments of the present application, the application method is root irrigation and / or spraying.

[0121] Example

[0122] Example 1 Screening and Identification Process of Serratia marcescens AHPC29

[0123] In a clean bench, rinse the larvae from which intestinal bacteria are to be removed in 75% alcohol and sterile water for about 3 seconds respectively; use sterile dissecting scissors to slowly cut open the larval body wall, remove the intestine, and place it in a sterile centrifuge tube; add 1 mL of sterile water to grind; dilute the grinding solution by 10 6 Then take 10 μL of the dilution and spread it on the plate, and place the plate in a 28°C incubator in the dark for 2 to 3 days.

[0124] When a single colony grows on the culture medium, a small amount of mycelium is picked with a sterile toothpick to continue culturing on a plate to obtain a purified strain. The obtained pure strain is classified, numbered and counted according to the colony size, color, thickness, transparency and texture on the same culture medium. The single colony color of the strain described in this application is red. After purification, a red single colony is picked, genomic DNA is extracted, and PCR amplification and sequencing are performed as a template. The sequencing results are compared with the model strains in the EzBioCloud database and the sequences in the GenBank database, and a phylogenetic tree is constructed to show that the isolated strain belongs to Serratia marcescens.

[0125] The strain obtained by screening was named Serratia marcescens AHPC29.

[0126] The 16S rDNA gene sequence of the strain obtained by screening is shown in SEQ ID NO:1.

[0127] The Serratia marcescens AHPC29 was deposited on March 22, 2022 at the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. The strain name is Serratia marcescens AHPC29 and the collection number is CGMCC24567.

[0128] Example 2

[0129] The optimal fermentation temperature and fermentation time of the Serratia marcescens strain isolated in Example 1 were determined.

[0130] The temperatures of 25°C, 28°C, 30°C, 35°C and 37°C were selected respectively to culture the Serratia marcescens strains for 48 hours, and then OD600 was measured. Figure 1 The results showed that the fermentation temperature directly affected the fermentation rate. 25℃, 35℃ and 37℃ all inhibited the growth of Serratia. The strongest inhibition was at 35℃, at which the OD600 value was only 0.228. The fermentation temperature of 30℃ was most conducive to the growth of Serratia, at which the OD600 value reached 0.479. Therefore, the optimal fermentation temperature was 30℃.

[0131] The composition and proportion of the culture medium are: 10 g / L trypsin, 5 g / L yeast extract, and 5 g / L sodium chloride.

[0132] The Serratia marcescens strain was cultured in LB medium at 30°C for 12, 24, 36, 48, 60, and 72 h, and the OD600 of the bacterial solution was measured to detect the effect of different fermentation times on bacterial growth. Figure 2 As shown in the figure, 12-24h belongs to the growth adjustment period of Serratia marcescens, and the growth is slow; 24-48h is in the logarithmic growth period, the growth rate is fast, and the growth starts to slow down after 48h. Therefore, in the actual production process, in order to save fermentation time, the optimal fermentation time is selected as 36h and 48h.

[0133] Example 3 Screening and identification process of Enterococcus sp. 10A

[0134] In a clean bench, rinse the larvae from which intestinal bacteria are to be removed in 75% alcohol and sterile water for about 3 seconds respectively; use sterile dissecting scissors to slowly cut open the larval body wall, remove the intestine, and place it in a sterile centrifuge tube; add 1 mL of sterile water to grind; dilute the grinding solution by 10 6 Then take 10 μL of the dilution and spread it on the plate, and place the plate in a 28°C incubator in the dark for 2 to 3 days.

[0135] When a single colony grows on the culture medium, a small amount of mycelium is picked up with a sterile toothpick and continued to be cultured on a plate to obtain a purified strain. The single colony of the strain described in this application is a slightly yellow, moist, opaque, round colony. After purification, a single colony is picked, and genomic DNA is extracted and used as a template for PCR amplification and sequencing. The sequencing results are compared with the model strains in the EzBioCloud database and the sequences in the GenBank database, and a phylogenetic tree is constructed to show that the isolated strain belongs to Enterococcus sp.

[0136] The strain obtained by screening was named 10A.

[0137] The 16S rDNA gene sequence of the strain obtained by screening is shown in SEQ ID NO:2.

[0138] The Enterococcus sp. 10A was deposited on March 22, 2022 at the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. The strain name is Enterococcus sp. 10A and the collection number is CGMCC24566.

[0139] Example 4 Morganella morganii CDHu screening and identification process

[0140] In a clean bench, rinse the larvae from which intestinal bacteria are to be removed in 75% alcohol and sterile water for about 3 seconds respectively; use sterile dissecting scissors to slowly cut open the larval body wall, remove the intestine, and place it in a sterile centrifuge tube; add 1 mL of sterile water to grind; dilute the grinding solution by 10 6 Then take 10 μL of the dilution and spread it on the plate, and place the plate in a 28°C incubator in the dark for 2 to 3 days.

[0141] When a single colony grows on the culture medium, a small amount of mycelium is picked up with a sterile toothpick and continued to be cultured on a plate to obtain a purified strain. The single colony of the strain described in this application is a moist, raised, non-spreading, grayish-white colony. After purification, a single colony is picked, genomic DNA is extracted, and PCR amplification and sequencing are performed as a template. The sequencing results are compared with the model strains in the EzBioCloud database and the sequences in the GenBank database, and a phylogenetic tree is constructed to show that the isolated strain belongs to Morganella morganii.

[0142] The strain obtained by screening was named CDHu.

[0143] The 16S rDNA gene sequence of the strain obtained by screening is shown in SEQ ID NO:3.

[0144] The Morganella morganii CDHu strain was deposited on March 25, 2022 at the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. The strain name is Morganella morganii CDHu and the collection number is CGMCC24593.

[0145] Example 5

[0146] The Serratia marcescens isolated in Example 1 (referred to as Serratia marcescens treatment) was used to study its growth-promoting effect on corn seedlings, and the isolated Pseudochrobactrum sp. (referred to as Pseudochrobactrum treatment) was used as a comparative strain, and the control group 1 was sprayed with distilled water or root irrigated on the corn seedlings. Pseudochrobactrum sp. was purchased commercially.

[0147] The Serratia marcescens isolated in Example 1 and the control strain Pseudochallotrichia coli isolated in Example 1 were cultured in LB medium at 28° C. for 48 h, and then 20 mL of the bacterial solution was added to 200 mL of distilled water for spraying or root irrigation on newly germinated corn seedlings. Each newly germinated corn seedling was sprayed with 10 mL, and the process was repeated 10 times. The plant height, leaf number, and leaf width of the corn seedlings were counted every other week for three consecutive weeks. Figure 3 The effect of spraying or root dredging with Serratia marcescens on the plant height of corn seedlings 1-3 weeks later is shown. Figure 4 The effect of spraying or root dredging with Serratia marcescens on leaf number in maize seedlings is shown. Figure 5 The effect of spraying or root dredging with Serratia marcescens on leaf width of maize seedlings is shown.

[0148] Figure 3-5 In the table, the treatment with Serratia marcescens spraying was recorded as Serratia marcescens spraying, the treatment with Serratia marcescens root irrigation was recorded as Serratia marcescens root irrigation, the treatment with Pseudomonas aeruginosa spraying was recorded as Pseudomonas aeruginosa spraying, the treatment with Pseudomonas aeruginosa root irrigation was recorded as Pseudomonas aeruginosa root irrigation, and the control group 1 was sprayed and root irrigated with distilled water.

[0149] Depend on Figure 3 It can be seen that the height of corn seedlings after spraying or root irrigation with Serratia marcescens for 1-3 weeks was higher than that of the control group 1 and the group treated with Pseudomonas pseudopallidum. Figure 4 It can be seen that the number of leaves of corn seedlings sprayed or root-irrigated with Serratia marcescens was significantly higher than that of the control group 1 and the group treated with Pseudomonas pseudopallidum; Figure 5 It can be seen that the leaf width of corn seedlings root-irrigated with Serratia marcescens was significantly higher than that of the control group 1 and the Pseudomonas pseudopalmatis treatment group.

[0150] In conclusion, Serratia marcescens strains have the potential to be developed into microbial preparations for crop fertilization and can be used as plant growth promoters.

[0151] Example 6

[0152] The Serratia marcescens isolated in Example 1 (denoted as Serratia marcescens treatment) was used for control and efficacy against fall armyworm, and Pseudochrobactrum sp. (denoted as Pseudochrobactrum treatment) was used for comparison. Control group 2 was spraying or root irrigation of corn seedlings with distilled water.

[0153] The Serratia marcescens and Pseudocalcinoma isolated in Example 1 were cultured in LB medium at 28°C for 48 h, and then 20 mL of the bacterial solution was added to 200 mL of distilled water for spraying or root irrigation on the newly germinated corn seedlings. Each newly germinated corn seedling was sprayed with 10 mL, and the process was repeated 10 times. Three weeks later, 5 fall armyworms were inoculated, and the degree of damage to the corn seedlings was observed, including the number of leaves eaten by the fall armyworm, the leaf size, and the mortality rate of the corn seedlings. Figure 6 The number of leaves that indicate maize leaves have been eaten as a proportion of the total number of leaves is shown; Figure 7 The figure shows the proportion of the leaf area of ​​maize leaves that are eaten to the total leaf area; Figure 8 Shown is the mortality of corn plants inoculated with Serratia marcescens after being inoculated with Spodoptera frugiperda.

[0154] Figure 6-8 In the experiment, the treatment with Serratia marcescens spraying was recorded as Serratia marcescens spraying, the treatment with Serratia marcescens root irrigation was recorded as Serratia marcescens root irrigation, the treatment with Pseudomonas aeruginosa spraying was recorded as Pseudomonas aeruginosa spraying, the treatment with Pseudomonas aeruginosa root irrigation was recorded as Pseudomonas aeruginosa root irrigation, and the control group 2 was sprayed and root irrigated with distilled water.

[0155] Depend on Figure 6 It can be seen that the proportion of the number of leaves eaten by Serratia marcescens in the corn leaves inoculated with the Pseudomonas pseudovitilis treatment group and the control group was 2% lower; Figure 7 It can be seen that the proportion of the leaf area eaten by Serratia marcescens to the total leaf area is less than that of the Pseudomonas pseudopallidum treatment group and the control group. Figure 8 It can be seen that the mortality rate of corn plants inoculated with Serratia marcescens after being inoculated with Spodoptera frugiperda was significantly lower than that of the Pseudomonas pseudopallidum treatment group and the control group 2.

[0156] In summary, the damage caused by fall armyworm to corn plants inoculated with Serratia marcescens was significantly reduced, and Serratia marcescens strains have the potential to be developed into microbial pesticides for controlling fall armyworm.

[0157] It can be seen from Example 5 and Example 6 that the growth of corn seedlings and the damage after being inoculated with fall armyworm were compared under two different application measures of root irrigation and spraying with bacterial solution on newly germinated corn seedlings. The results showed that the corn plant height, leaf number and leaf width of the Serratia marcescens root irrigation treatment group were greater than those of the Serratia marcescens spraying group, and the reduction of fall armyworm damage was better than that of the spraying group. Therefore, the root irrigation application method is preferred for Serratia marcescens in the application of promoting crop growth and preventing fall armyworm.

[0158] Example 7

[0159] After the Serratia marcescens strain was cultured in LB medium at 30° C. for 48 h, distilled water was added at a ratio of 1:10 (the volume ratio of the Serratia marcescens bacterial solution to water was 1:10), and then an aqueous solution of methyl methacrylate and methacrylic acid copolymer nanoparticles (referred to as the aqueous solution of auxiliary agent 1) or an aqueous solution of nanogold particles (referred to as the aqueous solution of auxiliary agent 2) was added to prepare a pesticide. The final concentration of auxiliary agent 1 or auxiliary agent 2 was 1% by mass percentage of the total weight of the pesticide. Only distilled water was added to the control group 3.

[0160] Fig. 9 In the test, a mixture containing only Serratia marcescens bacterial solution and water was recorded as Serratia marcescens + water, a mixture containing Serratia marcescens bacterial solution, water, and auxiliary agent 1 was recorded as Serratia marcescens + auxiliary agent 1, a mixture containing Serratia marcescens bacterial solution, water, and auxiliary agent 2 was recorded as Serratia marcescens + auxiliary agent 2, and a control group 3 contained only distilled water.

[0161] Irrigate the roots of each newly germinated corn seedling with 10 mL of water, repeat 5 times, and compare the corn plant heights after 20 days. Fig. 9 The impact of adjuvants on Serratia marcescens in crop growth promotion applications is shown.

[0162] Depend on Fig. 9 It can be seen that the best adjuvant for Serratia in the application of promoting crop growth is methyl methacrylate and methacrylic acid copolymer nanoparticles (i.e. adjuvant 1).

[0163] Example 8

[0164] After the Serratia marcescens strain was cultured in LB medium at 30°C for 48h, distilled water (referred to as Serratia marcescens + water) was added directly at a ratio of 1:10 (according to the volume ratio of Serratia marcescens to water of 1:10), and aqueous solutions of methyl methacrylate and methacrylic acid copolymer nanoparticles (i.e., aqueous solutions of adjuvant 1) of different concentrations were added to prepare pesticides, wherein the final concentrations of methyl methacrylate and methacrylic acid copolymer nanoparticles were 10% (referred to as Serratia marcescens + adjuvant 1 10%), 1% (referred to as Serratia marcescens + adjuvant 1 1%) and 0.1% (referred to as Serratia marcescens + adjuvant 1 0.1%), respectively, and distilled water was added to 4 of the control group. 10mL was irrigated to the roots of each newly germinated corn seedling, repeated 5 times, and the height of the corn plant was statistically compared after 20 days. Fig.10 The effects of different methyl methacrylate and methacrylic acid copolymer nanoparticle concentrations on promoting crop growth are shown.

[0165] Fig.10 In the test, a mixture containing only Serratia marcescens liquid and water was recorded as Serratia marcescens + water, a mixture containing Serratia marcescens liquid, water, and adjuvant 1 (the final concentration of adjuvant 1 was 10% by mass percentage of the total weight of the pesticide) was recorded as Serratia marcescens + adjuvant 1 10%, a mixture containing Serratia marcescens liquid, water, and adjuvant 1 (the final concentration of adjuvant 1 was 1% by mass percentage of the total weight of the pesticide) was recorded as Serratia marcescens + adjuvant 1 1%, a mixture containing Serratia marcescens liquid, water, and adjuvant 1 (the final concentration of adjuvant 1 was 0.1% by mass percentage of the total weight of the pesticide) was recorded as Serratia marcescens + adjuvant 1 0.1%, and the control group 4 contained only distilled water.

[0166] Depend on Fig.10 It can be seen that when the concentration of methyl methacrylate and methacrylic acid copolymer nanoparticles is 10%, 1% and 0.1% by mass percentage of the total weight of the pesticide, they can all play a role in promoting crop growth. Calculated by mass percentage of the total weight of the pesticide, the best methyl methacrylate and methacrylic acid copolymer nanoparticles is 1%.

[0167] Example 9

[0168] Serratia marcescens isolated in Example 1, Enterococcus sp. isolated in Example 3, and Morganella morganii isolated in Example 4 were taken to study their control potential against fall armyworm, that is, their direct insecticidal effect against fall armyworm.

[0169] After three bacteria, Serratia marcescens, Enterococcus sp. and Morganella morganii, were cultured in LB medium at 30°C for several days, they were respectively mixed with aqueous solution of methyl methacrylate and methacrylic acid copolymer nanoparticles (adjuvant 1) (the concentration of methyl methacrylate and methacrylic acid copolymer nanoparticles was 1 mg / mL) to form a 99:1 bacterial solution, and then prepared into a pesticide. The final concentration of adjuvant 1 was 1% by mass percentage of the total weight of the pesticide. The larvae of fall armyworm were sprayed, and 100 μL was sprayed on each fall armyworm. Three groups of experiments were repeated for each bacterial species, and each group had 6 fall armyworm larvae. The control group 5 was sprayed with water and an aqueous solution of methyl methacrylate and methacrylic acid copolymer nanoparticles in a mass ratio of 99:1. The mortality rate of fall armyworm was counted 12 hours after the spraying treatment. Fig.11 Shown is the mortality of S. frugiperda larvae after spraying with Serratia marcescens, Enterococcus sp. and Morganella morganii.

[0170] Fig.11 In the figure, those treated with spraying of Serratia marcescens and adjuvant 1 were recorded as Serratia marcescens, and the corresponding control group represented those treated with spraying of water and adjuvant 1; those treated with spraying of Enterococcus and adjuvant 1 were recorded as Enterococcus, and the corresponding control group represented those treated with spraying of water and adjuvant 1; those treated with spraying of Morganella and adjuvant 1 were recorded as Morganella, and the corresponding control group 5 represented those treated with spraying of water and adjuvant 1.

[0171] Depend on Fig.11 It can be seen that the mortality rate of Spodoptera frugiperda larvae after spraying with Serratia marcescens, Enterococcus and Morganella was significantly higher than that in the control group5.

[0172] Example 10

[0173] The three bacteria isolated in Example 1, Enterococcus sp. isolated in Example 3, and Morganella morganii isolated in Example 4 were cultured in LB medium at 30°C for several days and the weight parts were calculated based on the same OD 600The volume ratio is calculated based on the premise of , and the following three groups of experiments are set, wherein the first group of Serratia marcescens is 1 part by weight, Enterococcus is 1 part by weight, and Morganella is 1 part by weight, the second group of Serratia marcescens is 1 part by weight, Enterococcus is 1 part by weight, and Morganella is 10 parts by weight, and the third group of Serratia marcescens is 10 parts by weight, Enterococcus is 1 part by weight, and Morganella is 10 parts by weight, and then the mixed bacterial liquid of the aqueous solution of methyl methacrylate and methacrylic acid copolymer nanoparticles (the aqueous solution of adjuvant 1, wherein the final concentration of adjuvant 1 is 1% in terms of the mass percentage of the total weight of the pesticide) is respectively prepared into pesticides, sprayed on the larvae of fall armyworms and the mortality rate of fall armyworms is calculated after 12 hours. Three experiments were repeated for each ratio, and each experimental repetition had 6 fall armyworm larvae, each fall armyworm was sprayed with 100μL, and the control group 6 was sprayed with water and methyl methacrylate and methacrylic acid copolymer nanoparticles in a ratio of 99:1. Fig.12 Shown is the mortality rate of Spodoptera frugiperda larvae after spraying with microbial agents.

[0174] Fig.12 The first group of pesticides was recorded as 1:1:1, the second group of pesticides was recorded as 1:1:10, the third group of pesticides was recorded as 10:1:10, and the control group 6 represented spraying of an aqueous solution of the same adjuvant 1 concentration.

[0175] Depend on Fig.12 It can be seen that the mortality rate of fall armyworm larvae after spraying with the microbial agent, i.e. a mixture of three bacteria, Serratia marcescens, Enterococcus and Morganella, was significantly higher than that of the control group 6, and the insecticidal effect of the three bacteria, Serratia marcescens, Enterococcus and Morganella, was best at a ratio of 10:1:10.

[0176] Embodiment 11

[0177] The three bacteria, namely, the Serratia marcescens isolated in Example 1, the Enterococcus isolated in Example 3, and the Morganella isolated in Example 4, were cultured in LB medium at 30°C for several days, and then an aqueous solution of methyl methacrylate and methacrylic acid copolymer nanoparticles (the aqueous solution of adjuvant 1, calculated as a percentage by mass of the total weight of the pesticide, is 1%) were added respectively, and the pesticide was sprayed on the larvae of the fall armyworm; calculated in parts by weight, the first group of Serratia marcescens was 10 parts by weight, the Enterococcus was 1 part by weight, and the Morganella was 10 parts by weight, and then the mixture was added with an aqueous solution of methyl methacrylate and methacrylic acid copolymer nanoparticles (the aqueous solution of adjuvant 1, calculated as a percentage by mass of the total weight of the pesticide, is 1%), and 100uL was sprayed on each fall armyworm; the second group of Serratia marcescens indicated that at the same OD 600The pesticide prepared by adding methyl methacrylate and methacrylic acid copolymer nanoparticle aqueous solution (the aqueous solution of adjuvant 1, as a percentage of the total weight of the pesticide, adjuvant 1 is 1%) to Serratia marcescens under the premise of OD 100 μL was sprayed on each fall armyworm; the third group was Enterococcus, which was expressed at the same OD 600 The pesticide prepared by adding an aqueous solution of methyl methacrylate and methacrylic acid copolymer nanoparticles (the aqueous solution of adjuvant 1, calculated by mass percentage of the total weight of the pesticide, adjuvant 1 is 1%) to the enterococci under the premise of OD 100 μL was sprayed on each fall armyworm; the fourth group was Morganella, which means that at the same OD 600 Morganella under the premise of adding methyl methacrylate and methacrylic acid copolymer nanoparticles aqueous solution (the aqueous solution of adjuvant 1, in terms of mass percentage of the total weight of the pesticide, adjuvant 1 is 1%) to prepare the pesticide, and spray 100μL on each fall armyworm; the control group 7 was sprayed with water and methyl methacrylate and methacrylic acid copolymer nanoparticles in a ratio of 99:1. Each treatment was repeated 3 times, and each experimental repetition was repeated 6 fall armyworm larvae, and each fall armyworm was sprayed with 100uL. After 6 hours of treatment, the probability of fall armyworm cannibalism was counted. Fig.13 It shows the cannibalism rate of fall armyworm larvae after spraying with microbial agents.

[0178] Fig.13 In the formula, 10:1:10 means that 10 parts by weight of Serratia marcescens, 1 part by weight of Enterococcus, and 10 parts by weight of Morganella are mixed and then an aqueous solution of nanoparticles of methyl methacrylate and methacrylic acid copolymer (aqueous solution of adjuvant 1) is added to prepare the pesticide;

[0179] Serratia marcescens expressed at equivalent OD 600 A pesticide prepared by adding an aqueous solution of nanoparticles of methyl methacrylate and methacrylic acid copolymer (aqueous solution of adjuvant 1) to Serratia marcescens under the premise of

[0180] Enterococci are expressed at equal OD 600 The pesticide prepared by adding an aqueous solution of nanoparticles of methyl methacrylate and methacrylic acid copolymer (aqueous solution of adjuvant 1) to enterococci under the premise of

[0181] Morganella morganii is expressed at the same OD 600 The Morganella morganii under the premise of adding methyl methacrylate and methacrylic acid copolymer nanoparticles aqueous solution (aqueous solution of adjuvant 1) to prepare the pesticide;

[0182] Control group 7 represents an aqueous solution prepared with a 1% aqueous solution of nanoparticles of a copolymer of methyl methacrylate and methacrylic acid (aqueous solution of auxiliary agent 1).

[0183] Depend on Fig.13 It can be seen that the probability of cannibalism among the larvae of the fall armyworm increases after the three bacteria, Serratia marcescens, Enterococcus and Morganella, are mixed and sprayed.

[0184] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application. Sequence Listing <110> Institute of Zoology, Chinese Academy of Sciences <120> Serratia marcescens, microbial agent, pesticide and application thereof <130> PF02093 <160> 5 <170> PatentIn version 3.5 <210> 1 <211> 1442 <212> DNA <213> Artificial sequence <220> <223> Artificial sequence description: artificially synthesized sequence <400> 1 ttcacaaagt ggtaagcgcc ctcccgaagg ttaagctacc tacttctttt gcaacccact 60 cccatggtgt gacgggcggt gtgtacaagg cccgggaacg tattcaccgt agcattctga 120 tctacgatta ctagcgattc cgacttcatg gagtcgagtt gcagactcca atccggacta 180 cgacgtactt tatgaggtcc gcttgctctc gcgaggtcgc ttctctttgt atacgccatt 240 gtagcacgtg tgtagcccta ctcgtaaggg ccatgatgac ttgacgtcat ccccaccttc 300 ctccagttta tcactggcag tctcctttga gttcccggcc gaaccgctgg caacaaagga 360 taagggttgc gctcgttgcg ggacttaacc caacatttca caacacgagc tgacgacagc 420 catgcagcac ctgtctcaga gttcccgaag gcaccaatcc atctctggaa agttctctgg 480 atgtcaagag taggtaaggt tcttcgcgtt gcatcgaatt aaaccacatg ctccaccgct 540 tgtgcgggcc cccgtcaatt catttgagtt ttaaccttgc ggccgtactc cccaggcggt 600 cgatttaacg cgttagctcc ggaagccacg cctcaagggc acaacctcca aatcgacatc 660 gtttacagcg tggactacca gggtatctaa tcctgtttgc tccccacgct ttcgcacctg 720 agcgtcagtc ttcgtccagg gggccgcctt cgccaccggt attcctccag atctctacgc 780 atttcaccgc tacacctgga attctacccc cctctacgag actctagctt gccagtttca 840 aatgcagttc ccaggttgag cccggggatt tcacatctga cttaacaaac cgcctgcgtg 900 cgctttacgc ccagtaattc cgattaacgc ttgcaccctc cgtattaccg cggctgctgg 960 cacggagtta gccggtgctt cttctgcgag taacgtcaat tgatgaacgt attaagttca 1020 ccaccttcct cctcgctgaa agtgctttac aacccgaagg ccttcttcac acacgcggca 1080 tggctgcatc aggcttgcgc ccattgtgca atattcccca ctgctgcctc ccgtaggagt 1140 ctggaccgtg tctcagttcc agtgtggctg gtcatcctct cagaccagct agggatcgtc gcctaggtga gccattaccc cacctactag ctaatcccat ctgggcacat ctgatggcaa gaggcccgaa ggtccccctc tttggtcttg cgacgttatg cggtattagc taccgtttcc 1320 agtagttatc cccctccatc aggcagtttc ccagacatta ctcacccgtc cgccgctcgt 1380. cacccaggga gcaagctccc ccgtgctacc gctcgacttg catgtgttaa aggccccccc cg 1442 <210> 2 <211> 1475 <212> DNA <213> The snowstorm <220> <223> A snowstorm: a snowstorm <400> 2 aaaaaatcca gtgcgggtgc tatacatgc agtcgaacgc ttctttcctg ccgagtgctt gcactcactt ggaaagagga gtggcggacg ggtgagtaac acgtgggtaa cctgcccatc agggggat aacacttgga aacaggtgct aataccgcat aatagtcgac accgcatggt gttgatttga aagacgcttt cgggtgtcac tgatggatgg acccgcggtg cattagctag 240 ttggtgaggt aacggctcac caaggccatg atgcatagcc gacctgagag ggtgatcggc 300 caacactggga ctgagacacg gcccagactc ctacgggagg cagcagtagg gaatcttcgg 360 420 480 accagaaagc cacggctaac tacgtgccag cagccgcggt aatacgtagg tggcaagcgt 540 tgtccggatt tattgggcgt aaagcgagcg caggcggttt cttaagtctg atgtgaaagc 600 ccccggctca accggggagg gtcattggaa actgggagac ttgagtgcag aagaggag 660 tggaattcca tgtgtagcgg tgaaatgcgt agatatatgg aggaacacca gtggcgaagg 720 cgactctctg gtctgtaact gacgctgagg ctcgaaagcg tggggagagcaa acaggattag 780 ataccctggt agtccacgcc gtaaacgatg agtgctaagt gttggagggt ttccgccctt 840 cagtgctgca gcaaacgcat taagcactcc gcctggggag tacgaccgca aggttgaaac 900 tcaaaggaat tgacggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac 960 gcgaagaacc ttaccaggtc ttgacatcct ttgaccactc tagagataga gctttccctt 1020 cggggacaaa gtgacaggtg gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt 1080 taagtcccgc aacgagcgca acccttattg ttagttgcca tcattcagtt gggcactcta 1140 gcgagactgc cggtgacaaa ccggaggaag gtggggatga cgtcaaatca tcatgcccct 1200 tatgacctgg gctacacacg tgctacaatg ggaagtacaa cgagtcgcta ggccgcgagg 1260 tcatgcaaat ctcttaaagc ttctctcagt tcggattgta ggctgcaact cgcctacatg 1320 aagccggaat cgctagtaat cgcggatcag cacgccgcgg tgaatacgtt cccgggcctt 1380 gtacacaccg cccgtcacac cacgaaagtt tgtaacaccc gaagtcggtg aggtaacctt 1440 ttggagccag ccgcctaagg tggaagaaaa ttctc 1475 <210> 3 <211> 1457 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic sequence <400> 3 accgcttttt atcacaaagt ggtaagcgcc ctcccggagg ttaagctacc tacttctttt 60 gcaacccact cccatggtgt gacgggcggt gtgtacaagg cccgggaacg tattcaccgt 120 agcattctga tctacgatta ctagcgattc cgacttcatg gagtcgagtt gcagactcca 180 atccggacta cgacgtactt tatgagttcc gcttgccctc gcggggtcgc ttccctttgt 240 atacgccatt gtagcacgtg tgtagcccta ctcgtaaggg ccatgatgac ttgacgtcat 300 ccccaccttc ctccggttta tcaccggcag tctcctttga gttcccgcca tcacgcgctg 360 gcaacaaagg ataagggttg cgctcgttgc gggacttaac ccaacatttc acaacacgag 420 ctgacgacag ccatgcagca cctgtctcag agttcccgaa ggcaccaaag catctctgct 480 aagttctctg gatgtcaaga gtaggtaagg ttcttcgcgg tgcatcgaat taaaccacaa 540 gctccaccgc ttgtgcgggc ccccgtcaat tcatttgagt tttaaccttg cggccgtact 600 ccccaggcgg tcgacttaac gcgttagctc ccggaagcca cgcctcaagg gcacaacctc 660 caagtcgaca tcgtttacag cgtggactac cagggtatct aatcctgttt gctccccacg 720 ctttcgcacc tgagcgtcag tctttgtcca gggggccgcc ttcgccaccg gtattcctcc 780 acatctctac gcatttcacc gctacacatg gaattctacc cccctctaca agactctagc 840 tgaccagtat cagatgcaat tcccgggtta agcccgggga tttcacatct gactcaatca 900 accgcctgcg tgcgctttac gcccagtaat tcccgattaa cgcttgcaac ctccgtatta 960 ccgcggctgc tggcacggag ttagccggtg cttcttctgt cggtaacgtc aattgccaag 1020 gttattaacc ttgacacctt cctcccgact gaaagtactt tacaacccga aggccttctt 1080 catacacgcg gcatggctgc atcaggcttg cgcccattgt gcaatattcc cactgctgc 1140 ctcccgtagg agtctgggcc gtgtctcagt cccagtgtgg ctgatcatcc tctcagacca 1200 gctagggatc gtcgcctagg taagccgtta cctcacctac tagctaatcc catatgggtt 1260 catctgatgg cgcgaggccc ggaggtcccc cgctttggtc cgaagacatt atgcggtatt 1320 agctaccgtt tccagtagtt atcccccgcc atcaggcaga tccccataca ttactcaccc 1380 gtccgccgct cgtcagcaga gaagcaagct tctccctgtt accgcccgac ttgcatgtgt 1440 tagctgccgc aagttcc 1457 <210> 4 <211> 39 <212> DNA <213> Artificial sequence <220> <223> Artificial sequence description: artificially synthesized sequence <400> 4 gcggatccgc ggccgctgca gagtttgatc ctggctcag 39 <210> 5 <211> 37 <212> DNA <213> Artificial sequence <220> <223> Artificial sequence description: artificially synthesized sequence <400> 5 ggctcgagcg gccgcccggg ttaccttgtt acgactt 37

Claims

1. A Serratia marcescens, whose deposit number is CGMCC No.24567.

2. A microbial agent, comprising Serratia marcescens, wherein the deposit number of Serratia marcescens is CGMCC No. 24567.

3. A pesticide or a plant growth promoter, comprising the microbial agent according to claim 2.

4. The pesticide or plant growth promoter according to claim 3, wherein: The pesticide or plant growth promoter also includes adjuvants.

5. The pesticide or plant growth promoter according to claim 4, wherein: The auxiliary agent is selected from one or more of methyl methacrylate and methacrylic acid copolymer nanoparticles, nanogold particles, urea, wettable powder cuprous oxide, ferrous sulfate, sodium alkyl sulfonate, and soapberry extract.

6. The pesticide or plant growth promoter according to claim 5, wherein: The auxiliary agent is nanoparticles of a copolymer of methyl methacrylate and methacrylic acid.

7. The pesticide or plant growth promoter according to claim 6, wherein: Calculated by weight percentage of the total weight of the pesticide or plant growth promoter, the adjuvant is 0.1-10wt%.

8. A method for promoting plant growth or controlling fall armyworm, characterized in that: The method comprises applying the microbial agent according to claim 2 or the pesticide or plant growth promoter according to any one of claims 3 to 7 on a plant, wherein the plant is corn.

9. A microbial agent, comprising Serratia marcescens, Enterococcus and Morganella; The deposit number of the Serratia marcescens is CGMCC No.24567; The deposit number of the enterococcus is CGMCC No.24566, and the deposit number of the Morganella is CGMCC No.24593.

10. The microbial agent according to claim 9, wherein The bacterial agent consists of Serratia marcescens, Enterococcus and Morganella.

11. The microbial agent according to claim 9, wherein In parts by weight, the mass ratio of the Serratia marcescens, the Enterococcus and the Morganella is 10:1:

10.

12. A pesticide or a plant growth promoter, comprising the microbial agent according to any one of claims 9 to 11.

13. The pesticide or plant growth promoter according to claim 12, wherein: The pesticide or plant growth promoter also includes adjuvants.

14. The pesticide or plant growth promoter according to claim 13, wherein: The auxiliary agent is selected from one or more of methyl methacrylate and methacrylic acid copolymer nanoparticles, nanogold particles, urea, wettable powder cuprous oxide, ferrous sulfate, sodium alkyl sulfonate, and soapberry extract.

15. The pesticide or plant growth promoter according to claim 14, wherein: The auxiliary agent is nanoparticles of a copolymer of methyl methacrylate and methacrylic acid.

16. The pesticide or plant growth promoter according to claim 13, wherein: Calculated by weight percentage of the total weight of the pesticide or plant growth promoter, the adjuvant is 0.1-10wt%.

17. A method for promoting plant growth or controlling fall armyworm or promoting cannibalism of fall armyworm larvae, characterized in that: The method comprises applying the microbial agent according to any one of claims 9 to 11 or the pesticide or plant growth promoter according to any one of claims 12 to 16 on a plant, wherein the plant is corn.

18. The method according to claim 17, wherein: The application method is root dredging and / or spraying.

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