Bacillus velezensis LM3 and compsition for controlling plant desease or promoting plant growth compring them

Abstract

The present invention relates to a novel Bacillus velezensis LM3 strain (accession number: KACC 92538P) that induces resistance to plant bacterial and viral diseases and promotes plant growth, a composition for promoting plant growth or preventing plant diseases comprising said strain, a culture solution or a culture solution extract thereof as an active ingredient, a method for culturing said strain, and a method for promoting plant growth or preventing plant diseases using said composition.

Description

Technology Field

[0001] The present invention relates to a novel Bacillus velezensis strain that induces resistance to plant bacterial and viral diseases and promotes plant growth, and a composition for promoting plant growth or preventing plant diseases comprising said strain. Background Technology

[0002] Plant diseases inhibit normal plant growth, causing parts of the plant to necrose or, in severe cases, to wither and die. It is known that approximately 10 to 20 percent of the plants distributed on Earth experience reduced yields due to disease. This decrease in yield is not limited to merely affecting farmers' income. Potato blight, which occurred in Ireland between 1845 and 1846, caused a Great Famine that killed 1.5 million Irish people, leading more than 1 million to leave Ireland and emigrate to the New World. Coffee rust, which occurred in the Ceylon region of Sri Lanka in the late 19th century, shifted the primary cultivated plant from coffee to tea, giving birth to "Ceylon Tea" and enabling Sri Lanka to become the world's third-largest tea exporter as of 2019. In Korea as well, there is a case from 1978 where rice blast disease occurred in Tongilbyeo, a variety known for its high productivity that spearheaded the Green Revolution in solving absolute poverty, resulting in a severe decline in production.

[0003] The causes of plant diseases are broadly divided into abiotic and biotic factors. Abiotic causes include deficiencies or excesses of temperature, humidity, light, and nutrients, as well as environments unsuitable for plant growth, such as air pollution. Plant diseases caused by abiotic factors can be prevented by eliminating inappropriate environmental factors or improving the crop's stress resistance.

[0004] Plant diseases caused by biological factors are triggered by fungi, bacteria, viruses, parasitic plants, and pests. Globally, approximately 1,100 species of viruses that cause plant diseases have been isolated and reported, with about 120 species reported in Korea alone. Furthermore, as the volume of international trade in agricultural crops increases, damage caused by bacteria, viruses, and pests introduced from abroad—in addition to native species—is on the rise. To control plant diseases caused by biological factors, the use of pesticides utilizing synthetic organic compounds is the most commonly employed method because it is effective in protecting crops within a short period. It is reported that over 300 tons of antibiotic pesticides are currently produced annually in Korea to control bacterial plant diseases caused by pathogenic bacteria. The use of antibiotics on crops can have harmful effects on the environment, such as soil and water quality, and disrupt ecosystems; additionally, safety issues arise if antibiotic residues remain in crops. Moreover, the types of antibiotics applied to crops are limited, and control rates are declining due to the emergence of resistant pathogens; consequently, the use of antibiotic pesticides is on an upward trend every year. Six types of antibiotics—streptomycin, oxytetracycline, oxolinic acid, kasugamycin, validamycin, and polyoxin—are registered and used for agricultural purposes in Korea; however, these antibiotics can form novel substances with significantly higher toxicity during the degradation process in the soil. Therefore, there is a need for alternatives to delay the chemical pesticideization of these antibiotics and to reduce their usage. While plant viruses affect various economic crops, yields of chili peppers—one of Korea's important horticultural crops—are particularly declining due to viruses such as Cucumber Mosaic Virus (CMV), Tomato Mosaic Virus (TOMV), and Pepper Mild Mottling Virus (PMMOV). Since treatment for these viruses is nearly impossible after infection, physically removing the infected plants is the best method.

[0005] With concerns raised regarding the side effects of pesticides, such as pesticide residues and environmental pollution, research is currently underway to develop biological control methods utilizing microorganisms to control plant viruses. For example, studies are being conducted on methods to control bacterial or viral plant diseases by utilizing strains of species such as Serratia and Bacillus, or their culture solutions. As with human diseases, the most desirable approach to plant diseases is to prevent infection itself by promoting plant growth and increasing resistance in advance. Prior art literature

[0006] Korean Registered Patent 10-1163986 Korean Registered Patent 10-2307028 The problem to be solved

[0007] The present invention aims to provide a novel strain that promotes plant germination and growth, exhibits antimicrobial activity against plant disease-causing fungi, and provides systemic resistance to plant diseases.

[0008] Another objective of the present invention is to provide a composition for promoting plant growth or preventing plant diseases by utilizing the above strain's properties of promoting plant germination and growth, inducing systemic resistance to plant diseases, and antifungal properties.

[0009] The present invention also has another objective of providing a method for promoting plant growth or preventing plant diseases using the above composition.

[0010] The technical problem that the present invention aims to solve will be clearly understood by those skilled in the art, even if not mentioned above. Furthermore, in describing the invention, if it is determined that a detailed description of known technology related to the invention may unnecessarily obscure the essence of the invention, such detailed description will be omitted. means of solving the problem

[0011] The present invention for achieving the aforementioned purpose (Bacillus velezensis) Bacillus velezensisThis invention relates to the LM3 strain (Deposit No.: KACC 92538P). The strain was selected from the intestines of third-instar rhinoceros beetle larvae and was confirmed to contain 16S rRNA represented by the nucleotide sequence of SEQ ID NO. 1. As a result of identification based on 16S rRNA, it was confirmed that there is no strain that completely matches the gene of the strain of the present invention, and conventional Bacillus velezensis ( Bacillus velezensis ) showed 99.92% similarity with the 16S rRNA of CR-502. Accordingly, the selected strain Bacillus velezensis It was named LM3 and deposited at the National Institute of Agricultural Sciences Agricultural Genetic Resources Center on July 19, 2023, and was assigned accession number KACC 92538P on August 30, 2023.

[0012] The above strain is a pathogen that causes gray mold, damping-off, bacterial wilt, and anthracnose. Botrytis cinere , Rizoctonia solani , Fusarium oxysporum , Colletotrichum coccodes It exhibited antifungal activity against, promoted germination and growth of plants, especially rice, and induced systemic resistance to plant diseases.

[0013] Accordingly, the present invention (Bacillus velezensis ( B. velezensis The present invention provides a composition for promoting plant growth or preventing plant diseases, comprising LM3, a culture solution thereof, or a culture solution extract thereof as an active ingredient. The plant disease is caused by a plant disease pathogen, and the plant disease pathogen refers to fungi that live a parasitic life on plants (parasitic fungi), fungi that live a biocidal life on plants (biocidal fungi), facultative parasitic fungi that parasitize specific plants, etc. The plant may be rice, chili pepper, or Chinese cabbage, but is not limited thereto.

[0014] When rice seeds were treated with the composition of the present invention, the germination rate was improved and the average germination period was shortened compared to the chemical insecticide-treated group or the untreated group; furthermore, the growth of rice was significantly promoted even compared to another microbial strain, Bacillus amyloliquefaciens. In addition, the plant disease pathogen Pectobacterium carotoborum ( Pectobacterium carotovorum ), Xanthomonas juvesicatoria( Xanthomonas euvesicatoria Upon treatment with ) or cucumber mosaic virus, the expression of resistance genes induced during the systemic resistance response was significantly enhanced, thereby demonstrating excellent plant disease prevention effects in actual fields. The strain of the present invention is a pathogen that causes gray mold, damping-off, bacterial wilt, and anthracnose. Botrytis cinere , Rizoctonia solani , Fusarium oxysporum , Colletotrichum coccodes Since it is selected from strains having antifungal activity against, it can naturally be usefully used to prevent plant diseases caused by the said pathogen.

[0015] In the present invention, "culture medium" means the entire medium containing the strain, its metabolites, excess nutrients, etc., obtained by culturing the strain for a certain period in a medium supplied with nutrients to enable Bacillus velezensis LM3 to grow, or the medium from which the strain has been removed after cultivation.

[0016] In the present invention, "culture extract" refers to a fraction containing a component that exhibits antimicrobial activity against plant disease pathogens in the culture medium of Bacillus velezensis LM3. As long as the extract can exhibit antimicrobial activity, it is not limited thereto and may include an extract, a diluted solution, a concentrate, a dried product, a modified product, or a purified product. The extract may be prepared from the supernatant or dried product of the culture medium by a conventional extraction method known in the art using an extraction solvent, such as hot water extraction, cold maceration extraction, reflux cooling extraction, or ultrasonic extraction. As the extraction solvent, not only C1 to C4 alcohols but also polar / non-polar organic solvents such as chloroform, acetonitrile, ethyl acetate, and hexane may be used.

[0017] When the culture medium of the strain of the present invention contains maltose as a carbon source, not only the growth of the strain but also the spore conversion rate was excellent. In the case of carbon sources such as dextrose or sucrose, although they were advantageous for the growth of the strain, the spore conversion rate was low, which reduced their suitability as a culture medium. Accordingly, the present invention provides a method for culturing the strain as another embodiment, characterized by including sucrose as a carbon source in the medium. It is more preferable that the medium includes soybean fiber as a nitrogen source. As a specific example, the composition of the medium may be maltose 15 g / ℓ, soybean fiber 5 g / ℓ, KH2PO4 2.5 g / ℓ, MgSO4·7H2O 0.5 g / ℓ, CaCl2 0.5 g / ℓ, FeSO4·4H2O 0.5 g / ℓ, and MnSO4·H2O 0.5 g / ℓ.

[0018] To use the composition for controlling plant diseases of the present invention as a microbial pesticide, it may be used either as is or mixed with pharmaceutically acceptable carriers, excipients, diluents, etc., by methods known in the field of pesticides. The composition of the present invention may be formulated into various known forms, for example, in the form of liquids, granules, powders, emulsions, oils, wettable powders, or coatings. Any formulation method commonly used in the field of pesticides may be used for formulation.

[0019] Another aspect of the present invention relates to a method for promoting plant growth or preventing plant diseases using the strain LM3, its culture medium, or a culture medium extract.

[0020] The composition described above may be applied to the surrounding environment where the plant is growing, such as soil or irrigation, or directly to the plant. When applied directly to the plant, it may be sprayed, immersed, or drenched in an amount effective for one or more of the following: promoting plant growth, antifungal activity, and inducing systemic resistance, on seeds before germination, or on the surface of leaves, fruits, etc., of growing plants, but is not limited thereto. For example, it may be used not only for the plant itself but also for disinfecting agricultural tools that come into direct contact with the plant. Regarding the timing of application, applying it prophylactically before the occurrence of bacterial plant diseases may yield a higher preventive effect; in particular, greater effects can be observed when treating with a high concentration during the seedling stage before transplanting, followed by transplanting, and then drenching after transplanting.

[0021] Furthermore, the composition of the present invention may be used alone or in combination with a substance known in the art that promotes plant growth and / or controls plant diseases; when used in combination with other substances, it may be used sequentially or simultaneously. The concentration of the active ingredient included in the composition of the present invention can be appropriately adjusted by a person skilled in the art by considering the type of crop, the type of plant disease, the degree of plant growth, the cultivation environment, the incidence of plant disease, and the density of pathogens in the field. For example, in the above control method, the LM3 strain is 1×10 6 cfu / ㎖~1×10 8 It can be treated at a concentration of cfu / ml, but for control, 1×10 8 Since treatment at high concentrations of cfu / mL or higher is more effective, it is not limited to the above range.

[0022] The present invention relates to a fertilizer additive composition comprising the strain LM3, its culture solution, or a culture solution extract as an active ingredient. Unlike chemical additives, the composition of the present invention does not exhibit harmful effects such as environmental pollution or toxicity to the human body, promotes plant germination and growth, and can enhance crop disease resistance by inducing systemic resistance, thereby contributing to increased yield. Effects of the invention

[0023] As described above, the novel Bacillus velezensis LM3 strain of the present invention promotes plant growth and exhibits antifungal activity, while inducing systemic resistance to enhance resistance to plant diseases. Therefore, it can be usefully used for preventing plant diseases and promoting growth without concerns about environmental pollution, as it can replace chemical pesticides or fertilizers. Brief explanation of the drawing

[0024] Figure 1 is a photograph showing the antifungal activity of the Bacillus velezensis LM3 strain. FIG. 2 is the 16S rDNA sequence of the Bacillus velezensis LM3 strain of the present invention. FIG. 3 is a phylogenetic tree of the Bacillus velezensis LM3 strain of the present invention. Figure 4 is an electrophoretic image showing the expression of a systemic induced resistance gene by the Bacillus velezensis LM3 strain of the present invention. Figure 5 is a photograph showing the plant disease prevention effect by the induced resistance of the Bacillus velezensis LM3 strain of the present invention in packaging. Figure 6 is a graph showing the plant disease prevention effect by the induced resistance of the Bacillus velezensis LM3 strain of the present invention in packaging. Specific details for implementing the invention

[0025] The present invention will be explained in more detail below with reference to the attached examples. However, these examples are merely illustrative to easily explain the content and scope of the technical concept of the present invention, and the technical scope of the present invention is not limited or altered by them. It will be obvious to those skilled in the art that various modifications and changes are possible within the scope of the technical concept of the present invention based on these examples.

[0026] [Example]

[0027] Example 1: Isolation and identification of strains exhibiting antifungal activity against plant diseases

[0028] To select strains from soil exhibiting antifungal activity against plant diseases, the intestines of third-instar rhinoceros beetle larvae were isolated, ground, and used as a microbial extract source. 100 ml of sterile distilled water was added to 1 g of the ground sample and shaken at 150 rpm for 6 hours at 25°C. The shaken sample was diluted using the serial dilution method and plated onto TSA solid media to obtain the strains. The obtained strains were respectively identified as pathogens causing gray mold, damping-off, bacterial wilt, and anthracnose. Botrytis cinere , Rizoctonia solani , Fusarium oxysporum , Colletotrichum coccodes Microorganisms exhibiting antifungal activity were isolated by culturing with. Figure 1 shows photographs of the results, where (A) to (D) are, respectively Botrytis cinere , Rizoctonia solani , Fusarium oxysporum and Colletotrichum coccodes This is a photo of the replacement culture result for.

[0029] The 16S rDNA sequence of the isolated strain (Sequence No. 1, Fig. 2) was compared with the strain registered through the 16S-based ID of EZ-biocloud to investigate the phylogenetic tree and genetic similarity. As a result of comparative analysis of genetic information, the above strain is Bacillus velezensis It showed 99.92% similarity to the 16S rRNA of CR-502. The phylogenetic classification of the above strain is shown in Fig. 3, and the selected strain Bacillus velezensis It was named LM3 and deposited at the National Institute of Agricultural Sciences Genetic Resources Center on July 19, 2023 (Deposit No.: KACC 92538P).

[0030] Example 2: Bacillus velezensis Establishment of Optimal Culture Medium Conditions for LM3

[0031] Bacillus velezensis In order to establish optimal medium conditions for the growth and maximum spore conversion of LM3, the effects of various carbon and nitrogen sources on the growth and spore production of the LM3 strain were investigated.

[0032] 1. Selection of carbon sources

[0033] To a basic medium composed of yeast extract 5 g / ℓ, KH2PO4 2.5 g / ℓ, MgSO4·7H2O 0.5 g / ℓ, CaCl2 0.5 g / ℓ, FeSO4·4H2O 0.5 g / ℓ, and MnSO4·H2O 0.5 g / ℓ, glucose, glycerol, sucrose, maltose, dextrose, lactose, or soluble starch is added at 15 g / ℓ as a carbon source. Test media were prepared by adding [specific amounts]. Each test medium was inoculated with spawn and cultured at 30°C and 150 rpm for 48 hours. After culture, the viable cell count was checked using the serial dilution streaking method, and after standing at 60°C for 1 hour, the spore count was measured and the spore conversion rate was calculated. The results are listed in Table 1.

[0034]

[0035] Table 1 shows that maltose is the most effective carbon source, with excellent viable cell count and spore conversion rate. Dextrose, sucrose, soluble starch, and glycerol had excellent viable cell counts but low spore conversion rates.

[0036] 2. Selection of Nitrogen Sources

[0037] A test medium was prepared by adding 5 g / ℓ of yeast extract, peptone, tryptone, ammonium sulfate, or soy bean flour as a nitrogen source to a basic medium composed of glucose 10 g / ℓ, KH2PO4 2.5 g / ℓ, MgSO4·7H2O 0.5 g / ℓ, CaCl2 0.5 g / ℓ, FeSO4·4H2O 0.5 g / ℓ, and MnSO4·H2O 0.5 g / ℓ. Except for using the above medium or TSB medium as the test medium, the viable cell count and spore count were measured and the spore conversion rate was calculated by the same method as for carbon source selection.

[0038]

[0039] Table 2 above describes the results, and the soybean floor was the most effective in both viable cell count and spore conversion rate.

[0040] Accordingly, a medium composed of 15 g / ℓ of maltose, 5 g / ℓ of soybean flour, 2.5 g / ℓ of KH2PO4, 0.5 g / ℓ of MgSO4·7H2O, and 0.5 g / ℓ of CaCl2 was determined to be the optimal medium.

[0041] Example 3: Bacillus velezensis Confirmation of the rice growth-promoting effect of LM3

[0042] Bacillus velezensis To confirm the growth-promoting effect of LM3 on rice, Bacillus velezensis ( Bacillus velezensis ) LM3 or Bacillus amyloliquefaciens( Bacillus amyloliquefaciens Rice seeds (Sindongjin rice) were immersed in the IM1 diluted culture solution for 24 hours, then thoroughly washed with distilled water and sown by distributing them into 50-cell seedling trays filled with paddy soil. The seeds used in the following examples Bacillus velezensis LM3 strain or Bacillus amyloliquefaciens The diluted culture of IM1 was cultured in the optimal medium determined in Example 2, and then the OD 600 =0.1(10 7 It was prepared by diluting to a concentration of cfu / mL. As a control group, rice seeds were immersed for the same amount of time in an optimal medium that did not contain the strain instead of the diluted culture solution, and as a positive control group, rice seeds were immersed for the same amount of time in a 2,000-fold dilution of the chemical fungicide prochloraz emulsion in distilled water.

[0043] After sowing, the germination and growth of rice seeds were checked daily for one week in seedling pots, and the germination rate and average germination days ((number of days after sowing × number of germinations on the day of investigation) / total number of germinations) were checked. These were then transplanted into Wagner pots filled with paddy soil, and the length of the rice leaves after one week was recorded in Table 3. As can be seen in Table 3, the use of microorganisms increased the germination rate and shortened the average germination period, whereas chemical disinfectants significantly reduced the germination rate and delayed the average germination period. Bacillus velezensis LM3 strain and Bacillus amyloliquefaciens There was no significant difference in germination rate and average germination days for the IM1 strain treatment group, but the rice leaf length after 2 weeks was Bacillus velezensis It was confirmed that leaf growth was best in the LM3 strain treatment group, with the leaves growing more than twice as long.

[0044]

[0045] Example 4: Bacillus velezensis Confirmation of LM3's induction of resistance to CMV in chili peppers

[0046] To confirm systemic resistance (ISR) induced by microbial treatment, Bacillus velezensis ( Bacillus velezensis ) LM3 and Bacillus amyloliquefaciens( Bacillus amyloliquefaciens ) 50 ml of IM1 was applied to each individual, and the plants were cultivated at 25°C for one week under conditions of 16 hours of light and 8 hours of darkness. After one week, Cucumber mosaic virus (CV201215-5) isolated from Guri, Gyeonggi-do and obtained from the Rural Development Administration was inoculated onto the underside of the pepper leaves using a sterile cotton swab. The expression of resistance genes induced during the systemic resistance response and the priming effect were confirmed via RT-PCR. Specifically, three hours after CMV treatment, the pepper leaves were frozen in liquid nitrogen, finely ground, and total RNA was extracted using an RNA prep kit (Qiagen). cDNA was synthesized by reverse transcription using the extracted RNA, and then RT-PCR was performed under the conditions in Table 6 using the primers in Table 4 and the composition in Table 5, with the synthesized cDNA serving as a template.

[0047]

[0048]

[0049]

[0050] Figure 4 shows images of electrophoresis results after RT-PCR, where (A) represents the control group (untreated), (B) represents the LM3 and CMV treatment group, (C) represents the IM1 and CMV treatment group, and (D) represents the negative control group treated only with CMV. Figure 4 shows that the CaPR1 or CaPR4 genes were not expressed in the microbial-untreated group, but were expressed identically in the ISR-inducing microbial treatment group or the pathogen treatment group. The CaTin-1 gene is Bacillus velezensis ( Bacillus velezensis The expression was best in the LM3 treatment group.

[0051] Example 5: In packaging Bacillus velezensis Confirmation of LM3's plant disease resistance

[0052] Regarding crops in packaging, the present invention Bacillus velezensis Whether the LM3 strain induces systemic resistance in crops and has a preventive effect against plant diseases Pectobacterium carotovorum Cabbage soft rot caused by SCC1, Xanthomonas euvesicatoria We tested for bacterial disease in peppers caused by (KACC 18722) and cucumber mosaic virus disease in peppers caused by cucumber mosaic virus (CV201215-5). As for the positive control, Bacillus velezensis Instead of LM3 Bacillus amyloliquefaciens IM1 was processed.

[0053] 1. In-flight test for prevention of cabbage soft rot

[0054] For each individual cabbage plant with 5 to 6 true leaves Bacillus velezensis LM3 strain or Bacillus amyloliquefaciens 50 ml of the diluted IM1 culture solution was inoculated onto the roots via drenching. After inoculation, the plants were cultivated at 25°C for one week under conditions of 16 hours of light and 8 hours of darkness. After one week, the plants were cultured in LB medium at 30°C and 150 rpm for 24 hours. Pectobacterium carotovorum SCC1 OD 600The strain was diluted to 0.4 (108 CFU / ml) and spray-inoculated into the treatment groups (experimental group and positive control group) and the untreated group (control group). Each treatment group was repeated three times with 5 individuals, and the disease prevalence was investigated by determining symptoms after culturing for one week. Table 7, Figure 5 (A), and Figure 6 (A) illustrate the results, showing that the strain of the present invention exhibits resistance to cabbage soft rot in actual fields.

[0055]

[0056] 2. In vitro test for the prevention of bacterial spot disease in chili peppers

[0057] For each pepper plant with 6 to 7 true leaves Bacillus velezensis LM3 strain or Bacillus amyloliquefaciens 50 ml of the diluted IM1 culture solution was inoculated onto the roots via drenching. After inoculation, the plants were cultivated at 25°C for one week under conditions of 16 hours of light and 8 hours of darkness. After one week, the plants were cultured in TS medium at 30°C and 150 rpm for 48 hours to exhibit viscosity Xanthomonas euvesicatoria (KACC 18722) OD 600 =0.4(10 8 The strain was diluted to a concentration of cfu / mL and spray-inoculated onto the treatment groups (experimental groups and positive control groups) and the untreated group (control group). Each treatment group was repeated three times with 5 individuals, and the disease prevalence was investigated by determining symptoms after incubation for one week. Table 8, Figure 5 (B), and Figure 6 (B) show the results, demonstrating that the strain of the present invention exhibits resistance to bacterial spot disease in peppers and promotes growth in actual fields.

[0058]

[0059] 3. In-flight test for prevention of cucumber mosaic virus disease in chili peppers

[0060] For each pepper plant with 6 to 7 true leaves Bacillus velezensis LM3 strain or Bacillus amyloliquefaciens50 ml of the diluted culture of IM1 was inoculated onto the roots via drenching. After inoculation, the plants were cultivated at 25°C for one week under conditions of 16 hours of light and 8 hours of darkness. After one week, the Cucumber mosaic virus (CV201215-5) isolated from Guri, Gyeonggi Province, obtained from the Rural Development Administration, was inoculated onto the underside of the pepper leaves using a sterile cotton swab. Each treatment group was repeated three times with five individuals, and the prevalence rate was investigated by identifying symptoms based on differences in lesions appearing on the leaves after one week of incubation. Table 9, Figure 5 (C), and Figure 6 (C) show the results, demonstrating that the strain of the present invention exhibits resistance to the cucumber mosaic virus in peppers and promotes growth in the actual field.

[0061]

[0062] Depository Name: National Institute of Agricultural Sciences, Rural Development Administration, Microbial Bank (KACC) Accession Number: KACC92538P Date of Deposit: 2023-07-19

Claims

Claim 1 delete Claim 2 delete Claim 3 delete Claim 4 delete Claim 5 delete Claim 6 delete Claim 7 delete Claim 8 delete Claim 9 delete Claim 10 delete Claim 11 delete Claim 12 delete Claim 13 delete Claim 14 delete Claim 15 Bacillus velezensis, deposited under accession number KACC 92538P, cultured in a medium composed of 15 g / ℓ maltose, 5 g / ℓ soybean flour, 2.5 g / ℓ KH2PO4, 0.5 g / ℓ MgSO4·7H2O, 0.5 g / ℓ CaCl2, 0.5 g / ℓ FeSO4·4H2O, and 0.5 g / ℓ MnSO4·H2O ( Bacillus velezensis A rice growth-promoting composition characterized by comprising a culture solution or culture solution extract of the LM3 strain and having an effect of promoting germination and growth of rice. Claim 16 delete Claim 17 delete

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