CN117821339B - Bacillus bailii B2 and application thereof in preventing and controlling wheat leaf blight - Google Patents
Bacillus bailii B2 and application thereof in preventing and controlling wheat leaf blight Download PDFInfo
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- CN117821339B CN117821339B CN202410195641.0A CN202410195641A CN117821339B CN 117821339 B CN117821339 B CN 117821339B CN 202410195641 A CN202410195641 A CN 202410195641A CN 117821339 B CN117821339 B CN 117821339B
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- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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Abstract
The invention discloses bacillus beleiensis B2 and application thereof in preventing and controlling wheat leaf blight, and belongs to the technical field of crop biocontrol strains, wherein bacillus beleiensis (Bacillus velezensis) is preserved in China Center for Type Culture Collection (CCTCC) with a preservation number of CCTCC NO: m20231076. According to the invention, the bacillus beijerinus B2 is obtained through separation and screening, and the bacillus beijerinus B2 has an inhibition effect on various pathogenic bacteria. The bacillus belicus B2 provided by the invention can effectively reduce the growth amount of the mycelium of the micronodular mould, has good control effect on wheat leaf blight caused by the micronodular mould, has the control effect of up to 95.4% in a greenhouse, and has great potential for developing a commercial living fungus biocontrol agent.
Description
Technical Field
The invention relates to the technical field of crop biocontrol strains, in particular to bacillus bailii B2 and application thereof in preventing and controlling wheat leaf blight.
Background
Wheat is a main grain crop in China, leaf blight caused by micro-tubercle bacillus (Microdochium majus) is a disease which infects a plurality of growth parts of wheat, and a plurality of provinces occur in a main wheat producing area in China, so that the yield and quality of the wheat are seriously affected. The disease becomes one of important factors limiting the wheat yield in China. At present, chemical agent control is mainly used for controlling wheat leaf blight, but drug resistance is inevitably generated due to the large-scale use of chemical agents. In 2009, an-sophiwalker et al found that more than 50% of strains from m.majus in france had developed resistance to methoxy acrylic acid ester fungicides by detecting the bases of cytochrome b genes, and therefore, there was an urgent need to find a green, safe and reliable control method.
In recent years, the prevention and treatment of plant diseases by using beneficial microorganisms is one of research hotspots in the field of plant diseases, and certain achievements are achieved, and some biopesticides are also developed successively. Bacillus is widely used in the field due to its characteristics of fast propagation speed, strong stress resistance, etc. Wherein bacillus beleiensis (Bacillus velezensis) has proved to have stronger disease prevention effect as a biocontrol bacterium, chinese patent CN111254086A, bacillus beleiensis and application thereof in biocontrol illustrate the remarkable biocontrol effect of B.velezensis Bv-6 strain wettable powder on gray mold caused by B.cinerea; researches of pelargonic et al in 2022 show that the control effect of B.velezensis SM905 on dendrobium candidum colletotrichum Colletotrichum gloeosporioides is better than that of difenoconazole; LI XJ in 2023 and the like find that the control effect of B.velezensis Ba-0321 on tobacco root rot fungus Fusarium oxysporum reaches 81.0%.
The control effect of the biocontrol strain is influenced by the biological characteristics of the strain, soil environment, climate and geographic conditions, and the problems of short duration, unstable effect and the like of some biocontrol bacteria exist, so that the further screening of biocontrol strain resources with stronger adaptability has important significance.
Disclosure of Invention
The invention aims to provide bacillus beliensis B2 and application thereof in preventing and controlling wheat leaf blight, so as to solve the problems in the prior art, and on the basis of finding that micronodular mold can cause wheat leaf blight, the bacillus beliensis (Bacillus velezensis) B2 is separated and screened, and the pathogen resistance of the biocontrol strain and the inhibition effect thereof on the pathogen causing wheat leaf blight are subjected to deep research, so that green prevention and control of the wheat leaf blight are realized.
In order to achieve the above object, the present invention provides the following solutions:
The invention provides bacillus beijerinckii (Bacillus velezensis) B2 which is preserved in China center for type culture collection (China center for type culture collection) at the year 2023 and the month 6, wherein the preservation number is CCTCC NO: m20231076.
The invention also provides a microbial agent containing the bacillus bailii B2 or the fermentation liquor thereof.
Further, the viable bacteria concentration of the microbial agent is 1X 10 7～1×109 CFU/mL.
The invention also provides application of the bacillus bailii B2 or the microbial agent in inhibiting growth of pathogenic bacteria, wherein the pathogenic bacteria comprise micronodular mold, fusarium graminearum, alternaria tenuis, colletotrichum gloeosporioides, trichoderma viticola and botrytis cinerea.
Further, the plant disease is wheat leaf blight caused by sarcoidosis.
The present invention also provides a method for inhibiting the growth of pathogenic bacteria comprising the steps of:
Treating pathogenic bacteria by adopting the bacillus belicus B2 or the fermentation liquor thereof, thereby inhibiting the growth of the pathogenic bacteria;
The pathogenic bacteria include microtubercle mold, fusarium graminearum, alternaria tenuis, alternaria grub, pediopsilosis and Botrytis cinerea.
The invention also provides a method for preventing and controlling wheat leaf blight caused by the microtubercle mould, which comprises the following steps:
Treating wheat plants with the bacillus beljalis B2 or fermentation liquor thereof;
the concentration of viable bacteria in the fermentation liquor is 1X 10 7～1×109 CFU/mL.
Further, the processing is as follows: wheat plants were uniformly sprayed with a fermentation broth of Bacillus bailii B2 at a concentration of 1X 10 8 CFU/mL.
The invention also provides a biocontrol agent for preventing and treating wheat leaf blight caused by the microtubercle bacillus, and the biocontrol agent comprises the bacillus subtilis B2 or fermentation liquid thereof.
The invention discloses the following technical effects:
The invention discovers and verifies that the micronodular mould can cause wheat leaf blight for the first time. Aiming at wheat leaf blight caused by micronodular mould, the bacillus beijerinus (Bacillus velezensis) B2 is obtained through separation and screening, belongs to a biological agent, avoids the problems caused by the use of chemical control agents, and is beneficial to pollution-free green production of wheat. The bacillus belay B2 separated and identified by the invention has an inhibition effect on various pathogenic bacteria, can effectively reduce the growth amount of the mycelium of the micronodular mould (about the growth amount of the mycelium 80.03 percent), has a good prevention and control effect on wheat leaf blight caused by the micronodular mould (the prevention and control effect in a potting test is as high as 95.4 percent), can effectively control the infection of the micronodular mould on wheat leaves, achieves the good effect of preventing and controlling the wheat leaf blight, and has great potential for being developed into a commercial living bacterial biocontrol agent.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram showing symptoms of wheat leaf blight caused by Rhizoctonia cerealis and verification test results thereof; in the diagram a: leaf disease symptoms of field plants; b: culturing the morphological characteristics of the micro-tubercle mold colony for 5 days on a PDA culture medium at 25 ℃; c: the morphology of conidia of the micro-nodular mould produced on PDA medium, scale = 50 μm; D-E: the method verifies that the microtubercle mould is wheat leaf blight pathogen;
FIG. 2 shows colony morphology of Bacillus belicus B2; in the diagram a: colony morphology on LB plates; b: gram staining; c: morphological observation of thalli under a scanning electron microscope;
FIG. 3 is a constructed Bacillus phylogenetic tree; in the diagram a: bacillus phylogenetic tree constructed based on 16S rDNA sequence; b: constructing a bacillus phylogenetic tree based on the Gyr A gene sequence;
FIG. 4 shows the inhibition of B.bailii B2 against different pathogenic fungi; in the diagram a: micronodular mold (m.majus); b: fusarium graminearum (F. Graminearum); c: alternaria tenuissima (A.tenuissima); D-F is micronodular mold (M.majus), fusarium graminearum (F.graminearum) and Alternaria graminearum (A.tenuissima) added with Bacillus bailii B2 in sequence; g: anthrax (c.truncatum); h: trichoderma viride (p.portugallica); i: botrytis cinerea (b.cinerea); J-L is, in order, bacillus belicus B2-added Botrytis cinerea (C.truncatum), pelargonium gracilis (P.portugallolica), botrytis cinerea (B.cinerea);
FIG. 5 is a graph showing the effect of fermentation broths of Bacillus bailii B2 at different concentrations on inhibiting the growth of mycelial growth of Rhizoctonia cerealis; in the diagram a: PDA medium (control group); b: PDA medium +1% fermentation broth; c: DA medium+5% fermentation broth; d: PDA medium +10% fermentation broth; e: PDA medium +15% fermentation broth; f: PDA medium +20% fermentation broth;
FIG. 6 is a schematic diagram showing the result of controlling Bacillus belicus B2 in greenhouse potting of Rhizoctonia cerealis; in the diagram a: (negative control group); b: inoculating micronodular mould plus B2 fermentation liquor (treatment group); c: only the micro-nodular mold (positive control group) was inoculated.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
In the following examples, the pathogenic strain, micronodular mold (M.majus), was involved on wheat leaves from a modern agricultural science demonstration garden in yellow oka, hubei province suffering from leaf blight; bacillus belicus B2 was isolated from isotopy healthy wheat leaves.
The culture media used in the following examples: LB solid/liquid medium: 10g of tryptone, 5g of yeast extract powder, 20g of agar powder and 5g of NaCl, adding distilled water to 1L, adjusting the pH to 7.2, packaging, and sterilizing for later use (no agar powder is added into the liquid culture medium); PDA solid/liquid medium: 200g of potato, 18g of sucrose and 15g of agar powder, adding distilled water to 1L, and sterilizing for later use (the agar powder is not added into the liquid culture medium).
Test pathogenic bacteria: fusarium graminearum (F. Graminearum), alternaria tenuissima (A. Tenuissima), alternaria grub (C. Truncatum), pediopside (P. Portugalica), botrytis cinerea (B. Cinerea) are all offered by the microorganism laboratory of the university of yellow subject matter, the institute of biological and agricultural sciences.
In the examples below, the experimental results were both statistically and analytically plotted using Microsoft Excel 2021, origin.
Example 1 micro-nodular mold is a pathogenic bacterium of wheat leaf blight in Hubei province
In this example, it was found for the first time that micronodular mold (m.majus) can cause wheat leaf blight in the province of hubei, and the procedure was as follows:
in 2 months 2019, a serious wheat disease is found in a wheat planting base in a modern agricultural science and technology demonstration garden in Hubei yellow-okang, a large-area leaf blight-like symptom is found in 98% of plants, the disease is early gray brown small disease spots from bottom leaves to sword leaves of the wheat, yellow halos are arranged at disease-key junctions, and the disease spots rapidly spread into irregular long-strip-shaped large disease spots at proper temperature and humidity (A in fig. 1).
To obtain the pathogens, diseased wheat leaves were surface sterilized with 75% ethanol for 30s, soaked in 0.1% hgcl 2 for 30s, rinsed three times with sterile water, and cultured upside down on PDA medium at 25 ℃. The hyphae of the diseased tissue were subcultured on new PDA medium to obtain pure cultures. After 7d incubation at 20℃white colonies (FIG. 1B) formed and a large number of sickle shaped conidia (FIG. 1C) were produced on the medium, the size was about 19.53-26.22. Mu.m.times.2.90-5.12. Mu.m, which was consistent with the micro-tubercle fungi reported in the literature (wheat stem rot epidemiological study by micro-tubercle fungi Microdochium majus, taijia, 2019) based on the colony morphology and conidium microscopic features of the pathogenic bacteria.
To verify pathogenicity of the micronodular mould, the micronodular mould cakes (diameter 6 mm) were inoculated on healthy and disease-free wheat plants in the tillering stage, and the plants were inoculated with sterile PDA blocks as controls, 3 pots per group, and repeated 3 times. After all wheat plants were grown in the greenhouse at 20℃for 5D, typical leaf blight lesions formed on the leaves of the wheat of the experimental group and a large number of hyphal layers formed at the base of the stems of the wheat plants, whereas the plants of the control group did not show any symptoms (D and E in FIG. 1). Colonies isolated in the treated group of wheat leaves exhibited the same morphological characteristics as the inoculated strain, whereas target colonies were not isolated in the control group of wheat leaves.
The method is the first discovery of the wheat leaf blight caused by the micronodular mold in Hubei province of China, and provides an important basis for the identification of the wheat leaf blight and the green prevention and control of diseases.
EXAMPLE 2 isolation, screening and identification of Bacillus bailii B2
The embodiment is a screening and identifying process of bacillus bailii B2, which specifically comprises the following steps:
(1) Isolation of strains
10G of collected healthy wheat leaves are sterilized for 30s by using 75% ethanol, washed three times by using sterile water, then the leaves are put into a sterilized mortar to be rapidly ground into paste, 10mL of sterile water is added for dilution and mixing uniformly, 100 μL of the paste is taken by a micropipette and added into a 2mL centrifuge tube filled with 900 μL of sterile water for sequential dilution, three gradient dilutions of 10 -4～10-6 are obtained, 100 μL of coating plates are taken after shaking and shaking uniformly, inversion culture is carried out for 48h at 28 ℃, single colony is selected for purification and preservation, and isolated strain B2 is obtained.
(2) Morphological characterization of Strain B2
The strain B2 preserved in glycerol is streaked on LB medium and cultured for 48 hours at 28 ℃, and the characteristics of colony shape, color, luster, edge morphology and the like are observed. The colony is opaque, milky white, wrinkled and irregularly shaped on the flat plate as shown in figure 2; is a gram positive bacterium; the strains were observed under electron microscopy in rod or short rod form, single or multiple bacterial arrangements.
(3) Physiological and biochemical characteristics of strain B2
The physiological and biochemical assay of the strain B2 is carried out by combining the methods described in the handbook of identification of common bacterial systems and the handbook of Berger identification.
TABLE 1 physiological biochemical assay of Strain B2
Note that: "+" indicates positive reaction; "-" indicates reaction negative.
As is clear from Table 1, the strain B2 was purple in gram stain, positive, unable to grow under anaerobic conditions, unable to liquefy gelatin, methyl red reaction negative, indole reaction positive, hydrogen sulfide reaction negative, and V-P reaction positive. The nitrate reduction reaction is positive, citrate, lactose, glucose, D-xylitol and L-arabinose can be used, starch can be hydrolyzed, propionate and D-mannitol cannot be used, the growth can be realized under the condition of pH5.7, and the growth can not be realized under the condition of 7% NaCl. These physiological and biochemical characteristics are consistent with those of Bacillus and are substantially consistent with those reported for Bacillus bailii.
(4) Molecular characterization of Strain B2
And picking the purified single colony to LB liquid medium, and placing the single colony in a shaking table for culturing for 24 hours at 28 ℃ at 200 r/min. Genomic DNA of the strain was extracted using a genomic DNA extraction kit (Biotechnology Co., ltd.). The bacterial strain B2 to be tested was subjected to PCR amplification of the 16S rDNA sequence using the bacterial 16S rDNA universal primers 27F (5 '-AGAGTTTGATCMTGGCTCAG-3') and 1492R (5'-GGTACCTTGTTACGACTT-3') (specific methods reference Azad SA,Hossain KM,Rahman SMM,Mazid MFA,Barai P,Gazi MS.In ovo inoculation of duck embryos with different strains of Bacillus cereus to analyse their synergistic posthatch antiallergic potentialities[J].Veterinary Medicine and Science,2020,6(4):992-999.);GyrA gene sequence identification (specific methods reference BAVYKIN S G,LYSOV Y P,ZAKHARIEV V,et al.Use of 16S rRNA,23S rRNA,and Gyr A genes sequence Analyze to determine physiological relationships of Bacillus cereus group microorganisms[J].Journal of Clinical Microbiology,2004,42(8):3711-3730.); used gyrA-F (5'-CAGTCAGGAAATGCGTACGTCCTT-3') and reverse primers gyrA-R (5'-CAAGGTAATGCTCCAGGCATTGCT-3'), primer sequences and genomic DNA of the test bacteria were synthesized and sequenced by Shanghai Biotechnology.
Sequencing PCR amplified products, and comparing and analyzing 16S rDNA and Gyr A gene sequences in NCBI GenBank database; phylogenetic tree was constructed using the proximity-joining method (NJ) in MEGA 7.0 software. Phylogenetic tree was constructed by MEGAX based on the 16S rDNA and Gyr A gene sequences (A and B in FIG. 3). The results show that these 2 sequences are clustered together with members of Bacillus bailii Bacillus velezensis; the NCBI database comparison shows that the homology with XG-FX22 and NJ21 strains in B.velezensis reaches 100% and 99.89%, respectively, and the strain B2 is determined to be bacillus bailii (Bacillus velezensis) through molecular biology and morphological identification.
The bacillus belicus B2 is preserved, and the preservation number is CCTCC NO: m20231076, which is classified and named Bacillus velezensis, and the preservation date is 2023, 06 and 21 days, and the preservation unit is China Center for Type Culture Collection (CCTCC), and the preservation unit is Jiuqiu 299 university in Wuchang district of Wuhan, hubei province.
EXAMPLE 3 cultivation of Bacillus bailii B2 and preparation of fermentation broth thereof
This example is a preferred method for culturing bacillus bailii B2 obtained by screening in example 2 and a method for preparing a fermentation broth thereof, comprising the steps of:
streaking and inoculating Bacillus bailii B2 stored in glycerol at-80 ℃ on a solid LB culture medium, culturing for 24 hours in a constant temperature cabinet at 28 ℃, then picking up single bacterial colonies and inoculating in 100mL of liquid LB culture medium, and culturing for 48 hours under the conditions of 28 ℃ and 200r/min under shaking, so as to obtain a fermentation broth of the Bacillus bailii B2, wherein the viable bacteria concentration of the fermentation broth is 1 multiplied by 10 8 CFU/mL.
Example 4 broad-spectrum resistance verification of Bacillus bailii B2
This example demonstrates the broad-spectrum resistance of bacillus belgium B2 to pathogenic bacteria, which includes the steps of: the 6 pathogenic bacteria of the species Rhizopus (M.majus), fusarium graminearum (F.graminearum), alternaria tenuissima (A.tenuissima), alternaria gruous (C.truncatum), pelargonium gracilis (P.portugalulica), botrytis cinerea (B.cinerea) stored at 4℃were activated on PDA plates (25 ℃,3 d), round cakes were punched along the outer edges of the colonies with a punch (diameter 6 mm), the cakes were placed on both sides of new PDA plates, and 20. Mu.L of fermentation broth of Bacillus berensis B2 (prepared in example 3) was inoculated in the middle of the PDA plates, with sterile water as a control. The inhibition ratio (%) = (control group colony diameter-treatment group colony diameter)/(control group colony diameter-bacterial cake diameter) ×100) was calculated after 5d incubation at 25 ℃.
As shown in the figure 4, the strain B2 has good inhibition effect on 6 pathogenic fungi, and the inhibition rate ranges from 45.38% to 85.18%; the inhibition effect on botrytis cinerea is strongest, and is obviously higher than other pathogenic fungi, and the inhibition rate is up to 85.18%; and the second is the micronodular mould with the inhibition rate of 81.35 percent.
EXAMPLE 5 Flat plate antagonistic Effect of Bacillus bailii B2 fermentation broths at different concentrations on Rhizoctonia cerealis
This example demonstrates the plate antagonism of Bacillus belicus B2 broth against Rhizopus microtuberosus (described in literature: rhizopus microtuberosus Microdochium majus-induced wheat stem basal rot epidemics, taijia, 2019's Shuoshi paper), comprising the steps of:
The fermentation broths of bacillus belicus B2 (prepared in example 3) were added to the melted PDA medium at about 45 ℃ at concentrations of 1%, 5%, 10%, 15% and 20%, respectively, and the mixture was poured into a plate after being mixed uniformly. The micro-nodular mold stored in the refrigerator at 4℃was activated (25℃for 3 d) on PDA plates, round cakes were punched along the outer edges of the colonies with a punch (diameter 6 mm), and the cakes were inoculated in the middle of the PDA plates with different concentrations of B2 broth, with the PDA plates without B2 broth as controls. Colony diameters were recorded after 5d incubation at 20 ℃. The inhibition (%) was calculated by the same method as in example 4 by the inhibition evaluation. As shown in FIG. 5 and Table 2, the strain B2 can significantly inhibit the growth of the mycelium of the Rhizoctonia cerealis.
TABLE 2 analysis of strain B2 antagonistic micronodular mold
Example 6 greenhouse control of Bacillus bailii B2 against wheat leaf blight
This example demonstrates the greenhouse efficacy of Bacillus belicus B2 against leaf blight caused by Rhizopus microtuberosus (described in literature: rhizopus microtuberosus Microdochium majus, taijia, 2019's paper), comprising the steps of:
100mL of bacillus beijerinus B2 fermentation liquid (1.0X10 8 CFU/mL) is uniformly sprayed on healthy and disease-free wheat plants in the seedling stage, and the control group is sprayed with equal amount of clean water. 100mL of the prepared microspore liquid of the microtuberous fungus (1.0X10 6/mL) was then sprayed evenly on wheat plants, and each treatment was repeated 3 times. Culturing in sunlight greenhouse (25deg.C) for 5d, and investigating disease index and morbidity. The index rating criteria (Gu Jingyi et al, 2021) are shown in Table 3.
TABLE 3 index grading criteria for disease states
Degree of onset = incidence index of disease
Control effect = (positive control incidence-treatment group incidence)/positive control incidence x 100%
As can be seen from FIG. 6 (A: negative control; B: treatment group; C: positive control), strain B2 has a certain inhibitory effect on wheat leaf blight. After 5d inoculation at 25 ℃, as shown in table 4, the negative control was normal, no disease occurred; the incidence rate of the positive control is 100% and the disease index is 5; the incidence rate of the treatment group is 23%, the disease index is 1, and the control effect of the strain B2 in the greenhouse on wheat leaf blight is as high as 95.4%.
TABLE 4 greenhouse control Effect of Strain B2 on wheat leaf blight
As shown in the above examples, the invention discovers that the micronodular mould can cause wheat leaf blight for the first time, and screens out a bacillus beijerinus B2 strain. The strain B2 has obvious inhibition effect on the hypha growth speed of the micro-nodular mould, and when the strain B2 is opposite to a pathogenic bacteria micro-nodular mould flat plate, the strain B2 can stimulate the micro-nodular mould to produce pigment, the hypha is aged, and a large number of conidia are rapidly produced, probably because the pathogenic bacteria are in an adversity stress environment at the moment, so that the pathogenic bacteria are stressed to produce spores, rapidly age and accelerate the death process. On a PDA flat plate added with 20% of strain B2 fermentation liquor, the mycelium of the micronodular mould can hardly grow normally, wheat plants are sprayed by adopting the strain B2 fermentation liquor (1.0X10 8 CFU/mL), the control effect on the micronodular mould reaches 95.4%, and the bacillus berryis B2 has a good biocontrol effect on the wheat micronodular mould and can be used as antagonistic strain resources.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (9)
1. Bacillus beijerinckii (Bacillus velezensis) B2, which is characterized by being preserved in the China Center for Type Culture Collection (CCTCC) NO: m20231076.
2. A microbial agent comprising Bacillus bailii B2 or a fermentation liquid thereof according to claim 1.
3. The microbial agent of claim 2, wherein the viable bacteria concentration of the microbial agent is 1 x 10 7～1×109 CFU/mL.
4. Use of bacillus bailii B2 according to claim 1 or a microbial agent according to any one of claims 2 to 3 for inhibiting the growth of pathogenic bacteria, wherein the pathogenic bacteria are micro-tubercle mould, fusarium graminearum, alternaria tenuis, anthrax grunniens, trichoderma viride and botrytis cinerea.
5. Use of bacillus beljalis B2 according to claim 1 or of a microbial agent according to any one of claims 2 to 3 for controlling plant diseases, wherein the plant diseases are wheat leaf blight caused by micro-nodulation fungi.
6. A method for inhibiting the growth of pathogenic bacteria comprising the steps of:
Treating pathogenic bacteria with bacillus belgium B2 or a fermentation broth thereof according to claim 1, thereby inhibiting pathogenic bacteria growth;
The pathogenic bacteria are microtuberous fungus, fusarium graminearum, alternaria tenuis, alternaria grub, pediopsilosis and Botrytis cinerea.
7. A method for controlling leaf blight of wheat caused by micro-nodulation fungi, comprising the steps of:
Treating wheat plants with bacillus beljavensis B2 or a fermentation broth thereof according to claim 1;
The concentration of viable bacteria in the fermentation liquor is 1X 10 7～1×109 CFU/mL.
8. The method of claim 7, wherein the method of treating wheat plants comprises: wheat plants were uniformly sprayed with a fermentation broth of Bacillus bailii B2 at a concentration of 1X 10 8 CFU/mL.
9. A biocontrol agent for controlling wheat leaf blight caused by micro-nodulation fungi, characterized in that the biocontrol agent comprises bacillus bailii B2 or a fermentation broth thereof according to claim 1.
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| CN112980721A (en) * | 2021-01-29 | 2021-06-18 | 青岛农业大学 | Bacillus belgii and biocontrol preparation and application thereof |
| CN117025466A (en) * | 2023-08-10 | 2023-11-10 | 赣州市蔬菜花卉研究所 | Bacillus bailii and application thereof in controlling melon crop diseases |
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| CN112980721A (en) * | 2021-01-29 | 2021-06-18 | 青岛农业大学 | Bacillus belgii and biocontrol preparation and application thereof |
| CN117025466A (en) * | 2023-08-10 | 2023-11-10 | 赣州市蔬菜花卉研究所 | Bacillus bailii and application thereof in controlling melon crop diseases |
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