CN109609402A - Bacillus tequilensis XG18 and applications - Google Patents

Abstract

The invention discloses a kind of Te Jila bacillus Bacillus tequilensis XG18 and application, which is Te Jila bacillus Bacillus tequilensis XG18, CCTCC No:M2018831.The Te Jila bacillus Bacillus tequilensis XG18 is the bacterial strain for having antagonism to Xanthomonas campestris, pseudomonas syringae and Botrytis cinerea.The Te Jila bacillus can inhibit various bacterial disease and fungal disease simultaneously, have wide spectrum antagonism, the speed of growth is fast, substantially reduces incubation time for general bacterium, fungi, reduces production cost, with good development and application prospects.

Description

Bacillus tequilensis XG18 and applications

technical field

The invention relates to the field of plant protection, in particular to a Bacillus tequilensis XG18 and application.

Background technique

Bacteria of the genus Xanthomonas are a class of gram-negative plant pathogens that can infect more than 400 species of plants, including important food crops and commercial crops, such as rice, cotton, wheat, soybean, citrus, banana, etc. Lead to production reduction and significant economic losses, see (Leyns F, De Cleene M, Swings J, De Ley J. The host range of the genus Xanthomonas. The Botanical Reviews, 1984, 50(3):308-355). Campestris (X.campestris pv.Campestris, Xcc) can infect all cruciferous plants and cause black rot. Rice bacterial blight (X.oryzae pv.Oryzae, Xoo) can cause rice bacterial blight, and 2.7 billion people in the world eat rice as the staple food. Rice bacterial blight is the three most serious diseases to rice. For one, see (John M, Stephane G, Shimpei M, et al. Top 10 plant pathogenic bacteria in molecular plant pathology [J]. Molecular Plant Pathology, 2012, 13(6):614-629). Citrus canker is a bacterial disease caused by Xanthomonas axonopodis pv.citri, a pathogenic variety of citrus, which seriously damages citrus branches, leaves and fruits. Please refer to (Ren Jianguo, Huang Siliang, Cen Zhenlu, etc. 21 kinds of agents). Experiment on the control of citrus canker [J]. Fruit Trees in South China, 2005, 34(3):13-15), which is a worldwide quarantine object including China. The disease occurs in some citrus producing areas in my country, especially Guangdong, Guangxi, Hunan, Fujian and other provinces (regions) are seriously affected, causing fallen leaves, withered, weakened tree vigor, and serious fruit drop, affecting citrus yield and fruit quality For quality, please see (Fang Yimin, Zhang Yixu. Influence of the incidence of citrus canker on leaves, fruit drop, fruit size and quality [J]. Chinese Journal of Phytopathology, 1992, 22(4): 361-364).

Kiwifruit canker is a bacterial disease caused by Pseudomonas syringaepv. actinidiae, Psa (Pseudomonas syringaepv. actinidiae, Psa) (Li Yao, Cheng Heyuan, Fang Shumiao, et al. Preliminary study on the epidemic prediction of kiwifruit bacterial canker[ J]. Journal of Applied Ecology, 2001, 12(3):359-362.). Its occurrence has the characteristics of wide range, rapid spread, strong pathogenicity, and difficulty in control, etc. It can cause large-scale tree death in a short period of time, and is a devastating disease in the production and development of kiwifruit (Li Li, Zhong Caihong, Li David, et al. Research progress on bacterial canker of kiwifruit [J]. Journal of Huazhong Agricultural University, 2013, 32(5): 124-133.)

However, chemical pesticides are still the main method for the prevention and control of the above diseases, and chemical pesticides have disadvantages such as polluting the environment, being unsafe to humans and animals, and destroying the ecological environment. Biological control has the advantages of being environmentally friendly, easily degradable, and harmless to humans and animals, and is a feasible solution to control plant diseases. Among them, Bacillus (Bacilus) is an ideal biocontrol bacteria screening object, mainly including Bacillus subtilis (B.subtilis), Bacillus megaterium (B.megaterlum), Bacillus polymyxa (B.polymyxa), Bacillus pumilus (B.pumilus), Bacillus amyloliquefaciens (B.amyloliquefaciens) and Bacillus cereus (B.cereus) and so on.

The Chinese invention patent with the application number of 201610174472.8 discloses a peony endophytic Bacillus amyloliquefaciens, its isolation method and application. The invention isolates and screened an excellent peony endophytic Bacillus amyloliquefaciens, but this strain is only suitable for potted peony. Botrytis cinerea has a good disease prevention effect, a narrow application range, and a certain growth-promoting effect on peony; Chinese invention patent application number 201410735508.6 discloses a strain of Bacillus amyloliquefaciens Lh-1 in the prevention and control of cotton Verticillium wilt , Chinese cabbage soft rot, bacterial wilt, peach tree rot, apple rot, etc., it is necessary to use Bacillus amyloliquefaciens and chitosan oligosaccharide to form a microecological bacterial agent, compared with cotton Verticillium wilt The control effect is good; Tian Chengming et al. disclosed a bacterial strain that antagonizes plant pathogenic bacteria and its application. The antagonistic bacteria screened by the invention is Paenibacillus profundus XZQ-20, which can produce protease and can inhibit poplar The growth of plant pathogenic bacteria such as canker (Lonsdaleaquercina), kiwifruit canker (Pseudomonas syringae), cabbage soft rot (Erwiniacarotovora) and walnut black spot (Xanthomonas juglandis) can be used to control the growth of plant pathogenic bacteria such as European and American poplar canker, kiwifruit canker, Plant diseases caused by pathogenic bacteria such as cabbage soft rot fungus and walnut black spot fungus. However, this strain only has good antagonistic effect on plant pathogenic bacteria, but cannot antagonize pathogenic fungi.

The Chinese invention patent with the application number of 201410469787.6 discloses a strain of Pseudomonas fluorescens and its application in preventing and treating walnut black spot. The strain is Pseudomonas fluorescens strain GH2-1, which can not only efficiently To prevent and control walnut black spot disease, it can also effectively promote the growth of walnut, but the seed liquid of this strain needs to be shaken at 26°C for 3-5 days, and a large amount of solid fermentation needs to be cultured for 5-6 days. The preparation time of biological inoculants is long, and The antibacterial spectrum is narrow; there is also the use of walnut endophyte B. amyloliquefaciens Ht-q6 to control walnut anthracnose, pear brown rot, apple brown rot, eggplant Fusarium wilt, tomato gray mold, cotton Reports on blight blight, pear black spot, apple dry rot, watermelon anthracnose and bean sclerotinia, the Chinese invention patent application number 201510462700.7 discloses the use of watermelon endophyte Bacillus amyloliquefaciens XK Application of -2 in inhibiting watermelon fusarium wilt, rice sheath blight, and walnut root rot (application number: 201510462700.7), but in these two inventions, what Bacillus amyloliquefaciens antagonizes are pathogenic fungi, and there is no antagonism to pathogenic bacteria effect.

However, there are few research reports that Bacillus tequila can simultaneously treat a variety of phytopathogenic bacterial diseases and fungal diseases. The present invention will protect the bacteria in the effective prevention and treatment of pathogenic bacteria of the genus Xanthomonas and Pseudomonas syringae. Use in plant diseases caused by bacilli.

SUMMARY OF THE INVENTION

The purpose of the present invention is to overcome the deficiencies of the prior art, and provide a Bacillus tequila Bacillus equilensis XG18 and application. The application of the Bacillus tequilensis XG18 in the effective prevention and treatment of plant diseases caused by pathogenic bacteria of the genus Xanthomonas, Pseudomonas syringae and Botrytis cinerea is a biocontrol strain with broad-spectrum antagonism Bacillus fills the technical gap of a single microbial inoculant for simultaneous control of Xanthomonas, Pseudomonas syringae, Botrytis cinerea and other plant pathogenic bacteria and pathogenic fungi.

In order to achieve the first objective, the present invention designs a Bacillus tequilensis, which is characterized in that: the strain is Bacillus tequilensis XG18, CCTCC No: M 2018831.

The applicant screened a bacterium from the rhizosphere soil of orchids suffering from botrytis, and identified it as Bacillus tequilensis XG18 based on physiological and biochemical characteristics and molecular characteristics. The strain Bacillusequilensis XG18 has been deposited in the China Center for Type Culture Collection (CCTCC for short, Luojia Mountain, Wuchang City, Hubei Province) on November 26, 2018, and the deposit number is M2018831.

Further, the Bacillus tequilensis XG18 contains the nucleotide sequence shown in SEQ ID NO: 1.

The above-mentioned Bacillus tequilensis XG18 has the following characteristics: in terms of morphology and culture characteristics of the strain: it grows on LB solid medium to be milky white, opaque, the colony is nearly circular, and the single bacteria is observed under a microscope to be rod-shaped, leathery. The result of Lan's staining was positive.

The above-mentioned screening obtains bacterial strain culture, extracts DNA, and analyzes the 16S rDNA sequence in the genome as shown in SEQ ID NO: 1 (fragment length is 1435bp) and carries out BLAST analysis with the sequence in the GenBank database, and selects 7 strains and uses ClustalX 2.1 to carry out multiple sequence comparison. For the analysis, MEGA6.0 software was used to construct a phylogenetic tree and cluster analysis using the neighbor-joining method. The strain XG18 was clustered with the accession number JF411301.1 (Bacillus tequilensis strain km11) in GenBank, and the homology reached 99. %. According to the 16S rDNA classification standard, it is generally believed that the 16S rDNA sequence homology is greater than 97%, which can be considered to belong to the same species.

In order to achieve the second object, the present invention designs an application of Bacillus tequilensisXG18 in preventing and treating plant diseases caused by Xanthomonas pathogenic bacteria; wherein,

The plant disease is black rot of cabbage caused by Xanthomonas campestris pv. Campestris (Xcc);

Or, rice bacterial blight caused by X. oryzae pv. Oryzae (Xoo) and citrus canker caused by X. axonopodis pv. Citri (Xac),

The plant diseased plants include cruciferous crops, rice, tomato and citrus.

In order to achieve the third object, the present invention designs the application of Bacillus tequilensisXG18 in preventing and treating plant diseases caused by Pseudomonas syringae pv. actinidiae (Psa); wherein,

The plant diseases are kiwifruit canker, tomato bacterial leaf spot and cruciferous bacterial black spot.

The invention also provides an application of Bacillus tequilensis XG18 in preventing and treating plant diseases caused by Botrytis cinerea (BC);

The preparation of the above-mentioned biocontrol agent of Bacillus tequilensis XG18:

Pick a ring of XG18 colonies on the LB plate or slant and transplant it into a test tube for activation, culture on a shaker at 30°C for 16h, and rotate at 150rpm. The LB liquid medium in the test tube is: peptone 10g/L, yeast extract 5g/L, chlorinated Sodium 5g/L, pH 7.0; the activated bacterial solution is transplanted into a 250mL conical flask containing 100mL LB liquid medium according to the inoculum volume of 1%-3%, the rotation speed is 150rpm, and the culture is shaken at 35°C for 10h-24h. When the bacterial content is more than 2×10 ⁸ cfu/mL, the fermentation bacterial liquid can be obtained and stored as a biocontrol agent. The biocontrol agent is easy to make and has a fast growth rate. The liquid culture metabolites only need to be cultured in the liquid medium for 10-24 hours to obtain a good antagonistic effect. Compared with ordinary bacteria and fungi, the culture time is greatly shortened, and the Cost of production.

Beneficial effects of the present invention:

1. The biocontrol agent of the present invention is easy to manufacture and has a fast growth rate, and the liquid culture metabolites only need to be cultured in the liquid medium for 10-24h to obtain a good antagonistic effect, which greatly shortens the culture compared with general bacteria and fungi. time, reducing production costs.

2. The bacterium has strong colonization ability, and has a strong antagonistic effect on a variety of bacterial and fungal diseases at the same time, and has a good development and application prospect.

3. The antibacterial zone antagonism experiment showed that the Bacillus tequilensis XG18 had obvious inhibitory effect on X.oryzae pv.Oryzae(Xoo) disease.

Description of drawings

Figure 1 is a schematic diagram of the colony and microscopic morphology of Bacillus tequilensis XG18;

Figure 2 is a graph showing the effect of the inhibition zone of Bacillus tequilensis XG18 strain on X.oryzae pv.Oryzae (Xoo);

Fig. 3 is a graph showing the effect of the inhibition zone of Bacillus tequilensis XG18 strain on Pseudomonas syringae pv. actinidiae (Psa).

Detailed ways

The present invention will be further described in detail below with reference to specific embodiments, so as to be understood by those skilled in the art.

Example 1 Screening and identification of Bacillus tequilensis XG18

1. Screening of Bacillus tequilensis XG18

a. Isolation and purification of Bacillus:

Take 10g of orchid plant rhizosphere soil sample with gray mold and add it to a triangular flask containing 90mL sterile water and 30-50 glass beads to make a 1:10 concentration suspension, shake at 28°C and 180rpm After shaking in the bed for 20-30min, the bacterial suspension was placed in a water bath at 75-85°C for 12-18min. Transfer 1 mL of soil suspension into a test tube filled with 9 mL of sterile water to prepare 10 ^-2 bacterial suspension, and so on to obtain 10 ^-3 , 10 ^-4 , 10 ^-5 and 10 ^-6 bacterial suspension. After the gradient dilution, draw 100 μL of bacterial suspension with a dilution ratio of 10 ^-3 -10 ^-6 and spread it evenly on the separation medium - beef extract peptone medium. The medium formula is as follows (1L): beef extract 3.0g, peptone 10.0g, NaCl 5.0g, agar 15.0g, distilled water 1000mL, pH 7.2-7.4. The plate coated with the bacterial suspension was placed in a 30°C incubator for 1 week. After 24 hours, the state of the colonies on the separation medium was observed immediately, and the separation and purification were carried out with three-step streaking, and the uniform number was registered. Storage of strains: After inoculation of slants, it can be temporarily stored at 4°C. With 50% (V/V) glycerol as a protective agent, it can be stored at -80°C for a long time.

b. Primary screening of Bacillus:

First, using the spot connection method, 100 μL of Xanthomonas campestrispv. Campestris, Xcc) and Pseudomonas syringaepv. actinidiae (Psa) bacterial solutions (about 1×10 ⁷ cfu/ ml) evenly spread on the LB plate, and then use a sterile toothpick to spot four test strains on each plate (choose 4 strains in XG1, XG2, XG3...XG28 in turn), and set three replicates for each treatment . Incubate at 30°C and 37°C incubator for 20-28h. At the same time, the plate confrontation method was used to further screen the antagonistic bacteria that can antagonize Botrytis cinerea at the same time. The pathogenic bacteria cake with a diameter of 7 mm was placed in the center of the PDA plate, and then the bacteria to be tested were dipped with a sterile toothpick at an equal distance (1.5 cm from the center of the plate). ) were placed in a constant temperature incubator at 28°C for cultivation. After 5 days, the size of the antibacterial zone was observed, and each treatment was repeated three times to further screen strains with an antibacterial diameter greater than 5 mm.

c. Rescreening of Bacillus:

Re-screening by the tube-dish method: the initially screened strains with antagonistic effects were cultured in LB liquid medium at 150rpm for 20-28h, the fermentation broth was centrifuged at 8000rpm for 10min, and the supernatant was filtered through a 0.22μm microporous membrane until sterilization EP tube. LB agar plates coated with 100 μL of pathogenic bacteria (approximately 1×10 ⁷ cfu/mL) were punched equidistantly with a hole punch with a diameter of 7 mm. 50 μL of fermentation broth was inserted into each well, and sterile water was used as a control, and three replicates were set for each treatment. After culturing for 20-28 hours in a 30°C incubator, measure the diameter of the inhibition zone, and determine the degree of antagonism according to the value. The strains with antagonistic effect on Xanthomonas campestris and Pseudomonas syringae kiwifruit pathogenic varieties were screened out. At the same time, the primary screening antagonistic filtrate and the PDA solid medium of about 50 ° C were mixed at 1:19 and poured into a petri dish. After cooling, a Botrytis cinerea cake with a diameter of 7 mm was placed in the center of the flat plate. Water was the control, and each treatment was replicated three times. After 5 days, the diameter of the colony of pathogenic bacteria was measured, and the strains with antagonistic effect on Botrytis cinerea were screened out.

2. Identification and identification of Bacillus tequilensis XG18:

a. Morphological, physiological and biochemical characteristics of Bacillus tequilensis XG18:

The morphological and cultural characteristics of the strain: the cells are rod-shaped, with flagella around and with motility; the colony is milky white, nearly round, with irregular edges, and does not produce soluble pigments. As shown in Figure 1. In terms of physiological and biochemical identification: it belongs to Gram-positive bacteria, has contact enzyme activity, can use D-glucose, L-arabinose, D-xylose, D-mannose to produce acid, can use citrate, can hydrolyze Casein liquefies gelatin, has a positive V-P reaction, does not have lecithinase activity, cannot hydrolyze hippurate and tyrosine, and cannot use glucose to produce gas.

b. Molecular identification of Bacillus tequilensis XG18:

The above-mentioned screening obtains bacterial strain culture, extracts DNA, and analyzes the 16S rDNA sequence in the genome as shown in SEQ ID NO: 1 (fragment length is 1435bp) and carries out BLAST analysis with the sequence in the GenBank database, and selects 7 strains and uses ClustalX 2.1 to carry out multiple sequence comparison. For the analysis, MEGA6.0 software was used to construct a phylogenetic tree and cluster analysis using the neighbor-joining method. The strain XG18 was clustered with the accession number JF411301.1 (Bacillus tequilensis strain km11) in GenBank, and the homology reached 99. %. According to the 16S rDNA classification standard, it is generally believed that the 16S rDNA sequence homology is greater than 97%, which can be considered to belong to the same species.

Combined with the above morphological and some physiological and biochemical characteristics, it can be preliminarily determined that the antagonistic strain XG18 belongs to Bacillus tequilensis, named Bacillus tequilensis XG18.

Example 2

The method for preparing biocontrol agent by using Bacillus tequilensis XG18:

Pick a ring of XG18 colonies on the LB plate or slant and transplant it into a test tube for activation, incubate with a shaker at 35°C for 16h, and rotate at 150rpm. The LB liquid medium in the test tube is: peptone 10g/L, yeast extract 5g/L, chlorinated Sodium 5g/L, pH 7.0; the activated bacterial solution is transplanted into a 250mL conical flask containing 100mL LB liquid medium according to the inoculum volume of 1%-3%, the rotation speed is 150rpm, and the culture is shaken at 35°C for 10h-24h. When the bacterial content is more than 2×10 ⁸ cfu/mL, the fermentation bacterial liquid can be obtained and stored as a biocontrol agent.

Example 3 Antagonistic effect of Bacillus tequilensis XG18 on Xanthomonas pathogens

1. Take one ring of each single colony of the 3 species of Xanthomonas on the LB plate and place it in sterilized liquid LB medium, and culture it on a shaker at 28°C for 24-48h at a speed of 150rpm to obtain Xanthomonas campestris. Fermentation liquid of three pathogenic bacteria, Xanthomonas campestrispv.Campestris(Xcc), X.oryzae pv.Oryzae(Xoo), and X.axonopodis pv.Citri(Xac);

2. Take a single colony of Bacillus tequilensis XG18 on the LB plate and put it in a sterilized liquid LB medium, and culture it on a shaker at 35°C for 12 hours at a speed of 150 rpm, to obtain an antagonistic Bacillus tequilensis XG18 bacterial cell fermentation broth;

3. Take 100 μL of each of the 3 pathogenic bacteria fermentation broths in step 1, spread them on the LB solid medium, and then use a sterile puncher to punch 6 holes of 7 mm evenly on the LB solid medium, 3 of which are 50 μL of Bacillus tequilensis XG18 fermentation broth was added to the wells, and 50 μL of sterile water was added to the other three wells as blank control. size. Its antibacterial effect is shown in Table 1 and Figure 2.

The results showed that the Bacillus tequilensis XG18 was effective against Xanthomonas campestris pv.Campestris(Xcc), X.oryzae pv.Oryzae(Xoo), X.axonopodis pv The three subspecies of .Citri(Xac) have good antibacterial effect, and the antibacterial zone reaches more than 22mm, especially the antibacterial zone for rice bacterial blight reaches more than 30mm.

Table 1 Evaluation of the antibacterial effect of B.tequilensis XG18 fermentation broth on Xanthomonas pathogens

Note: The plants mentioned include cruciferous crops, rice, tomato and citrus etc.

Example 4 The effect of Bacillus tequilensis XG18 on Pseudomonas syringae pv. actinidiae (Psa) is the pathogenicity of Bacillus tequilensis XG18 on Pseudomonas syringae pv. actinidiae Variant inhibition zone experiment:

1. Take a single colony of Pseudomonas syringaepv. actinidiae (Psa) on the LB plate and place it in a sterilized liquid LB medium, culture it on a shaker at 28°C for 24-48h, and rotate at 150rpm. That is, the fermentation bacterial liquid of Pseudomonas syringae pv.actinidiae (Psa) is obtained;

2. Take a single colony of Bacillus tequilensis XG18 on the LB plate and put it in a sterilized liquid LB medium, and culture it on a shaker at 35°C for 12 hours at a speed of 150 rpm, to obtain an antagonistic Bacillus tequilensis XG18 bacterial cell fermentation broth;

3. Take 100 μL of the fermented bacterial liquid of pathogenic bacteria in step 1, spread it on the LB solid medium, and then use a sterile hole puncher to punch 6 holes of 7 mm evenly on the LB solid medium, and add 3 holes to them respectively. 50 μL of Bacillus tequilensis XG18 fermentation broth, and 50 μL of sterile water were added to the other 3 wells as blank control. After culturing at 28°C for 24 hours, the antagonistic bacteria were observed to inhibit the bacteria of Pseudomonas syringae kiwifruit pathogenic varieties. circle size.

The results showed that the Bacillus tequilensis XG18 had a good bacteriostatic effect on Pseudomonas syringae pathogenic varieties of kiwifruit, and the inhibition zone reached more than 22 mm (Table 2). The effect of the inhibition zone on Pseudomonas syringae pv.actinidiae (Psa) is shown in Figure 3. The inhibition zone of the antagonistic fermentation broth after dilution 10 times also reaches more than 22mm.

Table 2 Evaluation of the antibacterial effect of B. amyloliquefaciens D2WM fermentation broth on Xanthomonas pathogens

The plant diseases are kiwifruit canker, tomato bacterial leaf spot, cruciferous bacterial black spot and the like.

Example 5 Bacteriostatic effect of Bacillus tequilensis XG18 on Botrytis cinerea

1. Centrifuge the above-mentioned biocontrol agents at room temperature at 10,000 r/min for 20 minutes, and filter them with a 0.22 μm microporous membrane to remove the bacteria to obtain an antagonistic filtrate;

2. Take the antagonistic filtrate and the PDA solid medium at about 50 ° C after sterilization and pour it into a petri dish after mixing at 1:19. Water was the control, and each treatment was replicated three times. Pathogen colony diameters were measured after 5 days.

Sterile filtrate bacteriostatic rate=(control colony diameter-treated colony diameter)/(control colony diameter-0.7)×100%.

Table 3 shows that the bacteriostatic rate of Bacillus tequilensis XG18 against Botrytis cinerea all reached more than 90%.

Table 3 Bacteriostatic rate of Bacillus amyloliquefaciens D2WM sterile filtrate against major diseases of Camellia oleifera

Other parts that are not described in detail are the prior art. Although the above embodiment has made a detailed description of the present invention, it is only a part of the embodiments of the present invention, rather than all the embodiments. People can also obtain other embodiments according to the present embodiment without creativity. These embodiments All belong to the protection scope of the present invention.

sequence listing

<110> Hubei Institute of Engineering

<120> Bacillus tequilensis XG18 and applications

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1435

<212> DNA

<213> Bacillus tequilensis XG18 (Bacillus tequilensis XG18)

<400> 1

gctatacatg cagtcgagcg gacagatggg agcttgctcc ctgatgttag cggcggacgg 60

gtgagtaaca cgtgggtaac ctgcctgtaa gactgggata actccgggaa accggggcta 120

ataccggatg gttgtttgaa ccgcatggtt caaacataaa aggtggcttc ggctaccact 180

tacagatgga cccgcggcgc attagctagt tggtgaggta acggctcacc aaggcaacga 240

tgcgtagccg acctgagagg gtgatcggcc acactgggac tgagacacgg cccagactcc 300

tacgggaggc agcagtaggg aatcttccgc aatggacgaa agtctgacgg agcaacgccg 360

cgtgagtgat gaaggttttc ggatcgtaaa gctctgttgt tagggaagaa caagtaccgt 420

tcgaataggg cggtaccttg acggtaccta accagaaagc cacggctaac tacgtgccag 480

cagccgcggt aatacgtagg tggcaagcgt tgtccggaat tattgggcgt aaagggctcg 540

caggcggttt cttaagtctg atgtgaaagc ccccggctca accggggagg gtcattggaa 600

actggggaac ttgagtgcag aagaggagag tggaattcca cgtgtagcgg tgaaatgcgt 660

agagatgtgg aggaacacca gtggcgaagg cgactctctg gtctgtaact gacgctgagg 720

agcgaaagcg tggggagcga acaggattag ataccctggt agtccacgcc gtaaacgatg 780

agtgctaagt gttagggggt ttccgcccct tagtgctgca gctaacgcat taagcactcc 840

gcctggggag tacggtcgca agactgaaac tcaaaggaat tgacgggggc ccgcacaagc 900

ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc ttgacatcct 960

ctgacaatcc tagagatagg acgtcccctt cgggggcaga gtgacaggtg gtgcatggtt 1020

gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccttgatc 1080

ttagttgcca gcattcagtt gggcactcta aggtgactgc cggtgacaaa ccggaggaag 1140

gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg tgctacaatg 1200

gacagaacaa agggcagcga aaccgcgagg ttaagccaat cccaaaatc tgttctcagt 1260

tcggatcgca gtctgcaact cgactgcgtg aagctggaat cgctagtaat cgcggatcag 1320

catgccgcgg tgaatacgtt cccgggcctt gtacacaccg cccgtcacac cacgagagtt 1380

tgtaacaccc gaagtcggtg aggtaacctt taggagccag ccgccgaagg tgaca 1435

Claims (7)

1. A Bacillus tequila, characterized in that: the bacterial strain is Bacillus tequila Bacillustequilensis XG18, CCTCC No: M 2018831. 2 . The Bacillus tequila according to claim 1 , wherein the nucleotide sequence shown in SEQ ID NO: 1 is contained in the Bacillus tequila Bacillustequilensis XG18. 3 . 3. the application of Bacillus tequilensis XG18 described in claim 1 in preventing and treating the plant disease caused by Xanthomonas genus pathogenic bacteria; It is characterized in that: described plant disease is Xanthomonas campestris Xanthomonas Cabbage black rot caused by campestris pv. Campestris; Alternatively, rice bacterial blight caused by X. oryzae pv. Oryzae and citrus canker caused by X. axonopodis pv. Citri. 4. The application according to claim 3, wherein the plants with plant diseases include cruciferous crops, rice, tomato and citrus. 5. An application of Bacillus tequilensis XG18 according to claim 1 in preventing and treating plant diseases caused by Pseudomonas syringae kiwifruit pathogenic varieties. 6 . The application according to claim 5 , wherein the plant diseases are kiwifruit canker, tomato bacterial leaf spot and cruciferous bacterial black spot. 7 . 7. The application of Bacillus tequilensis XG18 described in claim 1 in preventing and treating plant diseases caused by Botrytis cinerea.