CN111073827A

CN111073827A - Bacillus beilesensis JTB8-2 and application thereof in biological control of orobanche meloidogyne

Info

Publication number: CN111073827A
Application number: CN201911122043.6A
Authority: CN
Inventors: 何伟; 许建军; 罗文芳; 高海峰
Original assignee: Institute Of Plant Protection Of Xinjiang Academy Of Agricultural Sciences
Current assignee: Institute Of Plant Protection Of Xinjiang Academy Of Agricultural Sciences
Priority date: 2019-11-15
Filing date: 2019-11-15
Publication date: 2020-04-28
Anticipated expiration: 2039-11-15
Also published as: CN111073827B

Abstract

The invention discloses a Bacillus belgiiBacillus velezensis) JTB8-2GDMCC No: 60755 and its application in biological control of herba Orobanches. Bacillus belgii (B) with the number of JTB8-2 provided by the inventionBacillus velezensis) Through serial physiological and biochemical experiments, systematic classification and systematic identification and analysis, the bacillus belgii is known to be a strain of bacillus belgii (B.) (Bacillus velezensis) The new strain has the inhibition rate of separated strain fermentation liquor and fermentation diluent on the germination of the seed of the orobanche cucurbitacearum up to 100 percent, the unearthed quantity of the orobanche cucurbitacearum treated by using the strain JTB8-2 strain fermentation liquor is reduced by 70.5 percent, the fresh weight of the orobanche cucurbitacearum is reduced by 66.6 percent, and the orobanche cucurbitacearum biological control is realizedThe jig has wide application value.

Description

Bacillus beilesensis JTB8-2 and application thereof in biological control of orobanche meloidogyne

Technical Field

The invention relates to the technical field of microbial strains and application thereof, in particular to the technical field of application of belius bacillus in biological control.

Background

Orobanche aegyptica (Orabanche aegyptica) is a highly parasitic plant widely distributed in Mediterranean areas, Western Asia and eastern Europe, has a great impact on local crop production, and is considered to be the most harmful parasitic weed worldwide. There are about 23 kinds of Guajoba in China, which are mainly distributed in 11 provinces such as Xinjiang, Jilin, Gansu, Heilongjiang, Hebei, Shandong, Shanxi, Liaoning, Qinghai, inner Mongolia and Sichuan. The broomrape mainly harms melons in Xinjiang, and for years, the broomrape threatens the production of Hami melons, after being parasitized, the sugar content of the melons is reduced, the melons are light and tasteless, and the melons are not grown seriously, and although general herbicides can kill the broomrape, the general herbicides also damage melon plants; the extraction damages the melon root.

In recent years, because a large number of tomato seeds are introduced from abroad and abroad in a disordered way, the plant quarantine work is lagged, so that the Guarangam is expanded and spread in a large range in Xinjiang, and the damage degree is increasingly serious. The root parasitic weed of orobanche cyrtophylla (Orobancheaegyptiaca) seriously harms the yield and quality of a plurality of commercial crops such as watermelon, melon and tomato. How to effectively control the Guarandum is still one of the key points of research at present.

In recent years, the method for preventing and controlling the orobanche cucurbitae by using the microorganisms is more and more emphasized, and is a currently accepted growth inhibiting method with low cost, good effect and safety. However, the inhibition abilities of different microbial strains on the growth of the citrulline cupreum are greatly different, and the screening of excellent microbial strains is an important step for researching the biological control of the growth of the citrulline cupreum. The currently reported strains for inhibiting the growth of the cucurbitacin are few in types and mainly comprise fusarium, alternaria, pythium and the like. Therefore, the screening of the new strain capable of effectively inhibiting the growth capacity of the cucurbitacin is of great significance.

The Bacillus beleisi is derived from various ecological environments such as plant rhizosphere, soil, plant interior, rivers and the like, and most of the Bacillus beleisi separated from the plant rhizosphere can colonize the plant root and play an important role in inhibiting pathogenic bacteria. In recent years, the Bacillus belgii separated at home and abroad is researched in aspects of plant growth, disease and insect resistance, induction system disease resistance and the like, and a plurality of Bacillus belgii strains play important roles in biological control and serving as grain yield increasing agents. However, no relevant literature report exists on the biological control of the citrullinia cupana by using the bacillus belgii.

Disclosure of Invention

Aiming at the technical situation that the application of the field of Bacillus velezensis in the aspect of biological control of Guaranga aegypti is not recorded in the prior art, the invention aims to provide a new strain with remarkable control effect for the Guaranga aegypti biological control technology, a Bacillus velezensis JTB8-2 strain is separated and screened in the rhizosphere soil of diseased Guaranga aegypti, the strain can be known as a novel Bacillus velezensis strain through classification and system identification and analysis, the application of the novel strain Bacillus velezensis (Bacillus velezensis) in the biological control of Guaranga aegypti can be utilized to obtain remarkable biological control effect, and the novel strain has wide application value in the technical field of microbial strain application.

The purpose of the invention is realized by the following technical scheme:

the technical scheme of the invention is as follows: the method comprises the steps of obtaining a Bacillus velezensis (hereinafter referred to as strain JTB8-2) with the number of JTB8-2 by separating and screening rhizosphere soil of diseased Guaranga aethiopica, determining the strain as a new strain through serial identification, preparing a Bacillus velezensis fermentation stock solution by using a liquid fermentation technology, obtaining a good effect by applying the fermentation stock solution and a fermentation stock solution diluent bacterium to have a remarkable inhibiting effect on the germination of the Guaranga aethiopica seeds, comprehensively solving the effect of the Bacillus velezensis JTB8-2 in the biological control of the Guaranga aethiopica, and being convenient and practical.

Further, the invention carries out gene sequencing on a new strain Bacillus velezensis JTB8-2, the sequence is shown in SEQ ID NO 1 provided after the sequence is attached, and the obtained sequence is compared and analyzed by a common website, so that the result shows that the homology between the 16S rRNA gene sequence of the strain JTB8-2 and Bacillus velezensis JS64Y is the highest and is 97.85 percent, and the genetic relationship between the strain JTB8-2 and Bacillus velezensis JS64Y is the closest. The method comprises the steps of establishing a phylogenetic tree by utilizing MEGA 5.0 software commonly adopted in the field through a Neighbor-Joining method, finding through comparison analysis that the credibility of a strain JTB8-2 and a strain Bacillus velezensis JS64Y from the same branch is 96%, indicating that the strain has extremely high support rate as a new strain, and has excellent stability in the phylogenetic tree, and determining that the strain JTB8-2 is a new species of Bacillus velezensis through serial strain identification, wherein the strain JTB8-2 has the characteristic of clear typicality of the new strain, and temporarily naming the Bacillus velezensis (Bacillus velezensis) JTB8-2 with the strain number of JTB8-2 from the taxonomic point of view.

Specifically, Bacillus belgii (Bacillus velezensis) with the strain number JTB8-2 has been deposited with the International depositary organization for microorganisms under the Budapest treaty: guangdong province culture Collection of microorganisms (GDMCC). Address: china guangzhou city, first furious zhou No. 100, guangdong province microbiology institute, postcode: 510070, with a preservation date of 09 and 4 days 2019 and a deposit number of GDMCC No: 60755.

a strain Bacillus velezensis JTB8-2 is used for culturing a rod-shaped cell on an NA culture medium, and the two ends of the rod-shaped cell are blunt and round. The surface of the single colony is smooth, nearly circular, milky white and rough, the near edge of the single colony is raised to be crater-shaped, and the edge of the single colony is incomplete. The strain can hydrolyze starch and gelatin by using sucrose, D-glucose, sucrose, lactose, maltose, D-mannitol and galactose, but can not hydrolyze cellulose. The physiological and biochemical tests such as catalase, V-P test, nitrate reduction, milk coagulation and peptonization and the like are positive, and the M.R. determination is negative. The salt demand and the salt tolerance test show that the salt concentration is 0 and 5 percent positive, and 10 percent and 20 percent negative. Therefore, the strain JTB8-2 has some differences from the common Bacillus velezensis, and has the distinctive characteristics of some new strains.

Specifically, the invention utilizes a new strain Bacillus licheniformis JTB8-2GDMCC No: the fermentation liquor 60755 has a remarkable inhibiting effect on the germination of the seed of the orobanche japonica, and has a good application prospect in the development of a biocontrol preparation for the orobanche japonica.

Meanwhile, the invention provides fermentation liquor using the Bacillus velezensis JTB8-2 strain.

Furthermore, the new strain Bacillus velezensis JTB8-2GDMCC No: 60755 strain fermentation liquor and 1-500 times of dilution liquor can effectively inhibit the germination of the seed of the orobanche cucurbitacearum.

Preferably, the new strain Bacillus velezensis JTB8-2GDMCC No: the 50-100 times diluted solution of 60755 strain fermentation liquor has extremely obvious effect on inhibiting the germination of the seed of the orobanum cucurbitacearum.

Meanwhile, the new strain Bacillus velezensis JTB8-2GDMCC No: 60755 application in the biological control of the orobanche cucurbitacearum proves that the new strain Bacillus belief (Bacillus velezensis) JTB8-2GDMCC No: 60755 has obvious application effect in the biological control of the orobanche coerulescens.

By implementing the specific technical scheme provided by the invention, the following beneficial effects can be achieved by implementing the content of the invention:

(1) the invention provides a new strain Bacillus velezensis JTB8-2 which can grow on a common NA culture medium, has the characteristics of simple culture condition and quick propagation, and has a rod-shaped cell and two blunt ends on a nutrient agar culture medium. The surface of the single colony is smooth, nearly circular, milky white and rough, the near edge is raised to be crater-shaped, and the edge is incomplete; the strain can hydrolyze starch and gelatin but not cellulose by using sucrose, D-glucose, sucrose, lactose, maltose, D-mannitol and galactose; the test is positive through physiological and biochemical tests such as catalase, V-P test, nitrate reduction, milk coagulation and peptonization, and the like, and the M.R. test is negative; the salt and salt tolerance tests show that the salt concentration is 0 and 5 percent positive, and the salt concentration is 10 percent and 20 percent negative. And MEGA 5.0 software commonly adopted in the field is utilized to establish a phylogenetic tree through a Neighbor-Joining method, the results are compared and analyzed, the credibility that the strain JTB8-2 and the strain Bacillus velezensis JS64Y are from the same branch is 96 percent, the strain JTB8-2 is proved to be a new strain of Bacillus velezensis (Bacillus velezensis) through series strain identification to have the characteristics of extremely high support rate as the new strain and extremely good stability in the phylogenetic tree, and the strain JTB8-2 has the characteristic of bright typicality of the new strain.

(2) The application of Bacillus velezensis JTB8-2 in the biological control of the Liaobang provided by the invention utilizes the separated strain fermentation liquid and fermentation diluent to inhibit the germination of the Liaobang seeds by 100%, reduces the emergence amount of the Liaobang treated by the strain JTB8-2 strain fermentation liquid by 70.5%, reduces the fresh weight of the Liaobang by 66.6%, and has wide application value in the biological control of the Liaobang.

Drawings

FIG. 1 shows the construction of a phylogenetic tree based on the 16SrDNA sequence.

FIG. 2 shows the strain Bacillus licheniformis (Bacillus velezensis) JTB8-2GDMCC No: 60755 crude extract is used for inhibiting germination of seed of herba Orobanches. In the figure, A is the strain Bacillus licheniformis (Bacillus velezensis) JTB8-2GDMCC No: 60755 fermenting liquid is used for treating the seed group of the cucurbiturils, and B is used for treating the seed group of the cucurbiturils by using sterilized water.

Detailed Description

The present invention will be described below by way of examples, but the present invention is not limited to the following examples. All raw and auxiliary materials selected for use in the present invention, as well as methods for culturing the selected bacterial species, are well known and used in the art, and all percentages referred to herein are by weight unless otherwise indicated.

The following basic culture media are adopted in the embodiment of the invention: strain isolation and purification of NA medium: 3g of beef extract, 7g of peptone, 5g of NaCl, 17g of agar, and adding distilled water to 1.0L, wherein the pH value is 7.0; NA liquid medium: 3g of beef extract, 7g of peptone and 5g of NaCl, and distilled water is added until the volume is 1.0L, and the pH value is 7.0.

DNA extraction kit: beijing Ding Guoshang Biotechnology Limited liability company.

The first embodiment is as follows: bacillus velezensis JTB8-2GDMCC No: 60755 separation, screening and identification

Separation and purification:

from the soil sample, 0.1g of the soil sample was weighed out and added to 100mL of sterile water, stirred uniformly with a glass rod, and then left to stand (10 in this case)^-3). Further performing gradient dilution with sterile water from 10^-4、10^-5、10^-6、10^-7150uL of each of the four dilutions was spread on a NA medium plate and incubated in a 28 ℃ incubator for 5 days. And (4) selecting a single colony from the plate, inoculating the single colony into a new separation plate for purification, and repeating the purification for 3-4 times. Inoculating the purified Bacillus belius (Bacillus velezensis) JTB8-2 on a culture medium, culturing for 3-4 days at a constant temperature in an incubator at 28 ℃, and storing at 4 ℃ for later use.

(II) classification and identification:

1. sequencing and analysis of 16S rDNA of Bacillus velezensis JTB8-2 (hereinafter referred to as strain JTB8-2) are carried out:

(1) extraction of PCR template DNA

Inoculating the strain JTB8-2 into an NA culture medium, culturing for 72h at 28 ℃ to obtain bacterial cells, and extracting the genomic DNA by adopting a novel plant genomic DNA rapid extraction kit.

(2) PCR amplification

Primer:

F16S-27:5’-AGAGTTTGATCCTGGCTCAG-3’；

R16S-1492:5’-CGGTTACCTTGTTACGACTTC-3’。

reaction system:

composition of	Volume/ul
		2XMix	25
27F(10μM)	1
		1492R(10μM)	1
Fungus sample template	1
		ddH2O	22

The PCR amplification conditions were: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30s, annealing at 54 ℃ for 30s, extension at 72 ℃ for 1min for 30s, 35 cycles; extension 72 ℃ for 10 min.

(3) Sequence determination

The PCR amplification product is detected by electrophoresis, purified and sequenced, and the sequence is shown as SEQ ID NO. 1. The obtained sequence is compared and analyzed at an NCBI website, and the comparison and analysis show that the homology of the 16S rRNA gene sequence of the strain JTB8-2 and Bacillus _ velezensis JS64Y is the highest and is 97.85 percent, and the genetic relationship of the strain JTB8-2 and Bacillus _ velezensis JS64Y is the closest. The method is characterized in that MEGA 5.0 software commonly adopted in the field is utilized to establish a phylogenetic tree through a Neighbor-Joining method, the phylogenetic tree is shown in an attached figure 1, the result is shown in a comparison analysis, the credibility of the strain JTB8-2 and the strain Bacillus _ velezeissis JS64Y, which are derived from the same branch, is 96 percent, the support rate of the strain serving as a new strain is extremely high, the excellent stability is realized in the phylogenetic tree, the strain JTB8-2 is determined to be a new species of the Bacillus velezensis through the molecular level identification of series strains, the characteristic of the typicality of the new strain is provided, and the strain is temporarily named as the Bacillus velezensis JTB8-2 from the taxonomic angle.

The above-mentioned strain Bacillus velezensis (Bacillus velezensis) JTB8-2 has been deposited with the International depositary organization for microorganisms under the Budapest treaty: guangdong province culture Collection of microorganisms (GDMCC). Address: china guangzhou city, first furious zhou No. 100, guangdong province microbiology institute, postcode: 510070, with a preservation date of 09 and 4 days 2019 and a deposit number of GDMCC No: 60755.

example two: bacillus velezensis JTB8-2GDMCC No: 60755 measurement of physiological and biochemical characteristics

(1) The growth condition research result shows that the growth temperature of the strain JTB8-2 is 15-45 ℃, and the optimal culture temperature is 28 ℃; the most suitable growth medium is an NA medium, can grow within the pH range of 6.0-10.0, and has the most suitable pH of 7.0.

(2) The physiological and biochemical characteristics of the strain JTB8-2 are shown in Table 1, and the strain JTB8-2 is gram-positive, and can hydrolyze starch and gelatin but not cellulose by using sucrose, D-glucose, sucrose, lactose, maltose, D-mannitol and galactose. The physiological and biochemical tests such as catalase, V-P test, nitrate reduction, milk coagulation and peptonization and the like are positive, and the M.R. determination is negative. The salt demand and the salt tolerance test show that the salt concentration is 0 and 5 percent positive, and 10 percent and 20 percent negative. According to the morphological characteristics and physiological and biochemical characteristics of the strain, the strain can be preliminarily identified as Bacillus (Bacillus).

Table 1: measurement of physiological and biochemical Properties

The single colony of the strain JTB8-2 has smooth surface, nearly circular shape, milky white color, rough surface, volcano-mouth-shaped raised near edge, incomplete edge, rod-shaped cells and blunt and round ends.

Example three: bacillus velezensis JTB8-2GDMCC No: 60755 fermentation stock solution has effect of inhibiting seed germination

The purified strain Bacillus velezensis JTB8-2GDMCC No: 60755 single colony is inoculated into NA liquid culture medium, and shake culture is carried out at 28 ℃ and 130 r.min for 24h to serve as seed culture solution. Transferring 5mL of the seed culture solution into a triangular flask filled with 100mL of NA liquid culture medium, and performing shake culture at 28 ℃ for 120h at 130 r.min to obtain a fermentation stock solution for later use. A piece of whatman filter paper (GA/A) was placed in the petri dish, wetted with sterile water, and 20 broomrape seeds were placed on the filter paper, with 4 replicates per treatment. 5mL GR24 and 5mL of broth were added to each dish in sequence, with the positive control treatment being 5mL GR24 and 5mL of sterilized water, and the negative control treatment (NBCK) being 5mL GR24 and 5mL of liquid medium. And (4) culturing for 7-10 days at the room temperature of 25 ℃ under a shading condition, and observing and recording the germination quantity of the broomrape seeds under a body microscope.

Test results show that the germination inhibition rate of the stock solution of the strain JTB8-2 on the seed of the broomrape is 100%.

Table 2: inhibitory effect of strain JTB8-2 fermentation stock solution on germination of seed of orobanche cucurbitacearum

Strain numbering	Germination Rate (%)	Inhibition ratio (%)	Correction inhibition ratio (%)
				JTB8-2	0	100	100
NBCK	85.97
				CK	93.68

Example four: bacillus velezensis JTB8-2GDMCC No: 60755 fermentation diluent has effect of inhibiting seed germination

The purified strain Bacillus velezensis JTB8-2GDMCC No: 60755 single colony is inoculated into NA liquid culture medium, and shake culture is carried out at 28 ℃ and 130 r.min for 24h to serve as seed culture solution. Transferring 5mL of the seed culture solution into a triangular flask containing 100mL of NA liquid culture medium, and performing shake culture at 28 ℃ for 120h at 130 r.min to obtain a fermentation stock solution. The stock solution was diluted with sterilized water to 50-fold, 100-fold, 250-fold and 500-fold, respectively. A piece of whatman filter paper (GA/A) was placed in the petri dish, wetted with sterile water, and 20 broomrape seeds were placed on the filter paper. The experiment set 9 treatments, 50 times liquid, 100 times liquid, 250 times liquid, 500 times liquid of fermentation liquid, 50 times liquid of culture medium dilution, 100 times liquid, 250 times liquid, 500 times and CK, each treatment was repeated 4 times. 5mL GR24 and 5mL broth dilutions were added to each dish in sequence, CK treatment was 5mL GR24 and 5mL sterile water, and negative control treatment was 5mL GR24 and 5mL liquid medium. And (4) culturing for 7-10 days at the room temperature of 25 ℃ under a shading condition, and observing and recording the germination quantity of the broomrape seeds under a body microscope.

Test results show that 50-fold and 100-fold dilution of the strain JTB8-2 has 100 percent and 93.6 percent of corrected inhibition rate on germination of the seed of the Orobanches.

Table 3: comparison of inhibition effects of different dilution concentrations of strain JTB8-2 fermentation liquid on germination of seed of Orobanches schlegelii

Example five: pot culture test for determining control effect of strain JTB8-2 fermentation liquid on cucurbitacin

And (3) adopting a substrate plug seedling, and transplanting the tomato seedlings into a plastic pot when the tomato seedlings have 5-6 leaves. The plastic basin has a diameter of 25cm and a height of 18cm, and has a hole at the bottom. The cultivation soil was a soil mixture (matrix: vermiculite: soil: 1:2) containing about 4kg of soil mixture per pot. Taking out the soil mixture of about 10cm above the middle layer of the plastic pot, putting 50mg (about 10000 granules) of broomrape seeds into a 600mL plastic bottle, filling a small amount of soil, uniformly scattering the seeds in the plastic pot after shaking, transplanting 1 tomato seedling into the plastic pot, and covering soil. The experiment was set up with 2 treatments, respectively a bacterial liquid treatment and a clear water control. Each treatment had 3 replicates per pot, 3 replicates were set. And respectively irrigating 400mL of strain fermentation liquor in each pot 10d, 25d and 40d after the tomato seedlings are transplanted, and irrigating 400mL of clear water in contrast. The potted tomato is placed in an open area after the seedlings of the potted tomato are delayed, and the watering and fertilizing are carried out by adopting a drip irrigation device. After the broomrape comes out of the soil for 15 days, the tomato plants are pulled out, the roots of the tomato plants are carefully washed, the parasitic broomrape quantity of the roots of the tomatoes is investigated, the fresh weight of the broomrape is weighed and recorded, and the variance analysis is carried out by adopting SPASS software.

Test results show that the emergence amount and fresh weight of the broomrape treated by the strain JTB8-2 fermentation liquor are both obviously lower than those of a blank control.

Table 4: comparing the unearthed amount of the orobanche coerulescens in different treatments with the fresh weight of the orobanche coerulescens

Treatment of	Amount of unearthed melon seeds (trunk/pot)	Fresh weight of melon broomrape (gram/basin)
			JTB8-2	2.78^a	15.30^a
CK	9.44^b	45.83^b

The above tests show that the Bacillus subtilis JTB8-2GDMCC No: 60755 has apparent biological control ability of herba Orobanches, utilize strain JTB8-2 strain fermented solution that isolate to process herba Orobanches unearthed quantity to reduce by 70.5%, the fresh weight of herba Orobanches reduces by 66.6%, have extensive using value to the biological control of herba Orobanches.

The above examples are merely illustrative for clearly illustrating the present invention and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications can be made while remaining within the scope of the present invention.

Sequence listing

<110> institute for plant protection of Sinkiang academy of agricultural sciences

<120> Bacillus beiLeisi JTB8-2 and application in biological control of Gualibanum

<160>1

<170>SIPOSequenceListing 1.0

<210>1

<211>1444

<212>DNA

<213> Bacillus belgii (Bacillus velezensis JTB8-2)

<400>1

gggtgctata ctgcaagtcg aggccacaga tgggtgcttg cagggtgatg ttaccgggcg 60

acgggtgagt aacacgtggg taacctgcct gtaagactgg gataactccg ggaaaccggg 120

gctaataccg gatggttgtt tgaaccgcat ggttcagaca taaaaggtgg caacggctac 180

cacttacaca tggacccgcg gcgcattagc tagttggtga ggtaacggct caccaaggcg 240

acgatgcgta gccgacctga gagggtgatc ggccacacac ggactgagac acggggcaga 300

ctcctacggg aggctccagt agggaatctt ccgcaatgga cgaaagtctg acggagcaac 360

gccgcgtgag tgatgaaggt tttcggatcg taaagctctg ttgttaggga agaacaagtg 420

ccgttcaaat agggcggcac cttgacggta cctaaccaga aagccacggc taactacgtg 480

ccagcagccg cggtaatacg taggtggcaa gcgttgtccg gaattattgg gcgtaaaggg 540

ctcgcaggcg gtttcttaag tctgatgtga aagcccccgg ctcaaccggg gagggtcatt 600

ggaaactggg gaacttgagt gcagaagagg agagtggaat tccacgtgta gcggtgaaat 660

gcgtagagat gtggaggaac accagtggcg aaggcgactc tctggtctgt aactgacgct 720

gaggagcgaa agcgtgggga gcgaacagga ttagataccc tggtagtcca cgccgtaaac 780

gatgagtgct aagtgttagg gggtttccgc cccttagtgc tgcagctaac gcattaagca 840

ctccgcctgg ggagtacggt cgcaagactg aaactcaaag gaattgacgg gggcccgcac 900

aagcggtgga gcatgtggtt taattcgaag caacgcgaag aaccttacca ggtcttgaca 960

tcctctgaca atcctagaga taggacgtcc ccttcggggg cagagtgaca ggtggtgcat 1020

ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt 1080

gatcttagtt gccagcattc agttgggcac tctaaggtga ctgccggtga caaaccggag 1140

gaaggtgggg atgacgtcaa atcatcatgc cccttatgac ctgggctaca cacgtgctac 1200

aatggacaga acaaagggca gcgaaaccgc gaggttaagc caatcccaca aatctgttct 1260

cagttcggat cgcagtctgc aactcgactg cgtgaagctg gaatcgctag taatcgcgga 1320

tcagcatgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc acaccacgag 1380

agtttgtaac acccgaagtc ggtgaggttt ggtttatgga gggtgccgcc gaacctgaca 1440

gagt 1444

Claims

1. Bacillus belgii (B.), (B.), (B.beijerinckii)Bacillus velezensis) JTB8-2, wherein said Bacillus belgii (B.beijerinckii) ((B.beijerinckii))Bacillus velezensis) The strain accession number of JTB8-2 is GDMCC No. 60755.

2. The Bacillus belgii of claim 1 (b)Bacillus velezensis) JTB8-2, characterized by the bacterium Bacillus belgii (B.), (Bacillus velezensis) The gene sequence of JTB8-2 is shown in SEQ ID NO. 1.

3. The Bacillus belgii bacterium of any of claims 1 or 2 (b.), (b.) (Bacillus velezensis) Application of JTB8-2 in biocontrol of Guaranga japonica.

4. The Bacillus belgii of claim 3 (b.), (b.,) and (c.,) thereofBacillus velezensis) The application of JTB8-2 in the biocontrol of the cucurbituril seed is characterized in that 1-500 times of diluent of strain JTB8-2 fermentation liquor is used for inhibiting the germination of the cucurbituril seed.

5. The Bacillus belgii of claim 3 (b.), (b.,) and (c.,) thereofBacillus velezensis) JTB8-2 atThe application of the citrulline broomrape in biocontrol is characterized in that 50-100 times of diluent of strain JTB8-2 fermentation liquor is used for inhibiting the germination of the citrulline broomrape seeds.