CN110343641A - A kind of salt bacillus SYW-1 and the application in cotton verticillium wilt prevention and treatment - Google Patents

Abstract

The present invention provide a kind of novel bacterial rhizosphere salt bacillus (Bacillus halotolerans) SYW-1 CGMCC No.15008 and its application in Biological Control of Cotton Verticillium Wilt.The present invention by separated in Cotton rhizosphere soil one plant of rhizosphere salt bacillus (Bacillus halotolerans) SYW-1, the features such as growth temperature isolated is wide, easy culture, cotton seeds are impregnated using SYW-1 bacteria suspension, it can promote cotton seeds budding, SYW-1 bacteria suspension is applied to many years continuous cropping grave illness cotton field, the effect of the cotton verticillium wilt of cotton development stage can be effectively prevented, adverse effect of the cotton verticillium wilt to Cotton Production is reduced to bottom line, be conducive to improve the yield and quality of cotton, with significant and stable technical effect, as the value being applicable in Biological Control of Cotton Verticillium Wilt with extensive.

Description

A kind of halobacillus SYW-1 and its application in the control of cotton verticillium wilt

technical field

The invention relates to the technical field of microbial strains and applications thereof, in particular to the technical field of a halobacillus SYW-1 and its application in the biological control of cotton verticillium wilt.

Background technique

A large number of bacteria exist in and around plant roots, some of which are rhizobacteria (Plant growth promoting rhizobacteria, PGPR) that can promote plant growth. Certain rhizosphere bacteria can inhibit plant soil-borne diseases and promote plant growth by competing, producing antibiotics, synthesizing siderophores, and inducing plant systemic resistance. In recent years, there have been reports on the biological control of cotton blight using rhizosphere bacteria. Trichoderma, Gliocladium, etc. have good biocontrol effects on cotton seedling diseases.

Cotton rhizosphere microorganisms are an important part of the cotton micro-ecosystem. Cotton rhizosphere microorganisms refer to soil microorganisms close to cotton roots, which are different from the surrounding soil in terms of physical, chemical and biological characteristics, and closely related to cotton. Li Jiashu and Wang Zunong found through isolation that the closer the rhizosphere bacteria are to the root, the greater the number, the growth period first increased and then decreased, and the flowering period was the highest. Zhang Yaping used the method of isolation and culture to study the microorganisms in the rhizosphere of cotton in northern Xinjiang, and screened out the antagonistic bacteria of cotton blight. Research by Zhang Fenghua and others pointed out that the impact of water treatment on the number of rhizosphere microorganisms in cotton field is more obvious than that of fertilization. The number of suitable water and fertilizer treatments is the largest during the flowering and boll stage, while the maximum amount of water and high fertilizer is during the boll opening period. Gu Meiying and others found that the total amount of soil microorganisms in the rhizosphere soil of Verticillium dahliae plants in Xinjiang was greater than that of healthy plants through isolation, especially the number of fungi increased significantly. Shen Qirong et al. used microbial organic fertilizers to regulate the microbial community structure of the rhizosphere soil, changing the flora of soil microorganisms, greatly increasing the number of bacteria, and significantly reducing the number of fungi.

The antagonistic bacteria isolated and screened from the cotton rhizosphere are suitable for colonization in the cotton rhizosphere, and the application effect of the antagonistic bacteria is more stable. Therefore, the use of cotton rhizosphere isolation to screen microbial strains suitable for the control of cotton verticillium wilt, the development of efficient microbial agents, and the solution to the micro-ecological balance between cotton plants and the environment have important practical value for the prevention and treatment of cotton diseases and the improvement of cotton yield. .

Contents of the invention

In view of the fact that there is no report on the use of Bacillus halotolerans in the prior art for the biological control of cotton verticillium wilt, the present invention provides a new strain of Bacillus halotolerans (Bacillus halotolerans) SYW -1CGMCC No.15008 and its application in biological control of Verticillium wilt in cotton. The present invention isolates a batch of rhizosphere antagonistic bacteria strains in the cotton rhizosphere soil, and screens out a strain of Bacillus halotolerans (Bacillus halotolerans) SYW-1CGMCC No.15008 strain from them. Cultivation and other characteristics, using SYW-1 bacterial suspension to soak cotton seeds can promote cotton seed germination, and SYW-1 bacterial suspension can be applied to cotton fields with continuous cropping and severe diseases for many years, which can effectively prevent and control cotton Verticillium wilt during the cotton growth period. Minimizing the adverse effects of cotton Verticillium wilt on cotton production is conducive to improving the yield and quality of cotton, has significant and stable technical effects, and has extensive and applicable value as a biological control of cotton Verticillium wilt.

Main technical scheme that the present invention adopts:

The invention specifically provides a new strain of Bacillus halotoler ans SYW-1CGMCC No.15008. By screening and isolating a strain of Bacillus halotolerans (Bacillus halotolerans) numbered SYW-1 from the cotton rhizosphere soil, the Bacillus halotolerans (Bacillus halotolerans) SYW-1 was identified through the current common strains It was learned by means that it is a new strain, and the isolated strain has the characteristics of wide growth temperature and easy cultivation, etc. The bacterial suspension of this strain soaks cotton seeds, which can promote the germination of cotton seeds. The effect of preventing and controlling cotton verticillium wilt in the cotton growth period can minimize the adverse effects of cotton verticillium wilt on cotton production, which is conducive to improving the yield and quality of cotton, and has a significant and stable technical effect. It has a distinct application value orientation.

Specifically, by cultivating and separating microbial strains from cotton rhizosphere soil, a batch of excellent strains were screened, and a strain of Bacillus halotolerans (Bacillus halotolerans) strain numbered SYW-1 was isolated and screened therefrom, According to microbiological classification and identification, it belongs to the genus Halobacillus rhizosphere. The optimal growth conditions for this strain are: the growth temperature range is 8-42°C, and the optimum growth temperature is 30°C; the pH growth range is 4.9-11.2, and the medium is pancreas Peptone Soy Agar Medium. Referring to the ninth edition of "Bergey's Systematic Bacteriological Identification Manual" and "Common Bacterial System Identification Manual", the morphological, physiological and biochemical tests were carried out on the strain numbered SYW-1, and it was determined that the SYW-1 strain was a member of the genus Halobacillus rhizosphere. Members of the genus Rhizobacillus, but have different characteristics from the common members of the genus Rhizobacillus, and have some characteristics of new strains.

Further, the present invention conducts gene sequencing on the above-mentioned bacterial species numbered SYW-1, and the sequence is shown in the attached SEQ ID NO: 1. The obtained sequences are often compared and analyzed on websites, and a phylogenetic tree is constructed. The analysis found that the 16S rDNA sequence of the strain SYW-1 had the highest homology with the standard model Bacillus halotolerans strain DSM 8802 ^T , with a sequence similarity of 96.01%. The phylogenetic tree of 8802 ^T is the closest, and the MEGA 5.0 software commonly used in this field was used to establish a phylogenetic tree through the Neighbor-Joining method. After comparison and analysis, it was found that the strain SYW-1 and Bacillus halotolerans ATCC 25096 ^T were in the smallest branch, and it was its Similar species, with a confidence value of 89, indicating that the strain has a high support rate as a new strain and shows good stability in the phylogenetic tree. The strain SYW-1 is determined to be Bacillus halotolerans after molecular identification of a series of strains The new species has the typical characteristics of new species, and it is tentatively named Bacillus halotolerans SYW-1 from the perspective of taxonomy.

Specifically, the strain SYW-1 has been deposited in the International Depository of Microorganisms under the Budapest Treaty: the Center for General Microorganisms of the China Committee for the Collection of Microorganisms (CGMC C). Address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Courtyard, Beichen West Road, Chaoyang District, Beijing, 100101. The preservation date was December 4, 2017, and the strain preservation number was CGMCCNo.15008. After microbiological identification, it was tentatively named Bacillus halotolerans SYW-1.

The new strain Bacillus halotolerans (Bacillus halotolerans) SYW-1 grows well on the tryptone soybean agar medium. After culturing at 30°C for 48 hours, visible colonies are formed. The colonies of this strain are 2-4mm smooth and semi Transparent, milky white, moist surface, neat edges, fast growth, Gram-positive bacteria; the optimum growth conditions for this strain are: the growth temperature range is 8-42°C, and the optimum growth temperature is 30°C; the pH growth range is 4. 9-11.2.

Further, the present invention provides the application of Bacillus halotolerans (Bacillus halotolerans) S YW-1CGMCCNo.15008 in promoting the germination of cotton seeds, which has the effect of promoting the germination of cotton seeds and preventing and controlling various types of cotton verticillium wilt in the growth period of cotton. Significant and stable technical effect, with a wide range of application value.

Provide above-mentioned specific technical scheme by implementing the present invention, implement content of the present invention, can reach following beneficial effect:

(1) Bacillus halotolerans (Bacillus halotolerans) SY W-1CGMCCNo.15008 provided by the present invention is a typical new bacterial species, and the optimal growth conditions of the bacterial strain are: the growth temperature range is 8-42 °C, the optimum The growth temperature is 30°C; the pH growth range is 4.9-11.2. Taking advantage of the characteristics of wide growth temperature and easy cultivation of the isolated strain, the bacterial suspension of this strain can promote the germination of cotton seeds, and can effectively prevent cotton in the growth period. Verticillium wilt, it is designated as a new species of Bacillus halotolerans (Bacillus h alotolerans), which has the typical characteristics of a new species, and is tentatively named Bacillus h alotolerans SYW-1 from a taxonomic point of view.

(2) The Bacillus halotolera ns SYW-1CGMCC No.15008 isolated and screened by the present invention is placed in a culture medium with corn steep liquor and yeast extract as main raw materials, and the fermentation conditions are: temperature 32°C, rotation speed 200rpm , aeration 1:0.3, cultured for 24 hours to obtain a bacterial suspension; soaking cotton seeds with 1×10 ⁹ cfu/mL bacterial suspension can promote germination, uniform seedling emergence, and thick leaf color, and field application of SYW-1 for years of continuous cropping of severely diseased cotton The bacterial suspension is 5000mL, the concentration is 7×10 ⁸ cfu/mL, and the control effect is 69.9%. It can effectively prevent and control cotton verticillium wilt in the cotton growth period, and minimize the adverse effects of cotton verticillium wilt on cotton production. , is conducive to improving the yield and quality of cotton, has significant and stable technical effects, and has extensive and applicable value as the biological control of cotton verticillium wilt.

Description of drawings

Figure 1 shows the colony morphology of Bacillus halotolerans SYW-1C GMCC No.15008.

Figure 2 shows the phylogenetic tree of Bacillus halotolerans SYW-1C GMCC No.15008.

Figure 3 shows the growth curve of Bacillus halotolerans SYW-1CG MCC No.15008.

Detailed ways

The following examples illustrate the present invention, but the present invention is not limited to the following examples. All the raw and auxiliary materials selected in the present invention, and the strain culture method selected are well-known in the art and selected, involving 0ml in the present invention; activation medium: peptone 10g, beef powder 3g, sodium chloride 5g, agar 15g, Distilled water 1000mL, pH7.3±0.1; seed medium: peptone 10g, beef extract powder 3g, sodium chloride 5g, distilled water 1000mL, pH7.2±0.2.

Main reagents involved in the present invention: glycerin, beef extract, yeast extract, sodium chloride, sodium hydroxide, glucose, magnesium sulfate, zinc sulfate, ferrous sulfate, sodium dihydrogen phosphate, disodium hydrogen phosphate, dihydrogen phosphate Potassium, agar, sucrose, sodium hypochlorite, PCR master mix (TaKaRa Biotechnology), and other reagents were of analytical grade.

Main instruments and reagents: TG328A analytical balance, AB204-N electronic balance, PHS-3TC (0.01 level) precision digital pH meter, 101-2 drying oven, UV-21 00 spectrophotometer, HYG-Ⅱ rotary type Shaker cabinet with constant temperature and speed regulation, LS-B50L vertical pressure steam sterilizer, ultra-clean workbench, 3K15 desktop refrigerated centrifuge.

Example 1: Isolation, screening and identification of Bacillus halotolerans SYW-1

(1) Separation and screening:

Several strains of bacteria were isolated from the cotton rhizosphere soil by gradient dilution method, and the plate confrontation method was used to judge whether they had antagonistic effect according to the presence or absence of the inhibition zone, and the strains with certain antagonistic ability to the cotton Verticillium dahliae pathogen were preliminarily screened out. Further tests are carried out on the strains with antagonistic effect, and the strains with stronger antagonistic ability are screened out for subsequent tests.

A batch of rhizosphere microbial strains were obtained by isolating, screening and culturing in cotton rhizosphere soil, and a strain numbered SYW-1 was screened out, which belonged to Bacillushalotolerans of the genus Bacillus through microbiological classification and identification.

(2) Classification and identification:

1. Physiological and biochemical assays

(1) The results of the growth condition study showed that Bacillus hal otolerans (Bacillus hal otolerans) SYW-1 is a Gram-positive bacterium. It grows well on tryptone soybean agar medium. After culturing at 30°C for 48 hours, visible colonies are formed. The colonies of this strain are 2-4mm smooth, translucent, milky white, moist on the surface, with neat edges, and grow fast. See As shown in Figure 1; the temperature range of bacterial growth is 8-42°C, and the optimum growth temperature is 30°C; the pH growth range is 4.9-11.2.

(2) The main nitrogen sources include but not limited to peptone, yeast powder; the main carbon sources used include but not limited to sucrose, mannose, xylose, galactose, pectin sugar; the inorganic components used include but not limited to chlorinated Potassium, Sodium Chloride, Monosodium Phosphate, Dipotassium Phosphate, Calcium Chloride Dihydrate, Magnesium Sulfate Heptahydrate, Ferrous Sulfate Heptahydrate.

(3) Through the morphological determination of the SYW-1 strain, and detection of the shape, size, physiological and biochemical reactions of the strain SYW-1, the biological characteristics of the SYW-1 strain of the present invention are shown in Table 1.

Table 1: Physiological and biochemical characteristics of strain SYW-1

project result project result glucose + Galactosidase + pectin sugar + Amylase + Xylose + citrate utilization + Sorbitol + dihydroxypyruvate production + Melibiose – Lysine decarboxylase + lactose – arginine hydrolase + Raffinose + Ornithine decarboxylase + Trehalose + 45°C +

Note: "+" means positive; "-" means negative.

Observation of bacterial morphology and culture characteristics and determination of physiological and biochemical indicators of Bacillus halotolerans SYW-1 in the upper rhizosphere: observation of bacterial morphology, observation of culture characteristics of strains, determination of aerobicity and motility, determination of growth temperature, Salt tolerance test, citrate utilization test, catalase test, sugar fermentation test, nitrate reduction test, and starch hydrolysis are all carried out with reference to the methods in the "Common Bacterial System Identification Manual".

2. Sequence determination and analysis of Bacillus halotolerans SYW-116S rDNA:

(1) Pick a small amount of single colonies of SYW-1 bacteria, put them into an EP tube filled with 25 μL of sterile water, boil at 100°C for 8-10 minutes, and then quickly put them into the ice-water mixture for 5 minutes. Centrifuge at 10000r/min for 5min, store at 4°C, and take the supernatant when used. 16SrRNA gene sequence determination and phylogenetic tree construction: extract the total DNA of bacterial strains according to conventional methods,

(2) PCR amplification of 16S rDNA was carried out using specific primers, and the primers were designed as follows:

PA: 5'-AGAGTTTGATCCTGGCTCAG-3';

PB: 5'-AAGGAGGTGATCCAGCCGCA-3'.

The total volume of the PCR reaction system is 50 μl, 5 μl of 10×PCR buffer, 20 pmol of each primer, 1 μl of template DNA (1000 ng/μl), 0.5 U of TaqTM, and 8 μl of dNTP. PCR amplification conditions: first pre-denature at 94°C for 2 minutes, then cycle 30 times at 98°C for 10 seconds, 55°C for 30 seconds, and 72°C for 1.5 minutes, and finally extend at 72°C for 10 minutes. Purification of PCR products: Take 8 μl of PCR products, electrophoresis in 1% agarose gel, use TaKaRa PCR Fragment Recovery Kit to recover the target fragment from the gel, and dissolve it in 20 μl of high-purity water. For the PCR product, PA(+) and PB(-) were used as sequencing primers to determine the 16S rDNA sequence, and the sequence is shown in the attached SEQ ID NO:1.

(3) Perform BLAST analysis on the 16S rDNA sequence obtained by sequencing and the nucleotide sequence in the GenBank database to obtain similar 16S rDNA sequences, and construct a phylogenetic tree with Clustal X software and MEGA 5.2 Neighbor-joining method, see the attached As shown in Figure 2, it was found through comparative analysis that the 16S rDNA sequence of the strain SYW-1 had the highest homology with the standard model Bacillus halotolerans strain DSM 8802 ^T , with a sequence similarity of 96.01%. The Nei ghbor-Joining method was used to establish a phylogenetic tree. After comparative analysis, it was found that the strain SY W-1 and Bacillus halotolerans ATCC 25096 ^T were in the smallest branch, and they were similar species with a confidence value of 89, indicating that the strain SY W-1 was a new strain. The possibility is higher, and it shows better stability in the phylogenetic tree. The strain SY W-1 can be determined as a new species of Bacillus halotolerans after a series of identifications at the molecular level. Typical characteristics, tentatively named from the taxonomic point of view as Bacillus halotolerans (Bacill ushalotolerans) SYW-1.

Comprehensive 16SrDNA sequence analysis, phylogenetic analysis, and microbiological characteristic analysis results show that the rhizosphere halotolerans (Bacillus halotolerans) SY W-1 provided by the present invention is a new strain of bacteria of the rhizosphere halotolerans genus. It was named as Bacillus halotolerans SYW-1. The strain was deposited in the General Microorganism Center (CGMC C) of the China Committee for the Collection of Microbial Cultures on December 4, 2017. Address: Datun Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, preservation number is CGMCC No.15008.

Embodiment two: the growth factor of cotton rhizosphere salt bacillus (Bacillus halotolerans) SYW-1

According to the characteristics of cotton rhizosphere bacillus halotoler ans (Bacillus halotoler ans) SYW-1CGMCC No.15008 provided by the foregoing examples, its specific growth factors are determined, bacterial strain SYW-1 is inoculated and cultivated, and the effects of temperature, pH, salt, and antibiotics on bacterial strain SYW are determined. -1 growth, the results are shown in Table 2.

Table 2: Effects of temperature, pH, salt, and antibiotics on the growth of strain SYW-1

Growth curve and culture conditions. The rhizosphere SYW-1 bacteria were cultured in the following manner. The culture conditions were: TSA solid medium, pH 7.0, cultured at 34°C for 28 hours. The growth curve is shown in Figure 3. From the above results, it can be concluded that the optimal culture time of SYW-1 as a seed is 20 hours, the optimum growth pH is 7.0, and the time for mass production of spores is 24 hours.

According to the above growth factors and physiological, physical and chemical experimental data for the rhizosphere SYW-1 bacteria, it can be known that the rhizosphere SYW-1 bacteria are completely different from the common Bacillus halotolerans, and compared with the common rhizosphere Bacillus halotolerans It has the attributes and characteristics of new strains.

Embodiment three: the fermentation process research of cotton rhizosphere halobacterium (Bacillus halotolerans) SYW-1

Inoculate the SYW-1 strain on the activation medium and culture it at 30°C for 2 days. According to a certain amount of inoculum, insert it into a 500mL Erlenmeyer flask containing 100mL seed medium, and culture it in a shaker flask at 30°C and 180rmp. 1d, then inoculate 2% of the inoculum into a 500mL Erlenmeyer flask containing 100mL of fermentation medium, and culture in a shaker flask for 3d under the conditions of 30°C and 180rmp; through the optimization of carbon, nitrogen source, pH value, etc. to achieve fermentation The purpose of SYW-1 bacterial suspension. Determination of OD value: Take 0.5mL of fermentation broth, add 1mL of 2.5M HCl, add 13.5mL of deionized water, mix well, and use a spectrophotometer to measure at a wavelength of 600nm.

(1) Determination of medium pH

According to the pH value of 6.0, 6.5, 7.0, 7.5, 8.0, ferment at 200r/min and temperature of 30°C, the optimum growth pH value is 7.0.

(2) Carbon source optimization

Continue to optimize the carbon source on the basis of the seed medium, the carbon source is glucose, sucrose, and corn steep liquor, and the other components are KH ₂ PO ₄ 0.1%, K ₂ HPO ₄ 0.1%, MgSO ₄ 7H ₂ O 0.05%, pH7 .0; all experiments were repeated three times. After culturing for 3 days, the OD value of the bacterial suspension was measured, and the test results are shown in Table 3.

Table 3: Effects of different carbon sources in fermentation medium on SYW-1 strain

(3) Optimization of nitrogen source

Continue to optimize the nitrogen source on the basis of the seed medium. The nitrogen source is peptone, yeast extract, (NH ₄ ) ₂ SO ₄ , and the other components are KH ₂ PO ₄ 0.1%, K ₂ HPO ₄ 0.1%, MgS O ₄ · 7H ₂ O 0.05%, pH 7.0. The experiments were repeated three times, and the optimal nitrogen source of the strain was obtained by detecting the OD value of different nitrogen sources. The test results are shown in Table 4.

Table 4: Effects of different nitrogen sources in fermentation medium on SYW-1 strain

According to the test results in Table 3 and Table 4, it is determined that the best carbon source is corn steep liquor, and the best nitrogen source is yeast extract.

Embodiment four: cotton rhizosphere salt bacillus (Bacillus halotolerans) SYW-1 fermentation tank fermentation test

In the culture medium with corn steep liquor and yeast extract as the main raw materials, the fermentation conditions are: temperature 32°C, rotation speed 200rpm, aeration 1:0.3, time 24h. The results are shown in Table 5 below:

Table 5: Fermentation test results of Bacillus halotolerans (Bacillus halotolerans) SYW-110L

strain fermentation time Dilution factor OD(600nm) SYW-1 48h 50 2.825

After the fermentation, the plate counting results showed that all the SYW-1 bacteria fermentation liquid survived, and after freeze-drying treatment, the bacteria body survival rate was higher than 90%.

Embodiment five: the effect of cotton rhizosphere salt bacillus (Bacillus halotolerans) SYW-1 on cotton seed germination

(1) Seed soaking germination test

Seeds were soaked with 10 ⁹ cfu/mL bacterial suspension for 3 minutes, placed in a petri dish lined with wet filter paper, incubated at 28°C for 3 days, 5 days, and 7 days, and the germination rate was detected. Seeds soaked in water were used as a control.

(2) Effect of antagonistic bacteria on cotton seed germination

The test results are shown in Table 6. The germination rate of cotton seeds treated with SYW-1 bacterial suspension for 3 days, 5 days and 7 days was significantly higher than that of the control group. It shows that a certain concentration of SYW-1 bacterial suspension can promote the germination of cotton seeds, and the cotton seeds soaked in SYW-1 bacterial suspension have early emergence, uniform emergence and thick leaf color.

Table 6: Effect of SYW-1 bacterial suspension soaking on cotton seedling emergence

Note: Capital letters indicate significant differences at P=0.01, respectively.

Embodiment six: the field biocontrol experiment of cotton rhizosphere salt bacillus (Bacillus halotolerans) SYW-1

(1) Determination of field control effect

From 2016 to 2018, field control efficacy testing was carried out in Bole, Xinjiang for three consecutive years. Select a cotton field that has been continuously cropped for many years, drip irrigation with water for 13 hours with an antagonistic antibacterial solution after sowing the cotton, and divide it into two treatments at the seedling stage and bud stage after emergence, and apply Bacillus halotol erans SYW-1 bacterial suspension per mu 5000mL, concentration 7×10 ⁸ cfu/mL, water treatment was used as control, each treatment was repeated 3 times, the area of each repetition was 6667m ² , and the control was scattered among the treatment groups. A survey was conducted in late August to record the number of diseased plants at all levels and calculate the disease index.

The investigation began in late August, and the disease was recorded according to the general 5-level standard, namely: level 0: healthy plant, no symptoms; level 1: 1 to 2 cotyledons with disease; level 2: 1 true leaf with disease; level 3: 2 More than one true leaf becomes diseased or falls off and only the heart leaf remains; Grade 4: The growing point of the plant or the whole plant dies. The disease index was calculated based on the survey results on the 25th day after inoculation, and the resistant and susceptible types of varieties were divided.

(2) Determination of field control effect test

The results in Table 7 show that Bacillus halotolerans (Bacillus halotolerans) SYW-1 can reduce the disease index of cotton Verticillium wilt, and the control effect is 69.9%. SYW-1 strain has a good control effect on the main cotton Verticillium wilt. There were significant differences between the strain SYW-1 and the control.

Table 7: Field test control effect of SYW-1 bacterial suspension on major cotton verticillium wilt

The Bacillus halotoler ans (Bacillus halotoler ans) SYW-1CGMCC No.15008 provided by the above-mentioned series of examples is a typical new bacterial species, and the optimal growth condition of the bacterial strain is: 8-42°C, and the optimal growth temperature is 30°C; pH growth range 4.9-11. 2, taking advantage of the characteristics of the isolated strains such as wide growth temperature and easy cultivation, soaking cotton seeds with SYW-1 bacterial suspension can promote cotton seed germination, SYW-1 bacterial suspension Applied to cotton fields with severe diseases in continuous cropping for many years, it can effectively prevent and control the cotton verticillium wilt in the cotton growth period, minimize the adverse effects of cotton verticillium wilt on cotton production, and help improve the yield and quality of cotton. And stable technical effect, as the biological control of Verticillium wilt in cotton has extensive and applicable value.

The above-mentioned embodiments are only examples for clearly illustrating the present invention, rather than limiting the implementation. For those of ordinary skill in the art, other changes or changes in different forms can be made on the basis of the above description. It is not necessary and impossible to exhaustively list all the implementation manners here. However, the obvious changes or changes extended therefrom still fall within the scope of protection of the present invention.

sequence listing

<110> Institute of Applied Microbiology, Xinjiang Academy of Agricultural Sciences (China Xinjiang-Armenia Bioengineering Research and Development Center)

<120> A kind of halobacillus SYW-1 and its application in the control of cotton verticillium wilt

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1091

<212>DNA

<213> Bacillus halotolerans

<400> 1

aggagcgtgg gctgcctaat catgcaagtc cgagcggaca gatgggagct tgctccctga 60

tgttagcggc ggacgggtga gtaacacgtg ggtaacctgc ctgtaagact gggataactc 120

cgggaaaccg gggctaatac cggatgcttg tttgaaccgc atggatcaaa cataaaaggt 180

ggcttcggct accacttaca gatggacccg cggcgcatta gctagttggt gaggtaatgg 240

ctcaccaagg caacgatgcg tagccgacct gagagggtga tcggccacac tgggactgag 300

acacggccca gactcctacg ggaggcagca gtagggaatc ttccgcaatg gacgaaagtc 360

tgacggatca acgccgcgtg agtgatgaag gttttcggat cgtaaagctc tgttgttagg 420

gaagaacaag taccgttcca atagggcggc accttgacgg tacctaacca gaaagccacg 480

gctaactacg tgccagcagc cgcggtaata cgtaggtggc aagcgttgtc cggaattatt 540

gggcgtaaag ggctcgcagg cggttcctta attctgatgt gaaagccccc ggctcaaccg 600

gggagggtca ttggaaactg gggaacttga gtgcagaaga ggagagtgga attccacgtg 660

tagcggtgaa atgcctagag atgtggagga acaccagtgg cgaatgcgac tctctggtct 720

gtaactgacg ctgaggagcg aaagcgtgag gagcgaacac gattatatac cctggtagtc 780

cacgcccgta aacgatgagt gctaagtgtt agggggtttc cgccccttac tgctgcagct 840

cacgcattac acactccgcc tggggagtac ggtcgcgaga ttgaaactca aaggaattga 900

cgggggcccg cacatgcggt ggagtatgtg gtttaaattc gaagcaacgc gaagaacctt 960

accagtcct ggacatcctc tgacattcgt agatgatgga cgtccccttt caggggcaga 1020

ctgatcatgg ggacgcatgt tggttcgttc agctacgtgg acttcttagt tgacctatcc 1080

attcagattg c 1091

Claims (4)

1. a kind of rhizosphere salt bacillus (Bacillus halotolerans) SYW-1, which is characterized in that the rhizosphere salt Bacillus (Bacillus halotolerans) SYW-1 culture presevation number be CGMCC No.15008.

2. rhizosphere salt bacillus as described in claim 1 (Bacillus halotolerans) SYW-1 gene order such as Shown in SEQ ID NO:1.

3. rhizosphere salt bacillus as claimed in claim 1 or 2 (Bacillus halotolerans) SYW-1 prevention and treatment cotton Spend the application in the cotton verticillium wilt of breeding time.

4. rhizosphere salt bacillus as claimed in claim 1 or 2 (Bacillus halotolerans) SYW-1 promote cotton Application in flower seed budding.