CN112662591B

CN112662591B - Microbial preparation

Info

Publication number: CN112662591B
Application number: CN202110081087.XA
Authority: CN
Inventors: 王晓莉; 祁红英; 王云鹏; 袁志香; 史雨琪; 张弯弯; 颜歆雯
Original assignee: Huaian Chaimihe Agriculture Technology Co ltd; Huaiyin Institute of Technology
Current assignee: Huaian Chaimihe Agriculture Technology Co ltd; Huaiyin Institute of Technology
Priority date: 2019-09-29
Filing date: 2019-09-29
Publication date: 2023-01-20
Anticipated expiration: 2039-09-29
Also published as: CN110669688B; CN112640916B; CN110669688A; CN112662591A; CN112760254B; CN112825864A; CN112646752B; CN112646752A; CN112825864B; CN112640916A; CN112760254A

Abstract

The invention relates to the field of agricultural microorganisms and discloses a microbial preparation which comprises an adsorbent and Paenibacillus jimela (Bacillus jimela)Paenibacillus jamilae) W51; wherein the total viable bacteria concentration of Paenibacillus jimmara W51 is 1 x 10 ⁸ ~1×10 ⁹ CFU/mL. The Paenibacillus jimira is preserved in the common microorganism center of China Committee for culture Collection of microorganisms at 26.26.10.2018, the preservation address is No. 3 of West Lu No.1 of Beijing, chaoyang, and the strain preservation number is CGMCC No.16639 at the institute of microorganism of Chinese academy of sciences. The microbial preparation has a good effect on preventing and treating tomato bacterial wilt, and the prevention effect reaches 79.35 percent.

Description

Microbial preparation

Description of the cases

The invention is a divisional application with application date of 2019, 9 and 29, application number of 2019109339644, and invention name of Paenibacillus jimila, microbial preparation thereof, preparation method and application thereof.

Technical Field

The invention relates to the field of agricultural microorganisms, and particularly relates to a microbial preparation.

Background

The tomato is one of the most important vegetable planting crops in the production and planting system of China, the production scale of the tomato is continuously enlarged in recent years, the first disease causing the yield reduction of the tomato is tomato bacterial wilt in the cultivation and planting process, and the pathogenic bacteria is solanaceous Lawsonia (L) in the solanaceaeRalstonia solanacearum). The bacterial wilt can invade and hide from the skin holes or wounds of the roots and stem bases of the tomatoes by means of rainwater, irrigation water, underground insect pests or operating tools, and the like, quickly propagate in the vascular bundles when the conditions are proper, and expand upwards along the conduit to invade adjacent parenchyma cell tissues, so that the conduit is blocked, and the plants cannot obtain the corresponding water and nutrition supply and wither and die. Because the disease is extremely high in infectivity, pathogenic bacteria mainly complete overwintering and infection in soil, and the pathogenic mode of the pathogenic bacteria in a plant vascular bundle makes the traditional prevention and control means untreatable, so that the disease is called as 'plant cancer'. The south of the Yangtze river in China, particularly the south of China, is a serious disaster area of the disease, once the disease is difficult to control, large-area withering and death of tomato plants are often caused, the yield of a serious disease field is seriously reduced, even the disease is completely lost, and the development of the tomato industry and the improvement of economic benefit are seriously restricted by the disease.

At present, a plurality of reports on the control of the tomato bacterial wilt exist, such as pesticide irrigation, crop rotation, resistant breeding and the like, but no stable and effective control measures exist. In recent years, more and more researchers begin to search for breakthrough in biological control, such as pseudomonas, bacillus, fungi, phage and the like, and since bacillus is an antagonistic bacterium of various pathogenic bacteria, the bacillus can prevent and control crop diseases and is generally considered to be an environment-friendly, economic and effective disease control way. However, although the current biological control preparations used at home and abroad have made a certain research progress on the control of tomato bacterial wilt, most biological control preparations are still in the experimental stage, and have many problems in practical application, such as unsatisfactory field control effect, short control time, low colonization ability of biological control bacteria, large environmental influence factors and the like.

Thus, suitable carrier materials are needed to enhance their adaptability and effectiveness. Clay minerals such as illite and the like are the main minerals constituting claystone and soil, and the large specific surface area, porous structure and strong adsorption property of clay particles have attracted increasing interest to researchers and researchers. Illite is a common clay mineral, and mainly comprises illite, and also contains a small amount of kaolinite, montmorillonite, chlorite, pyrophyllite and the like. Illite crystal structure belongs to 2: dioctahedral type with 1-type unit layers containing a large amount of metal cations such as Fe ²⁺ 、Mg ²⁺ 、Si ²⁺ Etc., can be bound to peptidoglycan on the surface of bacterial cell walls. In addition, the characteristics of large specific surface area of the illite crystal also enable the illite crystal to have good electrostatic adsorption performance, the characteristics enable the illite to gather bacterial thalli in water to form granular substances rich in the thalli, and most bacilli can form a bacterial membrane when being enriched, so that the thalli have higher survival rate in a poor environment than single thalli. In production and application, the survival rate of the biocontrol bacteria around the plant root system directly influences the biocontrol effect, so that the thallus adsorbed by the illite has higher survival rate compared with a single thallus, thereby having higher biological control effect.

The method has the advantages that the illite is utilized to adsorb the biocontrol bacteria to prepare the microbial preparation, and the microbial preparation is used for preventing and treating the tomato bacterial wilt, so that the survival rate of the biocontrol bacteria in the natural environment is expected to be improved, the colonization capability of the biocontrol bacteria around the tomato root system is improved, and the tomato disease influence caused by the bacterial wilt infection is prevented or alleviated. Therefore, the microbial preparation has considerable production and application prospects.

Disclosure of Invention

The purpose of the invention is as follows: aiming at the problems in the prior art, the invention provides a microbial preparation which has a high function of preventing tomato bacterial wilt.

The technical scheme is as follows: the invention provides Paenibacillus jimela, which is named by classificationPaenibacillus jamilaeW51 is preserved in China general microbiological culture Collection center in 2018, 10 months and 26 days, wherein the preservation address is No. 3 of West Lu No.1 of North Chen of the Korean-Yang district in Beijing, and the strain preservation number is CGMCC No.16639 at the institute of microbiology of China academy of sciences.

The invention also provides application of Paenibacillus jimila in preventing and treating tomato bacterial wilt.

The invention also provides a microbial preparation, which comprises an adsorbent and Paenibacillus jimila W51; wherein the total viable bacteria concentration of Paenibacillus jimela W51 is 1 × 10 ⁸ ~1×10 ⁹ CFU/mL。

Preferably, the adsorbent is i Li Danfen.

Preferably, the concentration of said i Li Danfen in said microbial preparation is 4%.

Preferably, the particle size of the illite powder is less than 2 μm, and the natural whiteness is greater than 85%.

The invention also provides a preparation method of the microbial preparation, which comprises the following steps: s1: inoculating seed bacteria liquid of Paenibacillus jimila W51 into a PDB fermentation culture medium according to the volume ratio of 1; the conditions of fermentation culture are as follows: carrying out shaking culture at 32 ℃ and 200-220 rpm for 18-32 h; s2: centrifuging at 4000 to 6000 rpm for 25 to 30min, and taking the precipitate to be re-suspended by using sterilized water so that the OD value range of the prepared bacterial suspension at 600 nm is 0.8 to 1.0; s3: adding 4% of sterilized illite powder into the bacterial suspension, and carrying out oscillatory adsorption at 32 ℃ and 200-220 rpm for 0.5-1.0 h to obtain the microbial preparation.

Further, in S1, a method for preparing a seed bacterial solution of paenibacillus jimila W51 is as follows: a strain of Paenibacillus jimmalaensis W51 is inoculated into YPGB culture solution at 32 ℃, 12 h is subjected to shaking culture at 200 rpm, the OD value of the strain at 600 nm is measured by sampling every 2 h in an ultra-clean workbench, and the culture is finished when the OD value is 0.8, and the strain is used as seed strain liquid.

The invention also provides application of the microbial preparation in preventing and treating tomato bacterial wilt.

The invention also provides a method for preventing and treating tomato bacterial wilt, which uses a microbial preparation to perform root irrigation treatment on tomatoes.

Has the advantages that: the invention is a microbial preparation specially developed for tomato bacterial wilt, and because the microbial preparation is a biological preparation, a series of problems caused by the use of chemical pesticides are completely avoided, and the microbial preparation is not easy to generate resistance; the method is safe to the environment, pollution-free and residue-free; strong specificity, high activity, low production cost and simple preparation process, thereby being beneficial to the pollution-free production of tomatoes; the illite powder is a cheap natural clay mineral resource, so that farmers can not use or reduce the dosage of other chemical pesticides, thereby not only saving the expenditure of the farmers, but also reducing the residue of chemical agents.

The greenhouse experiment structure shows that: the Paenibacillus jimela W51 (CGMCC No. 16639) microbial preparation can obviously reduce the occurrence of tomato bacterial wilt. The root irrigation treatment is carried out by utilizing the microbial preparation prepared from Paenibacillus jimila W51 and the illite powder, and the control effect on the tomato bacterial wilt can reach 79.35 percent.

Drawings

FIG. 1 is a photograph showing the effect of the microbial preparation according to embodiment 6 on the control of tomato bacterial wilt disease; wherein A is the disease condition of tomato bacterial wilt in a blank control group, and B is the disease condition of tomato bacterial wilt in a microbial preparation group.

Detailed Description

The present invention will be described in detail with reference to the accompanying drawings.

Embodiment 1:

the embodiment provides Paenibacillus jimila W51, and the screening method of the strain comprises the following steps:

paenibacillus jimirabilis W51 (CGMCC No. 16639) is separated from tomato root soil at wharf town of Huaian city, jiangsu province. Cutting tomato root into 1cm pieces, soaking in 1% sodium hypochlorite solution for 5 min, soaking in 75% ethanol for 90 s, and cleaning with sterile water for 3 times (taking 50 μ L of the last cleaning solution and spreading on YGPA)Culturing on a solid culture medium, and considering that the surface is disinfected and sterile if the culture grows aseptically after 48 h). Placing a sterilized sample of 5 g in a mortar, adding 45 ml sterile 0.85% NaCl solution, macerating the tissue, grinding, and diluting with sterile 0.85% NaCl solution in a gradient manner. Get 10 ^-1 、10 ^-2 、10 ^-3 50 μ L of each of the three dilutions was spread on YGPA solid plate medium and incubated at 32 ℃ for 36 h. The largest single colony was selected and inoculated onto fresh YGPA solid plate medium, and the purified strain was obtained by repeated transfer. The purified strain was stored in an ultra-low temperature refrigerator of-80 ℃ with 50% glycerol.

The identification method comprises the following steps: the strain is identified by morphological characteristics, physiological and biochemical experiments and 16S rDNA sequence analysis

Morphological characteristics: gram-positive bacteria, rod-shaped, spore-producing, round colony, milky white, glossy and smooth surface.

Gene amplification and sequence analysis Paenibacillus jimila W51 was cultured in YGPB medium at 32 ℃ to log phase, centrifuged at 13 000R/min for 10 min to collect the cells, DNA of strain W51 was extracted according to the protocol of bacterial DNA extraction kit (Promega, USA), and the extracted DNA product was used as a template for amplification by 27F and 1492R primers. The primer sequence is as follows:

27F：AGAGTTTGATCCTGGCTCAG

1492R：GGTTACCTTGTTACGACTT

A16S rDNA gene fragment was amplified from the genomic DNA. The PCR product is sent to Shanghai Meiji biological medicine science and technology Limited company for sequencing. The sequencing results were as follows:

CGGGCGGTGTGTACAAGACCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCAATTCCGACTTCATGTAGGCGAGTTGCAGCCTACAATCCGAACTGAGACCGGCTTTTCTAGGATTGGCTCCAcctCGcggcTTCGCTTCCCGTTGTACCGGCCATTGTAGTACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTGCTTAGAGTGCCCAGCTTGACCTGCTGGCAACTAAGCATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCTCCTCTGTCCCGAAGGAAAGGTCTATCTCTAGACCGGTCAGAGGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATACTCCACTGCTTGTGCGGGTCCCCGTCAATTCCTTTGAGTTTCAGTCTTGCGACCGTACTCCCCAGGCGGAATGCTTAATGTGTTAACTTCGGCACCAAGGGTATCGAAACCCCTAACACCTAGCATTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGCGCCTCAGCGTCAGTTACAGCCCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGCTCTCCAGTTTCCAGTGCGACCCGAAGTTGAGCCTCGGGATTAAACACCAGACTTAAAGAGCCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCCCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGGGCTTTCTTCTCAGGTACCGTCACTCttgTAGCAGTTACTCTACAAGACGTTCTTCCCTGGCAACAGAGCTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGGCTTTCGCCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTAGGCCTTTACCCCACCAACTAGCTAATGCGCCGCAGGCCCATCCACAAGTGACAGATTGCTCCGCCTTTCCTCCTTCTCCCATGCAGGAAAAGGATGTATCGGGTATTAGCTACCGTTTCCGGTAGTTATCCCTGTCTTGTGGGCAGGTTGCCTACGTGTTACTCA

homology comparisons of the determined 16S rRNA gene sequences were performed by BLAST software, the results are shown in Table 1, strain W51 and Paenibacillus jimela reported in GenBankP. jamilae(KCTC 13919) has the closest relationship and the homology is 99.65%, so that the strain is identified as Paenibacillus jimila (B.jimmalaensis) (C.jimmalaensis)Paenibacillus jamilae）。

TABLE 1 W51 sequencing alignment results

Bacterial strains	Most similar strains and accession numbers	Degree of similarity (%)
			W51	Paenibacillus jamilae KCTC13919	99.65

The Paenibacillus jimira W51 is classified and namedPaenibacillus jamilaeAnd the strain is preserved in China general microbiological culture Collection center (CGMCC) at 26 months and 10 months in 2018, the preservation address is No. 3 of the institute of microbiology of the national academy of sciences, no.1 of West Xilu, north Chengyang, beijing, and the preservation number is CGMCC No.16639.

Embodiment 2: determination of effective adsorption efficiency of illite adsorbent on W51

The Paenibacillus jimmalaensis W51 (CGMCC No. 16639) strain is inoculated into YPGB culture solution at 32 ℃, 12 h is subjected to shaking culture at 200 rpm, the OD value of the Paenibacillus jimmalaensis W51 strain at 600 nm is measured by sampling every 2 h in an ultra-clean workbench, and the culture is finished when the OD value is 0.8, and the strain liquid is used as seed strain liquid. Mixing a W51 seed bacterial solution in a ratio of 1:100 into PDB liquid culture medium, 32 ℃,200 rpm vibration culture 32 h, 4000 rpm centrifugation 30min, abandon the supernatant, precipitate with sterile water heavy suspension and make it in 600 nm absorbance value of 0.8. The adsorption effect of illite at different ratios on W51 cells was measured by sucrose density gradient method. Adding 4% illite powder (the particle size is less than 2 mu m, the natural whiteness is greater than 85%, and the natural whiteness is provided by a key laboratory of probiotic preparation in Jiangsu province), carrying out shaking culture at 32 ℃ and 200 rpm in a gas bath shaker by 0.5 h, taking the mixed solution into a test tube, slowly adding 60% of sucrose solution to the bottom of the mixed solution, standing for 2 h, precipitating thalli adsorbed on the illite to the bottom of the sucrose solution due to the fact that the mass of the thalli is increased, keeping the thalli not adsorbed on the upper layer of the sucrose solution, absorbing the upper layer of the liquid to dilute and coat the flat plate, and counting the number of single colonies after culturing for 12 h at 32 ℃, wherein the adsorption rate C is calculated by the following formula:

wherein: m- -single colony number of the mixture solution on the upper layer of the sucrose solution after adding the adsorbent

M- - - -Single colony count of bacteria suspension without adsorbent on the sucrose solution

The adsorption rate of W51 by 4% illite was calculated to be 90.39%.

Embodiment 3: preparation of Paenibacillus jimila W51 microbial preparation

The Paenibacillus jimmalaensis W51 (CGMCC No. 16639) strain is inoculated into YPGB culture solution at 32 ℃, 12 h is cultured in a shaking way at 200 rpm, the OD value of the Paenibacillus jimmalaensis W51 strain at 600 nm is measured by sampling every 2 h in an ultra-clean workbench, and the culture is finished when the OD value is 0.8, and the strain liquid is used as seed strain liquid. Inoculating the seed bacterial liquid into PDB fermentation culture solution at a volume ratio of 1<2μm, natural whiteness>85% provided by key laboratory of probiotic preparation in Jiangsu province), culturing at 32 deg.C and 210 rpm under shaking for 0.5 h to obtain microbial preparation with total viable bacteria concentration of 1 × 10 ⁸ ~1×10 ⁹ CFU/mL。

Embodiment 4: preparation of Paenibacillus jimila W51 microbial preparation

The Paenibacillus jimmalaensis W51 (CGMCC No. 16639) strain is inoculated into YPGB culture solution at 32 ℃, after shaking culture is carried out at 220 rpm for 10 h, the OD value of the Paenibacillus jimmalaensis at 600 nm is measured by sampling every 2 h in an ultra-clean workbench, and the culture is finished when the OD value is 0.8, and the strain liquid is used as seed strain liquid. Inoculating the seed bacterial liquid into PDB fermentation culture solution at a volume ratio of 1<2μm, natural whiteness>85%), 32 ℃, and 1.0 h by shaking culture at 200 rpm to obtain the microbial preparation, wherein the total concentration of viable bacteria in the finished microbial preparation is 1 multiplied by 10 ⁸ ~1×10 ⁹ CFU/mL。

Embodiment 5: greenhouse test Paenibacillus jie W51 microbial preparation for tomato bacterial wilt prevention effect (2018.04-2018.12)

A place: huaiyin institute of Industrial and scientific colleges greenhouse

Greenhouse pot experiment: the method comprises the steps of washing tomato seeds (the variety is Shanghai cooperative 903) twice with sterile water, washing the tomato seeds with 75% alcohol for two minutes, sterilizing the tomato seeds, washing the tomato seeds with the sterile water for three times repeatedly, culturing the sterilized tomato seeds in a culture dish to germinate, selecting the germinated seeds after 72 h, sowing the germinated seeds into a hole basin, selecting seedlings with consistent growth vigor when the tomato seedlings grow to 5-6 true leaves, transferring the seedlings into a large basin to culture, dividing experiments into a blank control treatment group and a microbial preparation treatment group, wherein each treatment group consists of 36 tomato seedlings, and repeating the experiments for three times. After 30 days, the tomato Miao Genbu of the microbial preparation treatment group is filled with the W51 microbial preparation 15 mL, the blank control group is filled with the sterilized water 15 mL, and the bacterial wilt bacteria (the concentration is 1 multiplied by 10) are inoculated after one week ⁷ CFU/mL, inoculum size of 30 mL), greenhouse test conditions were: 35. c, natural illumination. The disease of tomato bacterial wilt is continuously observed and investigated for 15 days after 7 d.

According to tomato bacterial wilt resistance identification technical specification (NY/T1858.4-2010), the research of tomato bacterial wilt incidence records and analyzes test data by combining the actual situation of the experiment.

Grade 0, no symptoms;

grade 1, 1 leaf withered;

grade 2, 2-3 leaves withered;

grade 3, except 2-3 leaves at the bottom, the other leaves wither;

4, withering the whole leaf;

the disease level of each strain is recorded and investigated, and a disease index is calculated.

Disease Index (DI) = ∑ (number of disease stage × number of diseased plants)/(number of highest disease stage × total number of plants) × 100%

Biocontrol effect = (control group disease severity-treatment group disease severity)/control group disease severity × 100%

Note: the disease index and control effect of different treatments were calculated and differential significance analysis and multiple comparisons were performed using IBM SPSS Statistics 23 data analysis software.

The disease onset in the control and experimental groups after 15 days of microbial preparation treatment is shown in Table 2.

TABLE 2 biocontrol effect of microbial agents on tomato bacterial wilt

The result shows that the W51 microbial preparation has better biological control effect on the tomato bacterial wilt under the greenhouse condition. The tomato seedlings treated by the W51 microbial preparation have the bacterial wilt disease index of 17.62 percent, and the biocontrol efficiency is 79.35 percent relative to 85.33 percent of the control group.

Embodiment 6: greenhouse test Paenibacillus jie W51 microbial preparation for tomato bacterial wilt prevention effect (2019.02-2019.07)

A place: huaiyin institute of Industrial and scientific colleges greenhouse

Greenhouse pot experiment: the method comprises the steps of washing tomato seeds (the variety is Shanghai cooperative 903) twice with sterile water, washing the tomato seeds with 75% alcohol for two minutes, sterilizing the tomato seeds, washing the tomato seeds with the sterile water for three times repeatedly, culturing the sterilized tomato seeds in a culture dish to germinate, selecting the germinated seeds after 72 h, sowing the germinated seeds into a hole basin, selecting seedlings with consistent growth vigor when the tomato seedlings grow to 5-6 true leaves, transferring the seedlings into a large basin to culture, dividing experiments into a blank control treatment group and a microbial preparation treatment group, wherein each treatment group consists of 24 tomato seedlings, and repeating the experiments for three times. After 45 days, the tomato Miao Genbu of the microbial preparation treatment group is filled with the W51 microbial preparation 10 mL, the blank control group is filled with the sterilized water 10 mL, and the bacterial wilt bacteria (the concentration is 1 multiplied by 10) are inoculated after one week ⁷ CFU/mL, inoculum size of 20 mL), greenhouse test conditions were: 35. c, natural illumination. After 5 d, the disease of tomato bacterial wilt was continuously observed and investigated for 15 days.

According to the tomato bacterial wilt resistance identification technical specification (NY/T1858.4-2010), the research on the onset of the tomato bacterial wilt records and analyzes test data by combining the actual situation of the experiment.

Grade 0, no symptoms;

grade 1, 1 leaf withered;

grade 2, 2-3 leaves withered;

grade 3, namely withering the leaves except 2-3 leaves at the bottom;

4, withering the whole leaf;

The disease onset of the control group and the experimental group after 15 days of the microbial preparation treatment are shown in fig. 1 and table 3.

TABLE 3 biocontrol effect of microbial agents on tomato bacterial wilt

The result shows that the W51 microbial preparation has better biological control effect on the tomato bacterial wilt under the greenhouse condition. The tomato seedlings treated by the W51 microbial preparation have the bacterial wilt disease index of 15.83 percent, and the biocontrol efficiency reaches 78.58 percent relative to 73.90 percent of a control group.

In conclusion, the microbial preparation adopted by the invention has good effect, can effectively prevent and treat the tomato bacterial wilt, and has wide market application prospect.

The above embodiments are only for illustrating the technical idea and features of the present invention, and the purpose of the embodiments is to enable those skilled in the art to understand the content of the present invention and implement the present invention, and not to limit the protection scope of the present invention by this means. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.

Sequence listing

<110> Huaiyin institute of Industrial and technology

<120> microbial preparation

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1304

<212> DNA

<213> Paenibacillus jie (Paenibacillus jamilae)

<400> 1

cgggcggtgt gtacaagacc cgggaacgta ttcaccgcgg catgctgatc cgcgattact 60

agcaattccg acttcatgta ggcgagttgc agcctacaat ccgaactgag accggctttt 120

ctaggattgg ctccacctcg cggcttcgct tcccgttgta ccggccattg tagtacgtgt 180

gtagcccagg tcataagggg catgatgatt tgacgtcatc cccaccttcc tccggtttgt 240

caccggcagt ctgcttagag tgcccagctt gacctgctgg caactaagca taagggttgc 300

gctcgttgcg ggacttaacc caacatctca cgacacgagc tgacgacaac catgcaccac 360

ctgtctcctc tgtcccgaag gaaaggtcta tctctagacc ggtcagaggg atgtcaagac 420

ctggtaaggt tcttcgcgtt gcttcgaatt aaaccacata ctccactgct tgtgcgggtc 480

cccgtcaatt cctttgagtt tcagtcttgc gaccgtactc cccaggcgga atgcttaatg 540

tgttaacttc ggcaccaagg gtatcgaaac ccctaacacc tagcattcat cgtttacggc 600

gtggactacc agggtatcta atcctgtttg ctccccacgc tttcgcgcct cagcgtcagt 660

tacagcccag agagtcgcct tcgccactgg tgttcctcca catctctacg catttcaccg 720

ctacacgtgg aattccactc tcctcttctg cactcaagct ctccagtttc cagtgcgacc 780

cgaagttgag cctcgggatt aaacaccaga cttaaagagc cgcctgcgcg cgctttacgc 840

ccaataattc cggacaacgc ttgcccccta cgtattaccg cggctgctgg cacgtagtta 900

gccggggctt tcttctcagg taccgtcact cttgtagcag ttactctaca agacgttctt 960

ccctggcaac agagctttac gatccgaaaa ccttcatcac tcacgcggcg ttgctccgtc 1020

aggctttcgc ccattgcgga agattcccta ctgctgcctc ccgtaggagt ctgggccgtg 1080

tctcagtccc agtgtggccg atcaccctct caggtcggct acgcatcgtc gccttggtag 1140

gcctttaccc caccaactag ctaatgcgcc gcaggcccat ccacaagtga cagattgctc 1200

cgcctttcct ccttctccca tgcaggaaaa ggatgtatcg ggtattagct accgtttccg 1260

gtagttatcc ctgtcttgtg ggcaggttgc ctacgtgtta ctca 1304

Claims

1. A microbial preparation comprising an adsorbent and Paenibacillus jimela (B.jimela)) (Paenibacillus jamilae) W51; wherein the total viable bacteria concentration of Paenibacillus jimmara W51 is 1 x 10 ⁸ ~1×10 ⁹ CFU/mL；

The Paenibacillus jie is preserved in China general microbiological culture Collection center in 26 months 10 and 2018, the preservation address is No. 3 of West Lu No.1 of the Kyowa region in Beijing, and the preservation number of the strain in the institute of microbiology of China academy of sciences is CGMCC No.16639.

2. The microbial preparation of claim 1 wherein said adsorbent is i Li Danfen.

3. The microbial preparation of claim 2, wherein the concentration of the illite powder in the microbial preparation is 4%.

4. The microbial preparation according to claim 2 or 3, wherein the illite powder has a particle size of <2 μm and a natural whiteness of >85%.