CN117866812A - Bacillus bailii bv105 and application thereof in preventing and controlling crop diseases - Google Patents
Bacillus bailii bv105 and application thereof in preventing and controlling crop diseases Download PDFInfo
- Publication number
- CN117866812A CN117866812A CN202311777406.6A CN202311777406A CN117866812A CN 117866812 A CN117866812 A CN 117866812A CN 202311777406 A CN202311777406 A CN 202311777406A CN 117866812 A CN117866812 A CN 117866812A
- Authority
- CN
- China
- Prior art keywords
- potato
- bacillus
- sunflower
- diseases
- controlling
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 89
- 201000010099 disease Diseases 0.000 title claims abstract description 88
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 28
- 244000061456 Solanum tuberosum Species 0.000 claims abstract description 70
- 235000002595 Solanum tuberosum Nutrition 0.000 claims abstract description 66
- 241000208818 Helianthus Species 0.000 claims abstract description 61
- 235000003222 Helianthus annuus Nutrition 0.000 claims abstract description 53
- 241001123668 Verticillium dahliae Species 0.000 claims abstract description 27
- 241000082085 Verticillium <Phyllachorales> Species 0.000 claims abstract description 24
- 241000223221 Fusarium oxysporum Species 0.000 claims abstract description 20
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 14
- 241000233622 Phytophthora infestans Species 0.000 claims abstract description 13
- 241000813090 Rhizoctonia solani Species 0.000 claims abstract description 9
- 244000000005 bacterial plant pathogen Species 0.000 claims abstract description 9
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 claims abstract description 8
- 241000223218 Fusarium Species 0.000 claims abstract description 8
- 241001558929 Sclerotium <basidiomycota> Species 0.000 claims abstract description 8
- 238000004321 preservation Methods 0.000 claims abstract description 6
- 238000002360 preparation method Methods 0.000 claims description 10
- 244000063299 Bacillus subtilis Species 0.000 claims description 3
- 241000894006 Bacteria Species 0.000 claims description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 2
- 244000000003 plant pathogen Species 0.000 claims 2
- 230000000694 effects Effects 0.000 abstract description 27
- 239000002689 soil Substances 0.000 abstract description 12
- 235000012015 potatoes Nutrition 0.000 abstract description 9
- 241000221696 Sclerotinia sclerotiorum Species 0.000 abstract description 7
- 206010027145 Melanocytic naevus Diseases 0.000 abstract description 5
- 208000007256 Nevus Diseases 0.000 abstract description 5
- 238000011081 inoculation Methods 0.000 description 31
- 238000000034 method Methods 0.000 description 24
- 241000196324 Embryophyta Species 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 19
- 244000052616 bacterial pathogen Species 0.000 description 17
- 238000002474 experimental method Methods 0.000 description 17
- 239000007788 liquid Substances 0.000 description 16
- 239000002609 medium Substances 0.000 description 16
- 239000007787 solid Substances 0.000 description 16
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- 238000012258 culturing Methods 0.000 description 12
- 230000001580 bacterial effect Effects 0.000 description 10
- 239000001963 growth medium Substances 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- 239000008223 sterile water Substances 0.000 description 9
- 238000012360 testing method Methods 0.000 description 7
- 241000221662 Sclerotinia Species 0.000 description 6
- 108020004566 Transfer RNA Proteins 0.000 description 6
- 238000001914 filtration Methods 0.000 description 6
- 238000005286 illumination Methods 0.000 description 6
- 238000011835 investigation Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 5
- 230000035764 nutrition Effects 0.000 description 5
- 230000008653 root damage Effects 0.000 description 5
- 208000024891 symptom Diseases 0.000 description 5
- 235000015099 wheat brans Nutrition 0.000 description 5
- 208000035240 Disease Resistance Diseases 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 4
- 238000009629 microbiological culture Methods 0.000 description 4
- 230000001717 pathogenic effect Effects 0.000 description 4
- 238000004382 potting Methods 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000000443 biocontrol Effects 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 235000013399 edible fruits Nutrition 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 208000006278 hypochromic anemia Diseases 0.000 description 3
- 238000003973 irrigation Methods 0.000 description 3
- 230000002262 irrigation Effects 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 235000020238 sunflower seed Nutrition 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 241000194107 Bacillus megaterium Species 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000001851 biosynthetic effect Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 108091008053 gene clusters Proteins 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000009331 sowing Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 235000019354 vermiculite Nutrition 0.000 description 2
- 239000010455 vermiculite Substances 0.000 description 2
- 229910052902 vermiculite Inorganic materials 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- WIIZWVCIJKGZOK-IUCAKERBSA-N 2,2-dichloro-n-[(1s,2s)-1,3-dihydroxy-1-(4-nitrophenyl)propan-2-yl]acetamide Chemical compound ClC(Cl)C(=O)N[C@@H](CO)[C@@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-IUCAKERBSA-N 0.000 description 1
- 108020005075 5S Ribosomal RNA Proteins 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 241001669263 Bacillus amyloliquefaciens DSM 7 Species 0.000 description 1
- 241001492467 Bacillus amyloliquefaciens LL3 Species 0.000 description 1
- 241000869594 Bacillus amyloliquefaciens TA208 Species 0.000 description 1
- 241000003114 Bacillus velezensis FZB42 Species 0.000 description 1
- 241000137465 Bacillus velezensis UCMB5033 Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 241000831652 Salinivibrio sharmensis Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 235000021186 dishes Nutrition 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000003958 fumigation Methods 0.000 description 1
- 238000011331 genomic analysis Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 108700022487 rRNA Genes Proteins 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000002352 surface water Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000012070 whole genome sequencing analysis Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- 238000004383 yellowing Methods 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the technical field of biological control, and provides bacillus belicus bv105 separated from potato rhizosphere soil for controlling diseases of crops such as potatoes, sunflowers and the like, wherein the preservation number of bacillus belicus bv105 is CGMCCNo.27540. Bacillus belicus bv105 has certain control effects on crop diseases such as verticillium wilt, potato fusarium wilt, potato late blight, potato black nevus, sunflower verticillium wilt, sunflower sclerotium disease and the like, can be used for controlling potato diseases or preparing products for controlling potato diseases, and can be used for controlling sunflower diseases or preparing products for controlling sunflower diseases; also has certain inhibiting effect on verticillium dahliae, fusarium oxysporum, phytophthora infestans, rhizoctonia solani, sclerotinia sclerotiorum and the like, and can be used for inhibiting plant pathogenic bacteria or preparing products for inhibiting plant pathogenic bacteria.
Description
Technical field:
the invention belongs to the technical field of biological control, and particularly relates to bacillus beijerinus bv105 and application thereof in controlling crop diseases.
The background technology is as follows:
control of crop disease is a very challenging task. The existing various prevention and control measures are summarized in the following aspects: breeding disease-resistant varieties, agricultural measures, biological fumigation, biological control and chemical control. Among them, biological control is a promising control measure. The biological control not only avoids the problems of residue, environmental pollution and the like caused by the large-scale use of chemical pesticides, but also is beneficial to promoting the environmental protection and the agricultural sustainable development, and has better application prospect. At present, people are increasingly concerned with and develop safer and more effective biocontrol bactericides for realizing the prevention and control of crop diseases.
The invention comprises the following steps:
the first object of the present invention is to provide bacillus beijerinus bv105 which can be used for controlling crop diseases.
A second object of the present invention is to provide the use of bacillus beijerinus bv105 for controlling potato diseases or for preparing a product for controlling potato diseases.
A third object of the present invention is to provide the use of bacillus beijerinus bv105 for controlling sunflower diseases or for preparing a product for controlling sunflower diseases.
A fourth object of the present invention is to provide the use of bacillus beijerinus bv105 for inhibiting plant pathogenic bacteria or for the preparation of a product for inhibiting plant pathogenic bacteria.
The first object of the invention is implemented by the following technical scheme:
bacillus belicus (Bacillus velezensis) bv105 is currently preserved in China general microbiological culture Collection center (China General Microbiological Culture Collection Center, CGMCC) with the preservation number of CGMCC No.27540 and the preservation date of 2023, national academy of sciences of China, national academy of sciences of North Chen West road No. 1, the area of Korea of Beijing.
The second object of the invention is implemented by the following technical scheme:
use of bacillus belicus (Bacillus velezensis) bv105 for controlling potato diseases or for preparing a product for controlling potato diseases.
Further, the potato disease is at least one of verticillium potato wilt, fusarium wilt, late blight and black mole.
The third object of the present invention is implemented by the following technical scheme:
use of bacillus beleidsi (Bacillus velezensis) bv105 for controlling sunflower diseases or for the preparation of a product for controlling sunflower diseases.
Further, the sunflower disease is at least one of verticillium wilt, fusarium wilt and sclerotium disease of sunflower.
The fourth object of the present invention is achieved by the following technical scheme:
use of bacillus beleiensis (Bacillus velezensis) bv105 for inhibiting a plant pathogenic bacterium or for the preparation of a product for inhibiting a plant pathogenic bacterium.
Further, the plant pathogenic bacteria are at least one of verticillium dahliae, fusarium oxysporum, phytophthora infestans, rhizoctonia solani and sclerotinia sclerotiorum.
The invention has the advantages that:
according to the invention, 1 bacillus belicus bv105 with antagonism to important plant pathogenic bacteria of potatoes and sunflowers is separated from rhizosphere soil of the potatoes, and can effectively prevent and treat crop diseases such as verticillium wilt of the potatoes, late blight of the potatoes, black nevus of the potatoes, verticillium wilt of sunflowers, sclerotinia of sunflowers and the like. Laying a foundation for controlling diseases of potatoes and sunflowers by using the biocontrol microbial inoculum.
Description of the drawings:
in order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a genomic analysis map of strain bv105 of example 1;
FIG. 2 shows the control effect of bv105 on potato verticillium by inoculation in the potting experiment of example 2;
FIG. 3 shows the effect of bv105 inoculation in the potting experiment of example 2 on the disease index of potato verticillium;
FIG. 4 shows the effect of bv105 inoculation in the potting experiments of example 2 on the relative pathogenic bacterial load of potato verticillium;
FIG. 5 shows the control effect of bv105 on potato verticillium by inoculation in the detoxified seedling experiment of example 2;
FIG. 6 shows the effect of bv105 inoculation in the detoxified seedling experiment of example 2 on the disease index of potato verticillium;
FIG. 7 shows the effect of bv105 inoculated in the detoxified seedling experiment of example 2 on the relative pathogenic bacterial load of potato verticillium;
FIG. 8 is a schematic diagram showing the bacteriostatic effect of bv105 on Fusarium oxysporum, rhizoctonia solani, and Phytophthora infestans in example 3;
FIG. 9 shows the control effect of bv105 inoculated in example 4 on verticillium of sunflower;
FIG. 10 shows the effect of bv105 on the disease index of verticillium in sunflower inoculated in example 4;
FIG. 11 shows the effect of bv105 inoculated in example 5 on controlling sunflower wilt;
FIG. 12 shows the effect of bv105 inoculation in example 6 on the disease index of sunflower wilt;
FIG. 13 shows the control effect of bv105 inoculated in example 7 on sclerotinia rot of sunflower;
FIG. 14 shows the effect of bv105 on the disease index of sunflower sclerotinia inoculated in example 8.
The specific embodiment is as follows:
the following description of the embodiments of the present invention will be made in detail and with reference to the accompanying drawings, wherein it is apparent that the embodiments described are only some, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The quantitative tests in the following examples were all set up in triplicate and the results averaged.
The formulation of the medium used in the examples below is as follows (the liquid medium is formulated to remove agar powder from the solid medium, and to keep the other components and concentrations unchanged);
LB medium (solid): 10g of sodium chloride, 10g of peptone, 5g of yeast extract, 12g of agar powder, adding water to 1000mL, and adjusting the pH to 7.0;
KB medium (solid): peptone 20g, K 2 HPO 4 1.5g, glycerin 8mL, mgSO 4 0.74g, 12g of agar powder, adding water to 1000mL, and adjusting the pH to 7.0;
PDA medium (solid): peeling 200g of potato, 20g of glucose, 15g of agar powder and adding water to 1000mL;
wheat bran culture medium: wheat bran 12g, water 1000mL, and boiling for 10-15min with strong fire; filtering with four layers of gauze to remove filter residues, and adding water into the filtrate to 1000mL;
rye medium (solids): 50g of rye seeds, 1000mL of water, soaking for 24-36 hours, sterilizing for 30 minutes, filtering through four layers of gauze to remove filter residues, adding water into the filtrate to 1000mL, and adding 20g of agar.
EXAMPLE 1 isolation and characterization of Strain bv105
1. Isolation of Strain bv105
1. Sample collection: samples of potato plot rhizosphere soil (40°48'61 "north latitude, 113°34' 50" east longitude, 1361 meter altitude) were collected from the city of cold Xu Maao village, wulan scout.
2. Sample treatment: root systems are picked from rhizosphere soil of potato plots and placed in 25mL phosphate buffer solution, vortex shaking is carried out for 15s, a 100 μm nylon net is used for filtering rhizosphere soil samples, and filtrate is stored in a 50mL centrifuge tube.
3. Separating strains: serially diluting rhizosphere soil sample by 10 times and 10 times 2 Multiple of 10 3 Multiple of 10 4 Multiple of 10 5 Doubling, setting 3 repeats; then 100. Mu.L of the diluted solution was placed on KB medium (solid) with 100. Mu.g/ml of LDelvocid added thereto and LB medium (solid) and cultured in a constant temperature incubator at 25℃to observe the colony growth state every day. The results show that when diluted 10 5 At the time of doubling, no colonies grew out on the plate, but at 10 4 Single colony can be grown under dilution multiple; will 10 4 Single colonies growing at dilution were picked and streaked on KB medium (solid) to give single colonies, which were designated bv105, and then grown in KB medium (liquid) for further strain identification.
2. Identification of Strain bv105
1. Morphological identification of Strain bv105
After the strain bv105 is cultured on KB culture medium (solid) for 24-48 hours, semitransparent light yellowish-brown colonies can be formed, and the colonies are circular in state, smooth in surface, neat in edge and not sticky.
2. Molecular characterization of Strain bv105
Extracting DNA of the strain bv105, sending the total DNA to Shanghai Meiji biological medicine technology Co., ltd for whole genome sequencing, and carrying out gene prediction of an assembly result by adopting Glimmer software (http:// ccb.jhu.edu/software/Glimmer/index. Shtml); tRNA included in the genome is predicted using tRNA scan-SE v2.0 software (http:// tRNA. Ucsc. Edu/software /); rRNA contained in the genome was predicted using Barrnap software (https:// gitsub. Com/tseemann/Barrnap). And gene function annotation is carried out based on a GO, KEGG, swiss-Prot, COG, NR, CAZY database and the like.
The average nucleotide identity (Average nucleotide identity, ANI) of the bv105 genome was calculated using NCBI prokaryotic genome annotation pipeline (PGAP v2020-07-09. Build4716). Biosynthetic gene clusters (Biosynthetic gene clusters, BGCs) of secondary metabolites were predicted using antissmash (v 5.0.0).
The results show that: the total number of Reads of the strain bv105 three-generation sequencing is 475531, the total base length is 1910057260bp, the reading length of the longest Reads is 155895bp, and the reading length of the average Reads is 4016.68bp. The whole genome of the strain bv105 consists of one circular chromosome, the genome size is 4087900bp, the GC content is 46.39%, and 4180 encoding genes are encoded. The total length of the genes is 3648345bp, the average length of the genes is 872.81bp, and the average density of the genes is 1.02, namely the number of the corresponding genes in each 1kb base in the genome. The genome contains 87 tRNAs in total, including 20 tRNAs types; the number of 16S rRNA was 9, the number of 23S rRNA was 9, and the number of 5S rRNA was 9 in 27 rRNAs (as shown in FIG. 1 and Table 1).
TABLE 1bv105 genomic information
| Genomic characteristics | Value of |
| Genome assembly size (bp) | 4087900 |
| GC content (%) | 46.39 |
| Number of genes encoding proteins | 4180 |
| Gene region encoding protein (bp) | 3648345 |
| tRNA/rRNA Gene quantity (number) | 87/27 |
As shown in Table 2, by comparing the whole genome sequence of the strain bv105 with the reported ANI values of the representative strains of Bacillus amyloliquefaciens, bacillus velezensis, bacillus subtilis and Bacillus megaterium, the ANI of the strain bv105 and Bacillus velezensis was 97.64 to 98.31%, wherein the ANI of the strain bv105 and the model strain B.velezensis FZB42 was highest (98.31%) and the comparison with other Bacillus spp was lower than 95%, so that the strain bv105 was determined to be Bacillus bailii (B.velezensis).
TABLE 2 alignment of strain bv105 with ANI results
| Alignment of strains | Strain numbering | Sequence number | ANI(％) |
| Bacillus amyloliquefaciens | DSM7 | FN597644.1 | 93.61 |
| Bacillus amyloliquefaciens | TA208 | CP002627.1 | 93.56 |
| Bacillus amyloliquefaciens | LL3 | CP002634.1 | 93.55 |
| Bacillus amyloliquefaciens | ATCC13952 | CP009748.1 | 93.56 |
| Bacillus amyloliquefaciens | FZB42 | CP000560.2 | 98.3 |
| Bacillus amyloliquefaciens | Trigocor1448 | CP007244.1 | 98.23 |
| Bacillus velezensis | UCMB5033 | HG328253.1 | 98.27 |
| Bacillus velezensis | WF02 | CP053376.1 | 97.64 |
| Bacillus velezensis | SRCM103616 | CP011937 | 98.2 |
| Bacillus subtilis | Bacillus subtilis | CP028202 | 80.19 |
| Bacillus megaterium | Bacillus megaterium | CP001982 | 79.98 |
Bacillus belicus (Bacillus velezensis) bv105 is currently preserved in China general microbiological culture Collection center (China General Microbiological Culture Collection Center, CGMCC) with the preservation number of CGMCC No.27540 and the preservation date of 2023, national academy of sciences of China, national academy of sciences of North Chen West road No. 1, the area of Korea of Beijing.
Example 2 application of Bacillus bailii bv105 in controlling potato verticillium wilt and inhibiting verticillium dahliae
1. Experimental method
1. Potato planting
Potted experiment potato plant acquisition: the tuber of Netherlands No. 15 is cut into pieces of about 3cm by a blade sterilized with 70% alcohol, and at least one bud eye is ensured on each piece. Individual potato pieces were planted in plastic pots (10 cm diameter x 10cm height) filled with a mixture of sterile nursery substrate, sand and soil (1:1:1) at a temperature of 26 ℃ and a relative humidity of 50% for 12 hours of illumination time. The potato starts to be inoculated after 6 leaves are grown.
Acquisition of detoxicated seedlings experiment potato detoxicated seedlings: since potato company purchased detoxified seedlings (Netherlands No. 15), cut off consistently grown potato seedlings 2.5cm from the top with a 70% alcohol sterilized blade, transplanted into sterilized vermiculite matrix for cultivation, and covered with film for moisture retention. After 5 days of growth, the film is uncovered, and after 15 days of transplanting, potato seedlings with consistent growth are selected for a biocontrol strain verification experiment. The survival rate of transplanted seedlings can reach more than 98%, and the seedlings grow neatly, thereby being beneficial to comparing and researching the changes of potato plants treated differently.
2. Pathogenic bacteria liquid preparation
Culturing Verticillium dahliae (Verticillium dahliae) which is a pathogenic bacteria of potato verticillium wilt on PDA culture medium (solid) for 14 days, taking 5 fungus cakes with diameter of 9mm, inoculating in wheat bran culture medium (liquid) for promoting spore culture; culturing at 25deg.C for 10 days, washing testa Tritici with distilled water, filtering with sterilized four layers of gauze to obtain spore solution, and adjusting the concentration of Verticillium dahliae bacteria solution to 1×10 by dilution plate counting method 7 spores/mL.
3. Manual inoculation and disease investigation
Inoculation method for potting experiment: when 4-6 leaves of potato grow, 100mL of cultured OD is taken 600 Bv105 of 1 was inoculated around potato plants, and after 3 days of inoculation, verticillium dahliae was inoculated around potato roots using root-wound inoculation. The specific method comprises the following steps: cutting the potato into soil vertically downwards by a fruit knife in a potato planting flowerpot for root injury treatment; then, the concentration was taken to be 1X 10 7 Root irrigation is carried out on 100mL of spore/mL of verticillium dahliae conidium liquid; each treatment was repeated 3 times for 12 seedlings; culturing in a sunlight greenhouse after inoculation, wherein the culture conditions are as follows: the illumination time is 12 hours, the temperature is 26 ℃, and the relative humidity is 50 percent. The experimental group was designated bv105+Vd8, and a control group (designated Vd8) was set, and bv105 was replaced with an equal amount of sterile water, with other methods and parameters unchanged.
The inoculation method of the detoxified seedling experiment comprises the following steps: transplanting potato seedlings grown in sterile vermiculite for 15 days, picking up seedlings with consistent growth, and taking 20mL of cultured OD 600 Inoculating bv105 bacterial liquid with the length of 1 around the roots of potato seedlings; three days after inoculation, inoculating equal amount of verticillium dahliae conidium liquid around roots again, and repeating each treatment for 12 seedlings for 3 times; inoculating, and culturing in artificial climate chamberCulturing for 16h/8h, wherein the illumination intensity is about 3600lx and the temperature is 25 ℃. The experimental group was denoted bv105+vd8 and a plurality of control groups were set: the blank control group (CK) is prepared by replacing bv105 and Verticillium dahliae with equal amount of sterile water, and other methods and parameters are unchanged; a pathogen-free control group (recorded as bv 105) without inoculating verticillium dahliae, and other methods and parameters are unchanged; the control group (noted as Vd 8) was not vaccinated, bv105 was not vaccinated, and other methods and parameters were unchanged.
After inoculating verticillium dahliae conidium liquid for 30d, counting the disease condition of the potatoes, and recording the disease degree according to the following grading standard:
according to the incidence of potato, the potato is classified into 4 grades of 0, 1, 2 and 3, and the grading standard is as follows:
level 0: healthy plants, and plant leaves do not fade and die;
stage 1: 0-33.33% of plant leaves show chlorosis;
2 stages: 33.33-66.66% of plant leaves show chlorosis, partial necrosis and wilting;
3 stages: 66.66-100% of plant leaves show dead or wilting.
And calculating the disease index according to the recorded disease grade and the formula 1) according to the counted disease number. Performing variance analysis on test data by adopting SAS9.1 software, and performing difference significance test by adopting Duncan's new complex polar difference method; and calculating the control effect by using the formula 2).
Equation 1): disease index = [ Σ (number of disease plants at each stage×corresponding grade)/total number of investigation×highest grade ] ×100
Equation 2): control effect = (control disease index-treatment disease index)/control disease index x 100%
4. Isolation of pathogenic bacteria from potato stem base
Taking 1cm stem segment at the stem base of the inoculated potato plant, soaking and sterilizing with 70% alcohol for 15min, transferring to 10% HClO, soaking and sterilizing for 15min, cleaning with sterile water for more than 3 times, and sucking off surface water. The stems were cut longitudinally into 4 parts and placed on PDA medium (solid) supplemented with 34mg/L chloramphenicol with light pressure to bring the cut surface into contact with the medium. Each treatment was set up for 3 replicates and the growth of pathogenic bacteria was observed.
5. Quantification of potato stem basal pathogenic bacteria
Potato stem RNA was extracted using Trizol Reagent, the extracted RNA was inverted to cDNA using reverse transcription kit, and the concentration was diluted to 100ng/μl and used as a template for real-time fluorescent quantitative PCR. RNA is extracted from a sample of stem basal tissue which is only inoculated with pathogenic bacteria and is used as a control, and a potato actin gene is used as an internal reference gene. And detecting the bacterial load of pathogenic bacteria planted in the basal stem tissue by taking the VertBut specific sequence of the verticillium dahliae as a primer. Each sample was set up with 3 replicates.
Primer sequence:
qSTActin-F：5’-CACCCTGTTCTGCTCACT-3’
qSTActin-R：5’-CAGCCTGAATAGCAACATCA-3’
VertBt-F：5’-AACAACAGTCCGATGGATAATTC-3’
VertBt-R：5’-GTACCGGGCTCGAGATCG-3’
with 2 -△△CT The method of (2) calculates the relative expression level of the gene.
2. Disease resistance effect statistics
The experimental results of the potted plant are shown in fig. 2-4, and compared with the experimental results of the control group, the experimental results of the potted plant are 78.89, the disease condition of verticillium wilt of the potato is obviously reduced after the experimental results of the verticillium wilt of the potato are 30, and the control effect of the bv105 in the embodiment is 61.97% calculated; and after the treatment of the microorganism bv105, the colonization quantity of the verticillium dahliae in the potato is also obviously reduced.
The test results of the detoxified seedlings are shown in figures 5-7, and the investigation result of the disease condition of potato verticillium wilt after 30 days of inoculation shows that the disease index of the inoculated strain bv105 after seedling pretreatment and the disease index of the inoculated verticillium dahliae after seedling pretreatment is 30, the control effect is 46.10 percent, and the disease index is obviously lower than that of the inoculated verticillium dahliae (Vd 8) alone. The quantitative determination result of the bacterial load in the inoculated plants by using RT-qPCR shows that the pretreatment of the bacterial strain bv105 can obviously reduce the biomass of pathogenic bacteria.
EXAMPLE 3 use of Bacillus belicus bv105 in inhibiting potato fusarium wilt pathogen (Fusarium oxysporum Fusarium oxysporum), potato black nevus pathogen (Rhizoctonia solani Rhizoctonia solani) and potato late blight pathogen (Phytophthora infestans Phytophthora infestans)
1. Experimental method
1. Placing freshly cultured fusarium oxysporum and rhizoctonia solani cakes on a PDA (personal digital assistant) culture medium (solid); the cake of fresh phytophthora infestans cultivated on rye medium was placed on 1/5PDA medium (solid). Culturing bv105 to OD 600 In the case of 1, 5 mu Lbv of the bacterial suspension was dropped on the edge of the culture dish inoculated with the pathogenic bacteria as a treatment group, and the culture dish without bv105 of the bacterial suspension was used as a control group. And (3) placing the culture dishes of the treatment group and the control group in a constant temperature incubator at 25 ℃ for culture, and observing whether a bacteriostasis zone can be generated when hypha of the culture dish of the control group completely covers the culture medium. Each treatment was repeated 3 times.
2. Bacteriostasis experiment
The bacteriostasis rate was calculated according to the following formula: antibacterial ratio (%) = (control colony diameter-treated colony diameter)/(control colony diameter-cake diameter) ×100%, cake diameter was 6mm.
2. Statistics of bacteriostatic effects
As shown in FIG. 8 and Table 3, the graph shows that bv105 has obvious antibacterial effect on main diseases of potatoes, such as potato fusarium wilt, potato black nevus, potato late blight and other pathogenic bacteria, namely fusarium oxysporum, rhizoctonia solani and phytophthora infestans.
TABLE 3 antibacterial Rate of bv105 against Fusarium oxysporum, rhizoctonia solani, phytophthora infestans
| Disease of potato | Pathogenic bacteria name | Bacteriostasis rate (%) |
| Wilt of potato | Fusarium oxysporum | 70.19 |
| Potato black nevus disease | Rhizoctonia solani | 53.85 |
| Late blight of potato | Phytophthora infestans | 87.33 |
EXAMPLE 4 application of Bacillus bailii bv105 in preventing and treating verticillium wilt of sunflower and inhibiting verticillium dahliae
1. Experimental method
1. Sunflower planting
Obtaining sunflower plants: sowing sunflower seeds into nutrition bowls with 13cm multiplied by 13cm filled with sterilized soil, planting 5 nutrition bowls for each variety, and sowing 6 seeds in each bowl; the planting conditions are as follows: 25 ℃,16h light/8 h darkness, and relative humidity of 50 percent. 3 replicates were set. After the seeds germinate and come out of the soil, 5 sunflower seedlings are ensured to be in each nutrition pot through thinning. When 4 true leaves are grown on sunflower seedlings, the sunflower seedlings are used for inoculation.
2. Pathogenic bacteria liquid preparation
Culturing Verticillium dahliae (Verticillium dahliae) which is pathogenic bacteria of sunflower verticillium wilt on PDA culture medium (solid) for 14 days, taking 5 fungus cakes with diameter of 9mm, inoculating into wheat bran culture medium (liquid), and culturing; culturing at 25deg.C for 10 days, washing testa Tritici with distilled water, filtering with sterilized four layers of gauze to obtain spore solution, and adjusting the concentration of Verticillium dahliae bacteria solution to 1×10 by dilution plate counting method 7 spores/mL.
3. Manual inoculation and disease investigation
When 4-6 leaves of sunflower grow, 200mL of cultured OD is taken 600 Bv105 of 1 was inoculated around sunflower plants, and after 3 days of inoculation, verticillium dahliae was inoculated around sunflower roots by root injury inoculation. The specific method comprises the following steps: cutting the sunflower seeds into soil vertically downwards in a sunflower planting nutrition pot by a fruit knife to perform root injury treatment; then, the concentration was taken to be 1X 10 7 Root irrigation is carried out on 100mL of spore/mL of verticillium dahliae conidium liquid. Each treatment was repeated 3 times for 30 seedlings; culturing in a sunlight greenhouse after inoculation, wherein the culture conditions are as follows: the illumination time is 16h, the temperature is 25 ℃, and the relative humidity is 50%. The experimental group was denoted bv105+vdgn3; setting a blank control group (CK), and replacing bv105 and verticillium dahliae with equal amount of sterile water, wherein other methods and parameters are unchanged; an uninoculated control group (noted VdGn 3) was set, and bv105 was replaced with an equal amount of sterile water, with other methods and parameters unchanged.
After inoculating the verticillium dahliae conidium liquid for 35d, counting the disease condition of sunflower plants, and recording the disease degree according to the following grading standard:
according to the incidence of sunflowers, the sunflowers are classified into 5 grades of 0, 1, 2, 3 and 4, and the grading standards are as follows:
level 0: healthy plants, no symptoms;
stage 1: less than 25% of the leaves show symptoms such as chlorosis, yellowing, wilting, dead and the like;
2 stages: 25% -50% of leaves show symptoms and plants are dwarfed;
3 stages: 51% -75% of leaves show symptoms and plants are dwarfed;
4 stages: over 75% of the leaves showed severe symptoms and the plants died from wilting.
Based on the number of the counted disease, the disease index was calculated according to equation 1) mentioned in example 2 using the recorded disease grade. Performing variance analysis on test data by adopting SAS9.1 software, and performing difference significance test by adopting Duncan's new complex polar difference method; the control effect was calculated using formula 2) mentioned in example 2.
2. Disease resistance effect statistics
As shown in fig. 9 and 10, experimental results of the disease condition of the sunflower verticillium wilt after 35 days of inoculation show that the disease index of the inoculated strain bv105 after seedling pretreatment and the verticillium dahliae after inoculation is 43.94, the control effect is 30.94%, and the disease index is obviously lower than that of the verticillium dahliae (the control group VdGn 3) only.
EXAMPLE 5 application of Bacillus bailii bv105 in preventing and controlling sunflower fusarium wilt and inhibiting Fusarium oxysporum (Fusarium oxysporum)
1. Experimental method
1. Planting sunflowers: the same method as in example 4 for sunflower planting is not described here.
2. Pathogenic bacteria liquid preparation
Culturing fusarium oxysporum (Fusarium oxysporum) which is a pathogenic bacterium of sunflower fusarium wilt on a PDA (personal digital assistant) culture medium (solid) for 10 days, taking 5 bacterial cakes with the diameter of 9mm, and inoculating the bacterial cakes into a wheat bran culture medium (liquid) for promoting spore culture; culturing at 25deg.C for 10 days, washing testa Tritici with distilled water, filtering with sterilized four layers of gauze to obtain spore solution, and adjusting concentration of fusarium oxysporum solution to 1×10 by dilution plate counting method 7 spores/mL.
3. Manual inoculation and disease investigation
When 4-6 leaves of sunflower grow, 200mL of cultured OD is taken 600 Bv105 of 1 was inoculated around sunflower plants, and after 3 days of inoculation fusarium oxysporum was inoculated around sunflower roots using root injury inoculation. The specific method comprises the following steps: cutting the sunflower seeds into soil vertically downwards in a sunflower planting nutrition pot by a fruit knife to perform root injury treatment; then, the concentration was taken to be 1X 10 7 Root irrigation is carried out on 100mL of spore/mL of fusarium oxysporum conidium liquid. Each treatment was repeated 3 times for 30 seedlings; culturing in a sunlight greenhouse after inoculation, wherein the culture conditions are as follows: the illumination time is 16h, the temperature is 25 ℃, and the relative humidity is 50%. The experimental group was denoted bv105+Fo1-2; setting a blank control group (CK), and replacing bv105 and fusarium oxysporum with equal amounts of sterile water, wherein other methods and parameters are unchanged; an uninoculated control group (designated Fo 1-2) was set, and bv105 was replaced with an equal amount of sterile water, with other methods and parameters unchanged.
After inoculation of fusarium oxysporum conidium solution for 35d, the disease condition of sunflower plants is counted, and the disease degree is recorded according to the grading standard described in example 4; and calculating disease index and preventing and treating effect.
2. Disease resistance effect statistics
As shown in the results of FIG. 11 and FIG. 12, the experimental results of the disease condition of sunflower wilt after 35 days of inoculation show that the disease index of inoculated strain bv105 after seedling pretreatment and then inoculated with Fusarium oxysporum is 21.67, and the control effect is 59.37%, which is significantly lower than that of inoculated Fusarium oxysporum alone (control group Fo 1-2).
EXAMPLE 6 application of Bacillus bailii bv105 in preventing and treating sclerotinia sclerotiorum of sunflower and inhibiting sclerotinia sclerotiorum (Sclerotinia sclerotiorum)
1. Experimental method
1. Planting sunflowers: the same method as in example 4 for sunflower planting is not described here.
2. Pathogenic bacteria liquid preparation
Sclerotium formed by pathogenic bacteria of sunflower sclerotium disease (Sclerotinia sclerotiorum) was collected for use after 7 days of culture on PDA medium (solid).
3. Manual inoculation and disease investigation
When 4-6 leaves of sunflower grow, 200mL of cultured OD is taken 600 1bv105 is inoculated around sunflower plants, 1g sclerotium is buried 3 days after inoculation, and the sunflower plants are placed in a sunlight greenhouse for culture under the following culture conditions: the illumination time is 16h, the temperature is 25 ℃, and the relative humidity is 50%. The experimental group was designated bv105+X-8; setting a blank control group (CK), and replacing bv105 and sclerotinia sclerotiorum with equal amount of sterile water, wherein other methods and parameters are unchanged; an uninoculated control group (designated X-8) was set, bv105 was replaced with an equal amount of sterile water, and the other methods and parameters were unchanged.
After 7d inoculation of sclerotium, the disease condition of sunflower plants was counted and the disease degree was recorded according to the following grading standard:
according to the incidence of sunflowers, the sunflowers are classified into 4 grades of 0, 1, 2 and 3, and the grading standards are as follows:
level 0: the disease is not happened, and the plants are healthy;
stage 1: the disease is mild, only water-immersed disease spots are generated at the rhizome parts, and plants do not produce the phenomenon of wilting;
2 stages: moderately onset, brown disease spots are generated locally on the roots and stems of plants, but the spread of the disease spots is smaller, the wilting phenomenon of only individual leaves of the plants occurs, and the wilting of the plants is less than 50%;
3 stages: severe disease, withering of more than 50% of the leaves, lodging of plants or death of whole plants.
Based on the number of the counted disease, the disease index was calculated according to equation 1) mentioned in example 2 using the recorded disease grade. Performing variance analysis on test data by adopting SAS9.1 software, and performing difference significance test by adopting Duncan's new complex polar difference method; the control effect was calculated using formula 2) mentioned in example 2.
2. Disease resistance effect statistics
As shown in the results of FIG. 13 and FIG. 14, the experimental results of the disease condition of sclerotinia of sunflower after 7 days of inoculation show that the disease index of sclerotinia of inoculated strain bv105 after seedling pretreatment is 52.22, the control effect is 25.4%, and the disease index is remarkably lower than that of sclerotinia of inoculated strain (control group X-8) only.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (7)
1. Bacillus belicus bv105 is characterized by having a preservation number of CGMCC No.27540.
2. Use of bacillus beljalis (bacillus velengens) bv105 according to claim 1 for controlling potato diseases or for the preparation of a product for controlling potato diseases.
3. The use according to claim 2, wherein the potato disease is at least one of potato verticillium wilt, potato late blight, potato black mole.
4. Use of bacillus beljavensis (bacillus heleiensis) bv105 according to claim 1 for controlling sunflower diseases or for the preparation of a product for controlling sunflower diseases.
5. The use according to claim 4, wherein the sunflower disease is at least one of verticillium wilt, fusarium wilt and sclerotium disease of sunflower.
6. Use of bacillus subtilis bv105 according to claim 1 for inhibiting a plant pathogen or for the preparation of a product for inhibiting a plant pathogen.
7. The use according to claim 6, wherein the plant pathogenic bacteria are at least one of verticillium dahliae, fusarium oxysporum, phytophthora infestans, rhizoctonia solani, sclerotium bacteria.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311777406.6A CN117866812A (en) | 2023-12-21 | 2023-12-21 | Bacillus bailii bv105 and application thereof in preventing and controlling crop diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311777406.6A CN117866812A (en) | 2023-12-21 | 2023-12-21 | Bacillus bailii bv105 and application thereof in preventing and controlling crop diseases |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117866812A true CN117866812A (en) | 2024-04-12 |
Family
ID=90580334
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311777406.6A Pending CN117866812A (en) | 2023-12-21 | 2023-12-21 | Bacillus bailii bv105 and application thereof in preventing and controlling crop diseases |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117866812A (en) |
-
2023
- 2023-12-21 CN CN202311777406.6A patent/CN117866812A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109749943B (en) | Trichoderma asperellum and application thereof | |
| CN111979149B (en) | Salt-tolerant bacillus SY1836 and application thereof | |
| CN112175888B (en) | Bacillus belgii Hsg1949 and application thereof | |
| CN114410481B (en) | Trichoderma asperellum and application thereof | |
| CN108048354B (en) | Bacillus subtilis and application thereof | |
| CN105543132A (en) | Bacillus methylotrophicus YB-F7 and application thereof in preventing plant diseases | |
| CN110669691B (en) | Bacillus megaterium for preventing and treating plant nematode diseases and application thereof | |
| CN112375688B (en) | Beauveria bassiana and application thereof in biological prevention and control of corn northern leaf blight | |
| CN109749953B (en) | Bacillus cereus, microbial inoculum and preparation method and application thereof | |
| CN111778173A (en) | Bacillus subtilis Pro1A2, microbial inoculum and preparation method thereof, and application of bacillus subtilis Pro1A2 in cultivation of melons | |
| CN108913625B (en) | Salt-tolerant streptomycete, microbial inoculum thereof and application of microbial inoculum thereof in promoting plant growth | |
| CN111763646B (en) | Long lysine bacillus, microbial agent, biocontrol agent, and preparation method and application thereof | |
| CN111363691B (en) | Paenibacillus polymyxa and application thereof | |
| KR100942228B1 (en) | Biological control of plant diseases using flavobacterium hercynium epb-c313 | |
| CN113755389A (en) | Bacillus belgii and application thereof | |
| CN117165494A (en) | Kiwi fruit canker biocontrol strain Wq-1 and application thereof | |
| CN115873770B (en) | Bacillus bailii and application thereof in preventing and controlling tomato diseases | |
| CN109055265B (en) | Biocontrol bacterium and application thereof in prevention and control of gummy stem blight of crops | |
| CN116676234A (en) | Salt-tolerant bacillus JK-25, microbial inoculum, preparation method and application thereof | |
| CN116536192A (en) | Ginger endophytic bacillus belicus and application thereof in disease control | |
| CN111548951B (en) | Bacillus subtilis Pro6A5, microbial inoculum and preparation method thereof, and application of bacillus subtilis Pro6A5 in cultivation of melons | |
| CN111471620B (en) | Biocontrol bacterium BV03, application thereof, microbial inoculum and preparation method of microbial inoculum | |
| CN111321094B (en) | Microbial agent M1 for preventing and treating stem basal rot of corn and application thereof | |
| CN117866812A (en) | Bacillus bailii bv105 and application thereof in preventing and controlling crop diseases | |
| CN109321476B (en) | Trichoderma asperellum PT-29 for preventing and controlling potato and sunflower verticillium wilt and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |