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A strain of Bacillus amyloliquefaciens and its application

A technology for dissolving amyloid spores and bacilli, applied in the directions of application, bacteria, fungicides, etc., can solve the problems such as the prevention and control methods that have not yet been reported, and achieve the effect of strong inhibitory effect

Active Publication Date: 2021-01-01
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Bacillus amyloliquefaciens (B.amyloliquefaciens) is a Gram-positive bacterium. It has been found that Bacillus amyloliquefaciens has broad-spectrum antibacterial activity against plant pathogenic bacteria, but its antibacterial effectiveness is still very specific. The pathogenic bacteria need to screen out specific biocontrol strains to achieve the best antibacterial effect
And the research that bacillus amyloliquefaciens is applied to the control of plant fungal diseases mostly concentrates on crops such as strawberry, tomato, peach, potato, and the bacillus amyloliquefaciens of specific control asparagus stem blight and the specific control method have not been reported yet

Method used

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  • A strain of Bacillus amyloliquefaciens and its application
  • A strain of Bacillus amyloliquefaciens and its application
  • A strain of Bacillus amyloliquefaciens and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment one: the screening of bacterial strain

[0037] 1. Soil sample collection: Soil samples were collected from the asparagus planting area in Mianyang.

[0038] 2. Preparation medium:

[0039] (1) Chitosanase screening medium (g / L): colloidal chitosan 2.0, K 2 HPO 4 0.7, KH 2 PO 4 0.3, (NH 4 ) 2 SO 4 5, MgSO 4 ·7H 2 O 0.5, FeSO 4 ·7H 2 0 0.01, agar 12.0, pH=7.0.

[0040] (2) Broth medium (g / L): peptone 10.0, beef extract 3.0, NaCl 5.0, pH=7.0.

[0041] 3. Preparation of reagents:

[0042] Preparation of colloidal chitosan: Weigh 10 g of fine powder chitosan, add appropriate amount of concentrated hydrochloric acid, and stir on a magnetic stirrer for 4 to 6 hours to fully dissolve, then repeatedly wash and centrifuge with deionized water (4000r / min), and finally A jelly was formed, and then NaOH was slowly added while stirring to adjust the pH to neutral to obtain colloidal chitosan, which was stored at 4°C for later use.

[0043] 3. Primary scr...

Embodiment 2

[0050] Embodiment two: bacterial strain identification

[0051] 1. 16SrDNA sequencing identification

[0052] The genomic DNA of the target strain was extracted, and the 16S rDNA was cloned from the genomic DNA with universal primers for bacterial 16S rDNA (27F: AGA GTT TGA TCCTGG CTC AG; 1492R: TAC GGT TAC CTT GTT ACG ACT T). The PCR results were sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing, and the sequencing results were compared with BLAST on the Gene Bank website (http: / / www.nicbi.nlm.nih.gov / ), and the gene sequences of the standard strains were found and downloaded. Using MEGA7.0 The software calculates the genetic distance between different strains and draws a phylogenetic tree of bacteria. The 16SrDNA sequencing result is shown in SEQ ID NO 1.

[0053] Compare the target strain with the 16SrDNA of the Bacillus type strain, and use MEGA7.0 to construct a phylogenetic tree, such as image 3 As shown, the genetic distances between strain d1 and Bacillu...

Embodiment 3

[0062] Embodiment three: the indoor test of using Bacillus amyloliquefaciens alone to inhibit the pathogenic bacteria of asparagus stem blight

[0063] 1. Prepare pathogenic bacteria: isolate Phomopsis asparagus, Pestalotiopsis strains, and Curvularia strains from asparagus stem blight diseased strains.

[0064] 2. Preparation of PDA medium (g / L): Cut 200g peeled potatoes into small pieces, boil for 30min, filter the residue with gauze, add glucose 20.0g / L, agar 15.0g / L, pH natural.

[0065] 3. Inoculate the above three types of pathogenic strains in the center of the PDA medium, and culture them in a constant temperature incubator at 28°C until the diameter of the fungal colony is 3 to 4 cm, and then separate them in the four directions of the colony and 0.5 cm from the edge of the colony. Labeled 1,2,3,4. Sites 1 and 3 were used as controls, inoculated with Bacillus subtilis 168 strain, Bacillus subtilis 168 strain, purchased from the German Culture Collection Center DSMZ, ...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to a strain of bacillus amyloliquefaciens and its application. The technical solution adopted is: to screen and isolate a strain of Bacillus amyloliquefaciens MY001 with chitin-degrading ability from the soil where asparagus is planted, and it was preserved in the General Microbiology Center of China Microbiology Culture Collection Management Committee on July 26, 2017. Species name: Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) MY001, deposit number: CGMCC NO.14460. The strain can be used for the inhibition of asparagus stem blight, and the specific method of use is as follows: at a concentration of 1×10 7 Adding 1% colloidal chitin to the cfu / mL bacterial agent of Bacillus amyloliquefaciens MY001 forms a composite bacterial agent, which is of great significance for the control of asparagus stem blight.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a strain of bacillus amyloliquefaciens and its application. Background technique [0002] Asparagus stem blight (Asparagus stem blight) is also known as the "cancer" of asparagus. Its symptoms are mainly concentrated on stems, side branches or leaves. It has the characteristics of rapid onset, easy infection, and serious losses. Large-scale spread, causing the entire asparagus plant to suddenly turn yellow and wither, and cannot effectively produce asparagus products. The main causative pathogen is Phomopsis asparagi. At present, chemical pesticides such as carbendazim and chlorothalonil are mainly used for the prevention and treatment of asparagus stem blight. However, long-term use of such chemical pesticides may easily lead to the development of drug resistance of the bacteria, and it is impossible to effectively control asparagus stem blight continuously. Therefore,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01N43/16A01P3/00C12R1/07
CPCA01N43/16A01N63/00C12N1/205C12R2001/07
Inventor 王刚刚苟艳谢天蒲莉
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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