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Bacillus thuringiensis strain YN-2-2 and application thereof

A technology of Bacillus chrysogenum and strains, applied in the field of bioengineering, can solve the problems affecting the appearance quality and commodity value of potatoes, the effect is not ideal, the yield decline, etc., and achieve the effect of rich carbon source types, short cycle and growth inhibition

Active Publication Date: 2020-05-22
NINGXIA UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It not only affects the appearance quality and commodity value of potatoes, but also delays seedling emergence in severe cases, resulting in a decrease in yield and causing great economic losses
The current prevention and control measures used in production are mainly chemical pesticide control, but the effect is not ideal and will cause environmental pollution

Method used

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  • Bacillus thuringiensis strain YN-2-2 and application thereof
  • Bacillus thuringiensis strain YN-2-2 and application thereof
  • Bacillus thuringiensis strain YN-2-2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Isolation and Identification

[0017] Bacillus thuringiensis strain YN-2-2 was isolated from soil samples collected from potato fields in Zhaotong, Yunnan. Take 1g of soil sample and put it into a 50mL centrifuge tube, add 10mL of sterile water to shake evenly, shake it on a shaker at 37°C and 200rpm for 30min, and let it stand until clarified. The prepared soil suspension was serially diluted in an ultra-clean workbench, and 100 μL of 10 -3 、10 -4 、10 -5 The double dilution was spread on LB solid medium, repeated 3 times, and incubated at 37°C for 24h. After 24 hours, colonies with different shapes were picked in the ultra-clean workbench and purified by streaking, cultured at 37°C for 24 hours, and the purification was repeated 3 times. Transfer the obtained pure single colony to the slant medium of the test tube, and store it in a refrigerator at 4°C for later use.

[0018] The morphology of the isolated strain YN-2-2 is as follows: figure 1 As shown, the colon...

Embodiment 2

[0022] Antibacterial test on potato scab pathogen 4.1765

[0023] Potato scab pathogen 4.1765 was cultured on potato medium at 28°C for 4-5 days; spores and mycelia were scraped with 0.2mL sterile water to make a certain concentration of pathogen suspension, requiring 100 μL volume to grow mycelia when coating a plate Finally, it can cover the surface of the flat plate.

[0024] Take the activated strain YN-2-2 in 50mL potato liquid culture medium at 28°C and culture it on a shaker for 2-5 days. After the fermentation broth was centrifuged at 12,000r / min for 1 minute, the supernatant was discarded, and the precipitate was suspended in the same volume of sterile water to prepare a cell suspension.

[0025] Prepare oat agar solid medium (OMA): add 30g of oatmeal, add water and boil for 15min, filter the residue through three layers of gauze, and set the volume to 1L, add 15g of agar to the solid medium until it is completely dissolved. Sterilize at 121°C for half an hour.

[...

Embodiment 3

[0029] Antibacterial test on potato scab pathogen 4.1765

[0030] The activated strains were inoculated in liquid LB medium, and cultured at 37° C. and 200 rpm for 48 hours. The fermentation broth was centrifuged at 8000 rpm for 10 min, and the supernatant was absorbed and filtered through a 0.22 μm sterile water filter membrane to obtain the bacteria-filtered supernatant. In the ultra-clean workbench, take 100 μL of the spore suspension and spread it on the Gaoshi No. 1 solid medium, and place an Oxford cup symmetrically in the center of the plate. Use a pipette to draw 200 μL of sterile LB liquid medium and filtered bacterial supernatant into an Oxford cup to observe the inhibition zone.

[0031] Such as image 3 The experimental results are shown, which show that the metabolites of Bacillus thuringiensis strain YN-2-2 can effectively inhibit the growth of the test pathogen 4.1765.

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Abstract

The invention discloses a bacillus thuringiensis strain YN-2-2 CGMCC No.19316, which is preserved in the China General Microbiological Culture Collection Center (CGMCC) on Jan.17,2020, wherein the accession number is CGMCC No.19316, the preservation date is Jan.17,2020, and the address of the preservation unit is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The bacillus thuringiensis strain YN-2-2 provided by the invention has the advantages of simple culture, short period, no ecological toxicity, abundant available carbon sources, high salt tolerance, wide tolerable PH range and capability of effectively inhibiting growth of pathogenic bacteria of potato deep scab.

Description

technical field [0001] The invention belongs to the technical field of bioengineering. Background technique [0002] In recent years, with the expansion of potato planting area and the limitation of arable land area, it has become more and more difficult to rotate crops. High-intensity planting and continuous cropping obstacles lead to the widespread occurrence of scab all over the country, and the lack of effective control measures has become an important bottleneck restricting the development of the potato industry. The disease is a soil-borne disease caused by Streptomyces scabica, which can infect potatoes, sugar beets, radishes and other plants. After infecting potatoes, it mainly damages tubers, and can form flat, raised and depressed shapes on its surface. Various types of brown corky lesions. It not only affects the appearance quality and commodity value of potatoes, but also delays emergence in severe cases, resulting in a decrease in yield and causing great econo...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/23A01P1/00C12R1/07
CPCA01N63/00C12R2001/075C12N1/205
Inventor 仲乃琴石莹莹赵盼宋双伟
Owner NINGXIA UNIVERSITY
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