A Bacillus amyloliquefaciens strain w1 for controlling plant spider mites and its application

A technology of amylolytic spores and plant leaves, which is applied in the fields of application, plant growth regulator, and microbial-based methods, and can solve the problems of poor development and utilization value, low activity, and poor control effect

Active Publication Date: 2018-09-28
MICROBIAL FERMENTATION ENG RES CENT CO LTD OF YUNNAN PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the technical problems that chemical control of spider mites is easy to produce drug resistance, the control effect is poor, and the acaricidal activity of botanical pesticides is low, the active ingredient is not clear, and the development and utilization value is poor. Bacillus amyloliquefaciens strain W1 with high mite control effect and its application

Method used

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  • A Bacillus amyloliquefaciens strain w1 for controlling plant spider mites and its application
  • A Bacillus amyloliquefaciens strain w1 for controlling plant spider mites and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1 The acquisition, identification, culture preservation and preservation of Bacillus amyloliquefaciens strain W1CGMCC No. 11949 (hereinafter referred to as: strain W1) of the present invention.

[0023] 1 Isolation and screening of strain W1

[0024] 1.1 Isolation of strain W1

[0025] Find the naturally dead Tetranychus urticae from the corn leaves, put the Tetranychus urticae into a sterile concave glass slide, disinfect the surface with 75% (v / v) alcohol for 1min, and wash it with sterile water for 5 times Use sterile filter paper to blot the water on the surface of Tetranychus urticae, put Tetranychus urticae into a sterile mortar, add sterile water, after grinding, suck the grinding solution onto the LB medium plate, and spread it evenly with a triangle At the same time, absorb the same amount of sterile water rinse solution for the third time and smear the plate to test whether the surface of Tetranychus urticae is thoroughly disinfected; finally, place t...

Embodiment 2

[0044] The optimization of embodiment 2 bacterial strain W1 fermentation conditions

[0045] 1. Single factor experimental design

[0046] Take out the strain W1 stored in the -80°C refrigerator, streak it on the LB solid plate for overnight culture and activation, pick a single colony the next day, inoculate it in the LB liquid medium with a pH of 7, and ferment at 37°C and 160rpm for 24 hours For fermentation, the fermented liquid was inserted into a 500mL fermentation bottle equipped with 150mL fermentation medium according to the inoculum amount of 5%, cultivated in a constant temperature shaker at 37°C and 160rpm for 24h, set three repetitions, and mixed the three repeated fermented liquids evenly. Determination of OD of bacterial culture 600 value. At the same time, the sample was serially diluted 10 times with sterile water, and 10 -6 cfu / ml, 10 -7 cfu / ml and 10 -8 cfu / ml concentration gradient dilution, transferred to the pre-poured LB solid plate (200 μl / dish), s...

Embodiment 3

[0053] Colonization ability determination of embodiment 3 bacterial strain W1

[0054] 1. GFP marker of strain W1

[0055]Take out the strain W1 stored in the -80°C refrigerator and streak it on the LB solid plate for overnight culture and activation. Pick a single colony the next day and inoculate it in 5mL GMI medium, shake it at 30°C and 150rpm overnight, and use 10% inoculum size Transfer to 5mL GMI medium, 37°C, 230rpm shaking culture for 3.5h, take the fermentation liquid and inoculate it into 5mL GMII medium according to the inoculum size of 5%, 37°C, 230rpm shaking culture for 1.5h, take 1mL culture, and centrifuge at room temperature at 5000rpm After 5 minutes, resuspend the bacterial pellet with 1 / 10 volume supernatant, which is the competent cell of Bacillus amyloliquefaciens strain W1. Add 10 μL of plasmid pEGFP-C1 (pEGFP-C1 was purchased from Bao Biological Engineering (Dalian) Co., Ltd.) into the prepared Bacillus amyloliquefaciens strain W1 competent cells, mix...

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Abstract

The invention discloses a bacillus amyloliquefaciens strain W1 for preventing and controlling plant spider mites and its application. The preservation number of the strain W1 is: CGMCC No.11949. The strain W1 is isolated from the body of Tetranychus urticae that died naturally. The lethality rate of the strain W1 reached 92.3% in the petri dish killing experiment. In the greenhouse control effect experiment, the control effect on spider mites on corn leaves, strawberries, tomatoes, and broad beans, as well as on apple trees and pear trees in the orchard, has reached more than 80%.

Description

technical field [0001] The invention relates to a strain of Bacillus amyloliquefaciens and its application in preventing and controlling plant spider mites. technical background [0002] In the current control of agricultural pests, herbivorous mites are recognized as one of the most difficult pest groups in the world. , Acari (Acari), True Acaria (Acariformes), Tetranychoidea (Tetranychoidea). Tetranychus are a worldwide pest. Because of their small size, short generations, short developmental cycle, rapid reproduction, strong adaptability, and high mutation rate, they are prone to produce resistance to pesticides and are extremely difficult to control, resulting in the intensification of crop mite damage. The situation of small insects becoming a catastrophe has caused great losses in agricultural production, affecting agricultural production and farmers' income, and has become an important and urgent problem in agricultural production. [0003] Chemical control is still...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/00A01P7/02C12R1/07
CPCA01N63/00C12N1/20C12N1/205C12R2001/07
Inventor 何月秋李兴玉何鹏飞吴毅歆何鹏搏王再强王强杨发祥
Owner MICROBIAL FERMENTATION ENG RES CENT CO LTD OF YUNNAN PROVINCE
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