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A protein bvp10 for preventing and controlling spider mites and its application

A technology of protein and two-spotted spider mite, applied in the field of agricultural microbiology, can solve the problems of accelerating pest mite resistance, economic loss of growers, heavy mite infestation, etc., and achieves an environmentally friendly effect

Active Publication Date: 2022-02-01
HUBEI BIOPESTICIDE ENG RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, due to the lack of alternatives to acaricide varieties, a large number of acaricides have been used repeatedly, which has accelerated the generation of resistance to harmful mites, resulting in more and more serious mite infestations, and caused great economic losses to growers.
[0005] At present, most of the acaricides developed in the world are chemical substances or macromolecular antibiotics that are insoluble in water, and there are few reports on small molecular protein substances. Therefore, based on the above background, it is necessary to develop protein preparations for preventing and controlling spider mites. In the field of spider mite biological control, it has important production practical significance

Method used

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  • A protein bvp10 for preventing and controlling spider mites and its application
  • A protein bvp10 for preventing and controlling spider mites and its application
  • A protein bvp10 for preventing and controlling spider mites and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Acquisition of the mite-killing BVP10 protein:

[0020] 1) According to the sequence shown in SEQ ID NO.1 (encoding the protein shown in SEQ ID NO.2), artificially synthesize the BVP10 protein gene fragment (Sangon Bioengineering (Shanghai) Co., Ltd.), and connect it to the E. coli expression vector pet28a On the plasmid, the recombinant expression plasmid pet28a-BVP10 was constructed.

[0021] 2) Expression and purification of acaricidal BVP10 protein in Escherichia coli BL21

[0022] The above-mentioned recombinant expression plasmid pet28a-BVP10 carrying the coding sequence was transformed into Escherichia coli BL21 to obtain the recombinant bacterium BL21 / pet28a-BVP10. The recombinant bacteria were inoculated in 5 mL of LB liquid medium, placed in a shaker at 37° C. and cultured until the OD600 was 0.6, and then 1.0 mmol / L of isopropyl-B-D thiogalactoside (i.e. IPTG, purchased from Sigma Corporation) was added. ) at 30°C for 3 hours. The above 50mL recombinant ba...

Embodiment 2

[0024] Application of BVP10 protein in preparation of spider mite insecticide:

[0025] 1) BVP10 protein kills two-spotted spider mite (Tetranychus urticae):

[0026] Refer to the standard method for measuring harmful mites recommended by FAO (Food and Agriculture Organization of the United Nations)-slide immersion method. Cut the double-sided tape into 2-3 cm long, stick it on one end of the microscope slide, use tweezers to peel off the paper on the tape, use a zero brush to select female adult mites with the same size, bright body color and lively action, Its back is glued on the double-sided adhesive tape (note: do not stick mite feet, mite beard and mouthparts), each sticks 4 rows, and each row sticks 10 heads. Place them in a biochemical incubator with a temperature of 25°C and a relative humidity of about 85% for 4 hours, observe with binoculars, and remove dead or inactive individuals. On the basis of the pre-test, dilute the medicine to 5-7 concentrations with water...

Embodiment 3

[0038] Application of BVP10 protein in field control of strawberry spider mite (Tetranychus cinnabarinus):

[0039] Test agent: 20% avi. spirodiclofen suspension concentrate (Hebei Xingbai Agricultural Technology Co., Ltd., dilute 3000 times before use); the effective protein concentration of biological acaricide BVP10 is 389.5 μg / mL.

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Abstract

The invention belongs to the technical field of agricultural microbiology, and specifically relates to a protein BVP10 for preventing and controlling spider mites and its application, and the protein is shown in SEQ ID NO.2. The half-lethal concentration of BVP10 protein against Tetranychus urticae was 19.07 μg / mL, the half-lethal concentration against Panonychus citrus was 58.05 μg / mL, the half-lethal concentration against Tetranychus cinnabarinus was 36.08 μg / mL, and the half-lethal concentration against spider mite The control effect is 79.53%~95.45%. The protein provides a new selectivity for the preparation of new acaricides.

Description

technical field [0001] The invention belongs to the technical field of agricultural microbiology, and in particular relates to a protein BVP10 for preventing and controlling spider mites and its application. Background technique [0002] Spider mite, also known as red spider mite, spider mite (spider mite), plant-eating mite of the subclass Acarina (Acarina) Tetranychidae (Tetranychidae). Feeds on leaves and fruit of houseplants and important agricultural plants, including fruit trees. The life of spider mites goes through five stages: eggs, larvae, early nymphs, late nymphs, and adults, and it takes about 3 weeks from egg to adult. The life cycle of spider mites is short, and they reproduce rapidly. Generally, one generation can be completed in a few to ten days, and several to ten generations can be completed in one year. They often cause extremely serious damage to crops. Adult mites are about 0.5 mm long, with red, green or brown bodies, and form a loose silk net on th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/00C12N15/11C12N15/70A01N47/44A01P7/02
CPCC07K14/00C12N15/70A01N47/44A01N63/50A01P7/02
Inventor 刘晓艳陈凌闵勇周荣华饶犇龚艳邱一敏朱镭廖先清陈伟曹治钢石丽桥杨靖钟
Owner HUBEI BIOPESTICIDE ENG RES CENT
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