Insecticidal activity of AMEP412 protein on trialeurodes vaporariorum and application of AMEP412 protein in prevention and treatment of trialeurodes vaporariorum

A whitefly and protein technology, applied in the application, insecticide, biocide and other directions, can solve the problem of low insecticidal protein in whitefly and achieve good control effect.

Active Publication Date: 2020-02-07
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a new application of Bacillus subtilis protein AMEP412, to overcome the current problem of too little insecticidal protein for whitefly, and to solve the problem of green control of whitefly

Method used

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  • Insecticidal activity of AMEP412 protein on trialeurodes vaporariorum and application of AMEP412 protein in prevention and treatment of trialeurodes vaporariorum
  • Insecticidal activity of AMEP412 protein on trialeurodes vaporariorum and application of AMEP412 protein in prevention and treatment of trialeurodes vaporariorum
  • Insecticidal activity of AMEP412 protein on trialeurodes vaporariorum and application of AMEP412 protein in prevention and treatment of trialeurodes vaporariorum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] This example illustrates the insecticidal effect of Bacillus subtilis AMEP412 protein artificially fed to whitefly.

[0024] The liquid artificial bait for feeding is sterilized by dissolving 5% yeast extract and 30% sucrose in distilled water, adding different concentrations of AMEP412 (1, 5, 10, 20, 40 and 80 μg / ml), and mixing for later use. The artificial bait without AMEP412 was used as the control. The 1-2 day-old whitefly adults are driven from the plant leaves into 50ml test tubes, and at least 50 whitefly adults are taken in each test tube. Cover the test tube cap with two layers of stretched parafilm, add bait (100 μl) between the two layers of film, and then cover the tube cap in reverse, so that the double film structure remains on the inside of the test tube and upwards. Place vertically. Each experiment was repeated three times. Mortality was measured 2 days after feeding, and mortality was recorded by counting dead whitefly adults at the bottom of the ...

Embodiment 2

[0029] This example illustrates the stability test of Bacillus subtilis AMEP412 protein.

[0030] In order to test the heat resistance of AMEP412 protein, protein samples were treated at 95°C for 15 minutes and 30 minutes, respectively. The cooled protein sample was prepared as artificial bait according to the method in Example 1, and the final concentration was adjusted to 60 μg / ml to test the insecticidal effect on whitefly. Each treatment was repeated three times with untreated protein samples as controls. In order to test the stability of AMEP412 protein against natural degradation, protein samples were placed in 1.5ml centrifuge tubes at room temperature 25°C for 24 hours and 48 hours respectively. Afterwards, artificial bait was prepared from the protein sample according to the method in Example 1, and the final concentration was adjusted to 60 μg / ml to test the insecticidal effect of whitefly. Each treatment was repeated three times with untreated protein samples as c...

Embodiment 3

[0033] This example illustrates the fluorescent localization of Bacillus subtilis AMEP412 protein in whitefly.

[0034]The AMEP412 protein was incubated with fluorescein FITC, and the FITC not bound to the AMEP412 protein was removed by molecular sieve chromatography according to the difference in molecular volume to obtain FITC-labeled AMEP412 protein. The FITC-labeled AMEP412 protein was subsequently fed to whitefly adults. The dead whitefly adults were washed with distilled water, and then the Olympus BX60 fluorescence microscope was used to excite green fluorescence at a wavelength of 495 nm for fluorescence microscopic observation. The results show( image 3 ), the green fluorescence is concentrated in the intestine of the abdominal cavity of whitefly adults, indicating that AMEP412 protein can be taken into the body of whitefly adults and localized in the intestine. This result suggests that AMEP412 plays an insecticidal role in the intestinal tract.

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Abstract

The invention relates to application of bacillus subtilis protein AMEP412 in prevention and treatment of whiteflies. The insecticidal activity of the AMEP412 protein on the trialeurodes vaporariorum and the application of the AMEP412 protein in prevention and treatment of the trialeurodes vaporariorum are discovered, and the protein can kill the trialeurodes vaporariorum at a low concentration, has heat resistance and natural degradation resistance stability, can be ingested into intestinal tracts by the trialeurodes vaporariorum, but does not inhibit protease activity in intestinal tracts; inaddition, the AMEP412 is sprayed in a greenhouse, so that a good prevention effect on the trialeurodes vaporariorum is achieved; according to the invention, a new material is accumulated for biological prevention and control work of the trialeurodes vaporariorum.

Description

technical field [0001] The invention belongs to the field of plant protection and biological control, and relates to the insecticidal activity of AMEP412 protein on whitefly and its application in controlling whitefly. Background technique [0002] Whitefly (Bemisia tabaci) is an important pest of various crops such as vegetables, cotton, and ornamental plants. It damages crops by eating plant stem and leaf juice and spreading plant viruses, resulting in huge losses in agricultural production and national economy. The main strategy for controlling powdery mildew is based on chemical insecticides. However, the whitefly has developed resistance to those overused and frequently used insecticides. In view of this, exploring new insecticides should become a new research hotspot. [0003] Compared with chemical insecticides, proteins with insecticidal activity have a relatively low probability of insect resistance due to their different modes of action, which has become a hot s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N37/46A01P7/04
CPCA01N37/46A01N63/22A01N63/50A01P7/04
Inventor 刘权肖莉杰殷奎德
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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