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Application of ABC transport protein to agricultural pest mite control and Bt resistance management

A transporter and agricultural technology, applied in the biological field, can solve problems such as off-target effects

Inactive Publication Date: 2020-09-15
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after screening the target gene to synthesize dsRNA, there may be an off-target effect, that is, the non-target gene is silenced; it may be partially silenced; or there may be no silencing effect after being introduced into the pest in different ways
It can be seen that it is difficult to screen out targets that are highly lethal to pests

Method used

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  • Application of ABC transport protein to agricultural pest mite control and Bt resistance management
  • Application of ABC transport protein to agricultural pest mite control and Bt resistance management
  • Application of ABC transport protein to agricultural pest mite control and Bt resistance management

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] In vitro synthesis of embodiment 1 Turkestan spider mite dsRNA-ABC

[0030] (1) Primer design for ABC gene dsRNA of Turkestan spider mite

[0031] Use Primer5.0 to design dsRNA synthesis primers for ABCA1, ABCA2, ABCA7, ABCC16, ABCC17, ABCC18 and ABCC22 genes respectively, add T7 promoter in front of the designed dsRNA synthesis primers, and add GG after it (GG can increase dsRNA synthesis quantity). See Table 1 for details.

[0032] Table 1 dsRNA synthesis primers of ABC gene of Turkestan spider mite

[0033]

[0034] Note: The italic letters in the primer sequence indicate the T7 promoter sequence.

[0035] (2) Synthesis of Turkestan spider mite dsRNA-ABC template

[0036] Using the ABC gene as a template, the primers synthesized by the dsRNA in step (1) were used for PCR amplification. The PCR reaction system is shown in Table 2.

[0037] Table 2 PCR reaction system

[0038] Reagent quantity 10x PCR Buffer (2.5mM) 2.5μL dNTP Mixture (...

Embodiment 2

[0045] Example 2 Sensitivity Analysis of Turkestan Tetranychus ABC Gene Silenced to Bt Activating Toxin

[0046] (1) Experimental objects: the sensitive strain Tetranychus turkestan, the strain resistant to Cry1Ac activated toxin and the strain resistant to Cry2Ab activated toxin;

[0047] The sensitive strain Turkestan spider mite is a descendant strain that has never been sprayed with any pesticide since 2010, and it has been raised on potted cowpea seedlings for 160 generations;

[0048] The Cry1Ac-activating toxin-resistant strain Turkestan spider mite and the Cry2Ab-activating toxin-resistant strain Turkestan spider mite were selected from the sensitive strain Turkestan spider mite through Cry1Ac-activating toxin and Cry2Ab-activating toxin respectively. 9.18 and 20.64, respectively.

[0049](2) Test items: Sensitivity to Cry1Ac activating toxin or Cry2Ab activating toxin after gene silencing of Tetranychus Turkistan ABCA1, ABCA2, ABCA7, ABCC16, ABCC17, ABCC18, ABCC22; ...

Embodiment 3

[0060] Example 3 Effect of dsRNA-ABC on hatching rate of Turkestan spider mite eggs

[0061] (1) Experimental method:

[0062] Pick 20 healthy female adults of Turkestan Tetranychus mites, put them into leaf discs (the preparation method is the same as the embodiment step (31)), put 1 head in each grid, allow them to lay eggs for 24 hours, and then remove the female adults and redundant eggs , a total of 60 eggs were retained. Add 0.2 μL of dsABCA1, dsABCA2, dsABCA7, dsABCC16, dsABCC17, dsABCC18 or dsABCC22 synthesized in Example 1 dropwise to each egg with a micropipette. CK control group was dripped with ddH 2 O, placed in a biochemical incubator to cultivate, observe and count the hatching rate of eggs and the survival rate of larvae after 72 hours of hatching. Each treatment had 3 biological replicates, and each replicate had 60 eggs of Turkestan spider mite, a total of 180 eggs.

[0063] (2) Experimental results: as shown in Table 4.

[0064] Table 4 The effect of ds...

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Abstract

The invention discloses an application of an ABC transport protein to agricultural pest mite control and Bt resistance management, and belongs to the technical field of biology. Compared with the prior art, the ABC transport protein has the following beneficial effects that the ABC transport protein is a special gene for specially interfering agriculture pest mites, and is safe to higher animals and human; the ABC transport protein has specificity and is safe to target biology; and the ABC transport protein is non-toxic and harmless to environment.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to the application of ABC transporter in the prevention and control of agricultural pests and Bt resistance. Background technique [0002] Agricultural pest mites belong to Arthropoda, Arachnida, Acari, small and micro animals of the order Eucaridae, widely distributed on various agricultural and forestry crops, and are the main pests on agricultural and forestry crops in the world today. Harm to cotton, forest fruit, corn and other crops. In recent years, the ecological environment of cotton fields and farmland has also gradually changed, resulting in the outbreak of pest mites year after year, which has become a key factor limiting the high yield and high quality of crops. [0003] For many years, the control of agricultural pest mites in my country has always been based on chemical control. Due to the frequent use of chemical insecticides, on the one hand, a large n...

Claims

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Application Information

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IPC IPC(8): C12N15/87A01K67/033
CPCA01K67/0337A01K2217/058A01K2227/706C07K14/43531C12N15/87
Inventor 赵伊英刘峰包建红
Owner SHIHEZI UNIVERSITY
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