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Method for detecting avermectin pesticide resistance produced by diamondback moth

An abamectin, diamondback moth technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as the frequency of occurrence of insect-resistant individuals, long test time, etc., which can not reflect the occurrence of insect resistance, and save labor. and resources, accurate resistance results, reducing the effect of intermediate processes

Inactive Publication Date: 2012-03-21
高希武 +2
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Problems solved by technology

[0003] However, there are obvious deficiencies in the above-mentioned traditional monitoring method for the resistance of Plutella xylostella xylostella to abamectin pesticides: (1) large-scale artificial breeding of test insects is required
(2) Long test time is required
The traditional biological test takes more than two days; (3) The traditional biological test results are only an overall judgment on the resistance of the tested insect population, and cannot reflect the frequency of insect resistant individuals

Method used

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Embodiment Construction

[0067] The preferred technical solutions of the present invention will be described in detail below.

[0068] A method for detecting the resistance of diamondback moth to avermectins pesticides, comprising the following steps:

[0069] In the first step, after picking back the diamondback moth from the field (it can be an alcohol-soaked specimen or a live worm), the extraction of the genome DNA of a single head is carried out, and the steps include,

[0070] (1) Place the Plutella xylostella sample in a pre-cooled homogenizer at -4°C, add 200 μL TE buffer for homogenization,

[0071] (2) Add 200 μL of cell lysate, mix well, transfer to a 1.5 mL centrifuge tube, then add 3 μL proteinase K, place at 55°C for 10 minutes, shake well 3 times during the period, until the sample is a translucent viscous liquid,

[0072] (3) Add 300 μL chloroform, mix gently,

[0073] (4) Add 250 μL of precipitation solution, place at room temperature for 2 minutes, and centrifuge at 10,000 rpm for ...

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Abstract

The invention discloses a method for detecting avermectin pesticide resistance produced by diamondback moth. The method comprises the following steps of 1, carrying out single-end genome DNA extraction of a diamondback moth sample, 2, carrying out specific gene fragment PCR amplification of a DNA template extracted by the step 1; 3, recovering a PCR amplified product DNA obtained by the step 2, 4, according to a sequencing result, detecting if an N-end of the recovered PCR amplified product DNA has a mutational site, and 5, according to a sequencing result obtained by the step 4, carrying out determination of avermectin pesticide resistance produced by the diamondback moth sample. The method does not need a mass of captive-bred experimental insects and saves labor forces and resources. Through the method, accurate resistance detection results are obtained and occurrence frequency of an insect individual with resistance is shown.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting the resistance of diamondback moth to avermectins pesticides. Background technique [0002] At present, the resistance detection of Plutella xylostella to avermectins pesticides is mainly carried out through the traditional bioassay method-leaf drug film method: take fresh and non-polluted cabbage leaves and soak them in serial concentrations of avermectins pesticide solutions for 10 Second, with distilled water (containing 0.01% Triton-X100) as a control, after indoor drying, 10 to 20 3rd instar larvae of the same size were connected, and each concentration was repeated 3 times. Check the results after 48 hours. Then use POLO or SAS software to process the data, calculate the LC50 value, and judge the resistance level of Plutella xylostella according to the comparison of the LC50 value. [0003] However, there are obvious deficiencies in the above-mentioned ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 高希武宋敦伦梁沛史雪岩尚庆利
Owner 高希武
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