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Molecule detection method for triazophos target resistance in rice borer

A triazophos target and molecular detection technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of long cycle, low sensitivity, and high material requirements, and achieve short cycle, high sensitivity, and accuracy high effect

Inactive Publication Date: 2009-02-11
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0005] Technical Problems The purpose of the present invention is to provide a molecular detection method for the resistance of the stem borer to the triazophos target in order to solve the problems of low sensitivity, long cycle time and high material requirements of the detection method for the resistance of the stem borer to the triazophos in the prior art, To achieve the purpose of high accuracy, short cycle and high sensitivity

Method used

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  • Molecule detection method for triazophos target resistance in rice borer
  • Molecule detection method for triazophos target resistance in rice borer
  • Molecule detection method for triazophos target resistance in rice borer

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Embodiment Construction

[0036] 1. Select 10 fourth-instar larvae from the field populations of Gaochun County, Nanjing City, Jiangsu Province, and Jiande County, Hangzhou City, Zhejiang Province, and extract genomic DNA (Shenggong UNIQ-10 Column Animal Genomic DNA Extraction Reagent) box):

[0037](1) Take a single fourth-instar C. borer, grind it into powder in liquid nitrogen, and transfer it to a 1.5ml centrifuge tube; (2) Then add 300 μl of ACL solution and 20 μl of proteinase K. (Incubate proteinase K at 37°C for 1 hour in advance, which will help activate the activity of proteinase K); (3) Shake and mix for 1 minute, then place at 55°C for 1-3 hours, during which time the mixture can be properly removed (4) Take out the sample, shake it gently when it cools down to room temperature, and then centrifuge at 12,000rpm for 5 minutes; (5) Pipette 300μl supernatant to UNIQ-10 column, and then pipette 300μl Put the AB solution into the UNIQ-10 column, mix up and down 2-3 times, and let stand at room ...

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Abstract

The invention relates to a molecular biology method for testing target enzyme resistance of chilo suppressalis which is rice insect pest for triazophos, belonging to the field of biotechnology. Auele Specific Primers which are F: 5'-GGGCAAGAAAGTAGACGCCTGGTT-3', and R: 5'- GTTCGTCAGCACTCTTCTTCCTTA-3' are used for expanding genome DNA of the chilo suppressalis, and PCR expanding products is digested by restricted enzyme MspA1 I; when the obtained segment is 534bp and 224bp, the chilo suppressalis individual is homozygote SS which is sensitive to the target enzyme of triazophos; when the obtained segment is 758bp, the chilo suppressalis individual is homozygote RR for the target enzyme resistance of triazophos; when the obtained segment is 758bp,534bp and 224bp, the chilo suppressalis individual is heterozygote RS for the target enzyme resistance of triazophos. The method has high stability, short period and high sensitivity, and is specially used for quickly testing the target enzyme resistance of the chilo suppressalis for triazophos with high sensitivity.

Description

(1) Technical field [0001] The invention relates to a molecular detection method for the resistance of Chilo suppressalis Walker to a triazophos target, belongs to the field of biotechnology, and is specially used for the high-sensitivity rapid detection of the resistance of Chilo suppressalis Walker to a triazophos target. (2) Background technology [0002] Rice is an important food crop for humans. About 50% of the world's population uses rice as a staple food, accounting for 1 / 3 of the global cereal crop planting area. Rice is also the main food crop in my country. 50% of the country's population takes rice as a staple food. The rice planting area accounts for 26.6% of the grain sown area, and the total rice output accounts for 43.6% of the total grain output. Rice production has an impact on my country's grain yield and security. maximum. Chilo suppressalis Walker (Chilo suppressalis Walker) is a major pest of rice and is widely distributed in all rice growing areas in m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 韩召军姜晓静
Owner NANJING AGRICULTURAL UNIVERSITY
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