Molecular detection method for resistance of chilo suppressalis to chlorantraniliprole and primers special for method

The invention relates to a technology for resistance to chlorantraniliprole, which is applied in the field of molecular detection of chlorantraniliprole resistance and its special primers, and can solve problems such as a large increase in the number of residual insects, environmental pollution, and a decline in field control effects. , to achieve the effect of delaying the development of resistance and high sensitivity

Inactive Publication Date: 2016-07-13
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the irrational use of chemical pesticides has led to the continuous increase in the level of borer resistance, the decline in field control efficiency, and the large increase in the number of residual insects, thereby increasing the rampant damage of borers.
At the same time, problems such as environmental pollution caused by chemical pesticides and excessive drug residues in agricultural products have become increasingly prominent

Method used

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  • Molecular detection method for resistance of chilo suppressalis to chlorantraniliprole and primers special for method
  • Molecular detection method for resistance of chilo suppressalis to chlorantraniliprole and primers special for method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Application of special primers in the detection of individuals resistant to Chilo borer in Xiaoshan, Zhejiang Province

[0018] Primer design

[0019] According to the gene sequence (sequence 1) of the nidine receptor in Chilo suppressalis, self-designed allele-specific PCR primers are as follows:

[0020] Primer 1: 5'-TACTAGACGTCGCTGACGG-3'

[0021] Primer 2: 5'-TACTAGACGTCGCTGACGA-3'

[0022] Primer 3: 5'-TGTTTCCCATTATGGGTGAC-3'

[0023] The primer design of the present invention does not rely on any primer design software, but is designed by itself according to the ryanodine receptor gene fragment to be amplified and the basic principles of primer design. Among them, primer 1 is an upstream primer dedicated to sensitivity, primer 2 is an upstream primer dedicated to resistance, and primer 3 is a downstream primer dedicated to sensitivity / resistance. The first nucleotide at the 3' end of primer 1 (see the double underline) is consistent with the nucle...

Embodiment 2

[0028] Example 2: Application of special primers in the detection of individuals resistant to Chilo borer in Jinhua, Zhejiang Province

[0029]Compared with Example 1, the primers and methods used are the same as those in Example 1 except that the regions where the individuals of Chilo suppressalis come from are different. The ryanodine receptors of 30 individuals of Chilo suppressalis collected from Jinhua area, Zhejiang Province in 2016 were amplified by PCR and verified by sequencing (the method is the same as in Example 1). The number of individuals with high resistance to chlorantraniliprole was 1, and the mutation frequency was 3.3%; the number of homozygous individuals without the G4946E mutation, that is, chlorantraniliprole-sensitive individuals was 13, and the frequency was 43.3%; the heterozygous individuals were The number of individuals between the chlorantraniliprole-sensitive type and the high-resistant type was 16, and the frequency was 53.3% (Table 1). In a...

Embodiment 3

[0030] Example 3: Application of special primers in the detection of individuals resistant to Chilo borer in Quzhou, Zhejiang Province

[0031] Compared with Example 1, the primers and methods used are the same as those in Example 1 except that the regions where the individuals of Chilo suppressalis come from are different. The ryanodine receptors of 30 Chilo suppressalis individuals collected from Quzhou, Zhejiang Province in 2016 were amplified by PCR and sequenced for verification (the method is the same as in Example 1). The number of individuals with high resistance to chlorantraniliprole was 0; the number of homozygous individuals without the G4946E mutation, that is, the number of chlorantraniliprole-sensitive individuals was 10, and the frequency was 33.3%; the heterozygous individuals were between chlorantraniliprole and The number of individuals between the amide-sensitive type and the high-resistant type was 20, and the frequency was 66.7% (Table 1). In addition,...

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Abstract

The invention discloses a molecular detection method for resistance of chilo suppressalis to chlorantraniliprole and primers special for the method.The invention provides a specific primer composition which is composed of single-chain DNA molecules as shown in a sequence 1, a sequence 2 and a sequence 3 in a sequence table.The specific primer composition can be used for detecting whether an action site ryanodine receptor of chlorantraniliprole is subjected to gene G4946E mutation or not, G4946E mutation of a ryanodine receptor gene of chilo suppressalis means that the 14837 nucleotide located at the 5' tail end of an open reading frame of the ryanodine receptor gene is mutated into A from G, and therefore the resistance of chilo suppressalis to chlorantraniliprole is detected.The specific primer composition and application thereof have important significance in monitoring a development trend of the resistance of chilo suppressalis field populations to chlorantraniliprole, guiding reasonable pesticide use, delaying resistance development, making a resistance management policy and adjusting a control strategy in time.

Description

technical field [0001] The invention relates to a molecular detection method for the resistance of Chilo suppressalis to chlorantraniliprole and special primers thereof. technical background [0002] Chilosuppressalis belongs to Lepidoptera Pyralidae of Lepidoptera. It is a rice borer pest, commonly known as core borer, and is the dominant species of borer in most rice production areas. The outbreaks in East Asia are mainly distributed in China, South Korea, and Japan, directly threatening agricultural production. The damage of rice stem borer at the seedling stage can cause dry sheaths and dead hearts, and damage at the booting and earing stages can cause dry booting, white ears, and insect damage to plants, etc., seriously affecting the quality and yield of rice. In recent years, with the adjustment of planting structure and farming system, the area of ​​hybrid rice has increased year by year, which has expanded the effective insect source area of ​​overwintering generati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2600/172C12Q2531/113
Inventor 鲁艳辉郑许松何晓婵周小军钟列权黄贤夫吕仲贤
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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