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
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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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