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Molecular detection method and special primer for resistance of sesamia inferens to chlorantraniliprole

A technology of resistance to the large borer, which is applied in the field of molecular detection of the resistance of the large borer to chlorantraniliprole and its special primers, can solve the problems affecting the safe production of rice, and achieve the effect of high sensitivity and delaying the development of resistance

Active Publication Date: 2016-07-06
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the use of fipronil has been banned in paddy fields since July 1, 2009, with the continuous expansion of single-double mixed planting and super rice planting area, the damage of the large borer has gradually become the main obstacle affecting the safe production of rice

Method used

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  • Molecular detection method and special primer for resistance of sesamia inferens to chlorantraniliprole
  • Molecular detection method and special primer for resistance of sesamia inferens to chlorantraniliprole
  • Molecular detection method and special primer for resistance of sesamia inferens to chlorantraniliprole

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Application of special primers in the detection of resistant individuals of the overwintering generation of the large borer in Quzhou, Zhejiang Province

[0016] Primer design

[0017] According to the gene sequence (SEQ ID NO: 4) of the nicotine receptor gene of the large borer, self-designed allele-specific PCR primers are as follows:

[0018] Primer 1: 5'-TGTTGGATGTCGCTG G -3'; (SEQ ID NO: 1)

[0019] Primer 2: 5'-TGTTGGATGTCGCTG G -3'; (SEQ ID NO: 2)

[0020] Primer 3: 5'-GAAGAAGTTGAACGCGATCA-3' (SEQ ID NO: 3)

[0021] 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...

Embodiment 2

[0025] Example 2: Application of special primers in the detection of resistance individuals of the overwintering generation of S. moerens in Jinhua, Zhejiang Province

[0026] Compared with Example 1, the primers and methods used are the same as those in Example 1 except that the region of origin of S. PCR amplification and sequencing of the ryanodine receptors of 30 overwintering individuals of S. chinensis collected from the Jinhua area of ​​Zhejiang Province in 2016 (the method is the same as in Example 1), the results show that the homozygous individuals with the G4924E mutation, namely The number of individuals highly resistant to chlorantraniliprole was 0; the number of homozygous individuals without the G4924E mutation, that is, the number of individuals sensitive to chlorantraniliprole was 12, and the frequency was 40.0%; the heterozygous individuals were between chlorantraniliprole and The number of individuals between the formamide-sensitive type and the high-resis...

Embodiment 3

[0027] Example 3: Application of special primers in the detection of overwintering resistant individuals of S. arborescens in Wenling, Zhejiang Province

[0028] Compared with Example 1, the primers and methods used are the same as those in Example 1 except that the region of origin of S. The ryanodine receptors of 30 overwintering individuals of S. chinensis collected from Wenling area, Zhejiang Province in 2016 were amplified by PCR and verified by sequencing (the method was the same as that in Example 1). The results showed that the individuals homozygous for the G4924E mutation, namely The number of individuals highly resistant to chlorantraniliprole was 5, and the mutation frequency was 16.7%; the number of homozygous individuals without the G4924E mutation, that is, the number of individuals sensitive to chlorantraniliprole was 8, and the frequency was 26.7%; heterozygous individuals The number of individuals between the chlorantraniliprole-sensitive type and the high-...

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Abstract

The invention discloses a molecular detection method and special primer for resistance of sesamia inferens to chlorantraniliprole, and provides a special primer composition which is composed of single-chain DNA molecules shown in the sequence 1, the sequence 2 and the sequence 3 in the sequence table. The special primer composition can be used for detecting whether gene G4924E mutation happens to an acting site, namely, a ryanodine receptor, of chlorantraniliprole or not. The gene G4924E mutation of the ryanodine receptor of sesamia inferens means that the 14771th nucleotide at the 5' end of an open reading frame of the ryanodine receptor gene mutates into A from G, and therefore resistance of sesamia inferens to chlorantraniliprole is detected. The special primer composition and application of the special primer composition have important significance in monitoring development trends of resistance of sesamia inferens field populations to chlorantraniliprole, guiding reasonable pesticide utilization, delaying resistance development, formulating resistance government policies and regulating prevention and control strategies in time.

Description

technical field [0001] The invention relates to a molecular detection method and special primers for the resistance to chlorantraniliprole of C. technical background [0002] Sesamia inferens (Walker) belongs to Lepidoptera Pyralidae. It is a borer pest with a wide range of hosts. It is a typical omnivorous insect and can feed on rice, wild rice stems, corn, sorghum, wheat, and sugarcane. and other grasses. The rice borer damages rice at the seedling stage can cause dry sheaths and dead hearts, and damages at the booting and earing stages can cause dry booting, white ears, and insect damage to plants, which seriously affects the quality and yield of rice. For a long time, Ceratocystis medinalis has been considered as a secondary pest on rice due to its miscellaneous feeding habits and other reasons, and has not received extensive attention and in-depth research. After the mid-1990s, with the evolution of rice cultivation systems, fipronil was used extensively to control th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2600/172C12Q2531/113
Inventor 鲁艳辉钟列权王国荣张发成郑许松吕仲贤
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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