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Primer pair, kit and detection method for detecting abamectin resistance of tetranychus urticae

A mite abamectin and abamectin technology, applied in the field of molecular detection, can solve the problems of large number of test insects, long time-consuming, difficult micro-operation of test insects, etc., to achieve high sensitivity, delay the development of resistance, and reasonable The effect of medication

Inactive Publication Date: 2013-12-18
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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Problems solved by technology

However, traditional spider mite resistance monitoring adopts slide immersion method. It is not easy to operate small test insects, and requires a large number of test insects (at least 700), and the test results can only be observed after at least 24 hours. By comparing with relatively sensitive strains , to know the level of resistance to the agent, so it takes a long time

Method used

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  • Primer pair, kit and detection method for detecting abamectin resistance of tetranychus urticae
  • Primer pair, kit and detection method for detecting abamectin resistance of tetranychus urticae
  • Primer pair, kit and detection method for detecting abamectin resistance of tetranychus urticae

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Effect test

Embodiment 1

[0036] Embodiment 1 is to the acquisition of Abamectin resistance and sensitive population detection specific primer

[0037] 1. Extraction of Tetranychus genomic DNA

[0038] The genomic DNA extraction method of a single female adult mite used the DNA extraction kit (solution type) of Beijing Biotech Co., Ltd., and the method was carried out according to the kit instructions. The concentration of the genomic DNA of the single-headed spider mite was measured by Nanodrop2000c to be 6.60ng / μl.

[0039] 2. PCR amplification and sequencing of the spider mite genome

[0040] According to the point mutation site and sequence of the glutamate-gated chloride ion channel (GluCl) of the two-spotted spider mite reported by Kwon et al (2010), a primer pair containing the GluCl fragment of this point mutation was designed: 5'405F: GGG GAA TAC AGC TGT CTC AAA, 3'405F: TGA ATC CTT GGC GGT GTC AAA, Amplification of genomic DNA from indoor and field resistant populations of Tetranychus urtic...

Embodiment 2

[0044] Embodiment 2 detects and analyzes to Abamectin resistance and sensitive population

[0045] 1. Bioassay of different populations of Tetranychus urticae

[0046] Table 1 Sensitivity determination results of the sensitive and resistant populations of Tetranychus urticae to abamectin

[0047] population

b±SE

LC 50 (mg / L)

95% confidence interval

Resistance multiple

sensitive species

1.12(±0.17)

0.093

0.0486-0.176

1.00

Huairou population

0.81（±0.14）

343.54

144.73-815.44

3693.98

Haidian population

1.61(±0.19)

201.13

142.60-283.68

2162.69

[0048] As can be seen from Table 1, the Huairou population and the Haidian population have produced extremely high levels of resistance to abamectin, respectively, and the LC50 values ​​for the female adult mites of the Huairou and Haidian populations were 343.54 mg / L and 201.13 mg / L respectively. L, compared with indoor relatively sensit...

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Abstract

The invention relates to the field of molecular detection, and in particular relates to a primer pair, a kit and a detection method for judging abamectin resistance of tetranychus urticae. The primer pair comprises (1) upstream primers for detecting resistant and susceptible populations, which are RF:5'-AAGCTGTTGACATTTGGACGA-3' and SF:5'-AAGCTGTTGACATTTGGACGG-3', and (2) a common downstream primer which is paired with the upstream primers for use. According to the primer pair, the kit and the detection method, the resistance or susceptibility of tetranychus urticae to abamectin is judged by detecting GluCl (Glutamate-gated chloride channel) gene mutation in tetranychus urticae. The primer pair, the kit and the detection method have the advantages of high speed, effectiveness, high sensitivity, capability of realizing high-flux detection of a large number of samples and the like, and have the effects of monitoring the resistance gene frequency and resistance development trend of the field population of tetranychus urticae to abamectin, timely adjusting a prevention and control strategy for tetranychus urticae, guiding rational drug use and delaying the resistance development.

Description

technical field [0001] The invention relates to the field of molecular detection, in particular to a primer pair, a kit and a detection method for detecting the abamectin drug resistance of spider mites. Background technique [0002] The two-spotted spider mite Tetranychus urticae belongs to the spider mite subclass Acariidae Tetranychus, commonly known as red spider, is an important agricultural pest worldwide, and damages a variety of hosts, including fruit trees, vegetables, cotton, wheat and other important crops. The adult and young nymphs of spider mites gather on the back of plant leaves for piercing and sucking, causing chlorotic spots on the leaves, and in severe cases, causing the leaves to dry and fall off, which greatly affects the yield and quality of crops. [0003] Due to the characteristics of small size, many generations, fast reproduction speed and short development period, the spider mite is very easy to develop drug resistance. Abamectin, as an antibioti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 王少丽唐小凤张友军吴青君谢文徐宝云
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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