Avermectin-resistant Neoseiulus barkeri molecular marker, and application and detetion method thereof

A technology of Neoseiid mite and abamectin, which is applied in the field of molecular biology, can solve the problems of the death of M. pasteurii and the inability to coordinate the development of chemical control and biological control, and achieves strong specificity and optimization. PCR system and amplification procedure, detection specificity effect

Active Publication Date: 2017-10-20
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The laboratory research shows that abamectin also has strong toxicity to Neoseiius pasteuri, and the indoor bioassay LC 50 The value is 50.03mg/L, so it i...

Method used

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  • Avermectin-resistant Neoseiulus barkeri molecular marker, and application and detetion method thereof
  • Avermectin-resistant Neoseiulus barkeri molecular marker, and application and detetion method thereof
  • Avermectin-resistant Neoseiulus barkeri molecular marker, and application and detetion method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Clone and SNP Analysis of Glutamate-gated Chloride Channel Gene of Neoseiusius mite Resistance and Sensitivity Strains

[0020] 1. Materials

[0021] 1.1 Source of tested mite

[0022] Mite source tested

[0023] Sensitive strain: Neoseius pasterei was collected from the lemon leaves around the orchard of the Citrus Research Institute of the Chinese Academy of Agricultural Sciences. After collection, it was reared indoors for multiple generations without contact with pesticides, and its toxicity was determined to determine its LC. 50 The value is at a low level, and there is no obvious resistance to abamectin.

[0024] Resistant strains: Using the sensitive strain of Neoseius pastereius as the raw material for resistance screening, after multiple generations of screening, the LC of Neoseius pastereius to abamectin 50 From 50.03mg / L to 20572.3mg / L, the resistant population increased by 411.2 times compared with the sensitive population. After that, it will ...

Embodiment 2

[0040] Example 2 Obtaining of molecular markers of Neoseiius pasteurii's resistance to abamectin

[0041] The sequence comparison software BioXM2.6 was used to compare the sequences of the target genes of the sensitive strain and the resistant strain. The comparison results are as follows: figure 1 As shown, it is found that there is a mutated base at the 1202 site, and the G base of the sensitive system is mutated into the A base of the resistant strain. At the same time, the mutation of the base eventually leads to the change of the amino acid sequence, such as figure 2 As shown, the base fragment was translated into an amino acid fragment by using BioXM2.6 software, and an amino acid mutation was found. The glycine (G) of the sensitive strain at the 401st position was mutated into the aspartic acid (D) of the resistant strain.

[0042] According to the position of the resistance mutation site, the resistance marker of Neoseiusius pasterei was carried out:

[0043]1) Prim...

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Abstract

The invention discloses an avermectin-resistant Neoseiulus barkeri molecular marker with the nucleotide sequence represented by SEQ ID No.3, and a primer 4F/4R for amplifying the marker, and also discloses an application of the molecular marker in the avermectin-sensitive line and avermectin-resistant line classification of the Neoseiulus barkeri m, and a detection method of the avermectin-resistant Neoseiulus barkeri molecular marker. The detection method comprises the following steps: extracting total DNA from Neoseiulus barkeri to be detected, carrying out PCR amplification by using the total DNA as a template and using the primer 4F/4R to obtain a NbGluCl gene fragment containing a mutation region, and carrying out sequence comparison on the obtained gene fragment and the avermectin-resistant Neoseiulus barkeri molecular marker with the nucleotide sequence represented by SEQ ID No.3 in order to judge whether a line is an avermectin-sensitive or avermectin-resistant line or not. The marker has a high specificity and a high stability, and can be used to analyze whether the sample to be detected is the avermectin-sensitive line or the avermectin-resistant line.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and relates to a molecular marker, in particular to a molecular marker for the resistance of Neoseiusius pasteri to abamectin and its application and detection method. Background technique [0002] Neoseiidi genus Acarina, Parasitica, Gamasinae, Phytoseiidae, and Neoseiidi genus, because of its short development period, low mortality rate, high oviposition rate, and spreading ability Strong advantages are considered to be one of the best biological control natural enemies. Neoseius pasteurii is a polyphagous predatory mite. In addition to preying on spider mites and thrips, it can also prey on aphids, psyllids, whiteflies, scale insects, springtails, tarsal mites, filamentous fungi and mosquitoes. larvae etc. [0003] Abamectin is a new biological insecticide and acaricide, and it is a widely used pesticide preparation. It is mainly used to control Panonychus citrus in citrus orchards....

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 冉春于士将罗忍潘琦王翠伦侯栋元丛林刘浩强李鸿筠
Owner SOUTHWEST UNIVERSITY
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