Rapid identification method of imported plant quarantine pest saperda octopunctata scopoli

A phytosanitary and pest technology, applied in the field of wedge beetle identification, can solve problems such as inability to identify accurate species, and achieve the effects of avoiding protein amino acid loss, good stability, and improving epidemic interception rate

Inactive Publication Date: 2017-05-10
海南出入境检验检疫局热带植物隔离检疫中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional quarantine identification method is to identify and judge whether it is a quarantine pest prohibited from entering the country according to the external morphological characteristics of the insect; When the adult or insect body is incomplete, it is impossible to identify the species accurately.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The rapid identification method of the imported phytosanitary pest Beetle longis, comprising the following steps:

[0022] A. Extracting the DNA of Beetleus octopus and Beetleus genus;

[0023] B, carrying out PCR amplification to the DNA of Beetleus octopus and Beetleus genus;

[0024] C. PCR products are stored for a period of time and then sequenced;

[0025] D. Sequence comparison to determine the species of Beetleus cuneiformis.

[0026] In this embodiment, the PCR amplification method in step B includes the following steps:

[0027] A. Add the target PCR to the template DNA 2ul and the colchicine solution, and shake the mixture;

[0028] B. Warm up the colchicine solution to 85°C to denature the mixture, and the reaction time is 50s; then modify it with streptavidin;

[0029] C. Rapid cooling of the modified mixed solution in step B to 50°C for annealing treatment; the annealing time is 30s;

[0030] D. The temperature of the mixed solution after the annealin...

Embodiment 2

[0034] The rapid identification method of the imported phytosanitary pest Beetle longis, comprising the following steps:

[0035] A. Extracting the DNA of Beetleus octopus and Beetleus genus;

[0036] B, carrying out PCR amplification to the DNA of Beetleus octopus and Beetleus genus;

[0037] C. PCR products are stored for a period of time and then sequenced;

[0038] D. Sequence comparison to determine the species of Beetleus cuneiformis.

[0039] In this embodiment, the PCR amplification method in step B includes the following steps:

[0040] A. Add the target PCR to the template DNA 2ul and the colchicine solution, and shake the mixture;

[0041] B. Warm up the colchicine solution to 95°C to denature the mixture, and the reaction time is 80s; then modify it with streptavidin;

[0042] C. Rapid cooling of the modified mixed solution in step B to 60°C for annealing treatment; the annealing time is 60s;

[0043] D. The temperature of the mixed solution after the annealin...

Embodiment 3

[0047] The rapid identification method of the imported phytosanitary pest Beetle longis, comprising the following steps:

[0048] A. Extracting the DNA of Beetleus octopus and Beetleus genus;

[0049] B, carrying out PCR amplification to the DNA of Beetleus octopus and Beetleus genus;

[0050] C. PCR products are stored for a period of time and then sequenced;

[0051] D. Sequence comparison to determine the species of Beetleus cuneiformis.

[0052] In this embodiment, the PCR amplification method in step B includes the following steps:

[0053] A. Add the target PCR to the template DNA 2ul and the colchicine solution, and shake the mixture;

[0054] B. Warm up the colchicine solution to 88°C to denature the mixture, and the reaction time is 60s; then modify it with streptavidin;

[0055] C. Rapid cooling of the modified mixed solution in step B to 52°C for annealing treatment; the annealing time is 35s;

[0056] D. The temperature of the mixed solution after the annealin...

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Abstract

The invention discloses rapid identification method of imported plant quarantine pest saperda octopunctata scopoli, 1772. The method comprises the following steps: A, extracting DNA of the saperda octopunctata scopoli and allied species of saperda; B, conducting PCR amplification on the DNA of the saperda octopunctata scopoli and allied species of the saperda; C, sequencing PCR products after the PCR products are preserved for a certain duration; and D, comparing sequences so as to determine saperda species. With the application of the rapid identification method provided by the invention, problem on detection and identification of a non-adult pest state or incomplete pest bodies can be effectively solved, and the saperda octopunctata scopoli and some allied species of the saperda can be rapidly identified, so as to protect China's farming and forestry production and ecological environment from being destroyed by pests which spread into China and to obviously improve an epidemic intercepting rate of the saperda octopunctata scopoli; and an adopted PCR amplification method is simple to operate, good in stability and high in expression amount and is capable of effectively protecting protein amino acids from getting lost.

Description

technical field [0001] The invention relates to the technical field of identification of the longhorn beetle, in particular to a rapid identification method for the imported plant quarantine pest, the longhorn beetle. Background technique [0002] Saperda octopunctata Scopoli, 1772, belongs to the class of Insecta, Coleoptera, Cyperidae, Subfamily of Cyceroptera, and the genus of Cyceroptera. Saperda spp. is a non-Chinese species of the genus Saperda spp., which is a phytosanitary pest imported into my country, and Saperda octopuntata Scopoli is a non-Chinese species of the genus Saperda that has not been distributed in China. Hosts: Broadleaf trees, mainly basswood (Tilia), but also poplar (Populus) and elm (Ulmus). Geographical distribution: Austria, Slovenia, Russia, Switzerland, Croatia, Bosnia and Herzegovina, Czech Republic, Slovakia, Maddavi, France, Ukraine, Albania, Poland, Romania, Yugoslavia, Italy, Hungary, Germany, Greece, Belgium, Spain. Biological learning: ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2537/143C12Q2563/143
Inventor 徐卫甘琴华敖苏尹新明袁永革蔡波潘英文
Owner 海南出入境检验检疫局热带植物隔离检疫中心
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