Identification method for L. sativae Blanchard, L. huidobrensis and L. trifolii

Abstract

The invention discloses an identification method for Liriomyza sativae. To be specific, the identification method comprises the following steps: respectively synthetizing specific primers of L. sativae Blanchard, L. huidobrensis and L. trifolii according to mtDNA (mitochondrial deoxyribonucleic acid) COI sequences of the L. sativae Blanchard, the L. huidobrensis and the L. Trifolii; performing PCR (Polymerase Chain Reaction) amplification on genome DNA of the three Liriomyza sativae adopted as templates for three times under the condition of the same template but different primers, thereby obtaining PCR products of the three Liriomyza sativae; detecting, staining, observing and imaging the PCR product of each Liriomyza sativae; if a bright amplification belt is formed in an electrophoresis result of the PCR product of the specific primer of the L. sativae Blanchard and the molecular weight of the PCR product ranges from 250bp to 500bp, determining that the Liriomyza sativae with the PCR product is the L. sativae Blanchard; if a bright amplification belt is formed in an electrophoresis result of the PCR product of the specific primer of the L. sativae Blanchard, and the molecular weight of the PCR product ranges from 550bp to 600bp, determining that the Liriomyza sativae with the PCR product is the L. Huidobrensis; and if a bright amplification belt is formed in an electrophoresis result of the PCR product of the specific primer of the L. trifolii, and the molecular weight of the PCR product ranges from 450bp to 550bp, determining that the Liriomyza sativae with the PCR product is the L. Trifolii. The identification method is simple to operate, strong in repeatability and high in sensitivity and has the advantages of stability and rapidness.

Description

technical field [0001] The invention relates to an identification method of specific primers for Liriomyza sativae, specifically, an identification method for Liriomyza sativae, Liriomyza sativae and Liriomyza clover. Background technique [0002] L. sativae Blanchard, also known as vegetable leafminer, American melon leaflet, and alfalfa leaflet, was first discovered in Argentina in 1938 and is now distributed in 29 provinces, autonomous regions, and municipalities directly under the central government in my country, such as Hainan, Guangdong, Fujian, Hubei, Hunan, Zhejiang, Anhui, Sichuan, Henan, Shandong, Beijing, etc. have caused serious harm to our country's fruits, vegetables, cotton, tobacco, flowers, etc., and have gradually become an important part of my country's agricultural production process. Outstanding issue. The feeding habit of Liriomyza sativae is omnivorous, with a wide range of hosts, which can harm more than 110 kinds of vegetables and fruits in more than...

AI Technical Summary

Problems solved by technology

Isoenzyme electrophoresis technology requires high professional skills of operators, and operators are required to be able to accurately interpret the electrophoretic zymogram; restriction fragment length polymorphism PCR technology can accurately distinguish related species of Liriomyza sativae, but the screening requirements are limited. The workload of the endonuclease species is relatively large; although the random amplification polymorphic DNA PCR is simple to operate, the reproducibility is poor and the requirements for experimental conditions are relatively high

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