关键词: 苹果蠹蛾, 香梨优斑螟, 梨小食心虫, 地老虎, 实时荧光定量PCR

Abstract: Laspeyresia pomonella, Euzophera pyriella Yang, Grapholitha molesta and Blank cutworm are important pets of fragrant pears and other fruits in Xinjiang, and they are also internationally important pests on inspection and quarantine. However, the identification relies mainly on morphological characters up to date. The lack of molecular detection methods greatly limits the international export trading of fragrant pears and related products. The 16S rDNA COI fragments from the four pests were cloned and sequenced. Then, specific primers and probes were designed according to the sequencing data, and the fluorescence Real-time quantification PCR detection system was established. The results demonstrates the limits of detection concentration of Laspeyresia pomonella, Euzophera pyriella Yang, Grapholitha molesta and Blank cutworm in this system are respectively 1.605×10^-2，0.729×10^-2，0.475×10^-2 and 0.818×10^-2ng/μL, meeting the demand of inspection and quarantine routine testing. To overcome the residual bodies of some insects and some pieces mixed samples, or several insects together mixed samples appeared in actual testing process, DNA extracted from single, multiple and mixed pest samples were detected and evaluated, further verifying the specificity of this method, the reliability and applicability.

Key words: Laspeyresia pomonella, Euzophera pyriella Yang, Grapholitha molesta, Blank cutworm, Real-time fluorescence quantitative PCR