Identification of three internal feeders from Korla fragrant pears by quantitative real-time polymerase chain reaction

Abstract

Cydia pomonella, Euzophera pyriella Yang, and Grapholitha molesta are destructive pest species of Korla fragrant pears in Xinjiang. They are also quarantine pests of concern in a number of countries. Identification of these small pest larvae by morphological characters is difficult, and misidentifications will influence appropriate quarantine decisions. Here, a 520 bp fragment of mitochondrial cytochrome oxidase subunit I (COI) was first amplified and sequenced from each species, and a diagnostic region was observed. Subsequently, the species-specific primer and probe sets of quantitative real-time PCR (qPCR) were designed, which amplified a 114–116 bp fragment of COI genes. This method was validated by amplification DNA extracted from single, multiple, and mixed pest samples. Results indicated that this method allows rapid discrimination and reliable identification of larvae, pupae, and adult specimens of all three species, which could help the international export trading of Korla fragrant pears and related products.

Keywords:

• Real-time fluorescence quantitative PCR
• quarantine pest
• Korla fragrant pears
• cytochrome oxidase subunit I

Disclosure statement

This work was supported by the Grant from the Science Research Project of Zhejiang Federation of Supply and Marketing Cooperative [number of 16SSY04]; the Science Research Project of Zhejiang Institute of Economic and Trade [number of 16QN02]; Korla Key Science and Technology Project. Authors do not have any conflicts of interest.

Additional information

Funding

This work was supported by the Grant from the Science Research Project of Zhejiang Federation of Supply and Marketing Cooperative [number of 16SSY04]; the Science Research Project of Zhejiang Institute of Economic and Trade [number of 16QN02]; Korla Key Science and Technology Project.