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.

Development of LAMP and droplet digital PCR methods for differentiation between Cydia pomonella (L.) and Grapholita molesta (Busck)

“…The conventional morphological identi cation of C. pomonella needs the related expert(s) and necessary equipment. Though the developed molecular methods for C. pomonella can overcome the shortcoming of the morphological method 13,15,16,17 , they are PCR-based method and still needs the complicated DNA extraction process and the laboratory setting. In recent years, the LAMP method has been recognized as a potential method to replace the PCR-based methods and has the potential to be used in the eld 20 .…”

“…Until now, the diagnostic methods to identify C. pomonella mainly include the morphological characteristics and molecular methods such as the mitochondrial COI gene 13,14,15 , real-time quantitative PCR 16,17 , specie-speci c COI 16,18 , and PCR-RFLP 19 . However, these molecular methods need a complicated DNA extraction process and a laboratory setting, which is not suitable for on-site diagnostics 20 .…”