title: 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.
authors.affiliation: a Department of Applied Engineering , Zhejiang Institute of Economic and Trade , Hangzhou , Zhejiang , P.R. China.
authors.forename: Fengjun
authors.initials: F
authors.lastname: Wang
authors.affiliation: b Institute of Seafood, Zhejiang Gongshang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Junli
authors.initials: J
authors.lastname: Feng
authors.affiliation: a Department of Applied Engineering , Zhejiang Institute of Economic and Trade , Hangzhou , Zhejiang , P.R. China.
authors.forename: Sudan
authors.initials: S
authors.lastname: Ye
authors.affiliation: c Xinjiang Environment Monitoring Center , Urumqi , Xinjiang , P.R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: a Department of Applied Engineering , Zhejiang Institute of Economic and Trade , Hangzhou , Zhejiang , P.R. China.
authors.forename: Hannian
authors.initials: H
authors.lastname: Huang
authors.affiliation: d Korla Entry-Exit Inspection and Quarantine Bureau , Korla , Xinjiang , P.R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Hua
mesh_terms: D000818:Animals; D003576:Electron Transport Complex IV / Q000235:genetics*; D019476:Insect Proteins / Q000235:genetics; D007915:Lepidoptera / Q000145:classification* / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D031989:Pyrus / Q000469:parasitology*; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D019476:Insect Proteins; D003576:Electron Transport Complex IV
keywords: Korla fragrant pears; Real-time fluorescence quantitative PCR; cytochrome oxidase subunit I; quarantine pest