Technique System For Molecular Identification Of Quarantine Fruit Flies In China

Fruit flies belong to the family of Tephritidae in Diptera. They are one of the most economically important pests around the world which attack a variety of tropical and subtropical fruits and vegetables. Fruit flies can be spread a long distance with their host, packaging and vehicle in the form of eggs, larvae or pupae by human. Because accurate identification based on morphology of immature stages is extremely difficult, molecular identification of fruit flies needs to be studied in plant quarantine and biological invasion areas.In this study, the research into molecular identification of quarantine fruit flies in China was carried out. The main results are as follows:(1) A DNA barcoding database of quarantine fruit flies in China was built. It included2252sequence records of181species in6genera from Asia, Oceania, Africa, Europe, North America and South America. The effectiveness of DNA barcoding for fruit flies identification was analyzed using Bactrocera spp. as a case, and the results showed that DNA barcoding can realize the accurate identification of fruit fly species except complex.(2) Twenty-seven pairs of species-specific primers for27species of quarantine fruit flies in China (Anastrepha ludens, A. obliqua, Bactrocera albistrigata, B. correcta, B. carambolae, B. dorsalis, B. latifrons, B. rubigina, B. tryoni, B. tuberculata, B. umbrosa, B. zonata, B. oleae, B. minax, B. tsuneonis, B. bezziana, B. cilifera, B. cucurbitae, B. scutellata, B. tau, Carpomya vesuviana, Ceratitis capitata, C. cosyra, C. rosa, Dacus bivittatus, Rhagoletis cerasi and R. pomonella) were designed and selected based on DNA barcodes, and the PCR technique was set up for identification of the main quarantine fruit flies in China.(3) Twenty-seven species-specific TaqMan-MGB probes for27species of quarantine fruit flies in China were designed and selected based on DNA barcodes, combined with27species-specific primer pairs, the real-time PCR technique was built for identification of the main quarantine fruit flies in China.(4) IFC chip was firstly introduced in molecular identification of entomology, and the technique for identification of the main quarantine fruit flies in China using IFC chip was established based on primers and probes obtained.(5) Fruit flies in immature stages from internal and external inspection were identified based on four techniques, and the results showed that they were B. correcta, C. cosyra, B. tau, B. minax and B. latifrons, respectively.(6) Technique system was built for molecular identification of quarantine fruit flies in China based on DNA barcoding, PCR, real-time PCR and IFC chip.This study will provide technical support for rapid identification of important quarantine fruit flies in China, and have important practical significance in plant quarantine and biological invasion areas.