A diagnostic multiplex polymerase chain reaction method to identify Japanese internal apple-feeding Lepidopteran pests: Grapholita molesta, Grapholita dimorpha (Lepidoptera: Tortricidae), and Carposina sasakii (Lepidoptera: Carposinidae)
- 254 Downloads
We present a multiplex polymerase chain reaction method to differentiate between three Japanese internal apple-feeding pests: the oriental fruit moth, Grapholita molesta (Busck), Grapholita dimorpha Komai, and the peach fruit moth, Carposina sasakii Matsumura. A 1,342-bp fragment of mitochondrial cytochrome oxidase subunit I (COI) was sequenced in each species. The three species showed consistent and diagnostic differences in the region of the COI gene, from which three species-specific forward primers were designed. The forward primers along with one universal reverse primer were used to selectively amplify DNA from specimens of diverse geographic origin for each corresponding target species. This method enabled easy, immediate, and accurate identification of internal feeding Lepidoptera in apples and other fruits.
KeywordsMultiplex polymerase chain reaction Identification Internal apple-feeding pests
We thank Nao Watanabe and Masao Kaneko (Nagano Fruit Tree Experiment Station); Yoichi Ishiguri, Masakazu Sato, and Tomoko Murai (Aomori Prefectural Industrial Technology Research Center); Ken Nakada (Tottori Prefectural Agriculture and Forest Research Institute); and Katsuhiko Yaginuma (National Institute of Fruit Tree Science) for providing the specimens.
- Kawashima K (2008) Bionomics of the peach fruit moth (Carposina sasakii Matsumura). Bull Apple Exp Stn Aomori Prefec Agric For Res Cent 35:1–51Google Scholar
- Komai F (1979) A new species of genus Grapholita Treitschke from Japan allied to the oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae). Appl Entomol Zool 14:133–136Google Scholar
- Muraji M, Hirai Y, Akino T, Wakamura S, Yasui H, Arakaki N, Nagayama A, Ando T, Tanizaki S, Hokama Y (2010) Relationship among local populations of the grub beetle, Dasylepida ishigakiensis (Coleoptera: Scarabaeidae), detected by phylogenetic analysis based on the mitochondrial DNA sequence. Appl Entomol Zool 45:289–296CrossRefGoogle Scholar
- Oku T, Ohira Y, Wakou M (1988) Preliminary notes on plum fruit moth, Grapholita dimorpha Komai (Lepidoptera: Tortricidae). Bull Fruit Tree Res Stn C 15:49–64 (in Japanese with English summary)Google Scholar
- Scheffer SJ, Wijesekara A, Visser D, Hallett RH (2001) Polymerase chain reaction-restriction fragment-length polymorphism method to distinguish Liriomyza huidobrensis from L. langei (Diptera: Agromyzidae) applied to three recent Leafminer invasions. J Econ Entomol 94:1177–1182PubMedCrossRefGoogle Scholar
- Simon C, Frati F, Beckenbach A, Crespi B, Liu H, Flook P (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and compilation of conserved polymerase chain reaction primers. Ann Entomol Soc Am 87:651–701Google Scholar
- Takanashi M, Haji T, Adachi Y, Arai T, Ihara F, Mochizuki F (2009) Occurrence and host plants of Grapholita dimorpha (Lepidoptera: Tortricidae) in central areas of Iwate prefecture. Ann Rept Plant Prot North Japan 60:253–258 (in Japanese with English summary)Google Scholar
- Tanaka M, Abe K, Saito N, Sato S, Hisataka M, Kaimai K, Okazaki K (2005) Survey on the occurrence of Grapholita inopinata (Heinrich) (Lepidoptera: Tortricidae) at some areas in Aomori, Fukushima and Miyagi prefectures. Res Bull Pl Prot Jpn 41:25–30 (in Japanese with English summary)Google Scholar
- Ueda S (2006) Simple and rapid detection by mtCOI PCR–RFLP to distinguish the Q biotype of Bemisia tabaci. Kyushu Pl Prot Res 52:44–48 (in Japanese with English summary)Google Scholar
- Watanabe K, Yuuki S (1993) Forecast of occurrence time of the oriental fruit moth adults, Grapholita molesta (Busck). Ann Rept Plant Prot North Jpn 44:164–166 (in Japanese with English summary)Google Scholar