Diagnostic molecular markers of six lepidopteran insect pests infesting apples in Korea
ABSTRACT Two molecular identification techniques for differentiating six lepidopteran pests infesting apples in Korea are presented. These six species include two internal fruit feeders (Grapholita molesta and Carposina sasakii), two leaf rollers (Adoxophyes sp. and Archips breviplicanus) and two leaf miners (Phyllonorycter ringoniella and Lyonetia prunifoliella). All species occur until near harvest and reduce apple production. A 489 bp fragment of mitochondrial cytochrome oxidase subunit I (COI) was sequenced in these six species. The sequence was used to select species-specific restriction enzyme sites and to design diagnostic polymerase chain reaction (PCR) primers, resulting in the development of restriction fragment length polymorphism (RFLP)-PCR and diagnostic PCR. These methods were reliable and rapid in the identification of these six species.
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- "The 180 sequences were aligned with Clustal W (Thompson et al. 1994) and were then checked for insertion and deletion (indel). The final 588-bp sequences (the alignment length) were used to analyze the lineage-specific restriction enzyme sites and the genetic structure of collection.DNA identification As precise identification of young and/or old larvae of C. sasakii and similar fruits-infesting species are difficult or may be impossible practically (Sony et al. 2009; Hada and Sekine 2011), we conducted a phylogenetic analysis using the COI sequences collected in the present study and the homologous GenBank sequences of C. sasakii as well allied species available by 1 July 2012. Based on the resulting phylogenetic tree, the haplotypes were involved in the monophyletic group of " C. sasakii " were retained and considered as C. sasakii. "
ABSTRACT: The genetic differentiation and genetic structure of the peach fruit moth, Carposina sasakii Matsumura (Lepidoptera: Carposinidae), was investigated in China, where the moth is native. The mitochondrial cytochrome c oxidase I (COI) gene of 180 individuals from 16 collections were sequenced and analyzed. The results showed that two sympatric and cryptic mtDNA lineages existed within C. sasakii in China. The genetic differentiation has significant correlation with the geographical distance, but has no evidence for host plant associations. Our results of haplotype distribution suggest that the C. sasakii individuals can naturally move between areas, while the movement of individuals between long-distance locations may be associated with human activities such as the transport of fruit. Finally, an mitochondrial COI gene PCR-RFLP method was developed to differentiate the two cryptic mtDNA lineages within C. sasakii, which provides rapid and reliable tool for the future research of the two lineages. © The Author 2015. Published by Oxford University Press on behalf of the Entomological Society of America.Journal of Insect Science 07/2015; 15(1). DOI:10.1093/jisesa/iev063 · 1.03 Impact Factor