Abstract
We examined the utility of a non-destructive DNA extraction method for identification of Bradysia odoriphaga Yang and Zhang, 1985 (Diptera: Sciaridae) based on PCR using species-specific primers and DNA barcoding targeting the mitochondrial cytochrome c oxidase subunit I gene. Using species-specific primers, PCR success rates for adults and pupae were more than 90%, while those for larvae and eggs were 51% and 13%, respectively. The success rates of PCR for DNA barcode region were slightly higher than those of PCR using species-specific primers. In all sequenced samples, the 658 bp of sequences in the DNA barcode region were identical to the sequences from B. odoriphaga that we previously obtained using a destructive DNA extraction method. The adults were successfully identified based on morphological characters after non-destructive DNA extraction. The non-destructive DNA extraction method examined in this study is useful for the identification of adults and pupae of B. odoriphaga collected in the fields.
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Acknowledgements
This research part of our project was financially supported by Regulatory Research Projects for Food Safety, Animal Health and Plant Protection (2905). We thank Ryuji Uesugi, Ren Iwaizumi, and Hideaki Matsuura for their useful suggestions.
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Arimoto, M., Hinomoto, N., Omata, R. et al. Non-destructive DNA extraction method for identification of Bradysia odoriphaga (Diptera: Sciaridae), a pest of Welsh onion, carrot, and Chinese chive in Japan. Appl Entomol Zool 55, 181–185 (2020). https://doi.org/10.1007/s13355-019-00661-x
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DOI: https://doi.org/10.1007/s13355-019-00661-x