Non-destructive DNA extraction method for identification of Bradysia odoriphaga (Diptera: Sciaridae), a pest of Welsh onion, carrot, and Chinese chive in Japan

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...

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Bibliographic Details
Published in:Applied entomology and zoology 2020-02, Vol.55 (1), p.181-185
Main Authors: Arimoto, Makoto, Hinomoto, Norihide, Omata, Ryosuke, Iwase, Ryozaburo, Sueyoshi, Masahiro, Yoshimatsu, Shin-ichi
Format: Article
Language:English
Subjects:
Agriculture
Applied Ecology
Biomedical and Life Sciences
Bradysia odoriphaga
Cytochrome-c oxidase
Deoxyribonucleic acid
Diptera
DNA
DNA barcoding
Eggs
Entomology
Environmental Management
Life Sciences
Mitochondrial DNA
Nucleotide sequence
Plant Pathology
Polymerase chain reaction
Primers
Sciaridae
Species
Technical Note
Zoology
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Summary: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.
ISSN:0003-6862
1347-605X
DOI:10.1007/s13355-019-00661-x