Identification and Phylogenetic Analysis of Dacini Fruit Flies in Thailand by <I>COX1</I> Gene
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Abstract
The fruit fly tribe Dacini is not only one of the most species-rich clades within the dipteran family Tephritidae, it is also one of most serious groups of horticultural pests and thus of particular biosecurity/quarantine concern. All Dacini members are notoriously difficult to identify by morphological characters. Molecular approaches, such as DNA ‘barcoding’ of the cytochrome c oxidase (COX1) gene, are commonly proposed to overcome this issue. Fresh materials of Dacini were collected from all regions of Thailand. Sequences were analyzed using a ‘barcode’ approach. Species were identified using standard nucleotide BLAST from GenBank database. Twenty species of Dacini were confirmed and their sequences were deposited in the GenBank database: Dacus (1 species): Dacus longicornis Wiedemann; Bactrocera (10 species): Bactrocera albistrigata de Meijere, B. carambolae Drew & Hancock, B. correcta (Bezzi), B. dorsalis Hendel, B. latifrons (Hendel), B. limbifera (Bezzi), B. nigrotibialis (Perkins), B. tuberculata (Bezzi), B. umbrosa (Fabricius), B. zonata (Saunders); and Zeugodaucs (9 species): Zeugodacus apicalis (de Meijere), Z. caudatus (Fabricius), Z. cilifer (Hendel), Z. cucurbitae (Coquillett), Z. diversa (Coquillett), Z. hochii (Zia), Z. incisus (Walker), Z. isolatus (Hardy), Z. platamus (Hardy) and Z. tau (Walker). A molecular phylogeny of these 20 species of Dacini was generated using COX1 gene. Maximum Likelihood and Bayesian analysis were used to investigate patterns of clustering of sequences with those of other pest species of fruit flies. Analyses of sequences gave consistent results. COX1 gene was proved to be effective in resolving Zeugodacus and Dacus from Bactrocera fruit fly species. The phylogeny showed that Zeugodacus clade is more closely related to Dacus rather than Bactrocera. This result supported raising Zeugodacus to the genus level. This study provided basic information on the number of species of fruit flies of the tribe Dacini present in Thailand. COX1 sequences were demonstrated as an effective tool in identifying Thai Dacini even from partial barcodes. Thus, we recommended that diagnostic and quarantine laboratories used the COX1 gene for the accurate identification of Dacini species.
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