基于细胞色素氧化酶I亚基(COI)基因序列的Kitanglad山部分地区飞蛾(鳞翅目:鞘科)分子鉴定

Michelle S. Suelo, Aprille Joy M. Luceno 2, Alma B. Mohagan, Reggie Y. Dela Cruz
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引用次数: 0

摘要

利用DNA条形码技术对菲律宾兰塔潘岛Kitanglad Lirongan山的sphingidae科植物进行了鉴定。共收集到13种,分别为:长尾绒螯虾、卷尾草、staudingeri、mindanaoamplypterus panopus、Daphnis hypothous、Gnathothlibus erotus erotus、brunneum Hippotion、echeclus、Psilogramma menephron、thetra nessus、thetra rhesus、manilae和sugii。使用QIAGEN血液和组织试剂盒分离基因组DNA。利用LepF1 (ATTCAACCAATCATAAAGATATTGG)和LepR1 (TAAACTTCTGGATGTCCAAAAAATCA)引物扩增线粒体细胞色素氧化酶(COI)基因,共获得656 ~ 666个碱基对。BLAST分析能够在物种水平上识别鞘鞘。10个物种的COI基因序列BLAST匹配度在95% ~ 99%之间。使用最大似然(ML)和贝叶斯推理(BI)来检查具有最高bootstrap值的COI的系统发育信号。鞘类飞蛾在进化支系的基础上形成了一个单系群。
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Molecular Identification of Hawkmoths (Lepidoptera: Sphingidae) in Selected Areas of Mt. Kitanglad Based on Cytochrome Oxidase Subunit I (COI) Gene Sequence
The study was conducted for the identification of selected species of family sphingidae through DNA Barcoding in Mt. Kitanglad Lirongan, Lantapan, Bukidnon, Philippines. Thirteen species were collected namely: Acherontia lachesis, Agrius convolvuli, Ambulyx staudingeri, Amplypterus panopus mindanaoensis, Daphnis hypothous, Gnathothlibus erotus erotus, Hippotion brunneum, Hippotion echeclus, Psilogramma menephron, Theretra nessus, Theretra rhesus, Theretra manilae and Theretra sugii. Isolation of the genomic DNA was carried out using the QIAGEN Blood & Tissue Kit. Mitochondrial cytochrome oxidase (COI) gene amplification was carried out using LepF1 (ATTCAACCAATCATAAAGATATTGG) and LepR1 (TAAACTTCTGGATGTCCAAAAAATCA) primers producing 656-666 base pairs were obtained from 30 samples of sphingid moth species. BLAST analyses were able to identify sphingid to the species level. BLAST hits of COI gene sequence of all 10 species ranged from 95%-99% similarity. Maximum likelihood (ML) and Bayesian inference (BI) was used to examine phylogenetic signals in COI with the highest bootstrap values. Sphingid moths formed a monophyletic group based on the clade.
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