为鉴定新疆阿拉尔苜蓿田的一种主要传粉昆虫，通过利用基因条码技术与形态特征分析，采用动物基因组DNA小量抽提试剂盒提取虫体基因组总DNA的方法，以鳞翅目通用引物扩增其线粒体细胞色素c氧化酶亚基Ⅰ(COⅠ)基因片段，切胶纯化后以试剂盒进行T-载体克隆，挑取阳性克隆送公司测序，获得序列在BOLD数据库中进行分子鉴定；同时根据形态特征进行常规鉴定。结果显示该基因片段序列与吉尔吉斯斯坦的2个黄斑蜂属未鉴定种虫标本相似度最高（99%以上），其次是希腊与俄罗斯的花黄斑蜂(Anthidium florentinum)标本相似度（96%以上），与其他切叶蜂标本序列相似度较低；形态学观察也发现与花黄斑蜂形态描述一致。因此，认定苜蓿田中这种常见黄斑切叶蜂为花黄斑蜂。
Abstract
The aim of the study was to identify a kind of common pollination insect in alfalfa field in Xinjiang based on DNA barcoding and morphological characteristics. Genomic DNA of the insect was extracted by Genomic DNA Mini Preparation Kit, mitochondrial cytochrome c oxidase subunit Ⅰ (COⅠ) gene fragment was amplified with a pair of universal primers of Lepidoptera insect. PCR product was purified by Gel Extraction Kit, cloned by the pUCm-T Vector Cloning Kit and then sent to a sequencing company. Finally the bee was identified based on morphological characteristics and the sequences in the BOLD database. The results showed that the sequences of gene fragments had the highest similarity (99%) with two unidentified specimens of Anthidium in Kyrgyzstan. Then were the two specimens of Anthidium florentinum in Greece and Russia, with the similarity of more than 96%. Sequences of the gene fragments had low similarity with other leaf cutting specimens in the BOLD database. Morphological characteristics also showed that this insect was the same with the description of A. florentinum. Therefore, this kind of leafcutting bee in alfalfa field in Alar, Xinjiang was identified as A. florentinum.
关键词
花黄斑蜂；COⅠ基因；基因条码
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Key words
Anthidium florentinum; COⅠ gene; DNA barcoding
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