云南省江城县五黑阿蠓形态学鉴定及ITS-1和COI基因序列分析  

Morphological identification and ITS-1 and COI gene sequence analysis of Alluaudomyia quinquepicina in Jiangcheng county,Yunnan province

作　　者：何于雯 李钊 孟锦昕 李楠 徐勇 元正菊 王静林 HE Yu-wen;LI Zhao;MENG Jin-xin;LI Nan;XU Yong;YUAN Zheng-ju;WANG Jing-lin(Yunnan Tropical and Subtropical Animal Viral Disease Laboratory,Yunnan Animal Science and Veterinary Institute,Kunming,Yunnan 650224,China)

机构地区：[1]云南省畜牧兽医科学院,云南省热带亚热带动物病毒重点实验室,云南昆明650224 [2]江城县曲水镇农业技术服务中心

出　　处：《中国国境卫生检疫杂志》2020年第5期321-324,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：云南省科技厅重点基金项目(2019FA015);国家自然科学基金项目(31660714);国家重点研发计划项目(2016YFD0500303);云南省科技人才和平台计划项目(2018HB046)。

摘　　要：目的对在云南省江城县采集的1种阿蠓进行形态学鉴定,并获得该蠓虫分子鉴定所需间隔区-1基因(ITS-1)和细胞色素氧化酶Ⅰ(COI)基因序列信息。方法 2019年5月用诱蚊灯在云南省江城县坝伞村养牛场采集蠓虫样本,提取蠓虫基因组DNA,用特异引物扩增蠓虫ITS-1基因和COI基因并测序,用生物信息学软件进行序列分析;同时对蠓虫进行压片和形态学鉴定。结果在江城采集到的1只雌性蠓虫BS9,具有以下主要形态学特征:大颚有齿20枚,端部9枚大而尖;翅面有明显17块暗斑。各足股节远端、胫节两端均有明显白环;第8腹节背板角质化深,覆盖住腹板。这些形态特征与五黑阿蠓相似,提示这只蠓虫为五黑阿蠓。遗传进化分析结果显示:在ITS-1基因和COI基因构建的系统进化树上,蠓虫BS9均位于独立的进化分支,与贝蠓(Bezzia)、阿蠓(Alluadomyia)和裸蠓(Atrichopogon)较近,核苷酸同源性在86.3%~89.8%之间。结论形态学鉴定提示江城采集的蠓虫BS9为阿蠓属多斑阿蠓种团的五黑阿蠓。获得五黑阿蠓分子鉴定所需要的ITS-1基因和COI基因序列信息,为今后五黑阿蠓的分子鉴定和遗传进化研究提供了序列信息。Objective To identify Alluaudomyia collected at Jiangcheng county,Yunnan province,and to obtain ITS-1 and COI gene sequence for molecular identification of the midge. Methods In May 2019,the midges were collected by using the trap lamp from a cattle farm in Basan village of Jiangcheng county,Yunnan province. The genomic DNA of the midge was extracted,and ITS-1 gene and COI gene of the midge were amplified by PCR with specific primers and sequenced. The analysis of the nucleotide and amino acid homology was carried out by bioinformatics software. The midge tablet and identification based on morphologic characters were carried out under the stereoscopic microscope. Results A female midge collected from Jiangcheng county had the following morphological characteris-tics:the mandibles had about 20 teeth,and the end had 9 big and sharp ones. There were 17 dark spots on the wing surface. Each foot was brown with obvious white ring at the distal end of femer and both ends of tibia. There was a nearly spherical spermathecal gland in the abdomen,and the 8 th tergum was deeply keratinized,covering the web. The phylogenetic analysis based on ITS-1 gene and COI gene show that BS9 was located in an independent evolutionary branch,and its genetic evolution was related to Bezzia,Alladomyia and Atrihopogon,and the nucleotide homology was 86.3% and 89.8% between them. Conclusion The morphological identification indicated that the midge BS9 collected in Jiangcheng was Alluaudomyia quinquepicina of maculipennis group of Alluaudomyia. The sequence of ITS-1 gene and COI gene needed for the molecular identification of Alluaudomyia quinquepicina was obtained,which will provide the sequence information for the rapid molecular identification and genetic evolution of Alluaudomyia quinquepicina in the future.

关 键 词：五黑阿蠓 细胞色素氧化酶Ⅰ基因 间隔区-1基因 分子鉴定 形态学鉴定 

分 类 号：S852.71[农业科学—基础兽医学]