高飞铗蠓的形态学和分子生物学鉴定

Morphological and molecular biological identification of Forcipomyia(Forcipomyia)praealtus

目的用形态学和分子生物学方法鉴定1只蠓虫,确定其种类并获得该蠓虫的COI和ITS-1基因序列。方法2021年9月在国内航班上采集1只蠓虫(编号meng-1),用组织消化液和蛋白酶K浸泡虫体,制作成玻片标本并进行形态学鉴定;从消化液中提取蠓虫DNA,扩增蠓虫COI和ITS-1基因,测序并分析。结果蠓虫meng-1的触角鞭节基部各节近似圆形;翅面淡色无斑,有大毛密布,前、中足淡色,后足股节端部和胫节基部浅棕色,其余淡色;有2个卵型基本等大的受精囊,殖下板宽环状。这些特征经检索表确定与铗蠓属铗蠓亚属高飞铗蠓Forcipomyia(Forcipomyia)praealtus Liu et Yu,2001一致。COI和ITS-1基因遗传进化分析也显示meng-1与铗蠓属蠓虫遗传关系最近,核苷酸同源性高于98%。结论形态学鉴定和分子生物学鉴定结果都提示这只飞机上采集的蠓虫meng-1为铗蠓属铗蠓亚属高飞铗蠓。同时提供了高飞铗蠓的COI基因和ITS-1基因序列,为今后快速、准确鉴定高飞铗蠓提供了分子靶标信息,为高飞铗蠓的遗传进化研究提供了序列信息。

Objective To systematically identify and classify one midge by using morphological and molecular biological identification,and to obtain COI and ITS-1 gene sequence of the midge.Methods In September 2021,one midge(No.meng-1)was collected on a domestic flight.The body of the midge was immersed in tissue digestive solution and proteinase K.The midge DNA was extracted from digestive supernatant,and the COI and ITS-1 genes of midge were amplified and sequenced.Results The midge(meng-1)had the following morphological characteristics:antenna basal flagellomeres were almost circular.Wing was pale without dark sport and covered with macrotrichia.Pre-legs and mid-legs were pale and femoral and tibial of hind-leg were partly brownish.There were two subequalsize spermathecae.Subgenital plate was cricoid.According to the key in the Forcipomyia,it was determined that the midge was Forcipomyia(Forcipomyia)praealtus Liu et Yu,2001.The phylogenetic analysis of COI and ITS-1 sequences showed that meng-1 had a closest relationship with Forcipomyia with nucleotide homology more than 98%.Conclusion The morphological and molecular biological identification suggested that this midge(meng-1)collected on flight was Forcipomyia(Forcipomyia)praealtus Liu et Yu,2001.At the same time,the sequences of COI gene and ITS-1 gene of Forcipomyia(Forcipomyia)praealtus were submitted,which provided molecular target information for rapid and accurate identification of Forcipomyia(Forcipomyia)praealtus,and sequence information for genetic evolution research of Forcipomyia(Forcipomyia)in the future.