国内未见分布蝇种:澳洲麻蝇的形态和DNA条形码鉴定

DNA barcoding and morphological identification of Sarcophaga australis-a non-recorded flesh fly species in China

目的应用DNA条形码与形态分类相结合的方法,快速、准确鉴定在中山口岸入境船舶截获的1只蝇类。方法 2014年4月30日中山出入境检验检疫局神湾办事处从澳大利亚入境载有鲜葡萄的货船上截获1只蝇类,做成针插标本,并拍照存档。进行形态学鉴定,然后取1条后足提取基因组DNA,采用动物DNA条形码的通用引物(LCO1490和HCO2198)扩增目的片段,并进行纯化、克隆测序和序列分析,与NCBI及BOLD中的序列进行比对,构建NJ树。结果形态学鉴定截获蝇类为雌性麻蝇科种类,由于缺少关于雌性麻蝇鉴定特征描述的参考资料,无法将该蝇鉴定到种,所获得的DNA条形码序列与BOLD中的澳洲麻蝇[Sarcophaga australis(Johnston et Tiegs,1921)]的序列相似度达100%。经过科技查新,确认是中国未见分布种。结论根据鉴定结果,判定所截获的蝇类为雌性澳洲麻蝇,是我国首次截获。应用DNA条形码与形态鉴定相结合可以准确快速鉴定雌性麻蝇种类。

Objective To identify the flesh fly from an entry ship rapidly and accurately with ＇Barcoding and Morphology＇ techniques. Methods A flesh fly specimen was collected by Zhongshan CIQ on an entry ship from Australia with grapes. Morphological identification was conducted, and genomic DNA were extracted from a hind-leg of the fly. Target fragments were amplified with animal DNA barcoding universal primers （LCO1490 and HCO2198）, then purified, cloned, sequenced and blasted in BOLD, a NJ tree was bulk based on the original data. Results From the morphological identification, it is a female Sarcophaga sp. Because of the lack of the references for female flesh flies, we could only identify it to a female Sarcophaga sp. Sequence of the DNA barcode of it was 100% identical to the published sequence of Sarcophaga australis in BOLD. ＇Science and technology novelty search＇ result showed that it is a non-recorded species in China. Conclusion Based on the results, it could be concluded that the fly was a female S. australia. It is intercepted for the first time at China port. ＇ B ＆ M＇ techniques is a good tool to identify the intercepted non-recorded fly species.