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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 quickly and accurately identify one flies intercepted by immigration vessels at the port of Zhongshan at the application of a combination of DNA barcodes and morphological classification. Method April 30, 2014 Zhongshan Exit Inspection and Quarantine Bureau, Shenwan Branch Intercepted 1 fly from a cargo ship carrying fresh grapes from Australia and stitched it into a needle for photographing. The morphological identification was carried out. Genomic DNA was extracted from one hind leg and amplified by PCR using general primers for animal DNA barcoding (LCO1490 and HCO2198), followed by purification, cloning and sequence analysis, and sequencing with NCBI and BOLD Compare and construct NJ tree. Results Morphological identification The flies were intercepted by females. Due to the lack of reference materials for characterization of females, the flies could not be identified and the DNA barcode sequences obtained were similar to those of Sarcophaga australis (Johnston et Tiegs, 1921)] sequence similarity of 100%. After the technology search, it is confirmed that there is no species in China. Conclusion According to the identification results, it is concluded that the intercepted flies are female Strix, the first interception in China. The combination of DNA barcoding and morphological identification can accurately and quickly identify the species of female flies.