基于ND2基因序列的燕窝DNA条形码鉴别

目的:利用DNA条形码鉴定技术,快速、准确地鉴别不同种类及产地的燕窝,为该药材的质量评价提供参考。方法:提取不同产地及品种燕窝的DNA,对烟酰胺腺嘌呤二核苷酸(NADH)脱氧酶亚单位2(ND2)部分序列进行扩增和测序,利用软件MEGA6.0进行序列比对,分析变异位点,计算Kimura-2参数遗传距离,构建邻接(neighbor-joining,NJ)系统发育树。结果:NJ系统发育树显示31个官燕窝样品均与爪哇金丝燕Aerodramus fuciphagus聚为1支;1个毛燕窝样品与大金丝燕A.maximus聚为1支。但官燕窝中经细胞色素b(Cytb)鉴定生物基原为爪哇金丝燕亚种(淡腰金丝燕A.fuciphagus germani)的8个样品单独聚为1支,支持率82%,并在该段ND2序列表现为2个位点的差异,表明该段ND2序列可鉴别爪哇金丝燕的亚种。结论:ND2条形码可用于快速、准确地鉴别燕窝的生物基原。

Molecular Identification of Edible Bird's Nest Based on ND2 DNA Barcode

Objective: To identificate different origins and categories of commodity edible bird's nest (EBN) by DNA barcoding technique fastly and acurrately. Method: Total genomic DNA was isolated from different origins and categories of commodity EBN. Nicotinamide adenine dinucleotide dehydrogenase subunit 2 (ND2) gene sequences were amplified and sequenced.Identification were performed using similar Blast from GenBank,nearest distance and neighbor-joining (NJ) methods. Result: NJ phylogenetic tree showed 31 white or silver EBN samples which were clustered with Aerodramus fuciphagus;a black EBN sample was clustered with A. maximus.However,among white or silver EBN samples,there were 8 samples clustered into a separate branch with support rate of 82%,which were identified by cytochrome b (Cytb) gene as the subspecies (A. fuciphagus germani),and sequence alignment showed 2 parsimony-informative sites.This result showed that segment of ND2 sequence can identify subspecies of A. fuciphagus. Conclusion: ND2 sequence is efficient barcode for identifying origin species of EBN.