基于COI和16S rRNA基因序列的鱼胶基原鱼种DNA条形码鉴定

DNA Barcoding identification on original species of isinglass through COI and 16S rRNA sequences

目的建立基于COI和16S rRNA基因序列的DNA条形码鱼胶基原鱼种鉴定方法。方法以汕头、厦门、莆田等地采集的28份鱼胶为研究对象,利用形态归类法对28份样品进行观察,再提取基因组DNA,分别利用6对引物扩增COI和16S rRNA基因序列并双向测序。聚合酶链式反应(polymerasechain reaction,PCR)产物测序分析后,输入Genbank中进行BLAST分析比对,再利用最大似然法(maximum likelihood,ML)进行系统发育分析。结果采集的28份鱼胶样品可分为11种类型,主要有萝卜型、“Y”字型、心型、纸片型和琵琶型等。BLAST分析比对共鉴定出26份鱼胶样品基原鱼种,隶属于5目6科9属12种。系统发育分析表明不同种类的鱼胶处于不同分支中。结论该方法可用于鱼胶基原鱼种的快速分子鉴定,也为鱼胶的真伪鉴定及溯源提供理论依据。

Objective To establish a method for the identification of the origin of isinglass by DNA barcoding.Methods A total of 28 experiment samples of isinglass were collected from Shantou,Xiamen and Putian.Samples were observed,genomic DNA were extracted from isinglass and PCR were performed with the COI and 16S rRNA primers.The amplified sequences were aligned in the Genbank database,ML-tree was built as well.Results All experiment isinglass samples were classified for 11 morphological types,including carrot-shaped,“Y”type,heart-shaped,piece of paper type and pipa type.Total 26 samples could be successfully distinguished by gene COI and 16S rRNA and belong to 5 orders 6 families 9 genes 12 species.The phylogenetic tree revealed that different types of isinglass were in different branches.Conclusion The COI and 16S rRNA sequences can be used to distinguish the origin of isinglass and they are efficient for tracing the origin species of isinglass.