基于多基因组片段构建枸杞属药用植物DNA条形码

DNA barcoding construction of medicinal plants in genus Lycium L. based on multiple genomic segments

目的采用核基因组核糖体DNA ITS区、叶绿体基因组9个DNA片段和线粒体基因组2个DNA片段构建国产枸杞属药用植物DNA条形码。方法采集枸杞属黑果枸杞Lycium ruthenicum、黄果枸杞L.barbarum var.auranticarpum、宁夏枸杞L.barbarum、枸杞L.chinense和新疆枸杞L.dasystemum 5个分类群标本及实验样品,测定各样品的核糖体ITS区,叶绿体mat K、rbc L、rpo C1、trn L(UAA)intron、psb A-trn H、atp B-rbc L、trn S(GCU)-trn G(UCC)、rpl20-rps12、trn L(UAA)-trn F(GAA)和线粒体nad1第2内含子、nad5第4内含子序列;以全萼秦艽Gentiana lhassica为外类群,进行系统学分析。结果共获得108条序列,ITS区变异位点最为丰富;筛选出6个叶绿体DNA片段;2个线粒体DNA片段可作为该属3个物种的条形码推荐序列;构建各物种多基因组多片段组合DNA条形码。结论 ITS区及筛选的6个叶绿体DNA片段均具有物种鉴别意义;2个线粒体DNA片段具有一定鉴别意义;为枸杞类物种鉴定及遗传背景分析提供基础资料。

Objective To construct the DNA barcoding of medicinal plants in genus Lycium L. from China based on sequences of ITS, nine chloroplast DNA segments, and two mitochondrial DNA segments. Methods Specimens and samples of Lycium ruthenicum, L. barbarum var. auranticarpum, L. barbarum, L. chinense, and L. dasystemum were collected and nuclear DNA ITS, chloroplast DNA mat K, rbc L, rpo C1, trn L（UAA） intron, psb A-trn H, atp B-rbc L, trn S（GCU）-trn G（UCC）, rpl20-rps12, trn L（UAA）-trn F（GAA）, and mitochondrial DNA nad1/b-c and nad5/d-e were sequenced from these samples. Gentiana lhassica was used as the external group to make statistical analysis. Results One hundred and eight sequences were obtained. The mutation site of ITS region was the most abundant; The six chloroplast DNA fragments were selected; Two mitochondrial DNA fragments could be used as the bar code recommendation sequences of the three species from the genus, and the multiple genomic and multi fragment combination DNA bar code of each species was constructed. Conclusion ITS region and the screening of six chloroplast DNA fragments have the significance of species identification; The two mitochondrial DNA fragments have a certain significance. The study may provide the reference for the species identification and genetic background of the plants in Lycium L. from China.