22种樟科植物DNA条形码分子鉴定  

Molecular Identification of 22 Species of Lauraceae by DNA Barcoding

作　　者：许丹芸 张辉菊 李可心 杨全[1] 高弘扬 XU Dan-yun;ZHANG Hui-ju;LI Ke-xin;YANG Quan;GAO Hong-yang(Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广东药科大学,广州510006

出　　处：《中国实验方剂学杂志》2021年第16期159-166,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广东省中医药局中医药科研项目(20201231);广东省普通高校青年创新人才类项目(2019KQNCX057);广东省基础和应用基础研究基金联合基金项目(2019A1515111043);罗定肉桂产业发展基础研究项目(2018-082);第四次全国中药资源普查项目(2017-152-006)。

摘　　要：目的:筛选适合樟科植物的DNA条形码,对采集的22种樟科植物进行鉴定分类。方法:以22种樟科植物为实验材料,提取其DNA,使用不同的DNA条形码引物进行聚合酶链式反应(PCR)扩增,电泳检测后进行双向测序。采用Codon Code Aligner软件对测序峰图和序列进行校对、拼接并去除正反向引物序列。将序列导入DNAMAN,进行多序列对比,查看并分析碱基变异位点。用MEGA计算不同种植物的遗传距离(K2P),根据遗传距离分析变异程度,基于邻接法构建系统进化树。结果:采用3对DNA条形码通用引物对22种樟科植物的DNA进行扩增测序后通过比较基因序列的特异碱基位点成功鉴别出其中20种植物。结论:matK序列能鉴定4个木姜子属植物;rbcL除粗脉桂外,对实验材料中楠属的3个物种都能进行区分;matK+rbcL+rpoB序列结合可鉴别20种樟科植物。其中matK序列对樟科植物鉴定效果最好,结果也表明将DNA条形码对植物进行组合鉴定,效果更佳。从分子角度研究樟科植物之间的亲缘关系,同时也为药用植物资源调查、栽培种植、开发保护利用提供依据。Objective: DNA barcodes suitable for Lauraceae plants were screened,and 22 Lauraceae plants were identified and classified. Method:The DNA of 22 species of Lauraceae was extracted and amplified by polymerase chain reaction(PCR)with different DNA barcoding primers,followed by electrophoresis and sequencing. Codon Code Aligner was used to proofread,splice,and remove the forward and reverse primer sequences. The sequence was imported into DNAMAN for multiple sequence alignment,and the base mutation sites were analyzed. The Kimura 2-Parameter(K2 P)distance of different plants was calculated by MEGA,and the variation degree was analyzed according to the genetic distance. The phylogenetic tree was constructed based on the adjacency method. Result:Three pairs of DNA barcoding primers were used to amplify and sequence the DNA of 22 species of Lauraceae,and 20 species were identified by comparing the specific base sites of gene sequences. Conclusion: Four Litsea plants could be identified by matK, three Phoebe plants except for Cinnamomum validinerve by rbcL,and 20 Lauraceae plants by the combination of matK,rbcL,and rpoB,which provided a theoretical basis for the identification and development of Lauraceae plants. Among them,matK was predominant in the identification of Lauraceae plants,and the results also showed that the combination of sequences for plant identification could achieve a better result in DNA barcoding. This study investigated the genetic relationship between Lauraceae plants at the molecular level,aiming at providing a basis for the investigation,cultivation,development,protection,and utilization of medicinal plant resources.

关 键 词：樟科 DNA条形码 鉴别 MATK RBCL RPOB 

分 类 号：R284.2[医药卫生—中药学] R289[医药卫生—中医学]