10种杓兰属植物rDNA ITS序列的克隆与分析  

Cloning and analysis of rDNA ITS sequences from ten Cypripedium plants

作　　者：蒋明[1] 李温平[1] 周晶[1] 陈贝贝[1] 

机构地区：[1]台州学院生命科学学院,浙江临海317000

出　　处：《浙江大学学报（理学版）》2012年第6期689-695,共7页Journal of Zhejiang University（Science Edition）

摘　　要：以10种杓兰属植物为材料,利用PCR技术从叶片DNA克隆ITS序列.测序结果表明,10种杓兰属植物ITS的长度为522-572bp,变异位点194个,其中信息位点99个,单核苷酸变异位点95个;5.8S序列的长度均为170bp,但变异位点十分丰富,其中信息位点30个,单核苷酸变异位点20个,利用这些位点可将10种杓兰属植物完全分开.系统发育分析结果表明,所研究的植物可分为三组,绿花杓兰、西藏杓兰、黄花杓兰和紫点杓兰聚为一组,高山杓兰、大花杓兰、褐花杓兰和杓兰聚为一组,斑叶杓兰和离萼杓兰聚为一组.序列已上传至NCBI,登陆号为JN018070-JN018079.本研究克隆和分析了10种杓兰属植物的ITS序列,为遗传多样性和系统发育研究奠定了基础.Ten Cypripedium plants were used as materials for internal transcribed spacer(ITS) sequence cloning by PCR methods.Sequencing results revealed that the full length ITS sequences of Cypripedium ranged from 522 to 572 bp with 194 variable sites,99 parsimony-informative sites and 95 single nucletide polymorphism sites.All the 5.8S sequences were 170 bp in length with rich variable sites including 30 parsimony-informative sites and 20 single nucletide polymorphism ones.The specific sites in ITS sequences could be used to identify these ten Cypripedium plants.Phylogenetic analysis results indicated that the plants studied constituted three groups: group 1 for C.henryi,C.flavum,C.tibeticum and C.guttatum,group 2 for C.himalaicum,C.macranthum,C.smithii and C.calceolus,and group 3 for C.margaritaceum and C.plectrochilum.In this study,ITS sequences of ten Cypripedium were cloned and analyzed providing the foundation for genetic diversity and phylogenetic analysis.

关 键 词：杓兰属 ITS序列 克隆 分子鉴定 

分 类 号：R287.7[医药卫生—中药学]