药用石斛的叶绿体matK基因序列分析及鉴别  

Analysis and authentication of chloroplast matK gene sequences of Herba Dendrobii

作　　者：刘静[1,2] 何涛[1] 淳泽[1] 

机构地区：[1]中国科学院成都生物研究所,四川成都610041 [2]中国科学院研究生院,北京100049

出　　处：《药学学报》2009年第9期1051-1055,共5页Acta Pharmaceutica Sinica

基　　金：国家科技支撑计划资助项目(2006BAI06A11-10);国家级星火计划项目资助项目(2008GA810003);四川省科技支撑计划资助项目(07FG001-005;2006Z08-014;2009FZ0077);四川省"十一五"育种攻关项目资助项目(2006YZGG12-3);四川省公益性研究计划资助项目(2008NG0007)

摘　　要：比较22份(包括待检材料4份)共计12种药用石斛及密花石豆兰(1份)的叶绿体DNA matK基因序列差异,从分子水平对药用石斛进行鉴定。采用改良的CTAB法提取石斛基因组DNA,PCR扩增,克隆测序法对叶绿体DNA matK基因全序列进行测定,运用BioEdit、MEGA4.0等软件分析序列,构建分子系统树。测得石斛属12种植物的叶绿体matK基因长1410bp,变异位点51个,信息位点11个。除了存在碱基替换的遗传变异外,还存在碱基的插入和缺失。石斛种间以及种内不同居群(品种)间均能在NJ树中明显分化开来,种间平均遗传距离为0.008,种间最大遗传距离为0.014,最小遗传距离为0.003,碱基相差8～20个。种内不同居群(品种)遗传距离为0.001,相差1～5个碱基。并成功地对4种待检种进行了鉴定。利用12种石斛的matK基因全序列数据库及遗传分析软件,通过对待检种的matK基因全序列进行测定,可以在分子水平对石斛不同种及种内不同居群(品种)进行鉴别,为石斛的分子鉴定提供依据。To compare the characteristic of chloroplast matK gene sequences of different Herba Dendrobium species and to authenticate inspected species, the matK gene sequences of 12 species （including 22 materials） and outgroup were amplified, cloned, and sequenced. Genomic DNA of Dendrobium plants was extracted using modified cetyltrimethyl ammonium bromide （CTAB） method. The matK gene sequences were about 1 410 bp in length. The variable sites were 51 while the parsim-informative sites were 11. There were nucleotides insertions and deletions in some species, in addition to transitions and transversions, such as in D.denneanum and D.chrysotoxum. Interspecies and different populations （variaties） of Dendrobium Could be distinguished on phylogeny tree. The average genetic distance was 0.008, and the maximal and minimal genetic distances between Dendrobium species were 0.014 and 0.003, respectively. There were 8-20 variable sites between Dendrobium species. The genetic distance between populations （variaties） was 0.001, and there were 1-5 variable sites. Moreover, the 4 inspected materials were successfully authenticated. The database of chloroplast matK gene sequences of 12 species of Herba Dendrobii and inspected species could be used for the molecular authentication between Dendrobium species and populations. The matK gene sequence could be used as molecular maker for authentication of Herba Dendrobium.

关 键 词：石斛 matK基因序列 分子鉴定 

分 类 号：R931.5[医药卫生—生药学]