摘要
目的:选取天麻3种变型,测定ITS-1序列并进行对比分析,找出变异信息位点,对不同变异类型亲缘关系作出判定。方法:采用改良的CTAB法提取DNA,利用合成的特异引物对ITS区进行扩增,并对目的片段测序分析。结果:ITS-1共计209bp,序列片段上有27个单核苷酸变异位点,ITS-1单核苷酸多态性以碱基转换为主,绿天麻最高达80.8%,乌天麻最低也达70%;黄天麻居中达71.4%。在多态性位点数及多态性频率上,绿天麻均表现为最高分别为26、8.04;黄天麻表现居中为21、9.95;乌天麻表现最低为20、10.45。结论:乌天麻是古老类型,绿天麻是较为进化的类型,黄、绿天麻是由乌天麻进化而来。
Objective： In order to identify the relationship of differences types of variation in three samples of Gastrodia clara, their ITS se- quences of rDNA genes were amplified. ITS - 1 was sequenced and comparatively analyzed and the variation information sites were identi- fied. Method：Genomic DNA from Gastrodia elata extracted by modified CTAB method were regarded as templates for PCR amplification, chining, and sequencing by synthetic specific primer the nrDNA ITS sequences. Result：The results showed that there were 27 single nucle- otide variable sites in the total of 209 bp. It were confirmed that there was a wealth of genetic diversity in C, astrodia data. The ITS - 1 sin- gle nucleotide polymorphisms base conversion in Gastrodia elata, G. elata BI. f. viridis Mak up to 80. 8% , G. elata B1. f. glauca S. Chow low- est 70% ,G. elata BI . f. flavida S. Chow centered at 71.4%. G. elata B1. f. viridis Mak was manifested in the top 26,8.04 respectively, G. elata B1 . f. flavida S. Chow centered at 21,9. 95, G. elata B1. f. glauca S. Chow lowest 20,10. 45 in Polymorphism of points and polymor- phism frequencies. Conclusion：There may be an ancient type. G. elata B1 . f. flavida S. Chow and G. elata B1. f. viridis Makyellow may be evolved from G. data B1. f. glauea S. Chow by 27 single nucleotide mutation sites.
出处
《生物技术》
CAS
CSCD
北大核心
2012年第5期48-51,共4页
Biotechnology
基金
云南省中药现代化项目("昭通天麻规范化种植技术(GAP)研究及示范基地建设项目"
2002ZY-1)
菏泽学院科研基金项目("天麻与蜜环菌分子标记技术研究"
XY06SW06)资助
关键词
天麻
ITS-1序列
测序
单核苷酸多态性
变异位点分析
Gastrodia elata
ITS - 1 sequence
sequencing
single nucleotide polymorphism
analysis of variable sites
作者简介
王德信（1975-），男，硕士，讲师，研究方向：植物资源开发与利用，发表论文10余篇，Tel：0530-6075996，Email：wangdexinl996@163．com。
