板桥党参基因组DNA提取方法的改进及其18S rRNA基因序列分析

目的:为研究道地药材板桥党参Codonopsis tangshen Oliv.的遗传多样性、种类鉴定等,提取高质量的基因组DNA;探讨板桥党参18S rRNA基因的序列特征,为板桥党参分子鉴定提供分子依据。方法:以板桥党参鲜嫩叶片为材料,采用自行改进的CTAB法提取出基因组DNA;采用PCR直接测序技术测定板桥党参的18S rRNA基因序列,并分析其序列组成。结果:使用改进的CTAB法从鲜叶中提取板桥党参的基因组DNA浓度较高;以其基因组DNA为模板对18S rRNA基因进行PCR克隆并测序,获得了板桥党参18S rRNA基因序列特征。结论:改进的CTAB法提取板桥党参基因组DNA效果较好;DNA测序技术可作为板桥党参基原鉴定准确而有效的分子鉴定方法。

Modifing on the Extraction Methods of the Genomic DNA and Analysising of 18S rRNA Gene Sequence from Codonopsis tangshen Oliv.

Objective：The research aimed to extract the high strength genomic DNA for studing the genetic diversion,the species identification and so on of Codonopsis tangshen Oliv. To identify the characteristics of 18S rRNA from the leaf of C. tangshen Oliv., it will provide molecular evidence to distinguish the C. tangshen Oliv.. Methods ： With fresh and tender leaves of C. tangshen Oliv. as tested materials, the genomic DNA was extracted from C. tangshen Oily. by using our modified CTAB method. To direct PCR sequencing detected the homology of 18S rRNA sequence of C. tangshen Oliv.. Meanwhile the character of 18S rRNA sequence also was analysised. Results ：There were high strength of the genomie DNA that was extracted from C. tangshen Oliv. by using modified CTAB method. The characteristics of ribosomal 18S rRNA gene sequence, were identified to detect direct PCR sequencing kill by using template of genomic DNA of C, tangshen Oliv.. Conclusion： The extraction effect of the genomic DNA from C. tangshen Oliv. by modified CTAB method was better. DNA sequencing can provide an accurate and reliable means for the identification of the origin and genuineness of C. tangshen Oliv..