摘要
[目的]研究道地药材板桥党参(Codonopsis tangshen Oliv.)叶nrDNA-ITS区的序列特征,为板桥党参分子鉴定提供依据。[方法]采用PCR直接测序技术测定板桥党参的nrDNA-ITS区核苷酸序列,并分析其序列组成。[结果]采用改良CTAB法提取得到样品的基因组DNA吸光值OD260/OD280比值在1.8～2.0,DNA条带清晰整齐,无明显的拖尾现象,表明所得DNA样品纯度和质量较好,符合试验要求。利用nrDNA-ITS序列通用引物进行PCR扩增反应,获得预期约800bp的双链产物。经过测序发现,板桥党参nrDNA-ITS1序列核苷酸序列长度为260bp,其G+C含量为52%,nrDNA-ITS2序列核苷酸序列长度为239bp,其G+C含量为60%;在GenBank上进行Blast对比分析,证实其属于党参科植物nrDNA-ITS序列,并已提交GenBank,注册登记号为GQ906566。[结论]DNA测序技术可作为准确而有效地鉴定板桥党参基原的分子鉴定方法,可为板桥党参分子鉴定提供基础性资料。
[Objective] To identify the characteristics of nrDNA-ITS from the leaf of famous-region drug Codonopsis tangshen Oliv.It will provide molecular evidence to distinguish the Codonopsis tangshen Oliv.[Method]Direct PCR sequencing was used to detect the homology of nrDNA-ITS seqμence of Codonopsis tangshen Oliv,and analyze its sequence composition. [Result] The ratio of OD260/OD280 of genome DNA absorption value in samples extracted by improved CTAB was 1.8-2.0,DNA strips were clear and tidy,no obvious tail phenomenon,indicating that purity and quality of DNA samples was better,met the test requirements. NrDNA-ITS sequences universal primers were used to PCR amplification reactions,the expected double-stranded product of about 800 bp were obtained. After sequencing,it revealed that the Nucleotide sequence length of Codonopsis tangshen Oliv. nrDNA-ITS1 sequence was 260 bp,its G ＋ C contents was 52%,the nucleotide sequence length of nrDNA-ITS2 sequence was 239 bp,its G＋C contents was 60%; Blast comparative analysis in GenBank confirmed that it belonged to Codonopsis plants of nrDNA-ITS sequences,and had been submitted to GenBank,registration No. was GQ906566. [Conclusion] DNA sequencing can provide an accurate and reliable mean for the identification of the origin and genuineness of C. tangshen Oliv,which also can provide the basic information for molecular identification of C. tangshen Oliv.
出处
《安徽农业科学》
CAS
北大核心
2010年第9期4427-4428,4431,共3页
Journal of Anhui Agricultural Sciences
基金
天然产物与利用湖北省重点实验室2007年科研资助项目(2007017)
2006年度湖北民族学院科研基金团队项目(9)
湖北民族学院科技学院教研课题(K200801)
关键词
分子鉴定
板桥党参
nrDNA-ITS
序列分析
Molecular identification
Codonopsis tangshen Oliv.
nrDNA-ITS
Sequence analysis
