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目的:对我国2个主产区的主要红景天(Rhodiola L.)品种r DNA-内转录间隔区(ITS)序列进行分析,为红景天种质资源的分子鉴别及进化关系提供依据。方法:提取核基因组DNA,聚合酶链反应(PCR)克隆ITS序列;经DNAstar软件拼接序列后,通过Clustal X软件进行比对,考察其变异位点及信息位点;用Mega 4.1计算品种之间的遗传距离,构建最大简约树(maximum parsimony,MP)和邻接树(neighbor-joining,NJ)。结果:5种红景天材料r DNA-ITS序列长度为603~604 bp,ITS1、5.8 S r DNA和ITS2的DNA长度分别为226 bp、164 bp和213~214 bp。ITS1和ITS2分别含有11和9个变异位点,其中,信息位点分别为6和2个;存在转换、颠换、缺失等现象;5.8 S r DNA序列含7个变异位点,2个信息位点;试验中红景天品种的遗传距离为0.018 2~0.627 3。大花红景天与柴胡红景天亲缘关系最近。结论:r DNA-ITS序列是鉴别红景天品种,研究遗传关系的良好方法。
OBJECTIVE: To analyze the sequence of rDNA-ITS region of two major Rhodiola species in two main producing areas in China, and provide the basis for the molecular identification and evolutionary relationship of Rhodiola rosea germplasm resources. Methods: Nucleotide genomic DNA was extracted and ITS sequences were cloned by polymerase chain reaction (PCR). DNA sequences were aligned by DNAstar software and compared with Clustal X software to investigate the variation sites and information sites. Mega 4.1 The maximum parsimony (MP) and neighbor-joining (NJ) were constructed. Results: The length of rDNA-ITS sequences of five Rhodiola species was 603-604 bp, and the lengths of ITS1, 5.8 S r DNA and ITS2 were 226 bp, 164 bp and 213-214 bp, respectively. ITS1 and ITS2 contain 11 and 9 mutation sites, respectively, of which 6 and 2 are located respectively; there are such phenomena as transformation, transversion and deletion; 5.8S rDNA sequence contains 7 variation sites and 2 information The genetic distance of Rhodiola rosea was 0.018 2 ~ 0.627 3. Rhodiola rosea and Bupleurum Rhodiola recently kinship. Conclusion: The rDNA-ITS sequence is a good way to identify Rhodiola rosea varieties and to study the genetic relationship.