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目的比较中国不同地区绞股蓝的nrDNAITS碱基序列的差异,以期分析绞股蓝的地理分布与ITS基因型的相关性,为我国不同地区绞股蓝的鉴别提供分子依据。方法PCR克隆测序,MegAlign(DNASTAR)软件对序列进行对位排列,PAUP4.0b10软件进行系统发育分析,构建最大简约树。结果绞股蓝ITS区长度为658～659bp,变异位点为8.48%,信息位点为2.72%。不同地区的样品在碱基的量、信息位点的碱基位置和遗传距离等方面存在一些差异。系统发育分析表明ITS基因型与绞股蓝的地理分布具有一定的相关性,但也有部分不一致,可能主要是因为绞股蓝种内复杂的倍性。结论ITS序列可作为中国不同地区绞股蓝的一种良好的分子标记,而要确定不同地区的绞股蓝合理的亲缘关系还需要结合其他方面的证据。
Objective To compare the differences of nrDNAITS in Gynostemma pentaphyllum from different regions in China in order to analyze the correlation between Gynostemma pentaphyllum geographical distribution and ITS genotype and to provide a molecular basis for the identification of Gynostemma pentaphyllum in different regions of China. Methods PCR cloning and sequencing, MegAlign (DNASTAR) software alignment of the sequence, PAUP4.0b10 software phylogenetic analysis, the construction of the largest parsimony tree. Results The length of ITS region of Gynostemma pentaphyllum was 658-659 bp, the variation locus was 8.48% and the information locus was 2.72%. There are some differences in the amount of bases, the base positions of information sites and genetic distance between samples in different regions. Phylogenetic analysis showed that there was some correlation between ITS genotypes and geographical distribution of Gynostemma, but there were some inconsistencies, probably due to the complex ploidy in Gynostemma. CONCLUSIONS: ITS sequences can be used as a good molecular marker for Gynostemma in different regions of China. To determine the reasonable genetic relationship of Gynostemma in different areas, other evidences need to be combined.