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目的对来自主要龙葵产区龙葵种质资源的转录间隔区(internal transcribed spacer,ITS)和trn H-psb A基因间隔区序列进行系统发育学分析,为龙葵资源的合理利用和道地性评价提供理论基础。方法用聚合酶链式反应(polymerase chain reaction,PCR)的方法扩增龙葵ITS区和trn H-psb A的DNA序列,并与美国国家生物技术信息中心(NCBI)数据库进行比对获取ITS1+ITS2和trn H-psb A的DNA序列。用邻接法(Neighbor joining,NJ)和最大简约法(maximum parsimony,MP)构建系统发育树,用Kimura two-parameter(K2-P)模型分析遗传距离,用Clustal X和DNAman软件进行多重比对。结果 17个龙葵样品的ITS1序列长度均为230 bp,ITS2序列长度均为206 bp,trn H-psb A序列长度为446 bp或447 bp,3个序列分别存在7个、2个和3个变异位点。系统发育的方法将中国龙葵种质资源聚为3个类群,这些类群与其所处的纬度存在一定的相关性。遗传距离的分析结果显示来自广东梅州的样品与来自北京和河北等地的样品存在最大的遗传距离。结论系统发育和变异位点的分析为中国龙葵资源的利用和进化研究,以及龙葵资源的道地性评价提供了理论基础,变异位点的分析还可应用于相关种质资源的鉴定。
OBJECTIVE: To carry out phylogenetic analysis of the spacer sequences of ITS and trn H-psb A in Solanum nigrum from the main Solanum nigrum L., Sex assessment provides the theoretical basis. Methods The DNA sequences of ITS and trn H-psb A were amplified by polymerase chain reaction (PCR) ITS2 and trn H-psb A DNA sequences. Phylogenetic tree was constructed by Neighbor joining (NJ) and maximum parsimony (MP), and genetic distance was analyzed by Kimura two-parameter (K2-P) model. Multiple alignment was performed by Clustal X and DNAman software. Results The sequences of ITS1 and ITS2 were all 230 bp and 174 bp, respectively. The length of trn H-psb A was 446 bp or 447 bp. There were 7, 2 and 3 Variation sites. Phylogenetic method will be China Solanum germplasm resources clustered into three groups, these groups and their latitudes there is a certain correlation. Analysis of genetic distances revealed that the samples from Meizhou, Guangdong, had the greatest genetic distance to samples from Beijing and Hebei. Conclusion The phylogenetic analysis and the analysis of the variation sites provide theoretical basis for the utilization and evolution of Solanum natans L. resources and the authenticity evaluation of Solanum nigrum resources. The analysis of the variation loci can also be applied to the identification of related germplasm resources.