基于AFLP分子标记和DNA条形码对无籽刺梨的鉴定

利用AFLP分子标记和DNA条形码技术,对采自贵州省兴仁县和贵州省安顺市的刺梨、无籽刺梨和光枝无籽刺梨共19份样本材料进行鉴定。结果表明:AFLP标记分析19个样本间的遗传相似系数在0.719 0 0.997 2之间,平均相似系数为0.936 5。采用UPGMA 法进行聚类分析,将19个样本聚类为2组,刺梨为单独1组,其他18份样本为1组。DNA条形码分析结果显示ITS无变异位点,将4种叶绿体序列片段(psbA-trnH、atpF-atpH、psbI-psbK、trnL-F)合并后以联合序列作为数据分析的基础,19份样本的平均误差为0.000 6,除刺梨外,其他18份样本之间的遗传距离均为0,而刺梨与无籽刺梨、光枝无籽刺梨之间的遗传距离均为0.005 8。两种方法的鉴定结果一致,无籽刺梨与刺梨是独立的2个种,兴仁县无籽刺梨与安顺市的光枝无籽刺梨为同一个种。本研究结果明确了无籽刺梨的系统地位,从分子水平和基因水平上解决无籽刺梨的分类分歧,为今后开展无籽刺梨种质资源的收集、保存、开发和应用等奠定了理论基础。

Identification of Rosa sterilis Based on AFLP Molecular Markers and DNA Barcodes

Nineteen Rosa roxbunghii, R. sterilis and R. sterilis var. leioclada specimen collected from Xing'ren and Anshun in Guizhou of China were identified by AFLP molecular markers and DNA barcodes. The results of AFLP showed that genetic similarity coefficient among the 19 specimen ranged from 0.719 0 0.997 2 and the average was 0.936 5. The result of UPGMA cluster analysis showed that the 19 specimen could be divided into two groups. R. roxbunghii was distinct from the others and formed one group separately, which the others fall into another group. The result of DNA barcodes showed that there was not variable site in ITS sequences and 4 cpDNA sequences showed highly closed affinity. Based on combined sequences, the average error was 0.000 6, while the genetic distance between R. roxbunghii and the others was 0.005 8. The identification results of two methods were consistent. R. sterilis and R. roxbunghii were two segregated species. The R. sterilis collected from Xing'ren and R. sterilis var. leioclada collected from Anshun belong to the same species. The results of this study determined the phylogeny status of R. sterilis at the level of molecular and gene and laid the theoretical foundation for further collection, preservation, exploitation and application of R. sterilis germplasm resources.