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目的:分析滇产糙苏属3种药用植物丽江糙苏、黑花糙苏和假秦艽野生居群的ITS区和matK基因片段碱基序列,为丽江糙苏、黑花糙苏和假秦艽野生资源的鉴别及保护提供分子依据。方法:采用ITS区特异引物ITS4/ITS5和matK基因特异引物matKXF/matK5R进行PCR扩增并测序。结果:在ITS1、ITS2和matK基因片段上,丽江糙苏、黑花糙苏和假秦艽的种间K2P最小遗传距离均大于种内K2P最大遗传距离,NJ系统发育树也显示这3种药用植物均可与Gen Bank数据库中糙苏属外源种植物鉴别开,其中丽江糙苏、黑花糙苏和假秦艽的ITS2序列上分别具有3个、2个和1个有效鉴别位点,ITS1序列上分别具有3个、3个和3个有效鉴别位点,假秦艽的matK序列具有3个有效鉴别位点,丽江糙苏和黑花糙苏的matK序列需多个变异位点结合方可进行有效鉴别。结论:本研究表明ITS1、ITS2和matK片段可用于丽江糙苏、黑花糙苏和假秦艽的分子鉴别。
OBJECTIVE: To analyze the nucleotide sequences of the ITS and matK gene fragments of three populations of the genus Polygonatum in Yunnan Province, which are the dominant species of R. solanacearum, P. nigrum and P. pseudoacacia The identification and protection of wild resources provide the molecular basis. Methods: ITS4 / ITS5 specific primer of ITS region and matKXF / matK5R of matK gene were used for PCR amplification and sequencing. Results: The minimum genetic distance (Kp) of K2P between species of Leguminosae, Pterocarya stenophyllum and Kochia scoparia was greater than the maximum genetic distance of K2P in ITS1, ITS2 and matK gene segments. NJ phylogenetic tree also showed that these three medicinal The plants can be distinguished from the genus Genus in genomic databases GenBank, among which 3, 2 and 1 valid discriminative sites were found on ITS2 sequences of Phyllostachys nigra, Gentiana macrophylla and Gentiana macrophylla, ITS1 There are 3, 3 and 3 valid discriminative sites in the sequence, respectively. There are three valid discriminating sites in the matK sequence of fake Gentiana macrophylla. The matK sequences of Phyllostachys pubescens and Gentiana macrophylla need to be combined with each other Effective identification. Conclusion: This study showed that ITS1, ITS2 and matK fragments can be used for molecular identification of Rhizoma.