基于ITS2序列鉴定甘遂种子  

Identification of Euphorbia kansui Seeds Based on ITS2 Sequence

作　　者：李振凯 雷燕 梁旺利 王红 彭励[1,2,3] Li Zhenkai;Lei Yan;Liang Wangli;Wang Hong;Peng Li(School of Life Sciences,Ningxia University,Yinchuan,750021;Key Lab of Ministry of Education for Protection and Utilization of Special Biologi-cal Resources in Western China,Ningxia University,Yinchuan,750021;Ningxia Summer Pharmaceutical Engineering Technology Center,Yinchuan,750021)

机构地区：[1]宁夏大学生命科学学院,银川750021 [2]宁夏大学,西部特色生物资源保护与开发利用教育部重点实验室,银川750021 [3]宁夏天然药物工程技术中心,银川750021

出　　处：《分子植物育种》2020年第16期5436-5443,共8页Molecular Plant Breeding

基　　金：国家自然基金项目(31260036);宁夏回族自治区重大专项(2015BY11103);宁夏回族自治区重点研发项目(2019BBF02029)共同资助。

摘　　要：为了对多形态甘遂种子的真实性进行鉴定,对收集到的3个省区共11份甘遂种子进行形态学观察,通过ITS2序列测序分析,并借助于中药材DNA条形码鉴定系统和NCBI数据库进行BLAST鉴定,应用MEGA 7软件计算遗传距离值、构建系统发育树,比较二级结构差异,并将结果进行田间试验验证。结果表明,ITS2序列测序分析共得到2条不同序列,2条序列存在明显的亲缘距离,GS序列Ⅰ与甘遂药材的ITS2序列相吻合,而GS序列Ⅱ与大戟属序列相近,其中GS序列Ⅰ对应种子批次均可发育成符合《中国药典》描述的呈椭圆形、长圆柱形或连珠形;此外在种子形态、田间生长势、植株形态等特征上2条序列对应种子也表现出明显的差异。由此表明基于ITS2序列分析的分子鉴定技术可以有效鉴定多形态甘遂种子的真实性,并明确了正品甘遂种子的形态特征,为甘遂种子生产、流通及种植提供理论依据和鉴定手段。In order to identify the authenticity of the seeds of Euphorbia kansui,morphological observation was carried out on the seeds collected from three provinces and regions.BLAST identification was carried out with the DNA barcode identification system and NCBI database of Chinese medicinal materials through ITS2 sequence analysis.The genetic distance value was calculated by MEGA 7 software,and phylogenetic tree was constructed to compare the two levels.The results showed that two different sequences were obtained by ITS2 sequencing analysis.The GS sequenceⅠwas highly consistent with the ITS2 sequence of E.kansui,while the GS sequenceⅡwas similar to that of the genus Euphorbia,and the two sequences were closely related to each other.The seed batches corresponding to GS sequenceⅠdeveloped into oval,long cylindrical or beaded tuber roots in accordance with the provisions of the Chinese Pharmacopoeia.In addition,there were significant differences between the two sequences in seed morphology,field growth potential and plant morphology.This indicates that based on ITS2 sequence analysis can effectively identify the authenticity of the seeds of Euphorbia kansui multimorphological,and clarify the morphological characteristics of the seeds of the original E.kansui.It provides a theoretical basis and identification method for seed production,circulation and planting of medicinal materials of E.kansui.

关 键 词：甘遂种子 ITS2序列 分子鉴定 种子真实性 

分 类 号：S567.239[农业科学—中草药栽培]