基于rbcL-a和ITS的枸杞鉴别、遗传关系分析及ITS假基因的发现

发布时间：2018-04-09 21:19

本文选题：枸杞　切入点：鉴别　出处：《生物技术通报》2017年05期

【摘要】：利用rbcL-a及ITS序列作为分子标记,对14个枸杞种或品种及1个伪品白刺进行了真伪鉴别及遗传关系分析。CTAB法提取总DNA,用通用引物对rbcL-a和ITS序列扩增、克隆、测序及分析,T克隆解决部分样品ITS无法成功测序问题。rbcL-a和ITS序列通用引物可在所有样品中成功扩增,rbcL-a序列全长643 bp,在14个枸杞种或品种中完全一致,与白刺具有45bp(7%)的碱基变异。ITS序列在14个枸杞种或品种中长度变异范围为513-692 bp。采用UPGMA法构建系统树,可区分白刺与枸杞,黑果枸杞和宁夏枸杞各品种分别聚为两支,云南枸杞、韩国枸杞及黄果枸杞则聚在宁夏枸杞大分支内。rbcL-a序列可有效进行枸杞真伪鉴别,ITS序列可用于枸杞品种鉴定及遗传关系分析。此外,本研究首次揭示了枸杞品种及个体内的ITS多态性,发现在枸杞中存在ITS假基因这一现象,为未来利用ITS序列进行枸杞系统进化研究提供一些新的参考。
[Abstract]:Using rbcL-a and ITS sequences as molecular markers, 14 species or varieties of Lycium barbarum and one fake Spurs were identified by identification and genetic relationship analysis. RbcL-a and ITS sequences were amplified and cloned by universal primer.Sequencing and analysis of T clone to solve the problem that partial ITS can not be successfully sequenced. RbcL-a and ITS universal primers can successfully amplify the full length of rbcL-a in all samples, which is completely consistent among 14 Lycium barbarum species or varieties.The length variation of its sequence in 14 Lycium barbarum species or varieties ranged from 513-692 BP.The UPGMA method was used to construct the system tree, which can distinguish the varieties of Chinese wolfberry, black fruit medlar and Ningxia medlar into two branches, Yunnan Lycium barbarum, Yunnan Lycium barbarum, and Chinese wolfberry, respectively.The rbcL-a sequence of Lycium barbarum and Lycium barbarum can be used to identify the true and false of Lycium barbarum and its sequence can be used to identify the varieties of Lycium barbarum and analyze the genetic relationship.In addition, the ITS polymorphism in Chinese wolfberry varieties and individuals was revealed for the first time, and the existence of ITS pseudogenes in Lycium barbarum was found, which provides some new references for the systematic evolution of Lycium barbarum in the future using ITS sequences.
【作者单位】：中国科学院过程工程研究所生化工程国家重点实验室;北京林业大学生物科学与技术学院;
【基金】：青海省基本科技计划项目(2013-N-505)
【分类号】：S567.19

【相似文献】