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植物研究 ›› 2014, Vol. 34 ›› Issue (3): 403-408.doi: 10.7525/j.issn.1673-5102.2014.03.018

• 论文 • 上一篇    下一篇

山茶属植物ITS的扩增及其序列特征分析

尤欢1,3;周阿涛2,3;岳亮亮3;寸东义3;丁元明3*   

  1. 1.云南农业大学植物保护学院,昆明 650201;2.昆明理工大学生命科学与技术学院,昆明 650224;3.云南出入境检验检疫局,昆明 650228
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2014-05-20 发布日期:2014-05-20
  • 通讯作者: 丁元明
  • 基金资助:
     

Amplification and Sequence Characterization of ITS in Camellia

YOU Huan;ZHOU A-Tao;YUE Liang-Liang;CUN Dong-Yi;DING Yuan-Ming*   

  1. 1.College of Plant Protection,Yunnan Agricultural University,Kunming 650201;2.Faculty of Life Science and Technology,Kunming University of Science and Technology,Kunming 650224;3.Yunnan EntryExit Inspection and Quarantine,Kunming 650228
  • Received:1900-01-01 Revised:1900-01-01 Online:2014-05-20 Published:2014-05-20
  • Contact: DING Yuan-Ming
  • Supported by:
     

摘要: 以山茶属(Camellia)植物为实验材料,通过在PCR反应液中加入二甲基亚砜(DMSO)来改进ITS区的扩增效果,并对获得的序列进行了相关的比较分析以及系统发育分析。分析结果表明:(1)PCR反应液中添加4%的DMSO能明显提高ITS的扩增效率,并能有效防止ITS假基因的扩增;(2)与功能拷贝相比,ITS假基因序列的ITS1和ITS2区存在多处碱基缺失,GC含量明显偏低,5.8S区二级结构稳定性降低;(3)应用ITS假基因进行种水平的系统发育分析会对其真实的系统关系造成影响,提示进行系统进化研究时应警惕假基因的干扰。

关键词: DMSO, ITS, 山茶属, 假基因

Abstract: In this study, PCR amplification of nrDNA ITS of Camellia was improved by the addition of DMSO in PCR protocol, the sequence characteristic of ITS were analyzed. The results showed that: (1)The PCR amplification efficiency of ITS was improved by the addition of 4% DMSO, and the ITS pseudogenes were prevented from being amplified; (2)ITS pseudogenes exhibited a significant deletion in ITS1 and ITS2 region, and displayed lower GC content and unstable secondary structure; (3)ITS pseudogenes could not provided the real phylogenetic relationships, it showed the necessity of great care when used the ITS sequence in evolutionary studies in genus Camellia.

Key words: DMSO, ITS, Camellia, pseudogene

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