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植物研究 ›› 2005, Vol. 25 ›› Issue (1): 69-73.doi: 10.7525/j.issn.1673-5102.2005.01.021

• 论文 • 上一篇    下一篇

干旱胁迫下刚毛柽柳消减文库的构建及分析

刘桂丰1,2, 侯英杰2, 王玉成2, 褚延广2   

  1. 1. 北京林业大学生物科学与技术学院, 北京 100083;
    2. 东北林业大学林学院, 哈尔滨 150040
  • 收稿日期:2004-07-27 出版日期:2005-03-15 发布日期:2016-06-14
  • 作者简介:刘桂丰(1960-), 男, 教授, 主要从事分子遗传及林木育种方面的研究工作。
  • 基金资助:
    国家973计划资助项目(G1999016000)

Construction and analysis of Tamarix hispida suppression subtractive hybridization library under drought stress

LIU Gui-Feng1,2, HOU Ying-Jie2, WANG Yu-Cheng2, CHU Yan-Guang2   

  1. 1. School of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083;
    2. Forest College Northeast Forestry University, Harbin 150040
  • Received:2004-07-27 Online:2005-03-15 Published:2016-06-14

摘要: 以干旱胁迫下的刚毛柽柳根部组织cDNA为tester,正常生长的刚毛柽柳根部组织cDNA为driver,利用抑制性消减杂交技术(SSH)构建了干旱胁迫下刚毛柽柳根部组织的消减文库。文库克隆的重组率为95%,插入片段大部分集中在250~600 bp之间。通过对文库阳性克隆的随机测序,获得了如Mn-SOD、myb相关蛋白、锌指蛋白等17种与干旱胁迫相关的基因,它们涉及了植物的渗透调节、信号传递、转录调控、活性氧清除等方面。所得EST序列已被GenBank收录。实验为抗逆基因克隆和系统研究干旱胁迫下柽柳根部基因的表达奠定了基础。

关键词: 干旱胁迫, 刚毛柽柳, 抑制性消减杂交, 表达序列标签

Abstract: Using cDNA from Tamarix hispida roots treated with drought stress as tester and cDNA from Tamarix hispida roots in normal growth as driver, suppression subtractive hybridization (SSH) was employed to construct cDNA subtracted library. In the library, the rate of recombination was 95%, the size of inserts was 250~600 bp. 17 drought stress-associated genes were obtained by DNA sequencing the positive clones picked randomly, such asMn-SOD myb-related protein zinc finger protein, etc. These genes included osmotic regulator signal component regulatory protein and antioxidant enzyme. GenBank had accepted all the sequences. The research had established a basis for cloning stress-resistance genes and further studying genes expression in Tamarix hispida roots under drought stress.

Key words: drought stress, Tamarix hispida, suppression subtractive hybridization, expressed sequence tags