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作物学报 ›› 2012, Vol. 38 ›› Issue (06): 971-979.doi: 10.3724/SP.J.1006.2012.00971

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

野生大豆GsGST19基因的克隆及其转基因苜蓿的耐盐碱性分析

王臻昱1,2,3，才华1，柏锡1，纪巍1，李勇1，魏正巍1，朱延明1,*   

  1. 1东北农业大学植物生物工程研究室，黑龙江哈尔滨150030；2 中国科学院东北地理与农业生态研究所，长春1300112； 3 中国科学院研究生院，北京100049
  • 收稿日期:2011-11-17 修回日期:2012-02-22 出版日期:2012-06-12 网络出版日期:2012-04-06
  • 通讯作者: 朱延明, E-mail: ymzhu2001@neau.edu.cn
  • 基金资助:

    本研究由国家自然科学基金项目(31171578), 黑龙江省高校科技创新团队建设计划(2011TD005), 国家高技术研究发展计划(863计划)项目(2008AA10Z153)和黑龙江省科技厅重大攻关项目(GA06B103)资助。

Isolation of GsGST19 from Glycine soja and Analysis of Saline-Alkaline Tolerance for Transgenic Medicago sativa

WANG Zhen-Yu1,2,3,CAI Hua1,BAI Xi1,JI Wei1,LI Yong1,WEI Zheng-Wei1,ZHU Yan-Ming1,*   

  1. 1 Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030, China; 2 Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Changchun 130012, China; 3 Graduate University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2011-11-17 Revised:2012-02-22 Published:2012-06-12 Published online:2012-04-06
  • Contact: 朱延明, E-mail: ymzhu2001@neau.edu.cn

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被引次数

24

摘要： 谷胱甘肽S-转移酶对植物抵御逆境胁迫和解除细胞毒素起着重要作用。本研究从野大豆盐碱胁迫基因表达谱中筛选并克隆得到GsGST19基因，将其转化苜蓿，获得超量表达的转基因苜蓿，并对转基因苜蓿进行耐盐碱性分析。结果显示在正常培养条件下，转基因苜蓿株系19-4和19-9的GST酶活性分别是非转基因株系的1.52倍和1.49倍。在100 mmol L–1 NaHCO3处理14 d后转基因株系生长状态良好，而非转基因对照株系明显萎蔫、失绿、甚至死亡；转基因株系的丙二醛含量和相对质膜透性显著低于非转基因株系(P<0.05)，而叶绿素含量和根系活力显著高于非转基因对照(P<0.05)，说明超量表达GsGST19基因增强了苜蓿的耐盐碱能力。

关键词: 谷胱甘肽S-转移酶(GST), 转基因苜蓿, 野大豆, 耐盐碱性

Abstract: Alfalfa (Medicago sativa L.) is one of the most important leguminous forage crops worldwide. Saline-alkaline stress significantly limits the productivity of alfalfa due to its adverse effects on growth, formation of nodules, and symbiotic nitrogen-fixation capacity, and resulting in the formation of reactive oxygen species (ROS). To protectplants from the toxicity ofreactiveoxygen, aerobic organisms are equipped with an array of defense mechanisms, including one based on glutathione S-transferases (GSTs). Glutathione S-transferases (GSTs) are ubiquitous enzymes that play a key role in stress tolerance and cellular detoxification. The GST gene GsGST19 isolated fromwild type soybean(Glycine soja) under saline-alkaline stress was transformed into alfalfa (Medicago sativa L.). Transgenic alfalfa plants showed 0.52–0.49 times higher levels of GST activity than wild type plants. Transgenic alfalfa grew well in the conditions of 100 mmol L–1 NaHCO3, while wild type plants exhibited discoloration and stunted growth, or even death. There were significantly changes in malondialdehyde content and relative membrane permeability caused by saline-alkaline stress in non-transgenic lines compared to transgenic lines (P<0.05). Moreover, compared with non-transgenic, transgenic alfalfa had higher levels of chlorophyll content and root activity under saline-alkali stress conditions. The results indicated that the gene GsGST19 could enhanceresistance to saline-alkaline in alfalfa.

Key words: Glutathione S-transferase, Transgenic alfalfa, Glycine soja, Saline-alkaline stress

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