《植物生理学报》 2013, 49(5): 493-500

过表达Mdip1基因提高番茄抗低温诱导的氧化胁迫能力

晋文娟¹, 张少杰¹, 陈双臣^1,*, 林晓民¹, 贺超兴²

1河南科技大学林学院, 河南洛阳471003; 2中国农业科学院蔬菜花卉研究所, 北京100081

通信作者：陈双臣；E-mail: lamont@126.com；Tel: 0379-64282345

摘　要：

采用农杆菌介导法获得了类萌发素蛋白(germin-like protein, Mdip1)超表达的转基因番茄植株, PCR和Southern杂交检测表明, 外源基因分别以1~3个拷贝整合到番茄基因组中。Real-time PCR检测表明, 在转基因个体中目的基因已经在受体基因组上完成整合, 且不同株系间的表达有所差异。分析了低温下转Mdip1基因株系和野生型植株中AsA、GSH含量及H2O2累计的差异。与未转基因植株相比, Mdip1超表达植株在低温胁迫下根干重、冠干重和叶绿素含量显著高于对照, 并显著提高了氧化还原信号AsA、GSH含量和AsA/DHA、GSH/GSSG以及GalLDH活性, 降低了低温胁迫下叶片H2O2、MDA的积累, 降低程度与表达量相关。亚细胞定位表明, H2O2的增加主要分布在质外体。表明Mdip1基因通过调控氧化还原信号在番茄抗氧化胁迫响应中起重要作用。

关键词：番茄; 氧化还原信号; 类萌发素蛋白; 活性氧; 低温胁迫

收稿：2013-01-17   修定：2013-04-09

资助：资助 国家科技支撑计划(2011BAD12B03)、国家自然科学基金(31101536)、山东省科技攻关计划(2012GNC011111)和河南省青年骨干教师计划(2011GGJS-075)。

Overexpression of Mdip1 Gene Enhanced Tolerance of Transgenic Tomato to Chilling Mediated Oxidative Stress

JIN Wen-Juan¹, ZHANG Shao-Jie¹, CHEN Shuang-Chen^1,*, LIN Xiao-Min¹, HE Chao-Xing²

¹Department of Forestry, Henan University of Science & Technology, Luoyang, Henan 471003, China; ²Institute of Vegetables and Flowers, CAAS, Beijing 100081, China

Corresponding author: CHEN Shuang-Chen; E-mail: lamont@126.com; Tel: 0379-64282345

Abstract:

The germin-like protein gene Mdip1 was introduced into tomato line Ailsa Craig via Agrobacterium-mediated method. Transgenic lines were confirmed for integration into the tomato genome using PCR, Southern blot hybridization. One to three copies of the transgene were integrated into the tomato nuclear genome. Tran-scription of Mdip1 in various transgenic lines was determined using real-time PCR. Performance test of resis-tance analyses to chilling stress with T1 generation transgenic tomato lines showed that the transgenic lines ex-hibited lighter symptoms of chilling injury and kept higher values of biomass accumulation and chlorophyll content than those of non-transformed plants. The resistance levels were related to expression levels of the transgene compared with wild-type plants, and the contents of AsA and AsA/DHA in both normal and low-temperature conditions were increased, while the H2O2 and MDA content were decreased in the transgenic plants. Mean-time, the contents of GSH and GSH/GSSG and GalLDH activity were also increased in the overexpression plants. Based on these results, it can be concluded that Mdip1 may play a pivotal role in increasing tomato tol-erance against chilling stress by increasing redox level and lowering H2O2 and lipid peroxidation accumulation.

Key words: tomato; redox signal; germin-like protein; reactive oxygen species; chilling stress