超表达番茄GDP-D-甘露糖焦磷酸化酶基因马铃薯对温度胁迫的响应  

Effect of Tomato GMPase Overexpression on Tolerance of Potato Plants to Temperature Stress

作　　者：李超汉[1,2] 李青竹[2] 史庆华[2] 白龙强[2] 郭晓青[2] 李霞[2] 于贤昌[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]山东农业大学园艺科学与工程学院/作物生物学国家重点实验室,山东泰安271018

出　　处：《中国农业科学》2011年第23期4952-4961,共10页Scientia Agricultura Sinica

基　　金：国家自然科学基金项目(30771473);国家科技支撑计划项目(2011BAD12B01);农业部园艺作物遗传改良重点开放实验室项目

摘　　要：【目的】研究番茄GDP-甘露糖焦磷酸化酶基因(GMPase)在马铃薯中的超表达对其抵御温度胁迫能力的影响。【方法】利用已经获得的超表达番茄GMPase的两个转基因马铃薯株系,用野生型马铃薯作对照,比较其在常温(25/20℃)、温度胁迫(10/5℃低温处理3 d,35/30℃高温处理3 d)及25/20℃恢复3 d后GMPase的相对表达量、抗坏血酸(AsA)含量及其相关生理指标的变化。【结果】在常温、温度胁迫与恢复阶段,与野生型植株相比,转基因马铃薯植株具有较高的GMPase相对表达量、GMPase活性、AsA和脱氢抗坏血酸(DHA)含量;高温和低温胁迫后,转基因马铃薯的脱氢抗坏血酸还原酶(DHAR)、单脱氢抗坏血酸还原酶(MDAHR)和抗坏血酸过氧化物酶(APX)活性均明显提高,丙二醛(MDA)和H2O2含量及细胞质膜透性均明显降低。【结论】番茄GMPase在马铃薯中的超表达可能有提高马铃薯植株抵御温度胁迫的能力。【Objective】 The aim of the study was to clarify the function of the GDP-D-mannose pyrophosphorylase（GMPase） in the L-galactose pathway of ascorbic acid（AsA） synthesis in plants,and to make sure the relation of GMPase overexpression with ascorbic acid content and temperature-stress tolerance ability.【Method】 Two transgenic potato strains of overexpressing tomato GMPase were obtained by agrobacterium-mediated transformation.The transgenic plants and wild-type plants were treated in temperature-stress conditions（10/5℃ as low-temperature stress,35/30℃ as high-temperature stress） for 3 days then in the recover-condition（25/20℃） for 3 days.Real time RT-PCR was used to analyze the expression of GMPase gene.And ascorbic acid content and relative index to temperature stress were also determined.【Result】Compared with wild-type plants,with the overexpression of GMPase gene in transgenic plants,the GMPase activity and contents of AsA and dehydroascorbate（DHA） in normal and temperature-stress conditions were increased as well as the activities of dehydroascorbate reductase（DHAR）,monodehydroascorbate reductase（MDHAR） and ascorbate peroxidase（APX）.Meanwhile,the malondialdehyde（MDA） and H2O2 contents and permeability were decreased in the transgenic plants.【Conclusion】 The overexpression of GMPase may increase potato tolerance to temperature stress by increasing AsA level and lowering reactive oxygen species（ROS）.

关 键 词：马铃薯 抗坏血酸 GDP-甘露糖焦磷酸化酶 转基因 温度胁迫 

分 类 号：S532[农业科学—作物学]