丛枝菌根真菌异形根孢囊霉14-3-3蛋白基因的克隆及功能验证  

Cloning and Function Verification of 14-3-3 Protein Genes from Arbuscular Mycorrhizal Fungus Rhizophagus irregularis

作　　者：徐丽娇 郝志鹏[1,2] 李涛 李芳[1,2] 李景龙 陈保冬 Xu Lijiao;Hao Zhipeng;Li Tao;Li Fang;Li Jinglong;Chen Baodong(State Key Laboratory of Urban and Regional Ecology,Research Center for Eco-Environmental Sciences,Chinese Academy of Science,Beijing 100085,P.R.China;University of Chinese Academy of Sciences,Beijing 100049,P.R.Chin)

机构地区：[1]城市与区域生态国家重点实验室(中国科学院生态环境研究中心),北京100085 [2]中国科学院大学,北京100049

出　　处：《东北林业大学学报》2018年第8期63-68,共6页Journal of Northeast Forestry University

基　　金：中国科学院战略性先导科技专项(XDB15030102);国家自然科学基金项目(41371264;41401281)

摘　　要：为证明丛枝菌根(AM)真菌在增强植物抗旱性方面的直接作用,克隆了AM真菌异形根孢囊霉(Rhizophagus irregularis)DAOM 197198中的14-3-3蛋白基因Ri14-3-3;通过设计特异性引物,在AM真菌中扩增到了该基因的全长片段。通过酿酒酵母(Saccharomyces cerevisiae)177 w异源表达实验验证了Ri14-3-3的功能。结果表明:在正常水分条件和干旱胁迫下,Ri14-3-3均提高了酵母的生长速率和生长量。正常水分条件下,Ri14-3-3上调了钙调蛋白基因(MAC1)、下调了水孔蛋白基因(AQY3)的表达;干旱胁迫下,Ri14-3-3上调了质膜H+-ATP酶基因(PMA1)、水孔蛋白基因(AQY1)、钙调蛋白基因(CMP)的表达。Ri14-3-3基因能够上调酵母质膜H+-ATP酶基因和水孔蛋白基因,说明干旱胁迫下AM真菌14-3-3蛋白可以调控干旱胁迫响应基因,促进真菌向植物运输水分,从而增强植物抗旱性。To prove direct involvement of Arbuscular mycorrhizal（ AM） fungi in plant drought tolerance,we cloned a 14-3-3 protein gene Ri14-3-3 from AM fungus Rhizophagus irregularis DAOM 197198. We amplified the length fragment of this gene by designing specific primers. The function of Ri14-3-3 was verified by heterologous expression in Saccharomyces cerevisiae 177 w. Ri14-3-3 increased the growth rate of yeast under both normal and drought stressed conditions. Under normal condition,the expression of calmodulin gene MAC1 was up-regulated and aquaporin gene AQY3 was down-regulated by Ri14-3-3. Under drought stress,the expressions of plasma membrane H＋-ATPase gene PMA1,aquaporin gene AQY1 and calmodulin gene CMP were up-regulated by Ri14-3-3. The up-regulation of plasma membrane H＋-ATPase and aquaporin genes of yeast by Ri14-3-3 indicated that AM fungal 14-3-3 proteins can regulate drought responsive genes to promote water transport from AM fungi to the plants,and consequently lead to enhanced plant drought tolerance under drought stress.

关 键 词：丛枝菌根真菌 14-3-3蛋白 抗旱性 基因功能验证 异源表达 

分 类 号：Q785[生物学—分子生物学]