平邑甜茶谷氨酸受体同源基因(MhGLR3.6)的克隆、表达及转化  

Cloning,Expression and Transformation of MhGLR3.6 Gene in Malus hupehensis var. pingyiensis

作　　者：主春福[1] 彭福田[1] 彭静[1] 姜远茂[1] 李光杰[1] 

机构地区：[1]山东农业大学园艺科学与工程学院国家作物生物学重点实验室

出　　处：《林业科学》2008年第9期48-53,共6页Scientia Silvae Sinicae

基　　金：国家自然科学基金资助项目(30571286);山东省自然科学基金资助项目(Y2007D08)

摘　　要：根据GenBank中检索到的苹果谷氨酸受体基因(GLRs)的EST序列,采用RACE方法克隆平邑甜茶谷氨酸受体同源基因MhGLR。该基因全长3600 bp,编码946个氨基酸,推测分子质量为107.809 ku。将MhGLR编码氨基酸与其他已知的谷氨酸受体氨基酸进行同源性比较,发现MhGLR属于谷氨酸受体第三亚家族(GLR3),且与拟南芥AtGLR3.6的同源关系最近,故将其命名为MhGLR3.6(GenBank注册号:EF432572)。亲水性分析表明,MhGLR3.6包含有动物离子型谷氨酸受体(iGLuRs)的6个具有重要功能的保守结构域。荧光定量PCR结果显示,MhGLR3.6在根、茎、叶中均有所表达,且在叶中的表达量最高;L-谷氨酸和IBA处理能够诱导根中MhGLR3.6的表达。成功构建了35S::MhGLR3.6反义表达载体,并对‘皇家嘎拉’苹果进行农杆菌介导的遗传转化。MhGLR gene was identified from Malus hupehensis var. pingyiensis by RACE on the basis of apple expressed sequence tag （EST） database. MhGLR cDNA was 3 600 bp in length and encoded a protein molecule with 946 amino acids, whose molecular mass was estimated of 107. 809 ku. Sequence alignment of MhGLR with other members of the GLR （glutamate receptor） family showed that MhGLR was closely related to clade Ⅲ Arabidopsis GLRs and was closest to AtGLR3.6. Therefore, we named it MhGLR3.6 （ GenBank accession No. ： EFg32572）. Hydropathy analysis indicated that MhGLR3.6 contained six signature domains of animal ionotropic GluRs. Quantitative real-time PCR analysis demonstrated that MhGLR3.6 was expressed in roots, stems and leaves. The expression level in leaves was higher than that in roots and stems. L-glutamate and IBA treatments were able to induce the expression of MhGLR3.6 in roots. To study the function of MhGLR3.6, we introduced the antisense MhGLR3.6 under the control of 35S promoter of Cauliflower mosaic virus into Malus domestica cv. Royal Gala plants.

关 键 词：平邑甜茶 MhGLR3.6 基因克隆 表达分析 转化 

分 类 号：S718.46[农业科学—林学] Q943.2[生物学—植物学]