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以‘嘎拉’苹果(Malus×domestica‘Royal Gala’)为材料,采用同源克隆和PCR技术分离了G蛋白偶联受体基因MdGCR1。其开放阅读框(ORF)为960 bp,编码含有319个氨基酸的蛋白。系统进化树分析显示,MdGCR1与梨GCR1亲缘关系最近,同源性最高。基因表达分析显示MdGCR1主要在苹果根、茎和叶中表达,在花和果实中的表达量较低;MdGCR1受多种逆境胁迫诱导,参与多种激素响应。构建了MdGCR1植物过表达载体,并通过农杆菌介导的遗传转化获得了转MdGCR1烟草。转基因材料抗性试验结果显示:超量表达MdGCR1显著降低烟草对干旱胁迫的抗性,表明苹果G蛋白偶联受体基因MdGCR1在响应植物抗旱胁迫中发挥重要作用。
The G protein-coupled receptor gene MdGCR1 was isolated from Malus × domestica’Royal Gala by homologous cloning and PCR. Its open reading frame (ORF) is 960 bp, encoding a 319 amino acid protein. Phylogenetic tree analysis showed that MdGCR1 had the closest genetic relationship with pear GCR1 and the highest homology. Gene expression analysis showed that MdGCR1 was mainly expressed in roots, stems and leaves of apple, but lower in flowers and fruits. MdGCR1 was induced by various stresses and was involved in many hormone responses. The MdGCR1 plant overexpression vector was constructed and transformed to MdGCR1 tobacco by Agrobacterium tumefaciens-mediated genetic transformation. Transgenic material resistance test results showed that over-expression of MdGCR1 significantly reduced tobacco resistance to drought stress, indicating that the apple G protein-coupled receptor gene MdGCR1 play an important role in response to plant drought stress.