园艺学报 ›› 2016, Vol. 43 ›› Issue (6): 1033-1043.doi: 10.16420/j.issn.0513-353x.2015-0719

• 果树 • 上一篇    下一篇


任怡然,赵 强,赵先炎,郝玉金,由春香*   

  1. (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,国家苹果工程技术研究中心,山东泰安 271018)
  • 出版日期:2016-06-25 发布日期:2016-06-25

Expression and Function Analysis of Apple MdDRB1 Gene

REN Yi-ran,ZHAO Qiang,ZHAO Xian-yan,HAO Yu-jin,and YOU Chun-xiang*   

  1. (College of Horticulture Science and Engineering,Shandong Agricultural University,State Key Laboratory of Crop Biology,National Research Center for Apple Engineering and Technology,Tai’an,Shandong 271018,China)
  • Online:2016-06-25 Published:2016-06-25


以‘嘎拉’苹果(Malus × domesticaRoyal Gala’)为试材,扩增MdDRB1基因,分析其序列结构,同时原核诱导MdDRB1蛋白并观察其亚细胞定位。利用qRT-PCR检测MdDRB1非生物胁迫下的表达量,通过遗传转化苹果苗鉴定MdDRB1在非生物胁迫中的功能。基因结构分析显示,MdDRB1具有2个内含子和3个外显子。亚细胞定位显示,MdDRB1主要定位于细胞核,少数定位在细胞质。原核诱导结果显示,MdDRB1融合蛋白以包涵体的形式存在。同时发现MdDRB1反义转基因愈伤中与抗性相关的miRNA的表达水平上调。在PEGABA、盐和低温处理的材料中MdDRB1的表达水平明显上调。另外,MdDRB1过量表达明显提高了转基因苹果组培苗的抗性。推测MdDRB1在抗逆胁迫响应中有重要作用

关键词: 苹果, MdDRB1, 原核表达, 亚细胞定位, 逆境响应


A gene named MdDRB1 cloned from Malus × domestica‘Royal Gala’was analyzed. To understand sequence characteristics of the gene,prokaryotic expression and subcellular localization were carried out. In addition,real-time quantitative PCR were performed to determine the expression levels of MdDRB1 in response to various abiotic stresses. Finally,MdDRB1 was genetically transformed into apple seedlings to identify its function. The analysis of gene structure revealed that there were two introns and three exons in genomic sequence of MdDRB1. Subcellular localization showed that MdDRB1 were mainly distributed in the nucleus,a small number distributed in the cytoplasm. Then,the induced protein showed that MdDRB1 fusion protein existed in the form of inclusion body. Furthermore,the miRNA related to resistance expression level raised in antisense transgenic apple callus. In addition,real-time quantitative PCR found that the MdDRB1 gene expression level significantly raised after treated with PEG,NaCl,ABA and 4 ℃. Overexpression of MdDRB1 remarkably increased the tolerance of transgenic apple seedlings to abiotic stresses. These results indicated that MdDRB1 might be involved in the response of apple to abiotic stresses.

Key words: apple, MdDRB1, prokaryotic expression, subcellular localization, stress esponse