| 苹果6–磷酸葡萄糖酸脱氢酶基因Md6PGDH1的克隆和功能分析 |
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| 引用本文： | 苏玲,刘鑫,安建平,李浩浩,赵锦,郝玉金,王小非,由春香.苹果6–磷酸葡萄糖酸脱氢酶基因Md6PGDH1的克隆和功能分析[J].园艺学报,2016,43(7):1225-1235. |
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| 作者姓名： | 苏玲 刘鑫 安建平 李浩浩 赵锦 郝玉金 王小非 由春香 |
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| 作者单位： | （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，农业部黄淮地区园艺作物生物学与种质创制重点实验室，山东泰安 271018） |
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| 基金项目： | 国家重点基础研究发展计划(‘973’)项目(2013CB945103)，国家自然科学基金项目(31272142 |
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| 摘 要： | 以‘嘎拉’苹果为材料,克隆了6–磷酸葡萄糖酸脱氢酶基因Md6PGDH1(序列号:MDP0000279299)全长。序列分析显示,该基因包含一个长为1 047 bp完整的开放阅读框,编码347个氨基酸,分子量为36.430 k D,预测等电点为9.24。同源性分析表明Md6PGDH1还有另外3个同源基因;功能域分析表明Md6PGDH蛋白含有两个保守的绑定域;亚细胞预测表明Md6PGDH定位存在差异。分析Md6PGDH1启动子发现存在多个响应非生物胁迫的顺式作用元件。定量分析显示,Md6PGDH1在苹果的不同组织中都有表达,且受非生物胁迫诱导。原核诱导Md6PGDH1蛋白并进行蛋白酶活的测定,为后续蛋白功能鉴定奠定了基础。Md6PGDH1在苹果愈伤组织中过量表达,提高其抗盐胁迫的能力。
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| 关 键 词： | 苹果 Md6PGDH 表达分析 Md6PGDH1 原核诱导 愈伤组织 |
Molecular Cloning and Functional Analysis of a 6-phosphoglueonate Dehydrogenase Gene Md6PGDH1 in Apple |
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| SU Ling,LIU Xin,AN Jian-ping,LI Hao-hao,ZHAO Jin,HAO Yu-jin,WANG Xiao-fei,YOU Chun-xiang.Molecular Cloning and Functional Analysis of a 6-phosphoglueonate Dehydrogenase Gene Md6PGDH1 in Apple[J].Acta Horticulturae Sinica,2016,43(7):1225-1235. |
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| Authors: | SU Ling LIU Xin AN Jian-ping LI Hao-hao ZHAO Jin HAO Yu-jin WANG Xiao-fei YOU Chun-xiang |
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| Institution: | （College of Horticulture Science and Engineering，Shandong Agricultural University，State Key Laboratory of Crop Biology，MOA Key Laboratory of Horticultural Crop Biology（Huanghuai Region）and Germplasm Innovation，Tai’an，Shandong 271018，China） |
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| Abstract: | A 6-phosphoglueonate dehydrogenase gene named Md6PGDH1（MDP0000279299）was cloned from‘Royal Gala’apple（Malus × domestica Borkh.）. Sequence analysis indicated that the length of Md6PGDH1 gene was 1 047 bp，which encoded 347 amino acids. It was predicted that the molecular mass of this protein was 36.430 kD，and pI was 9.24. Homology analysis showed that there were three other homologous genes；Analysis of functional domain showed that the Md6PGDHs protein included two conserved binding domains；the prediction of subcellular localization showed that there were differences in Md6PGDHs protein. In silico analysis suggested that the promoter sequence contained several cis-acting elements，including abiotic stresses responsive elements and hormone responsive elements. qRT-PCRs were performed to determine the expression levels of apple Md6PGDH1 in different tissues and in response to abiotic stresses. Prokaryotic expression and activity assay of the Md6PGDH1，which laid a foundation for protein function identification. Overexpression of Md6PGDH1 in apple callus increased the tolerance of transgenic apple callus to high salinity. |
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| Keywords: | Malus × domestica Md6PGDH expression analysis Md6PGDH1 prokaryotic expression callus |
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