| 苹果NAD–苹果酸酶基因的克隆及在不同组织和果实发育阶段的表达分析 |
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| 引用本文： | 董庆龙,余贤美,刘丹丹,王海荣,安淼,姚玉新,王长君.苹果NAD–苹果酸酶基因的克隆及在不同组织和果实发育阶段的表达分析[J].园艺学报,2013,40(4):739-747. |
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| 作者姓名： | 董庆龙 余贤美 刘丹丹 王海荣 安淼 姚玉新 王长君 |
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| 作者单位： | （1 山东省果树研究所，山东泰安 271000；2 山东农业大学园艺科学与工程学院，国家苹果工程技术研究中心，作物
生物学国家重点实验室，山东泰安 271018） |
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| 基金项目： | 国家自然科学基金项目，山东果树研究所所长科研基金项目 |
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| 摘 要： | NAD-malic enzyme（NAD-ME）是植物调控苹果酸代谢中的关键酶。以富士苹果（Malus ×
domestica Borkh.）叶片为试材，通过同源比对和RT-PCR 技术，克隆获得2 个NAD–苹果酸酶基因
（MdNAD-ME1 和MdNAD-ME2）。其ORF 分别为1 785 bp 和1 818 bp，推测其分别编码595 和605 个氨
基酸的多肽。氨基酸序列和结构分析显示，含有5 个保守的氨基酸区域（Ⅰ~Ⅴ），包含2 个功能结构域：
malic 和NAD_bind_1_malic_enz。进化树分析结果显示，MdNAD-ME1 属于α 组双子叶亚组，MdNAD-ME2
属于β 组双子叶亚组。荧光实时定量结果显示，MdNAD-ME 在被检测的组织中呈组成型表达，且在多种
组织中MdNAD-ME1 表达量高于MdNAD-ME2。在富士苹果果实不同发育阶段，MdNAD-ME1 与MdNAD-ME2
具有不同的表达模式。以上结果表明，克隆得到的MdNAD-ME1 和MdNAD-ME2 属于植物NAD-ME，在
苹果的生长和发育过程中MdNAD-ME1 和MdNAD-ME2 可能起到不同作用。
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| 关 键 词： | 苹果 苹果酸酶 克隆 序列分析 表达分析 |
Cloning of NAD-Malic Enzymes and Their Expression Analysis During Tissues and Fruit Development of Apple |
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| DONG Qing-Long,YU Xian-Mei,LIU Dan-Dan,WANG Hai-Rong,AN Miao,YAO Yu-Xin,WANG Chang-Jun.Cloning of NAD-Malic Enzymes and Their Expression Analysis During Tissues and Fruit Development of Apple[J].Acta Horticulturae Sinica,2013,40(4):739-747. |
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| Authors: | DONG Qing-Long YU Xian-Mei LIU Dan-Dan WANG Hai-Rong AN Miao YAO Yu-Xin WANG Chang-Jun |
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| Institution: | Department of Respiratory, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, China. |
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| Abstract: | NAD-malic enzyme is a key enzyme in malic acid metabolism pathway of plants. Two
apple NAD-ME genes（MdNAD-ME1 and MdNAD-ME2）were isolated from the‘Fuji’apple leaves
（Malus × domestica Borkh.）by homologous comparison and RT-PCR confirmation. MdNAD-ME1 and
MdNAD-ME2 contained 1 785 bp and 1 818 bp ORFs，which encoded 595- and 605-aa proteins，
respectively. Amino acid sequence and structure analysis indicated that MdNAD-MEs contained five
conservative amino acid areas（Ⅰ–Ⅴ）and two functional structure domains：Malic and NAD_bind_1_
malic_enz. The result of phylogenetic analysis showed that MdNAD-ME1 belonged to dicotyledon
subgroup of α group，while MdNAD-ME2 belonged to dicotyledon subgroup of β group. qRT-PCR result
showed that MdNADP-MEs were constitutively expressed in all examined tissues，and the expression level
of MdNAD-ME1 were higher than of MdNAD-ME2 during more tissues. MdNAD-ME1 and MdNAD-ME2
had different patterns of expression during fruit development of‘Fuji’apple. Taken together，the above
results indicated that MdNAD-MEs belonged to NAD-ME family，and MdNAD-ME1 and MdNAD-ME2
might play different roles in the growth and development of apple. |
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| Keywords: | apple malic enzyme cloning sequence analysis expression analysis |
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