园艺学报 ›› 2013, Vol. 40 ›› Issue (4): 739-.

• 研究报告 • 上一篇    下一篇


 董庆龙, 余贤美, 刘丹丹, 王海荣, 安 淼, 姚玉新, 王长君   

  1. (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)
  • 出版日期:2013-04-25 发布日期:2013-04-25

Cloning of NAD-Malic Enzymes and Their Expression Analysis During Tissues and Fruit Development of Apple

DONG  Qing-Long, YU  Xian-Mei, LIU  Dan-Dan, WANG  Hai-Rong, AN   Miao, YAO  Yu-Xin, WANG  Chang-Jun   

  1. (1Shandong Institute of Pomology,Tai’an,Shandong 271000,China;2College of Horticulture Science and Engineering,
    Shandong Agricultural University,National Research Center for Apple Engineering and Technology,State Key Laboratory
    of Crop Biology,Tai’an,Shandong 271018,China)
  • Online:2013-04-25 Published:2013-04-25

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-ME1MdNAD-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-ME1MdNAD-ME2
具有不同的表达模式。以上结果表明,克隆得到的MdNAD-ME1MdNAD-ME2 属于植物NAD-ME,在
苹果的生长和发育过程中MdNAD-ME1MdNAD-ME2 可能起到不同作用。

关键词: 苹果, 苹果酸酶, 克隆, 序列分析, 表达分析

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.

Key words: apple, malic enzyme, cloning, sequence analysis, expression analysis