Molecular cloning of three malic acid related genes MdPEPC, MdVHA-A, MdcyME and their expression analysis in apple fruits

Molecular cloning of three malic acid related genes MdPEPC, MdVHA-A, MdcyME and their expression analysis in apple fruits
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苹果酸相关基因MdPEPC、MdVHA-A、MdcyME的分子克隆及其在苹果果实中的表达分析

DOI:
10.1016/j.scienta.2009.05.033
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发表时间:
2009-10
影响因子:
4.3
通讯作者:
Wang, Dong-Mei
Wang, Dong-Mei
中科院分区:
农林科学2区
文献类型:
--
作者:
Hao, Yu-Jin;Li, Ming;You, Chun-Xiang;Zhai, Heng;Liu, Zhi;Yao, Yu-Xin;Wang, Dong-Mei

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苹果果实中的苹果酸(MA)是影响苹果口感、风味和果汁品质的主要有机酸。本研究从苹果果实中克隆了MdPEPC、MdVHA-A和MdcyME三个全长基因。它们分别编码胞质磷酸烯醇式丙酮酸羧化酶(MdPEPC,EC 4.1.1.31)、空泡H+-ATPase A亚基(MdVHA,EC 3.6.1.3)和胞质NADP依赖的苹果酸酶(MdcyME,EC 1.1.1.40),用于MA的合成、运输和降解。实时荧光定量聚合酶链式反应发现,三个基因的表达水平随着发育阶段的不同而不同,并且它们的表达模式在低酸(LA)和高酸(HA)基因型之间存在差异。此外,酶活性测定表明,在LA和HA果实发育过程中,PEPC和VHA对MA的积累都有正的贡献,而聚伞花序则对MA的积累有负的贡献。
Malic acid (MA) in apple fruit is the predominant organic acid associated with taste, flavour and juice quality. In this study, three full-length cDNAs of MdPEPC, MdVHA-A and MdcyME were cloned from apple fruit. They encoded cytosolic phosphoenolpyruvate carboxylase (MdPEPC, EC 4.1.1.31), subunit A of vacuolar H+-ATPase (MdVHA, EC 3.6.1.3) and cytosolic NADP-dependent malic enzyme (MdcyME, EC 1.1.1.40), respectively, for MA synthesis, transportation and degradation. Real-time quantitative PCR discovered that the expression levels of three genes varied with development stages, and that their expression patterns differed between low acid (LA) and high acid (HA) genotypes. In addition, enzyme activity assay showed that PEPC and VHA contributed positively to MA accumulation during fruit development both in LA and HA, while cyME did negatively.
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