Analysis of xyloglucan endotransglycosylase/hydrolase (XTH) genes and diverse roles of isoenzymes during persimmon fruit development and postharvest softening.
Analysis of xyloglucan endotransglycosylase/hydrolase (XTH) genes and diverse roles of isoenzymes during persimmon fruit development and postharvest softening.
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DOI：
10.1371/journal.pone.0123668
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Rao J
中科院分区：
文献类型：
--
作者：
Han Y;Zhu Q;Zhang Z;Meng K;Hou Y;Ban Q;Suo J;Rao J
木葡聚糖内糖基转移酶/水解酶(XTH)酶在半纤维素细胞壁重塑中起着重要作用。研究XTHs在柿(Diospyros kaki L.)在果实发育和采后软化过程中，克隆了5个cDNAs(DkXTH1~DkXTH5)，它们的推测蛋白含有保守的Xth的DEIDFEFLG基序。实时荧光定量聚合酶链式反应分析表明，DkXTH1、DkXTH4和DkXTH5基因在未成熟膨大果实中的表达达到高峰，并且在冷处理果实和硬度较高的品种果实中的表达水平较高。DkXTH2和DkXTH3的基因表达模式相反，随着果实软化程度的加深，DkXTH2和DkXTH3的基因表达谱达到最大值。同时，木葡聚糖内糖基转移酶(XET)在柿果实快速生长和成熟过程中起着重要作用。此外，重组的DkXTH1和DkXTH2蛋白显示出显著的XET活性，没有检测到任何XEH活性。然而，重组DkXTH2蛋白的XET活性与小受体分子的亲和力高于重组DkXTH1蛋白。前者可能更倾向于参与细胞壁重组，后者更倾向于参与细胞壁组装。此外，DKXTH蛋白还可以在信号肽的调控下通过靶向细胞壁发挥作用。结果表明，不同的DKXTHs可能表现出不同的表达模式，编码产物具有特定的酶特性，这与它们在柿果实生长和采后软化过程中的功能有关。
Xyloglucan endotransglycosylase/hydrolase (XTH) enzymes have played a role in the remodeling of cell wall hemicelluloses. To investigate the function of XTHs in persimmon (Diospyros kaki L.) fruit development and postharvest softening, five cDNAs (DkXTH1 to DkXTH5), whose putative proteins contained the conserved DEIDFEFLG motif of XTH, were cloned. Real time quantitative PCR analysis revealed that DkXTH1, DkXTH4, and DkXTH5 peaked in immature expanding fruit, and their higher expression was observed along with higher fruit firmness in cold-treated fruit or firmer cultivar fruit during storage. The opposite gene expression patterns were observed in DkXTH2 and DkXTH3, which reached maxima concomitance with pronounced fruit softening. Meanwhile, the xyloglucan endotransglycosylase (XET) enzymes play important roles in both the rapid growth and ripening of persimmon fruit. Furthermore, the recombined DkXTH1 and DkXTH2 proteins showed significant XET activity without any detected XEH activity. However, the XET activity of recombined DkXTH2 protein had a higher affinity for small acceptor molecules than that of recombined DkXTH1 protein. The former might prefer to participate in cell wall restructuring, and the latter is more inclined to participate in cell wall assembly. Besides, DKXTH proteins could function by targeting to the cell wall under regulation of a signal peptide. The data suggested that individual DKXTHs could exhibit different patterns of expression, and the encoded products possessed specific enzymatic properties conferring on their respective functions in growth and postharvest softening of persimmon fruit.
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