Cloning and Characterization of Functional Trehalose-6-Phosphate Synthase Gene in Maize
Cloning and Characterization of Functional Trehalose-6-Phosphate Synthase Gene in Maize
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DOI：
10.1007/s12374-010-9098-7
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发表时间：
2010-04-01
影响因子：
2.9
通讯作者：
Li, Wan-Chen
中科院分区：
文献类型：
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作者：
Jiang, Wei;Fu, Feng-Ling;Li, Wan-Chen
海藻糖是葡萄糖的一种不还原性双糖，在细菌、真菌和一些“复活植物”中作为一种相容性溶质，在高温和干燥胁迫下稳定生物结构。在植物界，海藻糖是由海藻糖-6-磷酸合成酶（TPS）和海藻糖-6-磷酸磷酸酶（TPP）合成的。据报道，外源和内源编码TPS和TPP的基因过表达可有效提高烟草、马铃薯、番茄、水稻和拟南芥的非生物胁迫耐受性。在生物信息学预测的基础上，我们从玉米中克隆了一个含有2820 bp开放阅读框的片段，该片段编码939个氨基酸的蛋白。系统发育分析表明该基因属于TPS基因家族I类亚家族。保守结构域分析显示存在一个TPS结构域和一个TPP结构域。酵母与TPS和TPP突变体的互补表明，该蛋白具有海藻糖-6-磷酸合成酶的活性。半定量RT-PCR和实时定量PCR结果表明，该基因在盐胁迫和冷胁迫下均表达上调。
Trehalose is a non-reducing disaccharide of glucose that functions as a compatible solute in the stabilization of biological structures under heat and desiccation stress in bacteria, fungi, and some "resurrection plants". In the plant kingdom, trehalose is biosynthesized by trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). Over-expression of exogenous and endogenous genes encoding TPS and TPP is reported to be effective for improving abiotic stress tolerance in tobacco, potato, tomato, rice, and Arabidopsis. On the basis of bioinformatics prediction, we cloned a fragment containing an open reading frame of 2,820 bp from maize, which encodes a protein of 939 amino acids. Phylogenetic analysis showed that this gene belongs to the class I subfamily of the TPS gene family. Analysis of conserved domains revealed the presence of a TPS domain and a TPP domain. Yeast complementation with TPS and TPP mutants demonstrated that this protein has the activity of trehalose-6-phosphate synthase. Semi-quantitative RT-PCR and real-time quantitative PCR indicated that the expression of this gene is upregulated in response to both salt and cold stress.