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麻疯树抗坏血酸过氧化物酶增强转基因拟南芥的耐盐性。

Ascorbate peroxidase from Jatropha curcas enhances salt tolerance in transgenic Arabidopsis.

作者信息

Chen Y, Cai J, Yang F X, Zhou B, Zhou L R

机构信息

College of Architecture and Environment, Sichuan University, Chengdu, Sichuan, China.

West China School of Pharmacy, Sichuan University, Chengdu, Sichuan, China.

出版信息

Genet Mol Res. 2015 May 11;14(2):4879-89. doi: 10.4238/2015.May.11.20.

Abstract

Ascorbate peroxidase (APX) plays a central role in the ascorbate-glutathione cycle and is a key enzyme in cellular H2O2 me-tabolism. It includes a family of isoenzymes with different character-istics, which are identified in many higher plants. In the present study, we isolated the APX gene from Jatropha curcas L, which is similar with other previously characterized APXs as revealed by alignment and phylogenetic analysis of its deduced amino acid sequence. Real-time qPCR analysis showed that the expression level of JcAPX transcript significantly increased under NaCl stress. Subsequently, to elucidate the contribution of JcAPX to the protection against salt-induced oxi-dative stress, the expression construct p35S: JcAPX was created and transformed into Arabidopsis and transcribed. Under 150-mM NaCl stress, compared with wild type (WT), the overexpression of JcAPX in Arabidopsis increased the germination rate, the number of leaves, and the rosette area. In addition, the transgenic plants had longer roots, higher total chlorophyll content, higher total APX activity, and lower H2O2 content than the WT under NaCl stress conditions. These results suggested that higher APX activity in transgenic lines increases the salt tolerance by enhancing scavenging capacity for reactive oxygen spe-cies under NaCl stress conditions.

摘要

抗坏血酸过氧化物酶(APX)在抗坏血酸-谷胱甘肽循环中起核心作用,是细胞H2O2代谢中的关键酶。它包括一个具有不同特性的同工酶家族,在许多高等植物中都有发现。在本研究中,我们从麻疯树中分离出APX基因,通过对其推导的氨基酸序列进行比对和系统发育分析,发现它与其他先前鉴定的APX相似。实时定量PCR分析表明,在NaCl胁迫下,JcAPX转录本的表达水平显著增加。随后,为了阐明JcAPX对抵御盐胁迫诱导的氧化应激的作用,构建了表达载体p35S:JcAPX并转化到拟南芥中进行转录。在150 mM NaCl胁迫下,与野生型(WT)相比,拟南芥中JcAPX的过表达提高了发芽率、叶片数量和莲座叶面积。此外,在NaCl胁迫条件下,转基因植株的根更长,总叶绿素含量更高,总APX活性更高,H2O2含量更低。这些结果表明,转基因株系中较高的APX活性通过增强NaCl胁迫条件下对活性氧的清除能力来提高耐盐性。

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