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单子叶盐生植物獐毛液泡膜Na(+)/H(+)逆向转运蛋白基因的鉴定与表达

Characterization and expression of a vacuolar Na(+)/H(+) antiporter gene from the monocot halophyte Aeluropus littoralis.

作者信息

Zhang Gao-Hua, Su Qiao, An Li-Jia, Wu Song

机构信息

Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116024, People's Republic of China.

出版信息

Plant Physiol Biochem. 2008 Feb;46(2):117-26. doi: 10.1016/j.plaphy.2007.10.022. Epub 2007 Oct 26.

Abstract

Plant vacuolar Na(+)/H(+) antiporter plays an important role in salt tolerance. In order to understand the molecular basis of vacuolar Na(+)/H(+) antiporter responded to salinity and reveal a possible role of salt tolerance in monocots, a vacuolar Na(+)/H(+) antiporter gene (AlNHX) was isolated by reverse transcription-PCR and RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE) based on the homology from Aeluropus littoralis (Gouan) Parl, a graminaceous halophyte. The AlNHX sequence contained 2706bp with an open read frame of 1623bp and the deduced transcripts encoding 540 amino acids shared a high homology with those putative vacuolar Na(+)/H(+) antiporters of higher plants. AlNHX was predicted containing ten putative hydrophobic regions, which was different with AtNHX1 and OsNHX1. DNA gel blot analysis indicated that there were two or three copies of AlNHX in the A. littoralis genome. The increased transcript levels of AlNHX were much higher in roots than that in shoots under salt stress. In addition, overexpression of AlNHX in tobacco conferred high salt tolerance to the transgenic plants. The analysis of ion contents indicated that under high salt stress for one month, the transgenic plants compartmentalized more Na(+) in the roots and kept a relative high K(+)/Na(+) ratio in the leaves compared with wild-type plants.

摘要

植物液泡Na(+)/H(+)逆向转运蛋白在耐盐性方面发挥着重要作用。为了了解液泡Na(+)/H(+)逆向转运蛋白对盐度作出反应的分子基础,并揭示单子叶植物耐盐性的可能作用,基于与禾本科盐生植物海滨碱蓬(Aeluropus littoralis (Gouan) Parl)的同源性,通过逆转录PCR和RNA连接酶介导的cDNA末端快速扩增(RLM-RACE)分离出一个液泡Na(+)/H(+)逆向转运蛋白基因(AlNHX)。AlNHX序列包含2706bp,开放阅读框为1623bp,推导的转录本编码540个氨基酸,与高等植物假定的液泡Na(+)/H(+)逆向转运蛋白具有高度同源性。预测AlNHX含有十个假定的疏水区域,这与AtNHX1和OsNHX1不同。DNA凝胶印迹分析表明,海滨碱蓬基因组中有两个或三个AlNHX拷贝。在盐胁迫下,AlNHX在根中的转录水平增加幅度远高于地上部分。此外,AlNHX在烟草中的过表达赋予转基因植物高耐盐性。离子含量分析表明,在高盐胁迫一个月后,与野生型植物相比,转基因植物将更多的Na(+)区室化在根部,并在叶片中保持相对较高的K(+)/Na(+)比率。

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