Rai Archana N, Tamirisa Srinath, Rao K V, Kumar Vinay, Suprasanna P
Plant Stress Physiology and Biotechnology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai, 400085, India.
Centre for Plant Molecular Biology, Osmania University, Hyderabad, 500007, India.
Plant Mol Biol. 2016 Mar;90(4-5):375-87. doi: 10.1007/s11103-015-0423-x. Epub 2015 Dec 28.
'Early responsive to dehydration' (ERD) genes are a group of plant genes having functional roles in plant stress tolerance and development. In this study, we have isolated and characterized a Brassica juncea 'ERD' gene (BjERD4) which encodes a novel RNA binding protein. The expression pattern of ERD4 analyzed under different stress conditions showed that transcript levels were increased with dehydration, sodium chloride, low temperature, heat, abscisic acid and salicylic acid treatments. The BjERD4 was found to be localized in the chloroplasts as revealed by Confocal microscopy studies. To study the function, transgenic Arabidopsis plants were generated and analyzed for various morphological and physiological parameters. The overexpressing transgenic lines showed significant increase in number of leaves with more leaf area and larger siliques as compared to wild type plants, whereas RNAi:ERD4 transgenic lines showed reduced leaf number, leaf area, dwarf phenotype and delayed seed germination. Transgenic Arabidopsis plants overexpressing BjERD4 gene also exhibited enhanced tolerance to dehydration and salt stresses, while the knockdown lines were susceptible as compared to wild type plants under similar stress conditions. It was observed that BjERD4 protein could bind RNA as evidenced by the gel-shift assay. The overall results of transcript analysis, RNA gel-shift assay, and transgenic expression, for the first time, show that the BjERD4 is involved in abiotic stress tolerance besides offering new clues about the possible roles of BjERD4 in plant growth and development.
“早期脱水响应”(ERD)基因是一组在植物胁迫耐受性和发育中具有功能作用的植物基因。在本研究中,我们分离并鉴定了一个芥菜型油菜的“ERD”基因(BjERD4),它编码一种新型RNA结合蛋白。在不同胁迫条件下分析ERD4的表达模式表明,脱水、氯化钠、低温、高温、脱落酸和水杨酸处理后转录水平升高。共聚焦显微镜研究显示,BjERD4定位于叶绿体。为了研究其功能,构建了转基因拟南芥植株并分析了各种形态和生理参数。与野生型植株相比,过表达转基因株系的叶片数量显著增加,叶面积更大,角果更大,而RNA干扰(RNAi):ERD4转基因株系的叶片数量减少、叶面积减小、呈现矮化表型且种子萌发延迟。过表达BjERD4基因的转基因拟南芥植株对脱水和盐胁迫的耐受性也增强,而在相似胁迫条件下,与野生型植株相比,基因敲除株系更敏感。凝胶迁移实验证明BjERD4蛋白能够结合RNA。转录分析、RNA凝胶迁移实验和转基因表达的总体结果首次表明,BjERD4除了为其在植物生长和发育中的可能作用提供新线索外,还参与非生物胁迫耐受性。