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Ectopic Expression of an Atypical Hydrophobic Group 5 LEA Protein from Wild Peanut Arachis diogoi Confers Abiotic Stress Tolerance in Tobacco

机译:花生中的非典型疏水性第5组LEA蛋白的异位表达赋予烟草非生物胁迫耐受性

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摘要

Late embryogenesis abundant (LEA) proteins are a group of hydrophilic proteins, which accumulate in plants under varied stress conditions like drought, salinity, extreme temperatures and oxidative stress suggesting their role in the protection of plants against these stresses. A transcript derived fragment (TDF) corresponding to LEA gene, which got differentially expressed in wild peanut, Arachis diogoi against the late leaf spot pathogen, Phaeoisariopsis personata was used in this study. We have cloned its full length cDNA by RACE-PCR, which was designated as AdLEA. AdLEA belongs to the atypical Group 5C of LEA protein family as confirmed by sequence analysis. Group 5C LEA protein subfamily contains Pfam LEA_2 domain and is highly hydrophobic. In native conditions, expression of AdLEA was upregulated considerably upon hormonal and abiotic stress treatments emphasizing its role in abiotic stress tolerance. Subcellular localization studies showed that AdLEA protein is distributed in both nucleus and cytosol. Ectopic expression of AdLEA in tobacco resulted in enhanced tolerance of plants to dehydration, salinity and oxidative stress with the transgenic plants showing higher chlorophyll content and reduced lipid peroxidation as compared to wild type plants. Overexpressed AdLEA tobacco plants maintained better photosynthetic efficiency under drought conditions as demonstrated by chlorophyll fluorescence measurements. These plants showed enhanced transcript accumulation of some stress-responsive genes. Our study also elucidates that ROS levels were significantly reduced in leaves and stomatal guard cells of transgenic plants upon stress treatments. These results suggest that AdLEA confers multiple stress tolerance to plants, which make it a potential gene for genetic modification in plants.
机译:晚期胚发生富集(LEA)蛋白是一组亲水蛋白,它们在各种胁迫条件下(例如干旱,盐度,极端温度和氧化胁迫)在植物中积累,表明它们在保护植物免受这些胁迫的作用。本研究使用了与LEA基因相对应的转录本衍生片段(TDF),该片段在野生花生Arachis diogoi中针对晚期叶斑病原体Phaeoisariopsis personata进行了差异表达。我们已通过RACE-PCR克隆了其全长cDNA,命名为AdLEA。经序列分析证实,AdLEA属于LEA蛋白家族的非典型5C组。 5C组LEA蛋白亚家族包含Pfam LEA_2结构域,并且高度疏水。在天然条件下,荷尔蒙和非生物胁迫治疗后AdLEA的表达明显上调,强调了其在非生物胁迫耐受性中的作用。亚细胞定位研究表明,AdLEA蛋白既分布在细胞核中,又分布在细胞质中。与野生型植物相比,AdLEA在烟草中的异位表达使植物对脱水,盐度和氧化胁迫的耐受性增强,转基因植物的叶绿素含量更高,脂质过氧化作用降低。叶绿素荧光测量表明,过表达的AdLEA烟草植物在干旱条件下保持了更好的光合作用效率。这些植物显示出一些胁迫响应基因的转录物积累增强。我们的研究还阐明,经过胁迫处理后,转基因植物的叶片和气孔保卫细胞中的ROS水平显着降低。这些结果表明AdLEA赋予植物多种胁迫耐受性,这使其成为植物遗传修饰的潜在基因。

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