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Profiling the Abiotic Stress Responsive microRNA Landscape of Arabidopsis thaliana

机译:对拟南芥的非生物胁迫响应性microRNA景观进行分析

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摘要

It is well established among interdisciplinary researchers that there is an urgent need to address the negative impacts that accompany climate change. One such negative impact is the increased prevalence of unfavorable environmental conditions that significantly contribute to reduced agricultural yield. Plant microRNAs (miRNAs) are key gene expression regulators that control development, defense against invading pathogens and adaptation to abiotic stress. Arabidopsis thaliana (Arabidopsis) can be readily molecularly manipulated, therefore offering an excellent experimental system to alter the profile of abiotic stress responsive miRNA/target gene expression modules to determine whether such modification enables Arabidopsis to express an altered abiotic stress response phenotype. Towards this goal, high throughput sequencing was used to profile the miRNA landscape of Arabidopsis whole seedlings exposed to heat, drought and salt stress, and identified 121, 123 and 118 miRNAs with a greater than 2-fold altered abundance, respectively. Quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) was next employed to experimentally validate miRNA abundance fold changes, and to document reciprocal expression trends for the target genes of miRNAs determined abiotic stress responsive. RT-qPCR also demonstrated that each miRNA/target gene expression module determined to be abiotic stress responsive in Arabidopsis whole seedlings was reflective of altered miRNA/target gene abundance in Arabidopsis root and shoot tissues post salt stress exposure. Taken together, the data presented here offers an excellent starting platform to identify the miRNA/target gene expression modules for future molecular manipulation to generate plant lines that display an altered response phenotype to abiotic stress.
机译:跨学科研究人员已经确定,迫切需要解决气候变化带来的负面影响。这种负面影响之一是不利环境条件的普遍存在,这极大地降低了农业产量。植物microRNA(miRNA)是控制发育,抵抗入侵病原体和适应非生物胁迫的关键基因表达调节剂。拟南芥(Arabidopsis)可以很容易地进行分子操作,因此提供了一个出色的实验系统,可以改变非生物胁迫响应性miRNA /靶基因表达模块的概况,从而确定这种修饰是否使拟南芥能够表达改变的非生物胁迫响应表型。为了实现这一目标,高通量测序用于分析暴露于高温,干旱和盐胁迫下的拟南芥整个幼苗的miRNA概况,并鉴定出121、123和118个miRNA的丰度变化分别大于2倍。接下来采用定量逆转录酶聚合酶链反应(RT-qPCR)来实验验证miRNA丰度变化,并记录确定为非生物胁迫响应的miRNA靶基因的相互表达趋势。 RT-qPCR还证明了在拟南芥全苗中确定为非生物胁迫响应的每个miRNA /靶基因表达模块均反映了盐胁迫暴露后拟南芥根和芽组织中miRNA /靶基因丰度的变化。综上所述,此处提供的数据为鉴定miRNA /靶标基因表达模块提供了一个极好的起始平台,可用于未来的分子操作,以产生对非生物胁迫表现出变化的反应表型的植物系。

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