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Transcriptional Responses in Root and Leaf of Prunus persica under Drought Stress Using RNA Sequencing

机译:利用RNA测序技术研究干旱胁迫下桃根和叶片的转录响应。

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摘要

Prunus persica L. Batsch, or peach, is one of the most important crops and it is widely established in irrigated arid and semi-arid regions. However, due to variations in the climate and the increased aridity, drought has become a major constraint, causing crop losses worldwide. The use of drought-tolerant rootstocks in modern fruit production appears to be a useful method of alleviating water deficit problems. However, the transcriptomic variation and the major molecular mechanisms that underlie the adaptation of drought-tolerant rootstocks to water shortage remain unclear. Hence, in this study, high-throughput sequencing (RNA-seq) was performed to assess the transcriptomic changes and the key genes involved in the response to drought in root tissues (GF677 rootstock) and leaf tissues (graft, var. Catherina) subjected to 16 days of drought stress. In total, 12 RNA libraries were constructed and sequenced. This generated a total of 315 M raw reads from both tissues, which allowed the assembly of 22,079 and 17,854 genes associated with the root and leaf tissues, respectively. Subsets of 500 differentially expressed genes (DEGs) in roots and 236 in leaves were identified and functionally annotated with 56 gene ontology (GO) terms and 99 metabolic pathways, which were mostly associated with aminobenzoate degradation and phenylpropanoid biosynthesis. The GO analysis highlighted the biological functions that were exclusive to the root tissue, such as “locomotion,” “hormone metabolic process,” and “detection of stimulus,” indicating the stress-buffering role of the GF677 rootstock. Furthermore, the complex regulatory network involved in the drought response was revealed, involving proteins that are associated with signaling transduction, transcription and hormone regulation, redox homeostasis, and frontline barriers. We identified two poorly characterized genes in P. persica: growth-regulating factor 5 (GRF5), which may be involved in cellular expansion, and AtHB12, which may be involved in root elongation. The reliability of the RNA-seq experiment was validated by analyzing the expression patterns of 34 DEGs potentially involved in drought tolerance using quantitative reverse transcription polymerase chain reaction. The transcriptomic resources generated in this study provide a broad characterization of the acclimation of P. persica to drought, shedding light on the major molecular responses to the most important environmental stressor.
机译:桃(Prunus persica L. Batsch)是最重要的农作物之一,在灌溉的干旱和半干旱地区得到了广泛的应用。但是,由于气候变化和干旱加剧,干旱已成为主要制约因素,在全世界范围内造成作物损失。在现代水果生产中使用耐旱的砧木似乎是缓解缺水问题的有用方法。然而,转录组变异和主要的分子机制是耐旱根系植物适应缺水的基础仍不清楚。因此,在这项研究中,进行了高通量测序(RNA-seq),以评估转录组的变化以及参与根组织(GF677砧木)和叶片组织(嫁​​接,Catherina变种)对干旱响应的关键基因。到16天的干旱压力。总共构建了12个RNA文库并进行了测序。这从两个组织总共产生了315 M的原始读取,这使得分别可以组装与根和叶组织相关的22,079和17,854个基因。鉴定了根中的500个差异表达基因(DEG)和叶中的236个子集,并用56个基因本体论(GO)术语和99个代谢途径在功能上进行了注释,这些主要与氨基苯甲酸酯的降解和苯丙烷的生物合成有关。 GO分析强调了根组织独有的生物学功能,例如“运动”,“激素代谢过程”和“刺激检测”,表明GF677砧木的应力缓冲作用。此外,揭示了参与干旱反应的复杂调控网络,其中涉及与信号转导,转录和激素调控,氧化还原稳态和一线屏障有关的蛋白质。我们在波斯假单胞菌中鉴定了两个特征较差的基因:可能与细胞膨胀有关的生长调节因子5(GRF5)和可能与根伸长有关的AtHB12。 RNA-seq实验的可靠性通过使用定量逆转录聚合酶链反应分析可能涉及干旱耐受性的34个DEG的表达模式进行了验证。在这项研究中产生的转录组资源提供了对P. persica适应干旱的广泛表征,从而阐明了对最重要的环境胁迫因子的主要分子反应。

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