Transcriptome analysis of Arabidopsis thaliana roots, H2O2 and Rose Bengal [20180920_172844_1O_H2O2]

In order to better understand the context of root water stress and its relationship to other stresses, I undertook an evaluation of osmotic water stress by transcriptome analyses. Whole transcriptome MARSeq (Diego Adhemar Jaitin, 2017) analysis can enable us to study the changes in gene expression (changes in transcript prevalence) in depth under different treatments. It is a cost effective method to develop transcriptional activity. It uses relatively small amounts of RNA and reads sequences from the 3'-polyA containing end. This method can reveal underlying trends that can help us understand the less obvious processes happening in our system. To carry this out I processed whole transcriptome data separately from WT seedling roots exposed to 5 mM H2O2 or 10 µM RB for 0, 1, 2, 3 h or 5 mM H2O2 or 10 µM for 2h . Overall design: WT Arabidopsis thaliana seedling roots were exposed to 5 mM H2O2 or 10 µM RB for 0, 1, 2, 3 h or 5 mM H2O2 and 10 µM for 2h ., and root RNA was extracted sequenced using the MARSeq protocol.

为深入解析根系水分胁迫的研究背景及其与其他胁迫的关联，本研究开展了基于转录组分析的渗透水分胁迫评估。全转录组MARSeq（Diego Adhemar Jaitin等，2017）分析可帮助我们深度探究不同处理条件下基因表达（转录本丰度变化）的动态变化。该方法具备出色的成本效益，仅需相对少量的RNA即可高效解析转录活性，且测序读段取自带有3'-polyA的末端区域。此方法能够揭示潜在的表达趋势，助力我们理解实验体系中较为隐蔽的生物学过程。为此，本研究针对野生型（WT）幼苗根系的全转录组数据进行了单独处理：将幼苗根系分别置于5 mM H₂O₂或10 μM RB中处理0、1、2、3小时，或采用5 mM H₂O₂与10 μM RB共同处理2小时。整体实验设计如下：将野生型拟南芥（Arabidopsis thaliana）幼苗根系分别置于5 mM H₂O₂、10 μM RB中处理0、1、2、3小时，或同时用5 mM H₂O₂与10 μM RB处理2小时，随后提取根系RNA并按照MARSeq实验流程完成测序。