研究目的
To identify and characterize a core set of ROS wave-associated transcripts involved in the systemic acquired acclimation response of Arabidopsis to excess light.
研究成果
The study reveals that the priming process of a systemic leaf to become acclimated to a potential stress event involves a rapid systemic transcriptomic response that is extensive and includes an early (2 min) and transient stage of transcripts expression. This early stage of expression is dependent on RBOHD and the function of the ROS/Ca2+ wave that originates in the stressed local leaf. A core set of transcripts is associated with the ROS/Ca2+ wave, suggesting that some of these transcripts could be involved in linking ROS with calcium signaling and initiate or amplify the ROS/Ca2+ wave.
研究不足
The study is limited to Arabidopsis thaliana and may not fully represent responses in other plant species. The rapid transcriptomic responses identified may vary under different environmental conditions or stress combinations.
1：Experimental Design and Method Selection:
Arabidopsis thaliana Col-0 and rbohD knockout plants were subjected to light stress, and local and systemic leaves were sampled at 0, 2, 4, and 8 min post light stress application. RNA-Seq analysis was performed to study transcriptomic responses.
2：Sample Selection and Data Sources:
Local and systemic leaves from 45-50 different plants were pooled for each time point, with the experiment repeated in three different biological replicates.
3：List of Experimental Equipment and Materials:
Gooseneck light source (ACE I; Schott), Trizol (Invitrogen Life Technologies), NucleoSpin RNA Clean-up kit (Macherey-Nagel), Bioanalyzer RNA 6000 Nano Kit (Agilent), Qubit RNA Broad Range Assay Kit (Invitrogen), TruSeq Stranded mRNA HT Library Prep Kit (Illumina), NextSeq High Output 1x75 Reagent Cartridges (Illumina).
4：Experimental Procedures and Operational Workflow:
Local leaves were exposed to light stress, and both local and systemic leaves were sampled at specified times. RNA was isolated, purified, and sequenced. Differential gene expression analysis was conducted using DESeq
5：Data Analysis Methods:
Transcripts expressing differentially in two or more conditions were identified by examining the difference in their abundance under the conditions. Differentially expressed transcripts were defined as those with a fold-change with an adjusted p-value < 0.05.
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