Transcriptome analyses of Pst DC3000-inoculated Arabidopsis thaliana Col-0 and 35S::CBP60g plants at normal and elevated temperature. Transcriptome analyses of Pst DC3000-inoculated Arabidopsis thaliana Col-0 and 35S::CBP60g plants at normal and elevated temperature

We performed RNA sequencing of Pseudomonas syringae pv. tomato (Pst) DC3000-infected A. thaliana Col-0 and 35S::CBP60g plants at normal (23C) and elevated (28C) temperatures. 4-week-old plants were pre-incubated at 23C and 28C for 48h and then leaves were syringe-infiltrated with DC3000 bacterial inoculum. Plants were incubated at their respective temperatures (23C or 28C) for another 24h post-inoculation before tissue collection for RNA extraction. RNA samples for submitted for RNA sequencing and we found different clusters of DC3000-regulated genes that were similarly or differentially regulated between Col-0 and 35S::CBP60g at elevated temperature. Temperature-downregulated DC3000-induced genes in Col-0 plants that were restored in 35S::CBP60g plants were enriched for immunity/defense-related genes, including those essential for host salicylic acid (defense hormone) biosynthesis and accumulation. Overall design: 4 treatments were used: (1) 23C DC3000 Col-0; (2) 23C DC3000 35S::CBP60g; (3) 28C DC3000 Col-0; and (4) 28C DC3000 35S::CBP60g. For each treatment, three individual plants (independent biological replicates) were sampled.

我们对被丁香假单胞菌番茄致病变种（Pseudomonas syringae pv. tomato, Pst）DC3000侵染的拟南芥（Arabidopsis thaliana）Col-0与35S::CBP60g植株，在正常（23℃）与高温（28℃）条件下开展了RNA测序。选取4周龄的植株，分别在23℃与28℃下预培养48小时，随后采用注射器渗透法将DC3000细菌接种液注入叶片。接种完成后，将植株继续置于对应温度（23℃或28℃）下培养24小时，再采集组织用于RNA提取。将提取的RNA样本提交进行RNA测序分析，结果显示存在多组受DC3000调控的基因，它们在高温条件下于Col-0与35S::CBP60g植株中呈现相似或差异的表达调控模式。进一步富集分析发现，Col-0植株中受高温下调的DC3000诱导基因，在35S::CBP60g植株中表达得以恢复，这类基因显著富集于免疫与防御相关通路，包括宿主水杨酸（防御激素）生物合成与积累所必需的基因。整体实验设计如下：共设置4组处理：(1) 23℃ DC3000 Col-0；(2) 23℃ DC3000 35S::CBP60g；(3) 28℃ DC3000 Col-0；(4) 28℃ DC3000 35S::CBP60g。每组处理均采集3株独立植株作为生物学重复。