Australian Research Council Centre of Excellence in Plant Energy Biology, Research School of Biology, Australian National University Canberra, Acton ACT 0200, Australia.
Department of Plant and Microbial Biology, University of Minnesota, Saint Paul, Minnesota 55108.
Plant Cell. 2017 Aug;29(8):1836-1863. doi: 10.1105/tpc.16.00828. Epub 2017 Jul 13.
Stress recovery may prove to be a promising approach to increase plant performance and, theoretically, mRNA instability may facilitate faster recovery. Transcriptome (RNA-seq, qPCR, sRNA-seq, and PARE) and methylome profiling during repeated excess-light stress and recovery was performed at intervals as short as 3 min. We demonstrate that 87% of the stress-upregulated mRNAs analyzed exhibit very rapid recovery. For instance, abundance declined 2-fold every 5.1 min. We term this phenomenon rapid recovery gene downregulation (RRGD), whereby mRNA abundance rapidly decreases promoting transcriptome resetting. Decay constants () were modeled using two strategies, linear and nonlinear least squares regressions, with the latter accounting for both transcription and degradation. This revealed extremely short half-lives ranging from 2.7 to 60.0 min for 222 genes. Ribosome footprinting using degradome data demonstrated RRGD loci undergo cotranslational decay and identified changes in the ribosome stalling index during stress and recovery. However, small RNAs and 5'-3' RNA decay were not essential for recovery of the transcripts examined, nor were any of the six excess light-associated methylome changes. We observed recovery-specific gene expression networks upon return to favorable conditions and six transcriptional memory types. In summary, rapid transcriptome resetting is reported in the context of active recovery and cellular memory.
应激恢复可能被证明是一种提高植物性能的有前途的方法,从理论上讲,mRNA 的不稳定性可能有助于更快地恢复。在重复的过量光照应激和恢复过程中,每隔很短的时间(短至 3 分钟)进行转录组(RNA-seq、qPCR、sRNA-seq 和 PARE)和甲基组谱分析。我们证明,分析的 87%应激上调的 mRNAs 表现出非常快速的恢复。例如,丰度每 5.1 分钟下降 2 倍。我们将这种现象称为快速恢复基因下调(RRGD),其中 mRNA 丰度迅速下降,促进转录组重置。使用两种策略(线性和非线性最小二乘回归)对衰减常数 () 进行建模,后者同时考虑转录和降解。这揭示了 222 个基因的半衰期极短,范围从 2.7 到 60.0 分钟。使用降解组数据进行核糖体足迹分析表明,RRGD 基因座经历共翻译降解,并在应激和恢复过程中鉴定了核糖体停滞指数的变化。然而,小 RNA 和 5'-3'RNA 降解对于所检查的转录物的恢复不是必需的,六个过量光照相关的甲基组变化也不是必需的。我们在返回有利条件时观察到恢复特异性基因表达网络和六种转录记忆类型。总之,在积极恢复和细胞记忆的背景下,快速转录组重置得到了报道。