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Analysis of Differential Expression Patterns of mRNA and Protein During Cold-acclimation and De-acclimation in Arabidopsis

机译:拟南芥冷驯化和去驯化过程中mRNA和蛋白质差异表达模式的分析

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摘要

Overwintering plants are capable of exhibiting high levels of cold tolerance, which is acquired through the process of cold acclimation (CA). In contrast to CA, the acquired freezing tolerance is rapidly reduced during cold de-acclimation (DA) and plants resume growth after sensing warm temperatures. In order to better understand plant growth and development, and to aid in the breeding of cold-tolerant plants, it is important to decipher the functional mechanisms of the DA process. In this study, we performed comparative transcriptomic and proteomic analyses during CA and DA. As revealed by shotgun proteomics, we identified 3987 peptides originating from 1569 unique proteins and the corresponding mRNAs were analyzed. Among the 1569 genes, 658 genes were specifically induced at the transcriptional level during the process of cold acclimation. In order to investigate the relationship between mRNA and the corresponding protein expression pattern, a Pearson correlation was analyzed. Interestingly, 199 genes showed a positive correlation of mRNA and protein expression pattern, indicating that both their transcription and translation occurred during CA. However, 226 genes showed a negative correlation of mRNA and protein expression pattern, indicating that their mRNAs were transcribed during CA and were stored for the subsequent DA step. Under this scenario, those proteins were specifically increased during DA without additional transcription of mRNA. In order to confirm the negative correlation of mRNA and protein expression patterns, qRT-PCR and western blot analyses were performed. Mitochondrial malate dehydrogenase 1 (mMDH1) exhibited a negative correlation of mRNA and protein levels, which was characterized by CA-specific mRNA induction and protein accumulation specifically during DA. These data indicate that the expression of specific mRNAs and subsequent accumulation of corresponding proteins are not always in accordance under low temperature stress conditions in plants.
机译:越冬植物能够表现出高水平的耐寒性,这是通过冷驯化(CA)过程获得的。与CA相比,在低温去适应(DA)期间,获得的抗冻性迅速降低,并且植物在感测到温暖的温度后恢复生长。为了更好地了解植物的生长发育并帮助耐寒植物的育种,重要的是破译DA过程的功能机制。在这项研究中,我们在CA和DA期间进行了比较转录组学和蛋白质组学分析。如shot弹枪蛋白质组学所揭示,我们鉴定了源自1569种独特蛋白质的3987个肽段,并分析了相应的mRNA。在1569个基因中,在冷驯化过程中在转录水平上特异性诱导了658个基因。为了研究mRNA与相应蛋白质表达模式之间的关系,分析了皮尔森相关性。有趣的是,199个基因显示出mRNA和蛋白质表达模式的正相关,表明它们的转录和翻译均在CA期间发生。然而,226个基因显示出mRNA和蛋白质表达模式的负相关,表明它们的mRNA在CA期间被转录并被存储用于随后的DA步骤。在这种情况下,那些蛋白质在DA期间特异性增加,而无需额外的mRNA转录。为了证实mRNA和蛋白质表达模式的负相关,进行了qRT-PCR和蛋白质印迹分析。线粒体苹果酸脱氢酶1(mMDH1)的mRNA和蛋白质水平呈负相关,其特征是CA特异的mRNA诱导和蛋白质在DA期间的蓄积。这些数据表明,在植物的低温胁迫条件下,特定mRNA的表达和随后相应蛋白质的积累并不总是一致的。

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