Nakaminami Kentaro, Matsui Akihiro, Nakagami Hirofumi, Minami Anzu, Nomura Yuko, Tanaka Maho, Morosawa Taeko, Ishida Junko, Takahashi Satoshi, Uemura Matsuo, Shirasu Ken, Seki Motoaki
From the ‡Plant Genomic Network Research Team, RIKEN Center for Sustainable Resource Science (CSRS), Yokohama, Kanagawa, 230-0045, Japan;
§Plant Proteomics Research Unit, RIKEN CSRS, Yokohama, Kanagawa, 230-0045, Japan;
Mol Cell Proteomics. 2014 Dec;13(12):3602-11. doi: 10.1074/mcp.M114.039081. Epub 2014 Oct 2.
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.
越冬植物能够表现出高水平的耐寒性,这种耐寒性是通过冷驯化(CA)过程获得的。与冷驯化相反,在冷脱驯化(DA)过程中,获得的冻融耐受性会迅速降低,植物在感知温暖温度后恢复生长。为了更好地理解植物的生长发育,并有助于耐寒植物的育种,解读冷脱驯化过程的功能机制非常重要。在本研究中,我们在冷驯化和冷脱驯化过程中进行了比较转录组学和蛋白质组学分析。通过鸟枪法蛋白质组学揭示,我们鉴定出了源自1569种独特蛋白质的3987种肽段,并对相应的mRNA进行了分析。在这1569个基因中,有658个基因在冷驯化过程中在转录水平上被特异性诱导。为了研究mRNA与相应蛋白质表达模式之间的关系,我们分析了皮尔逊相关性。有趣的是,199个基因的mRNA和蛋白质表达模式呈正相关,表明它们的转录和翻译都发生在冷驯化过程中。然而,226个基因的mRNA和蛋白质表达模式呈负相关,表明它们的mRNA在冷驯化过程中被转录,并被储存用于随后的冷脱驯化步骤。在这种情况下,这些蛋白质在冷脱驯化过程中特异性增加,而无需mRNA的额外转录。为了证实mRNA和蛋白质表达模式的负相关性,我们进行了qRT-PCR和蛋白质免疫印迹分析。线粒体苹果酸脱氢酶1(mMDH1)的mRNA和蛋白质水平呈负相关,其特征是冷驯化特异性mRNA诱导和蛋白质在冷脱驯化过程中特异性积累。这些数据表明,在植物低温胁迫条件下,特定mRNA的表达和相应蛋白质的后续积累并不总是一致的。