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在拟南芥体细胞胚胎发生过程中,转录因子转录组的广泛调节。

Extensive modulation of the transcription factor transcriptome during somatic embryogenesis in Arabidopsis thaliana.

机构信息

Department of Genetics, University of Silesia, Katowice, Poland.

出版信息

PLoS One. 2013 Jul 17;8(7):e69261. doi: 10.1371/journal.pone.0069261. Print 2013.

Abstract

Molecular mechanisms controlling plant totipotency are largely unknown and studies on somatic embryogenesis (SE), the process through which already differentiated cells reverse their developmental program and become embryogenic, provide a unique means for deciphering molecular mechanisms controlling developmental plasticity of somatic cells. Among various factors essential for embryogenic transition of somatic cells transcription factors (TFs), crucial regulators of genetic programs, are believed to play a central role. Herein, we used quantitative real-time polymerase chain reaction (qRT-PCR) to identify TF genes affected during SE induced by in vitro culture in Arabidopsis thaliana. Expression profiles of 1,880 TFs were evaluated in the highly embryogenic Col-0 accession and the non-embryogenic tanmei/emb2757 mutant. Our study revealed 729 TFs whose expression changes during the 10-days incubation period of SE; 141 TFs displayed distinct differences in expression patterns in embryogenic versus non-embryogenic cultures. The embryo-induction stage of SE occurring during the first 5 days of culture was associated with a robust and dramatic change of the TF transcriptome characterized by the drastic up-regulation of the expression of a great majority (over 80%) of the TFs active during embryogenic culture. In contrast to SE induction, the advanced stage of embryo formation showed attenuation and stabilization of transcript levels of many TFs. In total, 519 of the SE-modulated TFs were functionally annotated and transcripts related with plant development, phytohormones and stress responses were found to be most abundant. The involvement of selected TFs in SE was verified using T-DNA insertion lines and a significantly reduced embryogenic response was found for the majority of them. This study provides comprehensive data focused on the expression of TF genes during SE and suggests directions for further research on functional genomics of SE.

摘要

植物全能性调控的分子机制在很大程度上尚不清楚,而体细胞胚胎发生(SE)的研究——即已经分化的细胞逆转其发育程序并变得胚胎发生的过程——为解析调控体细胞发育可塑性的分子机制提供了一种独特的手段。在体细胞胚胎发生过程中,转录因子(TFs)是各种必需因素之一,它们是遗传程序的关键调节因子,被认为在其中发挥着核心作用。在此,我们使用定量实时聚合酶链反应(qRT-PCR)鉴定了拟南芥体外培养诱导 SE 过程中受影响的 TF 基因。在高度胚胎发生的 Col-0 品系和非胚胎发生的 tanmei/emb2757 突变体中,评估了 1880 个 TF 的表达谱。在 SE 诱导的 10 天培养期间,我们发现了 729 个 TF 的表达发生变化;在胚胎发生和非胚胎发生的培养物中,有 141 个 TF 的表达模式存在明显差异。SE 的胚胎诱导阶段发生在培养的前 5 天,与 TF 转录组的剧烈变化相关,即在胚胎发生培养中活跃的绝大多数(超过 80%)TF 的表达急剧上调。与 SE 诱导相反,胚胎形成的高级阶段显示出许多 TF 的转录水平减弱和稳定。总的来说,有 519 个 SE 调节的 TF 被功能注释,与植物发育、植物激素和应激反应相关的转录本最为丰富。使用 T-DNA 插入系验证了所选 TF 参与 SE,发现它们中的大多数的胚胎发生反应显著降低。本研究提供了关于 SE 过程中 TF 基因表达的综合数据,并为 SE 的功能基因组学研究提供了方向。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7bc9/3714258/68e22e22ad9e/pone.0069261.g001.jpg

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