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拟南芥早期胚胎中细胞类型特异性转录组分析

Cell type-specific transcriptome analysis in the early Arabidopsis thaliana embryo.

作者信息

Slane Daniel, Kong Jixiang, Berendzen Kenneth W, Kilian Joachim, Henschen Agnes, Kolb Martina, Schmid Markus, Harter Klaus, Mayer Ulrike, De Smet Ive, Bayer Martin, Jürgens Gerd

机构信息

Department of Cell Biology, Max Planck Institute for Developmental Biology, Tübingen 72076, Germany.

Department of Cell Biology, Max Planck Institute for Developmental Biology, Tübingen 72076, Germany Department of Developmental Genetics, Center for Plant Molecular Biology, University of Tübingen, Tübingen 72076, Germany.

出版信息

Development. 2014 Dec;141(24):4831-40. doi: 10.1242/dev.116459. Epub 2014 Nov 19.

Abstract

In multicellular organisms, cellular differences in gene activity are a prerequisite for differentiation and establishment of cell types. In order to study transcriptome profiles, specific cell types have to be isolated from a given tissue or even the whole organism. However, whole-transcriptome analysis of early embryos in flowering plants has been hampered by their size and inaccessibility. Here, we describe the purification of nuclear RNA from early stage Arabidopsis thaliana embryos using fluorescence-activated nuclear sorting (FANS) to generate expression profiles of early stages of the whole embryo, the proembryo and the suspensor. We validated our datasets of differentially expressed candidate genes by promoter-reporter gene fusions and in situ hybridization. Our study revealed that different classes of genes with respect to biological processes and molecular functions are preferentially expressed either in the proembryo or in the suspensor. This method can be used especially for tissues with a limited cell population and inaccessible tissue types. Furthermore, we provide a valuable resource for research on Arabidopsis early embryogenesis.

摘要

在多细胞生物中,基因活性的细胞差异是细胞分化和细胞类型确立的前提条件。为了研究转录组图谱,必须从给定组织甚至整个生物体中分离出特定的细胞类型。然而,开花植物早期胚胎的全转录组分析因其体积小和难以获取而受到阻碍。在此,我们描述了使用荧光激活核分选(FANS)从拟南芥早期胚胎中纯化核RNA,以生成整个胚胎、原胚和胚柄早期阶段的表达谱。我们通过启动子-报告基因融合和原位杂交验证了差异表达候选基因的数据集。我们的研究表明,就生物学过程和分子功能而言,不同类别的基因在原胚或胚柄中优先表达。该方法尤其可用于细胞群体有限和难以获取的组织类型。此外,我们为拟南芥早期胚胎发生的研究提供了宝贵的资源。

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