Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna Biocenter (VBC), Dr. Bohr-Gasse 3, 1030, Vienna, Austria.
Plant Reprod. 2019 Mar;32(1):77-91. doi: 10.1007/s00497-018-00357-2. Epub 2019 Jan 4.
Arabidopsis embryos possess unique transcriptomes relative to other plant tissues including somatic embryos, and can be partitioned into four transcriptional phases with characteristic biological processes. Cellular differentiation is associated with changes in transcript populations. Accurate quantification of transcriptomes during development can thus provide global insights into differentiation processes including the fundamental specification and differentiation events operating during plant embryogenesis. However, multiple technical challenges have limited the ability to obtain high-quality early embryonic transcriptomes, namely the low amount of RNA obtainable and contamination from surrounding endosperm and seed-coat tissues. We compared the performance of three low-input mRNA sequencing (mRNA-seq) library preparation kits on 0.1 to 5 nanograms (ng) of total RNA isolated from Arabidopsis thaliana (Arabidopsis) embryos and identified a low-cost method with superior performance. This mRNA-seq method was then used to profile the transcriptomes of Arabidopsis embryos across eight developmental stages. By comprehensively comparing embryonic and post-embryonic transcriptomes, we found that embryonic transcriptomes do not resemble any other plant tissue we analyzed. Moreover, transcriptome clustering analyses revealed the presence of four distinct phases of embryogenesis which are enriched in specific biological processes. We also compared zygotic embryo transcriptomes with publicly available somatic embryo transcriptomes. Strikingly, we found little resemblance between zygotic embryos and somatic embryos derived from late-staged zygotic embryos suggesting that somatic and zygotic embryo transcriptomes are distinct from each other. In addition to the biological insights gained from our systematic characterization of the Arabidopsis embryonic transcriptome, we provide a data-rich resource for the community to explore.
拟南芥胚胎相对于其他植物组织(包括体细胞胚胎)具有独特的转录组,并且可以分为四个具有特征性生物过程的转录相。细胞分化与转录群体的变化有关。因此,在发育过程中准确地定量转录组可以提供对分化过程的全面了解,包括在植物胚胎发生过程中起作用的基本特化和分化事件。然而,多种技术挑战限制了获得高质量早期胚胎转录组的能力,即获得的 RNA 量低,以及来自周围胚乳和种皮组织的污染。我们比较了三种低输入 mRNA 测序(mRNA-seq)文库制备试剂盒在 0.1 到 5 纳克(ng)总 RNA 分离自拟南芥(拟南芥)胚胎的性能,并确定了一种具有优异性能的低成本方法。然后,我们使用该 mRNA-seq 方法来描绘拟南芥胚胎在八个发育阶段的转录组。通过全面比较胚胎和胚胎后转录组,我们发现胚胎转录组与我们分析的任何其他植物组织都不相似。此外,转录组聚类分析揭示了存在四个独特的胚胎发生阶段,这些阶段富含特定的生物过程。我们还将合子胚胎转录组与公开的体细胞胚胎转录组进行了比较。引人注目的是,我们发现合子胚胎和体细胞胚胎之间几乎没有相似之处,这表明体细胞和合子胚胎转录组彼此不同。除了从我们对拟南芥胚胎转录组的系统描述中获得的生物学见解外,我们还提供了一个富含数据的资源供社区探索。