Manual Isolation of Living Early Embryos from Tobacco Seeds.
Manual Isolation of Living Early Embryos from Tobacco Seeds.
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DOI：
10.1007/978-1-0716-0342-0_8
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发表时间：
2020-01-01
期刊：
影响因子：
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通讯作者：
Sun, Meng-Xiang
中科院分区：
文献类型：
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作者：
Zhao, Peng;Zhou, Xuemei;Sun, Meng-Xiang
合子胚胎发生是可育种子发育的关键过程之一，因此在植物发育生物学领域受到了广泛的关注。然而，这一过程深深地嵌入母体组织中。早期胚胎的难以获取极大地阻碍了早期胚胎发生的研究，特别是限制了直接观察和精确的组学研究。为了研究现代技术调控胚胎发育的分子机制，有必要发展一种可靠的方法来分离不同阶段的活胚胎。为此，植物科学家们在过去的几十年里一直试图开发不同的方法来分离玉米、小麦、水稻和烟草等不同植物的合子和早期胚胎。烟草长期以来一直被认为是研究胚胎发生的理想模式真双子叶植物，其细胞分裂模式可追溯且可预测，从第一次合子分裂到成熟胚形成。在这里，我们提供了一个详细的协议，从受精卵到子叶胚分离活胚。分离的活合子和早期胚胎可用于几个重要的研究，如细胞类型特异性转录组构建和清晰的GFP观察。
Zygotic embryogenesis is one of key processes for fertile seed development and therefore has gained great attention for decades in the field of plant developmental biology. However, this process is deeply embedded in the maternal tissues. The inaccessibility of tiny early embryos has greatly hindered the study of early embryogenesis, especially limits direct observation and accurate omics investigations. In order to investigate the molecular mechanism regulating embryo development with modern technologies, it is necessary to develop a reliable method to isolate living embryos at different stages. For this purpose, plant scientists have been trying to develop different methods for isolating zygotes and early embryos in different plants such as maize, wheat, rice, and tobacco during past decades. Nicotiana tabacum has long been considered as an ideal model eudicot for the study of embryogenesis, which displays a traceable and predictable cell division pattern, spanning from the first zygotic division to the mature embryo formation. Here, we provide a detailed protocol for isolating living embryos from zygote to cotyledon embryo. Isolated living zygotes and early embryos could be used for several important studies such as celltype-specific transcriptome construction and clear GFP observation.