油菜裂外壁小孢子胚在分子水平上具有胚体/胚柄分化

早期合子胚取材困难, 难以开展相关研究。前人的工作表明, 油菜(Brassica napus)裂外壁小孢子胚胎发生系统能够较好地模拟合子胚的分化模式, 因而可替代早期合子胚胎作为研究材料。但目前尚缺乏该胚胎发生系统中胚胎具有胚体/胚柄分化的分子水平的证据。该文首次证明了油菜WOX家族基因能够用于标记胚体/胚柄的分化过程, 利用胚柄标记基因BnWOX8的表达模式, 从分子水平上证明了带胚柄的裂外壁小孢子胚的确存在胚体/胚柄的分化。研究结果为充分利用油菜裂外壁小孢子胚胎发生系统, 解决早期胚胎取材困难的问题奠定了坚实的基础。同时, 建立了活体激光切割分离特定细胞的技术, 结合用于少量细胞RNA提取的活体特异细胞RNA提取技术, 为鉴定少量特异分化细胞的基因表达模式提供了一个可行且明确的解决方案。

The Exine-dehisced Microspore Embryos Have the Proper- suspensor Differentiation in Molecular Level

Because of sampling difficulty, research of the early stages of zygotic embryos is difficult. Previous studies showed that the exine-dehisced microspore embryogenesis system can stimulate the differentiation pattern of the zygotic embryo; an alternative approach is to use exine-dehisced microspore embryos as a substitute for zygotic embryos for relevant research work. However, we lack adequate evidence for this approach at the molecular level. In this study, we verified that the WOX family of genes could be markers of the differentiation of the embryo proper or suspensor in Brassica napus and showed that the embryo proper/suspensor in the exine-dehisced microspore embryo with suspensor could be differentiated at the molecular level by the expression pattern of the suspensor marker gene BnWOX8. These results provide solid evidence to use the exine-dehisced microspore embryogenesis system to study the early stages of the zygotic embryo. Meanwhile, we established the efficiency of the living laser microdissection technique, combined with RNA extraction from a few cells. We provide a reliable and applicable solution for detecting gene expression patterns in specific cells.