Okamoto Takashi
Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa 1-1, Hachioji, Tokyo, 192-0397, Japan.
Methods Mol Biol. 2017;1669:251-263. doi: 10.1007/978-1-4939-7286-9_20.
In angiosperms, female gamete differentiation, fertilization, and subsequent zygotic development occur in embryo sacs deeply embedded in the ovaries. Despite their importance in plant reproduction and development, how the egg cell is specialized, fuses with the sperm cell, and converts into an active zygote for early embryogenesis remains unclear. Proteins enriched in each cell type are thought to reflect the biological function of the cells, since the composition of cellular proteins generally differs depending on cell type. Therefore, proteins enriched in gametes, egg and sperm cells, should have a role in reproductive and/or developmental processes, such as gamete differentiation, gamete fusion, egg activation, and early zygotic development. In this chapter, a method for identification of proteins enriched in rice gametes is first explained, and then functional analyses for these proteins using rice and Arabidopsis lines with mutations in genes encoding the putative gamete-enriched proteins are also described.
在被子植物中，雌配子分化、受精以及随后的合子发育发生在深埋于子房中的胚囊中。尽管它们在植物繁殖和发育中具有重要意义，但卵细胞如何特化、与精子细胞融合并转化为活跃的合子以进行早期胚胎发生仍不清楚。由于细胞蛋白质的组成通常因细胞类型而异，因此每种细胞类型中富集的蛋白质被认为反映了细胞的生物学功能。因此，在配子（卵细胞和精子细胞）中富集的蛋白质应该在生殖和/或发育过程中发挥作用，例如配子分化、配子融合、卵细胞激活和早期合子发育。在本章中，首先解释了一种鉴定水稻配子中富集蛋白质的方法，然后还描述了使用水稻和拟南芥品系对这些蛋白质进行功能分析的过程，这些品系在编码假定的配子富集蛋白的基因中存在突变。
