Plant Breeding Innovation Laboratory, RIKEN Cluster for Science, Technology and Innovation Hub, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan.
Department of Biological Sciences, Tokyo Metropolitan University, 1-1 Minami-osawa, Hachioji, Tokyo, 192-0392, Japan.
J Plant Res. 2022 Jul;135(4):627-633. doi: 10.1007/s10265-022-01393-w. Epub 2022 May 9.
Setaria viridis, the wild ancestor of foxtail millet (Setaria italica), is an effective model plant for larger C crops because S. viridis has several desirable traits, such as short generation time, prolific seed production and a small genome size. These advantages are well suited for investigating molecular mechanisms in angiosperms, especially C crop species. Here, we report a procedure for isolating gametes and zygotes from S. viridis flowers. To isolate egg cells, ovaries were harvested from unpollinated mature flowers and cut transversely, which allowed direct access to the embryo sac. Thereafter, an egg cell was released from the cut end of the basal portion of the dissected ovary. To isolate sperm cells, pollen grains released from anthers were immersed in a mannitol solution, resulting in pollen-grain bursting, which released sperm cells. Additionally, S. viridis zygotes were successfully isolated from freshly pollinated flowers. Isolated zygotes cultured in a liquid medium developed into globular-like embryos and cell masses. Thus, isolated S. viridis gametes, zygotes and embryos are attainable for detailed observations and investigations of fertilization and developmental events in angiosperms.
狗尾草（Setaria italica）的野生祖先绿色狗尾草（Setaria viridis）是较大 C 作物的有效模式植物，因为绿色狗尾草具有几个理想的特征，例如短世代时间、多产的种子生产和小的基因组大小。这些优势非常适合研究被子植物中的分子机制，尤其是 C 作物物种。在这里，我们报告了一种从绿色狗尾草花朵中分离配子和受精卵的程序。为了分离卵细胞，从未授粉的成熟花朵中收获卵巢并横向切割，这允许直接进入胚囊。此后，从解剖卵巢基部的切割端释放一个卵细胞。为了分离精子细胞，将从花药中释放的花粉粒浸入甘露醇溶液中，导致花粉粒爆裂，从而释放出精子细胞。此外，还成功地从新鲜授粉的花朵中分离出绿色狗尾草受精卵。在液体培养基中培养的分离受精卵发育成球状胚胎和细胞团。因此，可获得分离的绿色狗尾草配子、受精卵和胚胎，可用于详细观察和研究被子植物中的受精和发育事件。
