PT  - JOURNAL ARTICLE
AU  - Toda, Erika
AU  - Kiba, Takatoshi
AU  - Kato, Norio
AU  - Okamoto, Takashi
TI  - Isolation of gametes and zygotes from <em>Setaria viridis</em>
AID  - 10.1101/2021.11.01.466850
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.11.01.466850
4099  - http://biorxiv.org/content/early/2021/11/04/2021.11.01.466850.short
4100  - http://biorxiv.org/content/early/2021/11/04/2021.11.01.466850.full
AB  - Setaria viridis, the wild ancestor of foxtail millet (Setaria italica), is an effective model plant for larger C4 crops because it 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 C4 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.Competing Interest StatementThe authors have declared no competing interest.