In vitro ovule cultivation for live-cell imaging of zygote polarization and embryo patterning in Arabidopsis thaliana

Daisuke Kurihara, Yusuke Kimata, Tetsuya Higashiyama, Minako Ueda

研究成果: Article査読

7 被引用数 (Scopus)

抄録

In most flowering plants, the zygote and embryo are hidden deep in the mother tissue, and thus it has long been a mystery of how they develop dynamically; for example, how the zygote polarizes to establish the body axis and how the embryo specifies various cell fates during organ formation. This manuscript describes an in vitro ovule culture method to perform live-cell imaging of developing zygotes and embryos of Arabidopsis thaliana. The optimized cultivation medium allows zygotes or early embryos to grow into fertile plants. By combining it with a poly(dimethylsiloxane) (PDMS) micropillar array device, the ovule is held in the liquid medium in the same position. This fixation is crucial to observe the same ovule under a microscope for several days from the zygotic division to the late embryo stage. The resulting live-cell imaging can be used to monitor the real-time dynamics of zygote polarization, such as nuclear migration and cytoskeleton rearrangement, and also the cell division timing and cell fate specification during embryo patterning. Furthermore, this ovule cultivation system can be combined with inhibitor treatments to analyze the effects of various factors on embryo development, and with optical manipulations such as laser disruption to examine the role of cell-cell communication.

本文言語English
論文番号e55975
ジャーナルJournal of Visualized Experiments
2017
127
DOI
出版ステータスPublished - 2017 9 11
外部発表はい

ASJC Scopus subject areas

  • 神経科学(全般)
  • 化学工学(全般)
  • 生化学、遺伝学、分子生物学(全般)
  • 免疫学および微生物学(全般)

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