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

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article numbere55975
JournalJournal of Visualized Experiments
Volume2017
Issue number127
DOIs
Publication statusPublished - 2017 Sep 11
Externally publishedYes

Keywords

  • Arabidopsis thaliana
  • Developmental biology
  • Embryo
  • Issue 127
  • Live-cell imaging
  • Plant biology
  • Zygote

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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