Original video: https://youtu.be/Uo4Nah3TZ-0
Pollen embryogenesis provides exciting opportunities in the areas of breeding and biotechnology as well as representing a convenient model for studying the process of plant cell proliferation in general and embryogenesis in particular. A cell culture system was devised in which immature barley pollen could be cultured as a monolayer trapped between the bottom glass-cover slip of a live-cell chamber and a diaphanous PTFE membrane within a liquid medium over a period of up to 28 d, allowing the process of embryogenesis to be tracked in individual pollen. Z-stacks of images were automatically captured every 3min, starting from the unicellular pollen stage up to the development of multicellular, embryogenic structures. The method should prove useful for the elucidation of ultrastructural features and molecular processes associated with pollen embryogenesis.
The video shown here is representative of a total of eight individual trials lasting between 14 d and 28 d that were performed to prove the usefulness of the system. Every trial comprised between 4 and 15 observed pollen and showed similar patterns of pollen development and similar pollen types were identified. Without the possibility of agitating the culture medium, the rate of development was significantly delayed to the extent that it took as long as 28 d for embryogenic microcalli to emerge from the exine. In conventionally grown embryogenic pollen, exine breakage typically occurs in the third week of cultivation.
Citation: D. S. Daghma, J. Kumlehn, G. Hensel, T. Rutten, M. Melzer, Time-lapse imaging of the initiation of pollen embryogenesis in barley (Hordeum vulgare L.), Journal of Experimental Botany, Volume 63, Issue 16, October 2012, Pages 6017–6021, https://doi.org/10.1093/jxb/ers254
Keywords: Barley, chamber cover slip, live-cell microscopy, pollen embryogenesis
Published on: 24 September 2012
Attribution 4.0 International — CC BY 4.0 - Creative Commons
留言