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Production and Bioassay of a Diffusible Factor That Induces Gametophyte-to-Sporophyte Developmental Reprogramming in the Brown Alga Ectocarpus

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Citation

Yao, H., Scornet, D., Badis, Y., Peters, A., Jam, M., Hervé, C., et al. (2020). Production and Bioassay of a Diffusible Factor That Induces Gametophyte-to-Sporophyte Developmental Reprogramming in the Brown Alga Ectocarpus. Bio-protocol, 10(18): e3753. doi:10.21769/BioProtoc.3753.


Cite as: https://hdl.handle.net/21.11116/0000-000C-7BB7-C
Abstract
The brown alga Ectocarpus has a haploid-diploid life cycle that involves alternation between two multicellular generations, the sporophyte and the gametophyte. Life cycle generation is not determined by ploidy but by a genetic system that includes two different three amino acid loop extension homeodomain transcription factors called OUROBOROS and SAMSARA. In addition, sporophytes have been shown to secrete a diffusible factor into the medium that can induce gametophyte initial cells to switch from the gametophyte to the sporophyte developmental program. The protocol presented here describes how to produce sporophyte-conditioned medium containing the diffusible sporophyte-inducing factor and how to assay for activity of the factor using a meio-spore-based bioassay. The protocol, which describes how several steps of these procedures can be optimised, will represent a useful tool for future work aimed at characterising the diffusible factor and investigating its mode of action.