Yao Haiqin, Scornet Delphine, Badis Yacine, Peters Akira F, Jam Murielle, Hervé Cécile, Potin Philippe, Coelho Susana M, Cock J Mark
CNRS, Sorbonne Université, UPMC University Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, F-29688, Roscoff, France.
The Scottish Association for Marine Science, Scottish Marine Institute, Oban, Argyll PA37 1QA, United Kingdom.
Bio Protoc. 2020 Sep 20;10(18):e3753. doi: 10.21769/BioProtoc.3753.
The brown alga 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.
褐藻具有单倍体-二倍体生命周期，涉及两个多细胞世代（孢子体和配子体）之间的交替。生命周期世代不是由倍性决定的，而是由一个遗传系统决定的，该系统包括两种不同的具有三个氨基酸环延伸的同源结构域转录因子，称为衔尾蛇和轮回。此外，已证明孢子体可向培养基中分泌一种可扩散因子，该因子可诱导配子体初始细胞从配子体发育程序转变为孢子体发育程序。本文介绍的方案描述了如何制备含有可扩散孢子体诱导因子的孢子体条件培养基，以及如何使用基于 meio-孢子的生物测定法检测该因子的活性。该方案描述了如何优化这些程序的几个步骤，将成为未来旨在表征可扩散因子及其作用方式的工作的有用工具。
