Physcomitrium patens Protoplasting and Protoplast Transfection - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement
Journal Articles Methods in Molecular Biology Year : 2022

Physcomitrium patens Protoplasting and Protoplast Transfection

Florence Charlot
Guillaume Goudounet
Fabien Nogué

Abstract

Protoplast production with the moss Physcomitrium (Physcomitrella) patens has a long and successful history. As a tool, it has not only been the base of reverse genetic studies covering research fields as diverse as development, metabolism, or gene network regulation but also allowed its development as a bioengineering platform for protein production. We present here a standardized protocol for protoplast production from Physcomitrium (Physcomitrella) patens protonemata. Additionally, we detail procedures for their transfection, their plating for optimal regeneration, and three alternative selection approaches. To improve the consistency of protoplast regeneration, we describe a new option for protoplast embedding. The use of an alginate matrix to regenerate moss protoplast alleviates the use of warm agarized medium. Thus, it optimizes transformed protoplast survival without any morphological detrimental effect or impact on transfection efficiency.
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Dates and versions

hal-04437280 , version 1 (04-02-2024)

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Florence Charlot, Guillaume Goudounet, Fabien Nogué, Pierre-François Perroud. Physcomitrium patens Protoplasting and Protoplast Transfection. Methods in Molecular Biology, 2022, Methods in Molecular Biology, 2464, pp.3-19. ⟨10.1007/978-1-0716-2164-6_1⟩. ⟨hal-04437280⟩
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