Cellulase and Macerozyme-PEG-mediated Transformation of Moss Protoplasts

Rituraj Batth; Ilenuta Simina Cuciurean; Satish Kumar Kodiripaka; Sarah Skovlund Rothman; Caroline Greisen; Henrik Toft Simonsen

doi:10.21769/BioProtoc.3782

Cellulase and Macerozyme-PEG-mediated Transformation of Moss Protoplasts

Bio Protoc. 2021 Jan 20;10(19):e3782. doi: 10.21769/BioProtoc.3782.

Authors

Rituraj Batth¹, Ilenuta Simina Cuciurean¹, Satish Kumar Kodiripaka¹, Sarah Skovlund Rothman¹, Caroline Greisen¹, Henrik Toft Simonsen¹

Affiliation

¹ Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, 2800 Denmark.

Abstract

This protocol describes the generation of protoplasts from protonemal tissue of the moss Physcomitrium patens (syn. Physcomitrella patens), using Cellulase ONOZUKA R10 and Macerozyme R10, followed by polyethylene glycol (PEG) mediated transformation. The protonemal tissue grown in liquid suspension was harvested and treated with enzyme cocktails mix of 1.5% Cellulase ONOZUKA R10 and 0.5% Macerozyme R10 to generate 1,8 million protoplasts within 3 h.

Keywords: Cellulase; Macerozyme; Moss; Physcomitrium patens (syn. Physcomitrella patens); Protoplastation.