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

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

^*Corresponding author for this work

Technical University of Denmark

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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.

Original language	English
Article number	e3782
Journal	Bio-protocol
Volume	11
Issue number	2
Number of pages	12
ISSN	2331-8325
DOIs	https://doi.org/10.21769/BioProtoc.3782
Publication status	Published - 2021

Keywords

Moss
Physcomitrium patens (syn. Physcomitrella patens)
Protoplastation
Cellulase
Macerozyme

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title = "Cellulase and Macerozyme-PEG-mediated Transformation of Moss Protoplasts",

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 = "Moss, Physcomitrium patens (syn. Physcomitrella patens), Protoplastation, Cellulase, Macerozyme",

author = "Rituraj Batth and Cuciurean, {Ilenuta Simina} and Kodiripaka, {Satish Kumar} and Rothman, {Sarah Skovlund} and Caroline Greisen and Simonsen, {Henrik Toft}",

year = "2021",

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AU - Batth, Rituraj

AU - Cuciurean, Ilenuta Simina

AU - Kodiripaka, Satish Kumar

AU - Rothman, Sarah Skovlund

AU - Greisen, Caroline

AU - Simonsen, Henrik Toft

PY - 2021

Y1 - 2021

N2 - 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.

AB - 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.

KW - Moss

KW - Physcomitrium patens (syn. Physcomitrella patens)

KW - Protoplastation

KW - Cellulase

KW - Macerozyme

U2 - 10.21769/BioProtoc.3782

DO - 10.21769/BioProtoc.3782

M3 - Journal article

C2 - 33732756

SN - 2331-8325

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JO - Bio-protocol

JF - Bio-protocol

IS - 2

M1 - e3782

ER -

Cellulase and Macerozyme-PEG-mediated Transformation of Moss Protoplasts

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