Genetic Transformation of Protoplasts Isolated from Leave of Lolium temulentum and Lolium perenne

Huw R. Davies; Anne Maddison; Dylan Phillips; Huw Jones

doi:10.1007/978-1-4939-9865-4

Genetic Transformation of Protoplasts Isolated from Leave of Lolium temulentum and Lolium perenne

Huw R. Davies, Anne Maddison, Dylan Phillips, Huw Jones

Research output: Chapter in Book/Report/Conference proceeding › Chapter

Abstract

Transient expression of inserted recombinant DNA in plant protoplasts is a widely used tool for functional genomics research. Recently it has been utilized to screen potential sgRNA guides for CRISPR-mediated genome editing. However, little research has been conducted into the use of transient expression of protoplasts in Lolium perenne (a globally important pasture, hay, and amenity grass), and no studies have been conducted into Lolium temulentum (a weed in cereal crops but a potentially useful model species for Lolium research). In this chapter, we describe a methodology of protoplast extraction and transformation from 14-day-old leaf mesophyll cells from L. perenne and L. temulentum. We believe this is the first report of a procedure for obtaining high density, viable protoplasts from L. temulentum. The method of polyethylene glycol (PEG)-mediated transformation is also described to achieve genetic transformation of protoplasts

Original language	English
Title of host publication	Cereal Genomics
Editors	Luis M. Vaschetto
Publisher	Springer Nature
ISBN (Electronic)	978-1-4939-9865-4
ISBN (Print)	978-1-4939-9864-7
DOIs	https://doi.org/10.1007/978-1-4939-9865-4
Publication status	E-pub ahead of print - 21 Sept 2019

Publication series

Name	Methods in Molecular Biology
Volume	2072
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

protoplasts
lolium temulentum
lolium perenne
transient expression
PEG-mediated transformation

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10.1007/978-1-4939-9865-4

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BBSRC Core Strategic Programme in Resilient Crops: Grasslands Gogerddan
Armstead, I. (PI), Donnison, I. (CoI), Jones, H. (CoI), Skot, L. (CoI), Fernandez Fuentes, N. (CoI), Phillips, D. (PI), Kingston-Smith, A. (CoI) & Bosch, M. (CoI)
Biotechnology and Biological Sciences Research Council
01 Apr 2017 → 31 Mar 2020
Project: Externally funded research

Cite this

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title = "Genetic Transformation of Protoplasts Isolated from Leave of Lolium temulentum and Lolium perenne",

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PY - 2019/9/21

Y1 - 2019/9/21

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AB - Transient expression of inserted recombinant DNA in plant protoplasts is a widely used tool for functional genomics research. Recently it has been utilized to screen potential sgRNA guides for CRISPR-mediated genome editing. However, little research has been conducted into the use of transient expression of protoplasts in Lolium perenne (a globally important pasture, hay, and amenity grass), and no studies have been conducted into Lolium temulentum (a weed in cereal crops but a potentially useful model species for Lolium research). In this chapter, we describe a methodology of protoplast extraction and transformation from 14-day-old leaf mesophyll cells from L. perenne and L. temulentum. We believe this is the first report of a procedure for obtaining high density, viable protoplasts from L. temulentum. The method of polyethylene glycol (PEG)-mediated transformation is also described to achieve genetic transformation of protoplasts

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PB - Springer Nature

ER -

Genetic Transformation of Protoplasts Isolated from Leave of Lolium temulentum and Lolium perenne

Abstract

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Keywords

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Projects

BBSRC Core Strategic Programme in Resilient Crops: Grasslands Gogerddan

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