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

Allbwn ymchwil: Pennod mewn Llyfr/Adroddiad/Trafodion Cynhadledd › Pennod

Crynodeb

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

Iaith wreiddiol	Saesneg
Teitl	Cereal Genomics
Golygyddion	Luis M. Vaschetto
Cyhoeddwr	Springer Nature
ISBN (Electronig)	978-1-4939-9865-4
ISBN (Argraffiad)	978-1-4939-9864-7
Dynodwyr Gwrthrych Digidol (DOIs)	https://doi.org/10.1007/978-1-4939-9865-4
Statws	E-gyhoeddi cyn argraffu - 21 Medi 2019

Cyfres gyhoeddiadau

Enw	Methods in Molecular Biology
Cyfrol	2072
ISSN (Argraffiad)	1064-3745
ISSN (Electronig)	1940-6029

Mynediad at Ddogfen

10.1007/978-1-4939-9865-4

Cysylltiad parhaol

Dolen barhaus

1 Wedi Gorffen

BBSRC Core Strategic Programme in Resilient Crops: Grasslands Gogerddan
Armstead, I. (Prif Ymchwilydd), Donnison, I. (Cyd-ymchwilydd), Jones, H. (Cyd-ymchwilydd), Skot, L. (Cyd-ymchwilydd), Fernandez Fuentes, N. (Cyd-ymchwilydd), Phillips, D. (Prif Ymchwilydd), Kingston-Smith, A. (Cyd-ymchwilydd) & Bosch, M. (Cyd-ymchwilydd)
Biotechnology and Biological Sciences Research Council
01 Ebr 2017 → 31 Maw 2020
Prosiect: Ymchwil a ariannwyd yn allanol

Dyfynnu hyn

@inbook{afab1d66437b4ade871c3b13b5ec71ee,

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

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",

keywords = "protoplasts, lolium temulentum, lolium perenne, transient expression, PEG-mediated transformation",

author = "Davies, {Huw R.} and Anne Maddison and Dylan Phillips and Huw Jones",

year = "2019",

month = sep,

day = "21",

doi = "10.1007/978-1-4939-9865-4",

language = "English",

isbn = "978-1-4939-9864-7",

series = "Methods in Molecular Biology",

publisher = "Springer Nature",

editor = "Vaschetto, {Luis M.}",

booktitle = "Cereal Genomics",

address = "Switzerland",

}

TY - CHAP

T1 - Genetic Transformation of Protoplasts Isolated from Leave of Lolium temulentum and Lolium perenne

AU - Davies, Huw R.

AU - Maddison, Anne

AU - Phillips, Dylan

AU - Jones, Huw

PY - 2019/9/21

Y1 - 2019/9/21

N2 - 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

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

KW - protoplasts

KW - lolium temulentum

KW - lolium perenne

KW - transient expression

KW - PEG-mediated transformation

U2 - 10.1007/978-1-4939-9865-4

DO - 10.1007/978-1-4939-9865-4

M3 - Chapter

SN - 978-1-4939-9864-7

T3 - Methods in Molecular Biology

BT - Cereal Genomics

A2 - Vaschetto, Luis M.

PB - Springer Nature

ER -

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

Crynodeb

Cyfres gyhoeddiadau

Mynediad at Ddogfen

Cysylltiad parhaol

Ôl bys

Prosiectau

BBSRC Core Strategic Programme in Resilient Crops: Grasslands Gogerddan

Dyfynnu hyn