Chapter 6: An Improved Leaf Protoplast System for Highly Efficient Transient Expression in Switchgrass (Panicum virgatum L.)

Chien-Yuan Lin; Hui Wei; Bryon Donohoe; Melvin Tucker; Michael Himmel

doi:10.1007/978-1-0716-0195-2_6

Chapter 6: An Improved Leaf Protoplast System for Highly Efficient Transient Expression in Switchgrass (Panicum virgatum L.)

Chien-Yuan Lin, Hui Wei, Bryon Donohoe, Melvin Tucker, Michael Himmel

National Renewable Energy Laboratory

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

2 Scopus Citations

Abstract

As a robust perennial C4-type monocot plant and a native species to North America, switchgrass (Panicum virgatum) has been evaluated and designated as a strong candidate bioenergy crop by the U.S. DOE. Although genetic modifications of switchgrass have been used to successfully reduce the recalcitrance of switchgrass biomass for biofuel production, the generation of transgenic switchgrass is still a slow and laborious process. A transient protoplast system can provide an excellent platform to accelerate the selection of genes-of-interest for tailoring switchgrass biomass. However, partially due to the lack of the complete genomic information, the attempts to optimize the transient protoplast system for switchgrass remain scarce. In this chapter, we provide an improved protocol for switchgrass protoplast isolation, increased transformation efficiency using CsCl gradient ultracentrifugation-derived plasmid DNA and extended application of the transient switchgrass protoplast system to analyze protein expression using western blot.

Original language	American English
Title of host publication	Metabolic Pathway Engineering
Subtitle of host publication	Methods in Molecular Biology, Volume 2096
Editors	M. E. Himmel, Y. J. Bomble
Publisher	Humana Press Inc.
Pages	61-79
Number of pages	19
DOIs	https://doi.org/10.1007/978-1-0716-0195-2_6 https://doi.org/10.1007/978-1-0716-0195-2_6
State	Published - 2020

Publication series

Name	Methods in Molecular Biology
Volume	2096
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2020.

NREL Publication Number

NREL/CH-2700-74951

Keywords

CsCl gradient ultracentrifugation
GFP expression
Protoplast
Switchgrass
Transient expression system

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Lin, C.-Y., Wei, H., Donohoe, B., Tucker, M., & Himmel, M. (2020). Chapter 6: An Improved Leaf Protoplast System for Highly Efficient Transient Expression in Switchgrass (Panicum virgatum L.). In M. E. Himmel, & Y. J. Bomble (Eds.), Metabolic Pathway Engineering: Methods in Molecular Biology, Volume 2096 (pp. 61-79). (Methods in Molecular Biology; Vol. 2096). Humana Press Inc.. https://doi.org/10.1007/978-1-0716-0195-2_6, https://doi.org/10.1007/978-1-0716-0195-2_6

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Lin, C-Y, Wei, H , Donohoe, B, Tucker, M & Himmel, M 2020, Chapter 6: An Improved Leaf Protoplast System for Highly Efficient Transient Expression in Switchgrass (Panicum virgatum L.). in ME Himmel & YJ Bomble (eds), Metabolic Pathway Engineering: Methods in Molecular Biology, Volume 2096. Methods in Molecular Biology, vol. 2096, Humana Press Inc., pp. 61-79. https://doi.org/10.1007/978-1-0716-0195-2_6, https://doi.org/10.1007/978-1-0716-0195-2_6

Chapter 6: An Improved Leaf Protoplast System for Highly Efficient Transient Expression in Switchgrass (Panicum virgatum L.). / Lin, Chien-Yuan; Wei, Hui ; Donohoe, Bryon et al.
Metabolic Pathway Engineering: Methods in Molecular Biology, Volume 2096. ed. / M. E. Himmel; Y. J. Bomble. Humana Press Inc., 2020. p. 61-79 (Methods in Molecular Biology; Vol. 2096).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

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Lin CY, Wei H , Donohoe B, Tucker M, Himmel M. Chapter 6: An Improved Leaf Protoplast System for Highly Efficient Transient Expression in Switchgrass (Panicum virgatum L.). In Himmel ME, Bomble YJ, editors, Metabolic Pathway Engineering: Methods in Molecular Biology, Volume 2096. Humana Press Inc. 2020. p. 61-79. (Methods in Molecular Biology). doi: 10.1007/978-1-0716-0195-2_6, 10.1007/978-1-0716-0195-2_6

Chapter 6: An Improved Leaf Protoplast System for Highly Efficient Transient Expression in Switchgrass (Panicum virgatum L.)

Abstract

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Bibliographical note

NREL Publication Number

Keywords

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