Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts

Stéphanie Robert; Jan Zouhar; Clay Carter; Natasha Raikhel

doi:10.1038/nprot.2007.26

Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts

Stéphanie Robert
, Jan Zouhar
, Clay Carter
, Natasha Raikhel

Administration (CBS)

Research output: Contribution to journal › Article › peer-review

88 Scopus citations

Abstract

Vacuoles are very prominent compartments within plant cells, and understanding of their function relies on knowledge of their content. Here, we present a simple vacuole purification protocol that was successfully used for large-scale isolation of vacuoles, free of significant contamination from other endomembrane compartments. This method is based on osmotic and thermal disruption of mesophyl-derived Arabidopsis protoplasts, followed by a density gradient fractionation of the cellular content. The whole procedure, including protoplast isolation, takes approximately 6 h.

Original language	English (US)
Pages (from-to)	259-262
Number of pages	4
Journal	Nature Protocols
Volume	2
Issue number	2
DOIs	https://doi.org/10.1038/nprot.2007.26
State	Published - Mar 2007

Bibliographical note

Funding Information:
ACKNOWLEDGMENTS This work was supported by the U.S. Department of Energy, Division of Energy Biosciences (Grant DE-FG02-02ER15295 to N.R.).

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10.1038/nprot.2007.26

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abstract = "Vacuoles are very prominent compartments within plant cells, and understanding of their function relies on knowledge of their content. Here, we present a simple vacuole purification protocol that was successfully used for large-scale isolation of vacuoles, free of significant contamination from other endomembrane compartments. This method is based on osmotic and thermal disruption of mesophyl-derived Arabidopsis protoplasts, followed by a density gradient fractionation of the cellular content. The whole procedure, including protoplast isolation, takes approximately 6 h.",

author = "St{\'e}phanie Robert and Jan Zouhar and Clay Carter and Natasha Raikhel",

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AU - Zouhar, Jan

AU - Carter, Clay

AU - Raikhel, Natasha

N1 - Funding Information: ACKNOWLEDGMENTS This work was supported by the U.S. Department of Energy, Division of Energy Biosciences (Grant DE-FG02-02ER15295 to N.R.).

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N2 - Vacuoles are very prominent compartments within plant cells, and understanding of their function relies on knowledge of their content. Here, we present a simple vacuole purification protocol that was successfully used for large-scale isolation of vacuoles, free of significant contamination from other endomembrane compartments. This method is based on osmotic and thermal disruption of mesophyl-derived Arabidopsis protoplasts, followed by a density gradient fractionation of the cellular content. The whole procedure, including protoplast isolation, takes approximately 6 h.

AB - Vacuoles are very prominent compartments within plant cells, and understanding of their function relies on knowledge of their content. Here, we present a simple vacuole purification protocol that was successfully used for large-scale isolation of vacuoles, free of significant contamination from other endomembrane compartments. This method is based on osmotic and thermal disruption of mesophyl-derived Arabidopsis protoplasts, followed by a density gradient fractionation of the cellular content. The whole procedure, including protoplast isolation, takes approximately 6 h.

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ER -

Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts

Abstract

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