Uptake of a fluorescent dye as a swift and simple indicator of organelle intactness: import-competent chloroplasts from soil-grown Arabidopsis
Uptake of a fluorescent dye as a swift and simple indicator of organelle intactness: import-competent chloroplasts from soil-grown Arabidopsis
We developed a rapid and reliable technique for specifically staining intact chloroplasts using the fluorescent dye carboxyfluorescein diacetate. Intact, import-competent chloroplasts were isolated simply and rapidly from soil-grown Arabidopsis thaliana plants, with yields of 20 ± 5 µg chlorophyll per g FW, greater than previously reported yields from soil-grown Arabidopsis. Traditional chloroplast isolation buffers sometimes contain low concentrations (<10 mM) sodium ascorbate as a general-purpose anti-oxidant, but we found that only Arabidopsis chloroplasts isolated in the presence of high concentrations (50–100 mM) of sodium ascorbate in the initial grinding buffer were import-competent.
carboxyfluorescein diacetate, intact organelles, arabidopsis, chloroplast, glucosinolates, protein import
701-704
Schulz, Alexander
fd27e166-ea38-4127-b96c-0cbd24c20468
Knoetzel, Jürgen
5b9a2161-007f-4b9c-b9b7-7225c2a19e25
Scheller, Henrik V.
204a6d1c-a909-4e1c-81ab-cbf7204df715
Mant, Alexandra
63319e45-deeb-45ad-a30d-e05b42052a0d
2004
Schulz, Alexander
fd27e166-ea38-4127-b96c-0cbd24c20468
Knoetzel, Jürgen
5b9a2161-007f-4b9c-b9b7-7225c2a19e25
Scheller, Henrik V.
204a6d1c-a909-4e1c-81ab-cbf7204df715
Mant, Alexandra
63319e45-deeb-45ad-a30d-e05b42052a0d
Schulz, Alexander, Knoetzel, Jürgen, Scheller, Henrik V. and Mant, Alexandra
(2004)
Uptake of a fluorescent dye as a swift and simple indicator of organelle intactness: import-competent chloroplasts from soil-grown Arabidopsis.
Journal of Histochemistry and Cytochemistry, 52 (5), .
Abstract
We developed a rapid and reliable technique for specifically staining intact chloroplasts using the fluorescent dye carboxyfluorescein diacetate. Intact, import-competent chloroplasts were isolated simply and rapidly from soil-grown Arabidopsis thaliana plants, with yields of 20 ± 5 µg chlorophyll per g FW, greater than previously reported yields from soil-grown Arabidopsis. Traditional chloroplast isolation buffers sometimes contain low concentrations (<10 mM) sodium ascorbate as a general-purpose anti-oxidant, but we found that only Arabidopsis chloroplasts isolated in the presence of high concentrations (50–100 mM) of sodium ascorbate in the initial grinding buffer were import-competent.
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Published date: 2004
Keywords:
carboxyfluorescein diacetate, intact organelles, arabidopsis, chloroplast, glucosinolates, protein import
Identifiers
Local EPrints ID: 26596
URI: http://eprints.soton.ac.uk/id/eprint/26596
ISSN: 0022-1554
PURE UUID: c17f1127-5a30-4386-8d8f-26bfec8a9514
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Date deposited: 20 Apr 2006
Last modified: 09 Jan 2022 03:16
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Contributors
Author:
Alexander Schulz
Author:
Jürgen Knoetzel
Author:
Henrik V. Scheller
Author:
Alexandra Mant
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