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A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis

BACKGROUND: A decline in chloroplast DNA (cpDNA) during leaf maturity has been reported previously for eight plant species, including Arabidopsis thaliana. Recent studies, however, concluded that the amount of cpDNA during leaf development in Arabidopsis remained constant. RESULTS: To evaluate alter...

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Autores principales: Rowan, Beth A, Oldenburg, Delene J, Bendich, Arnold J
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2632658/
https://www.ncbi.nlm.nih.gov/pubmed/19128504
http://dx.doi.org/10.1186/1471-2229-9-3
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author Rowan, Beth A
Oldenburg, Delene J
Bendich, Arnold J
author_facet Rowan, Beth A
Oldenburg, Delene J
Bendich, Arnold J
author_sort Rowan, Beth A
collection PubMed
description BACKGROUND: A decline in chloroplast DNA (cpDNA) during leaf maturity has been reported previously for eight plant species, including Arabidopsis thaliana. Recent studies, however, concluded that the amount of cpDNA during leaf development in Arabidopsis remained constant. RESULTS: To evaluate alternative hypotheses for these two contradictory observations, we examined cpDNA in Arabidopsis shoot tissues at different times during development using several methods: staining leaf sections as well as individual isolated chloroplasts with 4',6-diamidino-2-phenylindole (DAPI), real-time quantitative PCR with DNA prepared from total tissue as well as from isolated chloroplasts, fluorescence microscopy of ethidium-stained DNA molecules prepared in gel from isolated plastids, and blot-hybridization of restriction-digested total tissue DNA. We observed a developmental decline of about two- to three-fold in mean DNA per chloroplast and two- to five-fold in the fraction of cellular DNA represented by chloroplast DNA. CONCLUSION: Since the two- to five-fold reduction in cpDNA content could not be attributed to an artifact of chloroplast isolation, we conclude that DNA within Arabidopsis chloroplasts is degraded in vivo as leaves mature.
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spelling pubmed-26326582009-01-29 A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis Rowan, Beth A Oldenburg, Delene J Bendich, Arnold J BMC Plant Biol Research Article BACKGROUND: A decline in chloroplast DNA (cpDNA) during leaf maturity has been reported previously for eight plant species, including Arabidopsis thaliana. Recent studies, however, concluded that the amount of cpDNA during leaf development in Arabidopsis remained constant. RESULTS: To evaluate alternative hypotheses for these two contradictory observations, we examined cpDNA in Arabidopsis shoot tissues at different times during development using several methods: staining leaf sections as well as individual isolated chloroplasts with 4',6-diamidino-2-phenylindole (DAPI), real-time quantitative PCR with DNA prepared from total tissue as well as from isolated chloroplasts, fluorescence microscopy of ethidium-stained DNA molecules prepared in gel from isolated plastids, and blot-hybridization of restriction-digested total tissue DNA. We observed a developmental decline of about two- to three-fold in mean DNA per chloroplast and two- to five-fold in the fraction of cellular DNA represented by chloroplast DNA. CONCLUSION: Since the two- to five-fold reduction in cpDNA content could not be attributed to an artifact of chloroplast isolation, we conclude that DNA within Arabidopsis chloroplasts is degraded in vivo as leaves mature. BioMed Central 2009-01-07 /pmc/articles/PMC2632658/ /pubmed/19128504 http://dx.doi.org/10.1186/1471-2229-9-3 Text en Copyright © 2009 Rowan et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Rowan, Beth A
Oldenburg, Delene J
Bendich, Arnold J
A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis
title A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis
title_full A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis
title_fullStr A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis
title_full_unstemmed A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis
title_short A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis
title_sort multiple-method approach reveals a declining amount of chloroplast dna during development in arabidopsis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2632658/
https://www.ncbi.nlm.nih.gov/pubmed/19128504
http://dx.doi.org/10.1186/1471-2229-9-3
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