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

BMC Plant Biology
Beth A RowanArnold J Bendich

Abstract

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. 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. 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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Citations

Dec 12, 2012·Planta·Delene J OldenburgArnold J Bendich
Nov 17, 2009·Molecular Genetics and Genomics : MGG·Uwe RauwolfReinhold G Herrmann
May 21, 2009·Journal of Experimental Botany·Beth A Rowan, Arnold J Bendich
Apr 22, 2010·Journal of Experimental Botany·Beth A RowanArnold J Bendich
Mar 24, 2011·Plant Physiology·Jean-Sébastien ParentNormand Brisson
Dec 14, 2011·PloS One·Martijn StaatsFreek T Bakker
Mar 29, 2014·The Plant Cell·Hieronim GolczykReinhold G Herrmann
Jan 24, 2016·Molecular Biotechnology·Hitomi S KikkawaRitsuko Sugita
Jan 12, 2016·Forensic Science International. Genetics·Karen L BellBerry J Brosi
Jan 14, 2012·The Plant Journal : for Cell and Molecular Biology·Lay Yin TangWataru Sakamoto
Jan 6, 2016·Plant, Cell & Environment·Srikanta Kaidala Ganesha DaniFrancesco Loreto
Dec 17, 2011·American Journal of Botany·Shannon C K StraubAaron Liston
Nov 19, 2015·Frontiers in Plant Science·Delene J Oldenburg, Arnold J Bendich
Sep 25, 2019·Plants·Stewart A MorleyBrent L Nielsen
Mar 16, 2018·Frontiers in Plant Science·Takeshi TakamatsuToshiaki Mitsui
Dec 20, 2019·The Plant Journal : for Cell and Molecular Biology·Stephan GreinerReinhold G Herrmann
Jun 4, 2018·Plant & Cell Physiology·Wataru Sakamoto, Tsuneaki Takami

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Methods Mentioned

BETA
PCR
fluorescence microscopy
fluorescence
electrophoresis
dot-blot
flow cytometry
restriction digest
light microscopy

Software Mentioned

NIH Image J
Opticon Monitor
Image J
Open lab

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