A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis
Open Access
- 7 January 2009
- journal article
- Published by Springer Science and Business Media LLC in BMC Plant Biology
- Vol. 9 (1), 3
- https://doi.org/10.1186/1471-2229-9-3
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.Keywords
This publication has 28 references indexed in Scilit:
- The Structure of Chloroplast DNA Molecules and the Effects of Light on the Amount of Chloroplast DNA during Development in Medicago truncatulaPlant Physiology, 2008
- White Leaf Sectors inyellow variegated2Are Formed by Viable Cells with Undifferentiated PlastidsPlant Physiology, 2007
- A high-throughput method for detection of DNA in chloroplasts using flow cytometryPlant Methods, 2007
- Constancy of organellar genome copy numbers during leaf development and senescence in higher plantsMolecular Genetics and Genomics, 2005
- Changes in the Structure of DNA Molecules and the Amount of DNA Per Plastid During Chloroplast Development in MaizeJournal of Molecular Biology, 2004
- Endosymbiotic gene transfer: organelle genomes forge eukaryotic chromosomesNature Reviews Genetics, 2004
- Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT MethodMethods, 2001
- A new mathematical model for relative quantification in real-time RT-PCRNucleic Acids Research, 2001
- Quantitative fluorescence microscopy on dynamic changes of plastid nucleoids during wheat developmentProtoplasma, 1986
- Use of the fluorochrome 4'6-diamidino-2-phenylindole in genetic and developmental studies of chloroplast DNA.The Journal of cell biology, 1979