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Changes in chloroplast DNA during development in tobacco, Medicago truncatula, pea, and maize

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Abstract

We examined the DNA from chloroplasts obtained from young and fully expanded leaves of tobacco (Nicotiana tabacum L.), Medicago truncatula, pea (Pisum sativum L.), and maize (Zea mays L.). The changes in plastid DNA content and structure were monitored by four independent methods: 4′,6-diamidino-2-phenylindole (DAPI) staining with intact chloroplasts, in situ DAPI staining of cytological sections, ethidium bromide staining at the single-molecule level after exhaustive deproteinization of lysed chloroplasts, and pulsed-field gel electrophoresis. During leaf development, we found a decline of chloroplast DNA (cpDNA) in all four plants. For tobacco, for which plants can readily be regenerated from somatic cells, cpDNA persisted longer than in the other three plants. We also found a striking progression from complex multigenomic DNA molecules to simple subgenomic molecules during plastid development. Although the decrease in molecular size and complexity paralleled the decrease in DNA content per plastid, 6% of the chloroplasts in a fully expanded tobacco leaf still contained DNA in complex branched structure, whereas no such complex structures were found in mature leaves for the hard-to-regenerate maize.

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Abbreviations

cpDNA :

Chloroplast DNA

DAPI :

4′,6-diamidino-2-phenylindole

IR :

Inverted repeat

References

  • Backert S, Dörfel P, Börner T (1995) Investigation of plant organellar DNAs by pulsed-field gel electrophoresis. Curr Genet 28:390–399

    Article  PubMed  CAS  Google Scholar 

  • Baumgartner BJ, Rapp JC, Mullet JE (1989) Plastid transcription activity and DNA copy number increase early in barley chloroplast development. Plant Physiol 89:1011–1018

    PubMed  CAS  Google Scholar 

  • Bendich A (1987) Why do chloroplasts and mitochondria contain so many copies of their genome? Bioessays 6:279–282

    Article  PubMed  CAS  Google Scholar 

  • Bendich A (1991) Moving pictures of DNA released upon lysis from bacteria, chloroplasts, and mitochondria. Protoplasma 160:121–130

    Article  Google Scholar 

  • Bendich A (2004) Circular chloroplast chromosomes: the grand illusion. Plant Cell 16:1661–1666

    Article  PubMed  CAS  Google Scholar 

  • Bookjans G, Stummann BM, Henningsen KW (1984) Preparation of chloroplast DNA from pea plastids isolated in a medium of high ionic strength. Anal Biochem 141:244–247

    Article  PubMed  CAS  Google Scholar 

  • Deng X-W, Wing RA, Gruissem W (1989) The chloroplast genome exists in multimeric forms. Proc Natl Acad Sci USA 86:4156–4160

    Article  PubMed  CAS  Google Scholar 

  • Fujie M, Kuroiwa H, Kawano S, Mutoh S, Kuroiwa T (1994) Behavior of organelles and their nucleoids in the shoot apical meristem during leaf development in Arabidopsis thaliana L. Planta 194:395–405

    Article  CAS  Google Scholar 

  • Kuroiwa T (1991) The replication, differentiation, and inheritance of plastids with emphasis on the concept of organelle nuclei. Int Rev Cytol 128:1–62

    CAS  Google Scholar 

  • Kuroiwa T, Suzuki T (1980) An improved method for the demonstration of the in situ chloroplast nuclei in higher plants. Cell Struct Funct 5:195–197

    Google Scholar 

  • Kuroiwa T, Suzuki T, Ogawa K, Kawano S (1981) The chloroplast nucleus: distribution, number, size, and shape, and a model for multiplication of the chloroplast genome during chloroplast development. Plant Cell Physiol 22:381–396

    Google Scholar 

  • Lamppa G, Bendich A (1979) Changes in chloroplast DNA levels during development of pea (Pisum sativum). Plant Physiol 64:126–130

    PubMed  CAS  Google Scholar 

  • Lamppa GK, Elliot LV, Bendich AJ (1980) Changes in chloroplast number during pea leaf development: an analysis of a protoplast population. Planta 148:437–443

    Article  CAS  Google Scholar 

  • Li B, Huang W, Bass T (2003) Shoot production per responsive leaf explant increases exponentially with explant organogenic potential in Nicotiana species. Plant Cell Rep 22:231–238

    Article  PubMed  CAS  Google Scholar 

  • Luo Y, Koop H-U (1997) Somatic embryogenesis in cultured immature zygotic embryos and leaf protoplasts of Arabidopsis thaliana ecotypes. Planta 202:387–396

    Article  PubMed  CAS  Google Scholar 

  • Miyamura S, Nagata T, Kuroiwa T (1986) Quantitative fluorescence microscopy on dynamic changes of plastid nucleoids during wheat development. Protoplasma 133:66–72

    Article  CAS  Google Scholar 

  • Nishimura Y, Misumi O, Kato K, Inada N, Higashiyama T, Momoyama Y, Kuroiwa T (2002) An mt+ gamete-specific nuclease that targets mt−chloroplasts during sexual reproduction in C. reinhardtii. Genes Dev 16:1116–1128

    Article  PubMed  CAS  Google Scholar 

  • Ogihara Y, Isono K, Kojima K, Endo A, Hanaoka M, Shiina T, Terachi T, Utsugi S, Murata M, Mori N, Takumi S, Ikeo K, Gojobori T, Murai R, Murai K, Matsuoka Y, Ohnishi Y, Tajiri H, Tsunewaki K (2002) Structural features of a wheat plastome as revealed by complete sequencing of chloroplasts. Mol Genet Genome 266:740–746

    Article  CAS  Google Scholar 

  • Oldenburg D, Bendich A (1996) Size and structure of replicating mitochondrial DNA in cultured tobacco cells. Plant Cell 8:447–461

    Article  PubMed  CAS  Google Scholar 

  • Oldenburg D, Bendich A (1998) The structure of mitochondrial DNA from the liverwort, Marchantia polymorpha. J Mol Biol 276:745–758

    Article  PubMed  CAS  Google Scholar 

  • Oldenburg D, Bendich A (2004a) Most chloroplast DNA of maize seedlings in linear molecules with defined ends and branched forms. J Mol Biol 335:953–970

    Article  PubMed  CAS  Google Scholar 

  • Oldenburg D, Bendich A (2004b) Changes in the structure of DNA molecules and the amount of DNA per plastid during chloroplast development in maize. J Mol Biol 344:1311–1330

    Article  PubMed  CAS  Google Scholar 

  • Rowan B, Oldenburg D, Bendich A (2004) The demise of chloroplast DNA in Arabidopsis. Curr Genet 46:176–181

    Article  PubMed  CAS  Google Scholar 

  • Scott NS, Possingham JV (1980) Chloroplast DNA in expanding spinach leaves. J Exper Bot 31:1081–1092

    Article  Google Scholar 

  • Seyer P, Kowallik KV, Herrman RG (1981) A physical map of Nictotiana tabacum plastid DNA including the location of structural genes for ribosomal RNAs and the large subunit of ribulose bisphosphate carboxylase/oxygenase. Curr Genet 3:189–204

    Article  CAS  Google Scholar 

  • Sodmergen, Kawano S, Tano S, Kuroiwa T (1991) Degradation of chloroplast DNA in second leaves of rice (Oryza sativa) before leaf yellowing. Protoplasma 160:89–98

    Article  CAS  Google Scholar 

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Acknowledgements

The project was supported by the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service, grant number#2002-35301-12021. We thank Marc Schleh for conducting the maize assays and Doug Ewing for assistance with growing plants.

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Authors and Affiliations

  1. Department of Biology, University of Washington, 355325, Seattle, WA, 98195-5325, USA

    Jeffrey M. Shaver, Delene J. Oldenburg & Arnold J. Bendich

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  1. Jeffrey M. Shaver
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  2. Delene J. Oldenburg
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  3. Arnold J. Bendich
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Correspondence to Arnold J. Bendich.

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Shaver, J.M., Oldenburg, D.J. & Bendich, A.J. Changes in chloroplast DNA during development in tobacco, Medicago truncatula, pea, and maize. Planta 224, 72–82 (2006). https://doi.org/10.1007/s00425-005-0195-7

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  • DOI: https://doi.org/10.1007/s00425-005-0195-7

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