Skip to main content
Log in

Uncoupling of chloroplast reproductive events from cell cycle division processes by 5-fluorodeoxyuridine in the algaScenedesmus quadricauda

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

FdUrd (5-fluorodeoxyuridine), a specific inhibitor of thymidylate synthase, was used to study the relationship between reproductive processes in chloroplast and nucleocytoplasmic compartments of the chlorococcal algaScenedesmus quadricauda. The courses of DNA replication and nuclear division in both the compartments were followed in populations synchronised by the alternation of light and dark periods. DAPI-staining of DNA-containing structures was used for their visualisation and quantification. In contrast with cellular reproductive events, those in chloroplasts were not substantially affected by the presence of FdUrd (25 μg/ml). It was shown that FdUrd specifically blocked nucDNA replication but not ptDNA replication. Thus, cells which had attained commitment to ptDNA replication, fission of pt-nuclei and chloroplast kinesis triggered and terminated these processes while the corresponding cellular processes were blocked. The courses of reproductive processes in chloroplasts were also substantially unaffected in cells grown in the presence of FdUrd for the whole cell cycle. This provided evidence that attainment of commitment to and termination of the entire sequence of reproductive events, including chloroplast fission, were controlled by different mechanisms than the reproductive processes in the nucleocytoplasmic compartment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

DAPI:

4′,6-diamidino-2-phenylindole

ptDNA DNA:

of chloroplast nuclei

nucDNA DNA:

in cell nuclei

FdUrd:

5-fluorodeoxyuridine

References

  • Bachmann B, Follmann H (1987) Deoxyribonucleotide biosynthesis in green algae: characterization of thymidylate synthase-dihydrofolate reductase inScenedesmus obliquus. Arch Biochem Biophys 256: 244–252

    PubMed  Google Scholar 

  • —, Hofmann R, Follmann H (1983) Tight coordination of ribonucleotide reduction and thymidilate synthesis in synchronous algae. FEBS Lett 152: 247–250

    Google Scholar 

  • Butterfass T (1995) Reproduction and continuity of chloroplasts in spermatophyta. Bot Rev 61: 1–27

    Google Scholar 

  • Decallonne JR, Weyns CJ (1976) A shortened procedure of the diphenylamine reaction for the measurement of deoxyribonucleic acid by using light activation. Anal Biochem 74: 448–456

    PubMed  Google Scholar 

  • Feller W, Schimpff-Weiland G, Follmann H (1980) Deoxyribonucleotide biosynthesis in synchronous algae cells. Eur J Biochem 110: 85–92

    PubMed  Google Scholar 

  • Follmann H (1983) Deoxyribonucleotide biosynthesis: a critical process for life. In: Pullman B, Jortner J (eds) Nucleic acids: the vector of life. D Reidel, Lancaster, pp 545–557

    Google Scholar 

  • Kuroiwa T (1990) The replication, differentiation, and inheritance of plastids with emphasis on the concept of organelle nuclei. Int Rev Cytol 128: 1–62

    Google Scholar 

  • —, 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 

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

    Google Scholar 

  • —, Nagashima H, Fukuda I (1989) Chloroplast division without DNA synthesis during the life cycle of the unicellular algaCyanidium caldarium M-8 as revealed by quantitative fluorescence microscopy. Protoplasma 149: 120–129

    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

    Google Scholar 

  • Nakamura S, Kuroiwa T (1989) Selective elimination of chloroplast DNA by fluorodeoxyuridine causing no effect on preferential digestion of male chloroplast nucleoids inChlamydomonas. Eur J Cell Biol 48: 165–173

    Google Scholar 

  • Nemoto Y, Kawano S, Kondoh K, Nagata T, Kuroiwa T (1990) Studies on plastid nuclei (nucleoids) inNicotiana tabacum. III. Isolation of chloroplast nuclei from mesophyll protoplasts and identification of chloroplast DNA binding proteins. Plant Cell Physiol 31: 767–776

    Google Scholar 

  • — —, Nagata T, Kuroiwa T (1991) Studies on plastid nuclei (nucleoids) inNicotiana tabacum L. IV. Association of chloroplast DNA with proteins at several specific sites in isolated chloroplast nuclei. Plant Cell Physiol 32: 131–141

    Google Scholar 

  • Santi DW, McHenry CS (1972) 5-fluoro 2′-deoxyuridylate: covalent complex with thymidylate synthetase. Proc Natl Acad Sci USA 69: 1855–1857

    PubMed  Google Scholar 

  • Šetlík I, Zachleder V (1984) The multiple fission cell reproductive patterns in algae. In: Nurse P, Streiblová E (eds) The microbial cell cycle. CRC Press, Boca Raton, pp 253–279

    Google Scholar 

  • Wurtz EA, Boynton JE, Gillham NW (1977) Perturbation of chloroplast DNA amounts and chloroplast gene transmission inChlamydomonas reinhardtii by 5-fluorodeoxyuridine. Proc Natl Acad Sci USA 74: 4552–4556

    Google Scholar 

  • Zachleder V (1984) Optimization of nucleic acids assay in green and blue-green algae. Extraction procedures and the light-activated reaction for DNA. Algolog Stud 36: 313–328

    Google Scholar 

  • — (1994) Effectof hydroxyurea and fluorodeoxyuridine on the cell cycle events in the chlorococcal algaScenedesmus quadricauda (Chlorophyta) 1. J Phycol 30: 274–279

    Google Scholar 

  • — (1995) Regulation of growth and reproductive processes during the cell cycle of the chlorococcal algaScenedesmus quadricauda under block of DNA replication. J Phycol 31: 941–947

    Google Scholar 

  • —, Cepák V (1987a) Visualization of DNA containing structures by fluorochrome DAPI in those algal cells which are not freely permeable to the dye. Algolog Stud 47: 157–168

    Google Scholar 

  • — — (1987b) The effect of light on the number of chloroplast nucleoids in daughter cells of the algaScenedesmus quadricauda. Protoplasma 138: 37–44

    Google Scholar 

  • — — (1987c) Variations in chloroplast nucleoid number during the cell cycle in the algaScenedesmus quadricauda grown under different light conditions. Protoplasma 141: 74–82

    Google Scholar 

  • —, Šetík I (1982) Effect of irradiance on the course of RNA synthesis in the cell cycle ofScenedesmus quadricauda. Biol Plant 24: 341–353

    Google Scholar 

  • —, Šetlík I (1990) Timing of events in overlapping cell reproductive sequences and their mutual interactions in the algaScenedesmus quadricauda. J Cell Sci 97: 631–638

    Google Scholar 

  • —, Kawano S, Kuroiwa T (1995) The course of chloroplast DNA replications and their relationship to other reproductive processes in chloroplast and nucleocytoplasmic compartment during the cell cycle of the algaScenedesmus quadricauda. Protoplasma 188: 245–251

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zachleder, V., Kawano, S. & Kuroiwa, T. Uncoupling of chloroplast reproductive events from cell cycle division processes by 5-fluorodeoxyuridine in the algaScenedesmus quadricauda . Protoplasma 192, 228–234 (1996). https://doi.org/10.1007/BF01273894

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01273894

Keywords

Navigation