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Spontaneous chromosome doubling results from nuclear fusion during in vitro maize induced microspore embryogenesis

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Abstract

A multidisciplinary study was carried out to analyse the chromosome doubling process during the early stages of in vitro maize microspore embryogenesis. The main stages (microspore derivatives) that were formed in the course of the culture were analysed. Chromosome number was determined from squashed cells, and DNA content was measured by cytometry. In parallel, an ultrastructural analysis of the microspore derivatives demonstrated the occurrence of a nuclear fusion process. It seems likely that nuclear fusion ensures chromosome doubling at early stages of induced microspore embryogenesis. It occurs precisely at the 5/7 day stage in the embryonic domain and probably leads to polyploidy in the endosperm domain of the microspore derivatives. As a conclusion a scheme summarises the results and proposes an interpretation of the sequence of chromosome doubling events during early maize microspore embryogenesis. Understanding of this process will be important for future efforts to increase the percentage of homozygous plants for crop improvement.

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References

  • Antoine-Michard S, Beckert M (1997) Spontaneous versus colchicine-induced chromosome doubling in maize anther culture. Plant Cell Tissue Organ Culture 48:203-207

    Article  Google Scholar 

  • Barnabas B, Pfahler PL, Kovacs (1991) Direct effect of colchicine on the microspore embryogenesis to produce dihaploid plants in wheat ( Triticum aestivum L.). Theor Appl Genet 81:675-678

    CAS  Google Scholar 

  • Beaumont VH, Widholm JM (1993) Ploidy variation of pronamide-treated maize calli during long term culture. Plant Cell Rep 12:648-651

    CAS  Google Scholar 

  • D’amato F (1984) Role of polyploidy in reproductive organs and tissues. In: Johri BM (ed) Embryology of Angiosperms. Springer-Verlag, New York, pp 519–566

  • Gaillard A, Vergne P, Beckert M (1991) Optimization of maize microspore isolation and culture conditions for reliable plant regeneration. Plant Cell Rep 10:55–58

    Google Scholar 

  • Gaillard A, Matthys-Rochon E, Dumas C (1992) Selection of microspore derived embryonic structures in maize related to transformation potential by microinjection. Bot Act 105:313–318

    Google Scholar 

  • Georgiev S (1987) Cytogenetic studies of the nature of the Sphaerococcum mutation in genus Triticum L. Genet Breed 20:95–100

    Google Scholar 

  • Ghosh S, Paweletz N (1984) Synchronous DNA synthesis and mitosis in multinucleate cells with one chromosome in each nucleus. Chromosoma 89:197–200

    CAS  PubMed  Google Scholar 

  • Giménez-Martín G, Risueno MC, López-Saez F (1965) Nuclear fusion in somatic cells: observations with the electron microscope. Fiton 22:173–175

    Google Scholar 

  • Gomez E, Royo J, Guo Y, Thompson R, Hueros G (2002) Establishment of cereal endosperm expression domains: identification and properties of a maize transfer cell-specific transcription factor, ZmMRP-1. Plant Cell 14:599–610

    Article  CAS  PubMed  Google Scholar 

  • Gonzàlez-Melendi P, Testillano PS, Ahmadian P, Reyes J, Risueno MC (2000) Immunoelectron microscopy of PCNA as an efficient marker for studying replication times and sites during pollen development. Chromosoma 109:397–409

    PubMed  Google Scholar 

  • Goralski G, Lafitte C, Bouazza L, Matthys-Rochon E, Przywara (2002) Influence of sugars on isolated microspore development in maize ( Zea mays L.). Acta Biol Crac 44:203–212

    Google Scholar 

  • Grafi G, Larkins BA (1995) Endoreduplication in maize endosperm: involvement of M phase-promoting factor inhibition and induction of S phase-related kinases. Science 269:1262–1264

    CAS  Google Scholar 

  • Hansen NJP, Andersen SB (1998) In vitro chromosome doubling with colchicine during microspore culture in wheat ( Triticum aestivum L.). Euphytica 102:101–108

    CAS  Google Scholar 

  • Henry Y (1998) Origin of microspore-derived dihaploid and polyploid in vitro plants. Plant Tissue Culture Biotechnol 4:127–134

    Google Scholar 

  • Joubès J, Chevalier C (2000) Endoreduplication in higher plants. Plant Mol Biol 43:735–745

    Article  CAS  PubMed  Google Scholar 

  • Kasha KJ, Hu TC, Oro R, Simion E, Shim YS (2001) Nuclear fusion leads to chromosome doubling during mannitol pretreatment of barley ( Hordeum vulgare L.) microspores. J Exp Bot 52:1227–1238

    Article  CAS  PubMed  Google Scholar 

  • Lin BY (1977) Ploidy variation in maize endosperm. J Hered 68:143-149

    Google Scholar 

  • López-Saez F, Risueno MC, Giménez-Martín G (1966) Inhibition of cytokinesis in plant cells. J Ultrastruct Res 14:85–94

    Google Scholar 

  • Magnard JL, Le Deunff E, Domenech J, Rogowsky PM, Testillano PS, Rougier M, Risueno MC, Vergne P, Dumas C (2000) Genes normally expressed in the endosperm are expressed in early stages of microspore embryogenesis in maize. Plant Mol Biol 44:559–574

    CAS  PubMed  Google Scholar 

  • Mogensen HL, Leduc N, Matthys-Rochon E, Dumas C (1995) Nuclear DNA amounts in the egg and zygote of maize ( Zea mays L.). Planta 197:641–645

    CAS  Google Scholar 

  • Neuffer MG, Coe EH, Wessler SR (1997) Mutants of maize. Cold Spring Harbor Laboratory Press, New York, p 30

  • Niebel A, de Almeida Engler J, Hemerly A, Ferreira P, Inzé D, Van Montagu M, Gheysen G (1996) Induction of cdc2a and Cyc1At expression in Arabidopsis thaliana during early phases of nematode-induced feeding cell formation. Plant J 10:1037–1043

    Article  CAS  PubMed  Google Scholar 

  • Olsen OA, Linnestad C, Nichols SE (1999) Developmental biology of the cereal endosperm. Trends Plant Sci 4:253–257

    PubMed  Google Scholar 

  • Risueno MC, Giménez Martín G, López-Saez F (1968) Experimental analysis of plant cytokinesis. Exp Cell Res 49:136-147

    CAS  PubMed  Google Scholar 

  • Testillano PS, Gonzales-Melendi P, Fadon B, Risueno MC (1995) The immunolocalisation of nuclear antigens during the pollen developmental program and the induction of pollen embryogenesis. Exp Cell Res 221:41–54

    CAS  PubMed  Google Scholar 

  • Testillano PS, Coronado MJ, Segui JM, Domenech J, Gonzàlez-Melendi P, Raska I, Risueno MC (2000) Defined nuclear changes accompany the reprogramming of the microspore to embryogenesis. J Struct Biol 129:223–232

    Article  CAS  PubMed  Google Scholar 

  • Testillano PS, Ramirez C, Domenech J, Coronado MJ, Vergne P, Matthys-Rochon E, Risueno MC (2002) Young microspore-derived maize embryos show two domains with defined features also present in zygotic embryogenesis. Int J Dev Biol 46:1035–1047

    CAS  PubMed  Google Scholar 

  • Traas J, Hülskamp M, Gendreau E, Höfte H (1998) Endoreduplication and development: rule without dividing? Cur Opin Plant Biol 1:498–503

    Article  CAS  Google Scholar 

  • Vergne P, Delvallée I, Dumas C (1987) Rapid assessement of microspore and pollen development stage in wheat and maize using DAPI and membrane permeabilization. Stain Technol 62:299–304

    CAS  PubMed  Google Scholar 

  • Wan Y, Duncan DR, Rayburn AL, Petolino JF, Widholm JM (1991) The use of antimicrotubule herbicides for the production of double haploid plants from anther derived maize callus. Theor Appl Genet 81:205–211

    CAS  Google Scholar 

  • Wong CK (1989) A new approach to chromosome doubling for haploid rice plants. Theor Appl Genet 77:149–151

    Google Scholar 

Download references

Acknowledgements

The authors thank Dr Christian Chevalier (Unité Mixte de Recherche de Physiologie et Biotechnologie Végétales, INRA-Bordeaux) for critically reading the manuscript. They are grateful to Mrs. L.Bouazza, F. Rozier and Mr. H. Leyral for their excellent technical work. This work was partly supported by PAI-Picasso contract (Nr04262VK) in the frame of France/Spain European exchanges.

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Authors and Affiliations

  1. CSIC, Velasquez 144, 28006, Madrid, Spain

    P. Testillano, M. J. Coronado & M. C. Risueno

  2. Sofia University Faculty of Biology, 8 Dragan Tsaunov clvd 1421, Sofia, Bulgaria

    S. Georgiev

  3. Department of Biological Sciences, Box 5640, Arizona University, Flagstaff, AZ 86011, USA

    H. L. Mogensen

  4. Reproduction et Développement des Plantes ENS Lyon, UMR 5667CNRS/INRA/ENS/LYON 1, 46 Allée d’Italie, 69364 Lyon Cédex 07, France

    C. Dumas & E. Matthys-Rochon

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  1. P. Testillano
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  2. S. Georgiev
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  3. H. L. Mogensen
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  4. M. J. Coronado
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  5. C. Dumas
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  6. M. C. Risueno
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  7. E. Matthys-Rochon
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Correspondence to E. Matthys-Rochon.

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Communicated by G. Almouzni

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Testillano, P., Georgiev, S., Mogensen, H.L. et al. Spontaneous chromosome doubling results from nuclear fusion during in vitro maize induced microspore embryogenesis. Chromosoma 112, 342–349 (2004). https://doi.org/10.1007/s00412-004-0279-3

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  • DOI: https://doi.org/10.1007/s00412-004-0279-3

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