Microspore Embryo Induction and Development in Higher Plants: Cytological and Ultrastructural Aspects

  • Christophe Clément
  • Rajbir S. Sangwan
  • Brigitte Sangwan-Norreel
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 56)


Zygotic Embryo Anther Culture Microspore Culture Pollen Wall Microspore Embryogenesis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alché JD, Castro AJ, Solymoss M, Timar I, Barnabas B, Rodríguez-García MI (2000) Cellular approach to the study of androgenesis in maize anthers: immunocytochemical evidence of the involvement of the ubiquitin degradative pathway in androgenesis induction. J Plant Physiol 156:146–155Google Scholar
  2. Binarova P, Hause G, Cenklova V, Cordewener JHG, van Lookeren Campagne MM (1997) A short severe heat shock is required to induce embryogenesis in late bicellular pollen of Brassica napus L. Sexual Plant Reprod 10:200–208CrossRefGoogle Scholar
  3. Cai Q, Szarejko I, Polok K, Maluszynski M (1992) The effect of sugars and growth regulators on embryoid formation and plant regeneration from barley anther culture. Plant Breed 109:218–226Google Scholar
  4. Caredda S, Clément C (1999) Androgenesis and albinism in Poaceae: influence of genotype and carbohydrates. In: Clément C, Pacini E, Audran JC (eds) Anther and pollen: from biology to biotechnology. Springer, Berlin Heidelberg New York, pp 211–226Google Scholar
  5. Caredda S, Devaux P, Sangwan RS, Clément C (1999) Differential development of plastids during microspore embryogenesis in barley. Protoplasma 208:248–256CrossRefGoogle Scholar
  6. Caredda S, Doncoeur C, Devaux P, Sangwan RS, Clément C (2000) Plastid differentiation during androgenesis in albino and non albino-producing cultivars of barley (Hordeum vulgare L.). Sexual Plant Reprod 13:95–104CrossRefGoogle Scholar
  7. Caredda S, Devaux P, Sangwan RS, Proult I, Clément C (2004) Plastid ultrastructure and DNA related to albinism in androgenetic embryos of various barley (Hordeum vulgare L.) cultivars. Plant Cell Tissue Organ Cult 76(1):35–43CrossRefGoogle Scholar
  8. Chu CC, Hill RD, Brule-Babel A (1990) High frequency of pollen embryoid formation and plant regeneration in Triticum aestivum L. on monosaccharide containing media. Plant Sci 66:255–262CrossRefGoogle Scholar
  9. Cistué L, Ramos A, Castillo AM, Romagosa I (1994) Production of large number of doubled haploid plants from barley anthers pretreated with high concentrations of mannitol. Plant Cell Rep 13:709–712Google Scholar
  10. Cordewener JHG, Custers JBM, Dons HJM, van Lookeren Campagne M (1996) Molecular and biochemical events during the induction of microspore embryogenesis. In: Jain MS, Sopory SK, Veilleux RE (eds) In vitro production of higher plants, vol 1. Kluwer, Dordrecht, pp 111–124Google Scholar
  11. Custers JBM, Cordewener JHG, Nöllen Y, Dons JJM, van Lookeren Campagne M (1994) Temperature controls both gametophytic and sporophytic development in microspore cultures of Brassica napus. Plant Cell Rep 13:267–271CrossRefGoogle Scholar
  12. Devaux P (1992) Haploidy in barley and wheat improvement. In: Dattée A (ed) Reproductive biology and plant breeding. Springer, Berlin Heidelberg New York, pp 139–151Google Scholar
  13. Devaux P, Zivy M, Kilian A, Kleinhofs A (1996) Doubled haploids in barley. In: Scoles G, Rossnagel B (eds) Proc 5th Int Oat Conf and 7th Int Barley Genet Symp. University Extenson Press, Saskatoon, Saskatchewan, Canada, pp 213–222Google Scholar
  14. Eady C, Lindsey K, Twell D (1995) The significance of microspore divisions and division symmetry for vegetative cell specific transcription and generative cell differentiation. Plant Cell 7:65–74PubMedCrossRefGoogle Scholar
  15. Garrido D, Vicente O, Heberle-Bors E, Rodriguez-Garcia I (1995) Cellular changes during the acquisition of embryogenic potential in isolated pollen grains of Nicotiana tabacum. Protoplasma 186:220–230CrossRefGoogle Scholar
  16. Goralski G, Matthys-Rochon E, Vergne P, Przywara L (1999) Androgenic development: a fascinating embryo formation process. Acta Biol Cracov 41:51–65Google Scholar
  17. Guha S, Maheshwari SC (1964) In vitro production of embryos anther of Datura. Nature 204:497–498Google Scholar
  18. Guha S, Maheshwari SC (1966) Cell division and differentiation of embryos in the pollen of Datura in vitro. Nature 212:97–98Google Scholar
  19. Hoekstra S, van Zijdervzld MH, Louwerse JD, Heidelkamp F, van der Mark F (1992) Anther and microspore culture of Hordeum vulgare L. Plant Sci 8:89–96Google Scholar
  20. Honys D, Twell D (2003) Comparative analysis of the Arabidopsis pollen transcriptome. Plant Physiol 132:640–652PubMedCrossRefGoogle Scholar
  21. Hu T, Kasha KJ (1999) A cytological study of pretreatments used to improve isolated microspores cultures of wheat (Triticum aestivum L.) cv. Chris. Genome 42:432–441CrossRefGoogle Scholar
  22. Ilic-Grubor K, Attree M, Fowke LC (1998) Comparative morphological study of zygotic and microspore-derived embryos of Brassica napus L. as revealed by scanning electron microscopy. Ann Bot 82:157–165Google Scholar
  23. Indrianto A, Barinova I, Touraev A, Heberle-Bors E (2001) Tracking individual wheat microspores in vitro: identification of embryogenic microspores and body axis formation in the embryo. Planta 212:163–174PubMedCrossRefGoogle Scholar
  24. Jacquard C, Wojnarowiez G, Clément C (2003) Anther culture in barley. In: Kasha K, Maluszinski M (eds) Doubled haploid manual. Springer, Berlin Heidelberg New York, pp 21–28Google Scholar
  25. Jähne A, Lörz H(1995) Cereal microspore culture. Plant Sci 109:1–12Google Scholar
  26. Jardinaud MF, Souvré A, Alibert G, Beckert M (1995a) uidA gene transfer and expression in maize microspores using the biolistic method. Protoplasma 187:138–143CrossRefGoogle Scholar
  27. Jardinaud MF, Souvré A, Alibert G, Beckert M (1995b) Optimisation of DNA transfer and transient ß-glucuronidase expression in electroporated maize (Zea mays L.) microspores. Plant Cell Rep 15:55–58CrossRefGoogle Scholar
  28. Kao KN (1993) Viability, cell division and microcallus formation of barley microspores in culture. Plant Cell Rep 12:366–369CrossRefGoogle Scholar
  29. Kasha KJ, Yao Q, Simion E, Hu T, Oro R (1995) Production and applications of haploids in crops. Use of induced mutation and molecular techniques of crop improvement. International Atomic Energy Agency, Vienna, Austria, pp 23–37Google Scholar
  30. Knox RB (1971) Pollen wall proteins: localization, enzymic and antigenic activity during development in Gladiolus. J Cell Sci 9:209–237PubMedGoogle Scholar
  31. Kumlehn J, Lörz H (1999) Monitoring sporophytic development of individual microspores of barley (Hordeum vulgare L.). In: Clément C, Pacini E, Audran JC (eds) Anther and pollen: from biology to biotechnology. Springer, Berlin Heidelberg New York, pp 183–190Google Scholar
  32. Lee JY, Lee DH (2003) Use of serial analysis of gene expression technology to reveal changes in gene expression in Arabidopsis pollen undergoing cold stress. Plant Physiol 132:517–529PubMedGoogle Scholar
  33. Loeb TA, Reynolds TL (1994) Transient expression of the uidA gene in pollen embryoids of wheat following microprojectile bombardment. Plant Sci 104:81–91CrossRefGoogle Scholar
  34. Luckett DJ, Darvey NL (1992) Utilisation of microspore culture in wheat and barley improvement. Aust J Bot 40:807–828CrossRefGoogle Scholar
  35. Luckett DJ, Smithard RA (1992) Doubled haploid production by anther culture for Australian barley breeding. Aust J Agric Res 43:67–78Google Scholar
  36. Lyne RL, Bennett RI, Hunter CP (1986) Embryoid and plant production from cultured barley anthers. In: Withers LA, Alderson PG (eds) Plant tissue culture and its agricultural application. Butterworths, London, pp 405–410Google Scholar
  37. Mackenzie A, Heslop-Harrison J, Dickinson HG (1967) Elimination of ribosomes during meiotic prophase. Nature 215:997–999PubMedGoogle Scholar
  38. 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 at early stages of microspore embryogenesis in maize. Plant Mol Biol 44:559–574PubMedCrossRefGoogle Scholar
  39. Mogensen HL (1996) The hows and whys of cytoplasmic inheritance in seed plants. Am J Bot 83:383–404Google Scholar
  40. Mordhorst AP, Lörz H (1993) Embryogenesis and development of isolated barley (Hordeum vulgare L.) microspores are influenced by the amount and composition of nitrogen sources in culture media. J Plant Physiol 142:485–492Google Scholar
  41. Norreel B (1970) Etude cytologique de l’androgenèse expérimentale chez Nicotiania tabacum et Datura innoxia. Bull Soc Bot Fr 117:461–478Google Scholar
  42. Paire A, Devaux P, Lafitte C, Dumas C, Matthys-Rochon E (2003) Proteins produced by barley microspores and their derived androgenic structures promote in vitro zygotic maize embryo formation. Plant Cell Tissue Organ Cult 73:167–176CrossRefGoogle Scholar
  43. Pickering RA, Devaux P (1992) Haploid production: approaches and use in plant breeding. In: Shewry PR (ed) Barley: genetics, biochemistry, molecular biology and biotechnology. CAB International, Oxford, pp 519–547Google Scholar
  44. Raghavan V (ed) (1976a) Experimental embryogenesis in vascular plants. Academic Press, LondonGoogle Scholar
  45. Raghavan V (1976b) Role of generative cell in androgenesis in henbane. Science 191:388–389Google Scholar
  46. Salmenkallio-Marttila M, Kauppinen V (1995) Efficient regeneration of fertile plants from protoplasts isolated from microspore cultures of barley (Hordeum vulgare L.). Plant Cell Rep 14:253–256CrossRefGoogle Scholar
  47. Sangwan RS (1986) Formation and cytochemistry of nuclear vacuoles during meiosis in Datura. Eur J Cell Biol 40:210–218Google Scholar
  48. Sangwan RS, Camefort H (1982) Ribosomal bodies specific to both pollen and zygotic embryogenesis in Datura. Experientia 38:395–397CrossRefGoogle Scholar
  49. Sangwan RS, Camefort H (1983) The tonoplast, a specific marker of embryogenic microspores of Datura cultured in vitro. Histochemistry 78:473–480PubMedCrossRefGoogle Scholar
  50. Sangwan RS, Camefort H (1984) Cold-treatment related structural modifications in the embryogenic anthers of Datura. Cytologia 49:473–487Google Scholar
  51. Sangwan RS, Sangwan-Norreel BS (1980) Biochemical and structural aspects of induction and development of pollen-embryos in Datura. Bull Soc Bot Fr 127:109–122Google Scholar
  52. Sangwan RS, Sangwan-Norreel BS (1987a) Biochemical cytology of pollen embryogenesis. Int Rev Cytol 107:221–272Google Scholar
  53. Sangwan RS, Sangwan-Norreel BS (1987b) Ultrastructural cytology of plastids in pollen grains of certain androgenic and nonandrogenic plants. Protoplasma 138:11–22CrossRefGoogle Scholar
  54. Sangwan RS, Mathivet V, Vasseur G (1989) Ultrastructural localization of acid phosphatase during male meiosis and sporogenesis in Datura: evidence for digestion of cytoplasmic structures in the vacuoles. Protoplasma 149:38–46CrossRefGoogle Scholar
  55. Sangwan RS, Ducrocq C, Sangwan-Norreel BS (1993) Agrobacterium-mediated transformation of pollen embryos in Datura innoxia and Nicotiana tabacum: production of transgenic haploid and fertile homozygous dihaploid plants. Plant Sci 95:99–115CrossRefGoogle Scholar
  56. Sangwan-Norreel BS (1978) Cytochemical and ultrastructural peculiarities of embryogenic pollen grains and of young androgenic embryos in Datura innoxia. Can J Bot 56(7):805–817Google Scholar
  57. Sangwan-Norreel BS (1981) Evolution in vitro du contenu en ADN nucléaire et de la ploïdie des embryons polliniques du Datura innoxia. Can J Bot 59(4):508–517Google Scholar
  58. Sangwan-Norreel BS (1983) Male gametophyte nuclear DNA content evolution during androgenic induction in Datura innoxia Mill. Z Pflanzenphysiol 111:47–54Google Scholar
  59. Shim YS, Kasha KJ (2003) The influence of pretreatment on cell stage progression and the time of DNA synthesis in barley (Hordeum vulgare L.) uninucleate microspores. Plant Cell Rep 21:1065–1071PubMedCrossRefGoogle Scholar
  60. Testillano PS, Gonzalez-Melendi P, Ahmadian P, Fadon B, Risueno MC (1995) The immunolocalization of nuclear antigens during the pollen developmental programme and the induction of pollen embryogenesis. Exp Cell Res 221:41–54PubMedCrossRefGoogle Scholar
  61. Testillano PS, Ramirez C, Domenech J, Coronado MJ, Vergne P, Matthys-Rochon E, Risueno MC (2002) Young microspore-derived embryos show two domains with defined features also present in zygotic embryogenesis. Int J Dev Biol 46:1035–1047PubMedGoogle Scholar
  62. Touraev A, Vicente O, Heberle-Bors E (1997) Initiation of microspore embryogenesis by stress. Trends Plant Sci 2:297–302CrossRefGoogle Scholar
  63. Vazart B (1973) Formation d’embryoídes à partir de microspores de tabac: évolution de l’infrastructure des cellules au cours de la première semaine de culture. Mémoire de la Société Botanique Française, Colloque de Morphologie, Paris, pp 243–260Google Scholar
  64. Wang M, van Bergen S, Lamers GEM, Oppedjik BJ, Schilperroort RA (1999a) Programmed cell death during androgenesis in Hordeum vulgare L. In: Clément C, Pacini E, Audran JC (eds) Anther and pollen: from biology to biotechnology. Springer, Berlin Heidelberg New York, pp 201–209Google Scholar
  65. Wang M, Hoekstra S, van Bergen S, Lamers GEM, Oppedijk BJ, de Priester W, Schilperroort RA (1999b) Apoptosis in developing anthers and the role of ABA in this process during androgenesis in Hordeum vulgare L. Plant Mol Biol 39:489–501PubMedCrossRefGoogle Scholar
  66. Wojnarowiez G, Jacquard C, Devaux P, Sangwan RS, Clément C (2002) Influence of copper sulfate on anther culture in barley (Hordeum vulgare L.). Plant Sci 162:843–847CrossRefGoogle Scholar
  67. Xie JH, Gao MW, Cai QH, Cheng X, Shen Y, Liang ZQ (1995) Improved isolated microspore culture efficiency in medium with maltose and optimized growth regulator combination in japonica rice (Oryza sativa). Plant Cell Tissue Organ Cult 42:245–250CrossRefGoogle Scholar
  68. Yao QA, Simion E, William M, Krochko J, Kasha KJ (1997) Biolistic transformation of haploid microspores of barley (Hordeum vulgare L.). Genome 40:570–581Google Scholar
  69. Zarsky V, Rihova L, Tupy J (1990) Biochemical and cytological changes in young tobacco pollen during in vitro starvation in relation to pollen embryogenesis. In: Van der Plas LHW, vanAartri JK (eds) Progress in plant cellular and molecular biology. Kluwer, Dordrecht, pp 228–233Google Scholar
  70. Zarsky V, Garrido D, Rihova L, Vincente O, Heberle-Bors E (1992) Derepression of the cell cycle by starvation is involved in the induction of tobacco pollen embryogenesis. Sexual Plant Reprod 4:189–194Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2005

Authors and Affiliations

  • Christophe Clément
    • 1
  • Rajbir S. Sangwan
    • 2
  • Brigitte Sangwan-Norreel
    • 2
  1. 1.UFR Sciences, Biologie et Physiologie VégétalesUniversité de Reims Champagne ArdenneReims Cedex 2France
  2. 2.Androgenése et BiotechnologiesUniversité de Picardie Jules VerneAmiensFrance

Personalised recommendations