Wheat pp 435-447 | Cite as

Culture of Isolated Pollen of Wheat (Triticum aestivum L.)

  • S. K. Datta
  • I. Potrykus
  • M. Bolik
  • G. Wenzel
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 13)


Pollen culture provides a system with haploid single cells and thus is a promising haplopahase tool for basic and applied biotechnological breeding programs. Anther culture (Nitzsche and Wenzel 1977; de Buyser and Henry 1980, 1986; Maheshwari et al. 1980; Bajaj 1983; Schaeffer et al. 1984; Wenzel and ForoughiWehr 1984; Bajaj and Gosal 1986; Dunwell 1986; Han 1986; Keller et al. 1987) is one of the methods of haploid induction most commonly used now for crop improvement. Culture of whole spikes (Wilson 1977; Brettel et al. 1980, Ozias-Akins and Vasil 1982; Datta 1987) may simplify this technique. The third way is the culture of isolated microspores. However, in most reports the microspores are cultured within the anthers. Only a rather limited number of reports have been published on the successful use of isolated microspore culture in cereals (Köhler and Wenzel 1985; Wei et al. 1986; Datta and Wenzel 1987, 1988; Datta and Potrykus 1988). While regeneration of plants from single cells of cereals is still a problem, plant regeneration from in vitro cultures originating from multicellular explants has been achieved from all the major cereal crops (Vasil 1983; Lörz et al. 1988). Plant regeneration from microspores of wheat follows the two general pathways of in vitro culture, viz organogenesis and embryogenesis. The latter one is preferred for its unicellular origin and production of non-chimeric plants.


Somatic Embryogenesis Plant Regeneration Anther Culture Isolate Microspore Culture Pollen Culture 
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  1. Bajaj YPS (1978) Regeneration of haploid tobacco plants from isolated pollen grown in drop culture. Indian J Exp Biol 16: 407–409Google Scholar
  2. Bajaj YPS (1983) In vitro production of haploids. In: Evans DA, Sharp WR, Ammirato PV, Yamada Y (eds) Handbook of plant cell culture, vol 1: Techniques for propagation and breeding. MacMillan, New York, pp 228–287Google Scholar
  3. Bajaj YPS, Gosal SS (1986) Biotechnology of wheat improvement. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 2: Crops 1. Springer, Berlin Heidelberg New York, Tokyo, pp 3–38Google Scholar
  4. Bajaj YPS, Reinert J, Heberle E (1977) Factors enhancing in vitro production of haploid plants in anthers and isolated microspores. In: Gautheret RJ (ed) La culture des tissus et des cellules des végétaux. Masson, Paris New York, pp 47–58Google Scholar
  5. Brettell RIS, Wernicke W, Thomas E (1980) Embryogenesis from cultured immature inflorescence of Sorghum bicolor. Protoplasma 104: 141–148CrossRefGoogle Scholar
  6. Chen Y, Wang RF, Tian WZ, Zuo QX, Zheng SW, Ludeyang ZG (1980) Studies on pollen culture in vitro and induction of plants in Oryza sativa subsp. Keng. Acta Genet Sin 7: 46–54Google Scholar
  7. Chu CC (1978) The N6 medium and its application to anther cultures of cereal crops. In: Proc Symp Plant tissue culture. Science Press, Beijing, pp 45–50Google Scholar
  8. Chuang CC, Ouyang TW, Chia H, Chou SM, Ching CK (1978) A set of potato media for wheat anther culture. In: Proc Symp Plant tissue culture. Science Press, Beijing, pp 51–56Google Scholar
  9. Clapham D (1971) In vitro development of callus from the pollen of Lolium and Hordeum. Z Pflanzenzücht65: 285–292Google Scholar
  10. Datta SK (1987) Plant regeneration by pollen embryogenesis from cultured whole spikes of barley (Hordeum vulgare). Theor Appl Genet 74: 121–124CrossRefGoogle Scholar
  11. Datta SK, Datta K, Potrykus I (1990) Embryogenesis and plant regeneration from mocrospores of both “Indica” and “Japonica” rice (Oryza saliva). Plant Sci 67: 83–88CrossRefGoogle Scholar
  12. Datta SK, Potrykus I (1988) Direct embryogenesis from pollen of cereals. Experientia 44: 43 (PLA 180 Abstr)Google Scholar
  13. Datta SK, Potrykus I (1989) Artificial seeds in barley: encapsulation of microspore-derived embryos. Theor Appl Genet 77: 820–824CrossRefGoogle Scholar
  14. Datta SK, Wenzel G (1987) Isolated microspore derived plant formation in Triticum aestivum L. Plant Sci 48: 49–54CrossRefGoogle Scholar
  15. Datta SK, Wenzel G (1988) Single microspore derived embryogenesis and plant formation in barley (Hordeum vulgare). Arch Züchtungsforsch 18: 125–131Google Scholar
  16. de Buyser J, Henry Y (1980) Comparison of different media used in culturing anthers in vitro in soft wheat. Can J Bot 58: 997–1000CrossRefGoogle Scholar
  17. de Buyser J, Henry Y (1986) Wheat. Production of haploids, performance of doubled haploids, and yield trials. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 2: Crops 1: Springer, Berlin Heidelberg New York Tokyo, pp 73–88Google Scholar
  18. Dunwell JM (1986) Pollen, ovule and embryo culture as a tool in plant breeding. In: Withers LA, Anderson PG (eds) Plant tissue culture and its agricultural applications. Butterworths, London, pp 375–404Google Scholar
  19. Foroughi-Wehr B, Mix G, Gaul H, Wilson HM (1976) Plant production from cultured anthers of Hordeum vulgare L. Z Pflanzenzücht 77: 198–204Google Scholar
  20. Fuji JAA, Slada DT, Redenbaugh K, Walker KA (1987) Artificial seeds for plant propagation. Trends in Biotech 5: 335–339CrossRefGoogle Scholar
  21. Ghandimathi H (1982) Direct pollen culture of Nicotiana tabacum. In: Fujimura A (ed) Plant tissue culture 1982. Maruzen, Tokyo, pp 527–528Google Scholar
  22. Guha-Mukherjee S (1973) Genotypic differences in the in vitro formation of embryoids from rice pollen. J Exp Bot 24: 139–144CrossRefGoogle Scholar
  23. Han H (1986) Wheat improvement through anther culture. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 2: Crops 1. Springer, Berlin Heidelberg New York Tokyo, pp 55–72Google Scholar
  24. Imamura J, Okabe E, Kyo M, Harada H (1982) Embryogenesis and plantlet formation through direct culture of isolated pollen of Nicotiana tabacum. cv.Google Scholar
  25. Samsun and Nicotiana rustica cv. Rustica. Plant Cell Physiol 23:713–716Google Scholar
  26. Jensen CJ (1974) Chromosome doubling techniques in haploids. In: Kasha KJ (ed) Haploids in higher plants — advances and potential. Univ Press, Guelph, pp 153–190Google Scholar
  27. Kao KN (1981) Plant formation from barley anther cultures with Ficoll media. Z Pflanzenphysiol 103: 437–443Google Scholar
  28. Keller WA, Arnison PG, Brian JC (1987) Haploids from gametophytic cells — recent developments and future prospects. In: Green CE, Somers DA, Hackett WP, Biesboer DD (eds) Plant tissue and cell culture. Liss, New York, pp 223–241Google Scholar
  29. Kitto S, Janick J (1985) Production of synthetic seeds by encapsulating asexual embryos of carrot. J Am Soc Hortic Sci 110: 277–282Google Scholar
  30. Kohlenbach HW (1978) Comparative somatic embryogenesis. In: Thorpé TA (ed) Frontiers of plant tissue culture. Univ Press, Calgary, pp 59–66Google Scholar
  31. Köhler F, Wenzel G (1985) Regeneration of isolated barley microspores in conditioned media and trials to characterize the responsible factor. J Plant Physiol 121: 181–191CrossRefGoogle Scholar
  32. Kyo M, Harada H (1985) Studies on conditiones for cell division and embryogenesis in isolated pollen culture of Nicotiana rustica. Plant Physiol 79: 90–94PubMedCrossRefGoogle Scholar
  33. Kyozuka J, Hayashi Y, Shimamoto Ko (1987) High frequency plant regeneration from rice protoplasts by novel nurse culture methods. Mol Gen Genet 206: 408–413CrossRefGoogle Scholar
  34. Lichter R (1982) Induction of haploid plants from isolated pollen of Brassica napus. Z Pflanzenphysiol 105: 427–434Google Scholar
  35. Lörz H, Göbel E, Brown P (1988) Advances in tissue culture and progress towards genetic transformation of cereals. Plant Breed 100: 1–25CrossRefGoogle Scholar
  36. Maheshwari SC, Tyagi AK, Malhotra K, Sopory SK (1980) Induction of haploidy from pollen grain in angiosperms — the current status. Theor Appl Genet 58: 193–206CrossRefGoogle Scholar
  37. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15: 473–497CrossRefGoogle Scholar
  38. Neuhaus G, Spangenberg G, Mittelsten Scheid O, Schweiger HG (1987) Transgenic rapeseed plants obtained by the microinjection of DNA into microspore derived embryoids. Theor Appl Genet 75: 30–36CrossRefGoogle Scholar
  39. Nitsch C (1974) Pollen culture a new technique for mass production of haploid and homozygous plants. In: Kasha KJ (ed) Haploids in higher plants — advances and potential. Univ Press, Guelph, pp 123–135Google Scholar
  40. Nitzsche W, Wenzel G (1977) Haploids in plant breeding. Parey, BerlinGoogle Scholar
  41. Olsen FL (1987) Induction of microspore embryogenesis in cultured anthers of Hordeum vulgare. The effect of ammonium nitrate, glutamine and asparagine as nitrogen source. Carlsberg Res Commun 52: 393–404CrossRefGoogle Scholar
  42. Ozias-Akins P, Vasil IK (1982) Plant regeneration from cultured immature embryos and inflorescences of Triticum aestivum L. (wheat): evidence for somatic embryogenesis. Protoplasma 110: 95–105CrossRefGoogle Scholar
  43. Pelletier G (1973) Les conditions et les premiers standes de l’androgenèse in vitro chez Nicotiana tabacum. Mem Soc Bot Fr 261–268Google Scholar
  44. Potrykus I (1988) Direct gene transfer to plants. In: Application of plant cell and tissue culture (Ciba Foundation Symposium 137 ). John Wiley & Sons, New York, pp 144–162Google Scholar
  45. Potrykus I, Paszkowski J, Saul M, Kruger-Lebeus, Muller T, Schocher R, Negrutiu I, Kunzler P, Shillito R (1985) Direct gene transfer to protoplasts: an efficient and generally applicable method for stable alteration of plant genomes. In: Freeling M (ed) Plant genetics. Liss, New York, pp 181–199Google Scholar
  46. Redenbaugh K, Brian DP, Nichol JW, Kossler ME, Viss PR, Walker KA (1986) Somatic seeds: encapsulation of asexual plant embryos. Bio/technology 4: 643–645CrossRefGoogle Scholar
  47. Reinert J, Bajaj YPS (eds) (1977) Applied and fundamental aspects of plant cell, tissue, and organ culture. Springer, Berlin Heidelberg New YorkGoogle Scholar
  48. Schaeffer GW, Lazar MD, Baenziger PS (1984) Wheat. In: Sharp WR, Evans DA, Ammirato PV, Yamada Y (eds) Handbook of plant cell culture, vol 2: Crop species. MacMillan, New York, pp 108–136Google Scholar
  49. Sharp WR (1972) The use of nurse culture in the development of haploid clones in tomato. Planta 104: 357–361CrossRefGoogle Scholar
  50. Sopory SK (1977) Development ofembryoids in isolated pollen culture ofdihaploid Solanum tuberosum. Z Pflanzenphysiol 84: 453–457Google Scholar
  51. Sunderland N, Huang B (1985) Barley anther culture–the switch of programme and albinism. Hereditas Suppl 3: 27–40Google Scholar
  52. Swanson EB, Coumans MP, Wu SC, Barsby TL, Beversdorf WD (1987) Efficient isolation of microspore and the production of microspore-derived embryos from Brassica napus. Plant Cell Rep 6: 94–97Google Scholar
  53. Vasil IK (1983) Regeneration of plants from single cells of cereals and grasses. In: Lurquin PF, Kleinhops A (eds) Genetic engineering in eukaryotes. Plenum, New York, pp 232–252Google Scholar
  54. Vasil IK (1986) Relative genetic stability of embryogenic cultures of the Gramineae and uniformity of regenerated plants. In: Semal J (ed) Somaclonal variations and crop improvement. Nijhoff, Dordrecht Boston Lancaster, pp 108–116Google Scholar
  55. Wang RF, Zuo QX, Zheng SW, Tian WZ (1979) Induction of plantlets from isolated pollen culture in rice (Oryza sativa subsp. Keng ). Acta Genet Sin 6: 7Google Scholar
  56. Wei ZM, (1982) Pollen callus culture in Triticum aestivum. Theor Appl Genet 63: 71–73CrossRefGoogle Scholar
  57. Wei ZM, Kyo M, Harada H (1986) Callus formation and plant regeneration through direct culture of isolated pollen of Hordeum vulgare cv. Sabarlis. Theor Appl Genet 72: 252–255Google Scholar
  58. Wenzel G, Foroughi-Wehr (1984) Anther culture of cereals and grasses. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants, vol 1. Academic Press, New York London, pp 311–327Google Scholar
  59. Wenzel G, Hoffmann F, Potrykus I, Thomas E (1975) The separation of viable rye microspores from mixed populations and their development in culture. Mol Gen Genet 138: 293–297CrossRefGoogle Scholar
  60. Wetherell DF, Dougall DK (1976) Sources of nitrogen supporting growth and embryogenesis in cultured wild carrot tissue. Physiol Plant 37: 97–103CrossRefGoogle Scholar
  61. Wilson HM (1977) Culture of whole barley spikes stimulates high frequencies of pollen calluses in individual anthers. Plant Sci Lett 9: 233–238CrossRefGoogle Scholar
  62. Xu ZH, Sunderland N (1981) Glutamine, inositol and conditioning factor in the production of barley pollen callus in vitro. Plant Sci Lett 23: 161–168CrossRefGoogle Scholar
  63. Ye JM, Kao KN, Harvey BL, Rossnagel BG (1987) Screening salt tolerant barley genotypes via Fl anther culture in salt stress media. Theor Appl Genet 74: 426–429CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1990

Authors and Affiliations

  • S. K. Datta
    • 1
  • I. Potrykus
    • 1
  • M. Bolik
    • 2
  • G. Wenzel
    • 2
  1. 1.Swiss Federal Institute of TechnologyETH, Institute of Plant SciencesZürichSwitzerland
  2. 2.Institute for Resistance GeneticsBBAGrünbach-BockhornGermany

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