Wheat (Triticum aestivum): In Vitro Production and Utilization of Doubled Haploids

  • E. Picard
  • A. Rode
  • G. Doussinault
  • M. Rousset
  • M. Rives
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 12)


The literature ofthe last 15 years on hexaploid wheat (Triticum aestivum L. Thell. em.) on in vitro techniques and more precisely on anther culture, has been discussed in four chapters of a second Volume of this Series (Bajaj and Gosal 1986; Kudirka et al. 1986; Hu Han 1986; de Buyser and Henry 1986). There has been tremendous progress in haploid production by anther culture since the first wheat pollen plants were produced simultaneously in China by Ouyang et al., and in France by Picard and de Buyser in 1973. For example, special lines used in Chinese programs such as Ciano, Orofen, and Huapei no. 1, are now currently yielding very high rates of embryos and regenerated plantlets. Ouyang et al. (1983) reported % EMB (percentage of embryos related to the number of cultivated anthers) reaching 120% and % GRPL (percentage of green plantlets related to the total cultivated anthers) to be as high as 10–70%. Using European genetic material, likewise, the results in % GRPL sometimes reached 2–10% as was the case for a F9 line B described by Henry and de Buyser (1985), or for the cross Highbury-5B Chinese SpringxSicco-5B Chinese Spring reported by Snape et al. (1986). In most cases, however, the European wheat material gave rather lower frequencies of DH lines with an average yield of 0.05% to 0.1 % GRPL. Important progress remains to be made in the understanding of the genetic and physiological components of the phenomenon in order to increase the efficiency of anther culture in this cereal.


Triticum Aestivum Double Haploid Anther Culture Recurrent Selection Cold Pretreatment 
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. Bajaj YPS (1977) In vitro induction of haploids in wheat (Triticum aestivum L.). Crop Improv 4:54 - 64Google Scholar
  2. Bajaj YPS (1984) The regeneration of plants from frozen pollen embryos and zygotic embryos of wheat and rice. Theor Appl Genet 67: 525 - 528CrossRefGoogle Scholar
  3. Bajaj YPS, Gosal SS (1986) Biotechnology of wheat improvement. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry 2. Crops I. Springer, Berlin Heidelberg New York Tokyo, pp 3 - 38Google Scholar
  4. Belliard G, Vedel F, Pelletier G (1979) Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature (Lond) 281: 401 - 403CrossRefGoogle Scholar
  5. Bennet MD, Hughes WG (1972) Additional mitosis in wheat pollen induced by Ethre!. Nature (Lond) 240:566 - 568Google Scholar
  6. Buyser de J, Henry Y (1986) Wheat: Production of haploids, performance of doubled haploids and yield trials. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry 2. Crops I. Springer, Berlin Heidelberg New York Tokyo, pp 73 - 85Google Scholar
  7. Buyser de J, Picard E (1975) Observation de divisions supplementaires dans les grains de pollen de plantes homozygotes de Ble tendre (Triticum aestivum L.) obtenues par androgenese in vitro. CR Acad Sci Paris, Ser 0 281: 1153 - 1156Google Scholar
  8. Buyser de J, Henry Y, Thleb G (1985) Wheat androgenesis: cytogenetical analysis and agronomic performance of doubled haploids. Z Pflanzenziicht 95: 23 - 34Google Scholar
  9. Buyser de J, Henry Y, Lonnet P, Hertzog R, Hespel A (1987) "Florin": A doubled haploid wheat variety developed by the anther culture method. Plant Breed 98:53 - 56Google Scholar
  10. Charmet G, Branlard G (1985) A comparison of androgenetic doubled haploid and single seed descent lines in Triticale. Theor Appl Genet 71: 193 - 200Google Scholar
  11. Charmet G, Vedel F, Bernard M, Bernard S, Mathieu C (1985) Cytoplasmic variability in androgenetic doubled haploid lines of triticale. Agronomie 5:709-717 Choo TM, Reinbergs E (1979) Doubled haploids for estimating genetic variances in presence of linkage and gene association. Theor Appl Genet 55: 129 - 132Google Scholar
  12. Choo TM, Christie BR, Reinbergs E (1979) Doubled haploids for estimating variances and a scheme for population improvement in self-pollinated species. Theor Appl Genet 54: 267 - 271CrossRefGoogle Scholar
  13. Collins GB, Genovesi AD (1982) Anther culture and its application to crop improvement. In: TomesGoogle Scholar
  14. DR, Ellis BE, Harney PM, Kasha KJ, Peterson RL (eds) Application of plant cell and tissue culture to agriculture and industry. University of Guelph pub!" pp 1-24Google Scholar
  15. Datta SK, Wenzel G (1987) Isolated microspore derived plant formation via embryogenesis in Triticum aestivum L. Plant Sci 48 (1): 49 - 54CrossRefGoogle Scholar
  16. Day A, Ellis THN (1984) Chloroplast DNA deletions associated with wheat plants regenerated from pollen: possible basis for maternal inheritance of chroplasts. Cell 39: 359 - 368PubMedCrossRefGoogle Scholar
  17. Deaton WR, Metz SG, Armstrong TA, Mascia PN (1987) Genetic analysis of the anther-culture response of three spring wheat crosses. Theor Appl Genet 74:334- 338Google Scholar
  18. Dhillon SS, Wernsman EA, Miksche JP (1983) Evaluation of nuclear DNA content and heterochromation changes in anther-derived dihaploids of tobacco (Nicotiana tabacum) cv. "Coker 139". Can J Genet CytoI25: 169 - 173Google Scholar
  19. Doussinault G, nottet M (1981) Application de la selection recurrente aux especes autogames ou allogames non strictes: allogamisation des autogames. Sel Fran~ais 29: 25 - 33Google Scholar
  20. Dunwell JM (1978) Division and differentiation in cultured pollen. In: Thorpe TA (ed) Frontiers of plant tissue culture 1978. Univ Press, Calgary, Canada, pp 102 - 112Google Scholar
  21. Evans DA, Sharp WR, Medina-Filho HP (1984) Somoclonal and gametoclonal variation. Am J Bot 71: 759 - 774CrossRefGoogle Scholar
  22. Foroughi-Wehr B, Friedt W, Wenzel G (1982) On the genetic improvement of androgenetic haploid formation in Hordeum vulgare L. Theor Appl Genet 62: 233 - 239Google Scholar
  23. Galiba G, Kovacs G, Sutka J (1986) Substitution analysis of plant regeneration from callus culture in wheat. Plant Breed 97: 261 - 263CrossRefGoogle Scholar
  24. Gallais A (1977) Amelioration des populations, methodes de selection et creation de varietes I. Ann Amelior Plant 27:281- 329Google Scholar
  25. Gallais A (1978a) Place de l'haploidie dans un schema. 8e1 Francais 26: 39 - 49Google Scholar
  26. Gallais A (1978b) Amelioration des populations, methodes de selection et creation de varietes II. Ann Amelior Plant 28: 637 - 666Google Scholar
  27. Gallais A (1979) The concept of varietal ability in plant breeding. Euphytica 28: 811 - 823CrossRefGoogle Scholar
  28. Gallais A (1987) Place de I'haploldisation dans les schemas de selection. Sel Franr;:cais 36: 47 - 58Google Scholar
  29. Gegenbach BG, Connelly JA, Pring DR, Conde MF (1981) Mitochondrial DNA variation in maize plants regenerated during tissue culture selection. Theor Appl Genet 59: 161 - 167CrossRefGoogle Scholar
  30. Griffing B (1975) Efficiency changes due to use of doubled-haploids in recurrent selection methods. Theor Appl Genet 46:367 - 386Google Scholar
  31. Grignac P, Poux J, Thmas A (1978) Comparaison de differentes methodes de selection utilisees pour I'amelioration des varietes de ble. Ann Amelior Plant 28 (4): 341 - 350Google Scholar
  32. Heberle-Bors E (1985) In vitro haploid formation from pollen: a critical review. Theor Appl Genet 71: 361 - 374CrossRefGoogle Scholar
  33. Heberle-Bors E, Odenbach W (1985) In vitro pollen embryogenesis and cytoplasmic male sterility in Triticum aestivum L. Z Pflanzenziicht 95: 14 - 22Google Scholar
  34. Henry Y, Buyser de J (1985) Effect of the 1 Bll R translocation on anther culture ability in wheat (Triticum aestivum L.). Plant Cell Rep 4: 307 - 310CrossRefGoogle Scholar
  35. Henry Y, Buyser de J, Guenegou T, Ory C (1984) Wheat microspore embryogenesis during in vitro anther culture. Theor AppJ Genet 67: 439 - 442Google Scholar
  36. Hu Daofen (1986) Jinghua no. 1, a winter wheat derived from pollen sporophyte. In: Hu Han, YangGoogle Scholar
  37. Hongyuan (eds) Haploids in higher plants in vitro. China Academic Publishers Beijing, Springer, Berlin Heidelberg New York Tokyo, pp 137-145Google Scholar
  38. Hu Han (1986) Wheat: Improvement through anther culture. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry 2. Crops I. Springer, Berlin Heidelberg New York Tokyo, pp 55 - 72Google Scholar
  39. Hu Han, Xi ZY, Jing JK, Wang XZ (1983) Production of wheat pollen-derived aneuploid plants through anther culture. Cell and tissue culture techniques for cereal crop improvement. Sci Press, Beijing, China, pp 173 -182Google Scholar
  40. Jones AM, Petolino JF (1987) Effects of donor plant genotype and growth environment on anther culture of soft-red winter wheat (Triticum aestivum L.). Plant cell, tissue and org cult 8 (3): 215 - 223CrossRefGoogle Scholar
  41. Kemble RJ, Flavell RB, Brettel RIS (1982) Mitochondrial DNA analyses of fertile and sterile maize plants derived from tissue culture with the texas male sterile cytoplasm. Theor Appl Genet 62: 213 - 217Google Scholar
  42. Knott DR, Kumar J (1975) Comparison of early generation yield testing and a single seed descent procedure in wheat breeding. Crop Sci 15: 295 - 299CrossRefGoogle Scholar
  43. Kudirka DT, Schaeffer GW, Baenziger PS (1986) Wheat: Genetic variability through anther culture. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry 2. Crops I. Springer, Berlin Heidelberg New York Tokyo, pp 39 - 53Google Scholar
  44. Larkin PJ, Scowcroft WR (1981) Somoclonal variation - a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60: 187 - 214CrossRefGoogle Scholar
  45. Lazar MD, Baenziger PS, Schaeffer GW (1985) The physical environment in relation to high frequency callus and plantIet development in anther cultures of wheat (Triticum aestivum L.) cv. Chris. J Plant Physiol 121 (2): 103 - 109CrossRefGoogle Scholar
  46. Lazar MD, Schaeffer GW, Baenziger PS (1984 a) Cultivar and cultivar x environments effects on the development of callus and polyhaploid plants from anther culture of wheat. Theor Appl Genet 67: 273 - 277Google Scholar
  47. Lazar MD, Baenziger PS, Schaeffer GW (1984 b) Combining abilities and heritability of callus formation and plantlet regeneration in wheat (Triticum aestivum L.) anther cultures. Theor Appl Genet 68:131-134 Marsolais AA, Kasha KJ (1983) The influence of cold pretreatments and plants genotypes on in vitro androgenesis in wheat. Agron Abstr Madison, Wisconsin, USA. Am Soc Agron 73Google Scholar
  48. Marsolais AA, Seguin-Swartz G, Kasha KJ (1984) The influence of anther cold pretreatments and donor plant genotypes on in vitro androgenesis in wheat (Triticum aestivum L.). Plant Cell, Tissue and Organ Culture 3 (1): 69 - 79CrossRefGoogle Scholar
  49. Mathias RJ, Fukui K (1986) The effect of specific chromosomes and cytoplasm substitutions on the tissue culture response of wheat (Triticum aestivum) callus. Theor Appl Genet 71: 797 - 800CrossRefGoogle Scholar
  50. McNay JW, Chourey PS, Pring DR (1984) Molecular analysis of genomic stability of mitochondrial DNA in tissue cultured cells of maize. Theor Appl Genet 67: 433 - 437CrossRefGoogle Scholar
  51. Moreas-Fernandes MIB, Picard E (1983) Variability of haploid production bYiUlther culture using brasilian wheat genotypes. Rev Brasil Genet VI 2: 261 - 277Google Scholar
  52. Ouyang JW (1986) Induction of pollen plants in Triticum aestivum. In: Hu Han, Yang Hongyan (eds) Haploids in higher plants in vitro. Springer, Berlin Heidelberg New York Tokyo, pp 26 - 38Google Scholar
  53. Ouyang JW, Hu H, Chuang CC, Tseng CC (1973) Induction of pollen plants from anthers of Triticum aestivum L. cultured in vitro. Sci Sin 16: 79 - 95Google Scholar
  54. Ouyang JW, Zhou SM, Jia SE (1983) The response of anther culture to culture temperature in Triticum aestivum. Theor Appl Genet 66: 101 - 109CrossRefGoogle Scholar
  55. Paepe de R, Pernes J (1978) Exemples de variations a hen:dite mendelienne induites au cours du developement des plantes. Physiol Veg 16 (2): 195 - 204Google Scholar
  56. Paepe de R, Bleton D, Gnangbe F (1981) Basis and extent of genetic variability among doubled haploid plants obtained by pollen culture in Nicotiana sylvestris. Theor Appl Genet 59 (3): 177 - 184CrossRefGoogle Scholar
  57. Paepe de R, Prat D, Huguet T (1982) Heritable nuclear DNA changes in doubled haploid plants obtained by pollen culture of Nicotiana sylvestris. Plant Sci Lett 28: 11 - 28Google Scholar
  58. Pan JL, Gao GH, Dan H (1983) Initial types of wheat pollen and their development in anther culture. Cell and tissue culture techniques for cereal crop improvement. Sci Press, Beijing, China, pp 117 - 129Google Scholar
  59. Parisi L, Picard E (1986) Disease response of doubled haploid lines and their original lines in wheat. Z Pflanzenzllcht 96:63 -78Google Scholar
  60. Picard E (1984) Contribution a I'etude de l'herMite et de I'utilisation de I'haploldisation par androgenese in vitro chez une cereale autogame: Triticum aestivum L. These de Doctorat d'Etat, Univ Paris XI, Orsay, France, 269 pp (no. 2902 )Google Scholar
  61. Picard E, Buyser de J (1973) Obtention de plantules haploldes de Triticum aestivum L. a partir de cultures d'antheres in vitro. CR Acad Sci Paris, Ser D 277:1463 -1466Google Scholar
  62. Picard E, Buyser de J (1977) High production of embryoids in anther culture of pollen derived homozygous spring wheats. Ann Amelior Plant 24(4):483 - 488Google Scholar
  63. Picard E, Buyser de J, Henry Y (1978) Technique de production d'haploldes de Ble par culture d'antheres in vitro. Sel Fran~ais 26:25 - 37Google Scholar
  64. Picard E, Rode A, Benslimane A, Parisi L (1986) Gametoclonal variations doubled haploids of wheat: biometrical and molecular aspects. In: Semal J (ed) Somaclonal variations and crop improvement. Sept 3-6, 1985, Gembloux (Belgium), Martinus Nijhoff, pp 136 - 147Google Scholar
  65. Picard E, Hours C, Gregoire S, Phan TH, Meunier JP (1987) Significant improvement of androgenetic haploid and doubled haploid induction from wheat plants treated with a chemical hybridization agent. Theor Appl Genet 74: 289 - 297CrossRefGoogle Scholar
  66. Picard E, Parisot C, Causse M, Brabant PH, Doussinault G, Trottet M, Rousset M (1988) Comparison of doubled haploid method with other breeding procedures in wheat (Triticum aestivum) when applied to populations. In: Miller TE, Koebner RMD (eds) Proc 7th Int Wheat Genet Symp, July 13-19,1988, Cambridge 2: 1155 - 1159Google Scholar
  67. Rives M, Picard E (1977) A case of genetic assimilation: selection through androgenesis or parthenogenesis of haploid producing systems (an hypothesis). Ann Amelior Plant 2 (4): 489 - 491Google Scholar
  68. Rode A, Hartmann C, Dron M, Picard E, Quetier F (1985) Organelle genome stability in anther-derived doubled haploids of wheat (Triticum aestivum L., cv. Moisson ). Theor Appl Genet 71: 320-324Google Scholar
  69. Rode A, Hartmann C, Benslimane A, Picard E, Quetier F (1987) Gametoclonal variation detected in the nuclear ribosomal DNA from doubled haploids lines of a spring wheat (Triticum aestivum L. cv. Cesar). Theor Appl Genet 74:31- 37Google Scholar
  70. Sarker P, Stebbins GL (1956) Morphological evidence concerning the origin of the B genome in wheat. Am J Bot 43: 297 - 304CrossRefGoogle Scholar
  71. Schmid J, Keller ER (1986) Effect of a gametocide on the induction of haploids in Triticum aestivum. Genetic manipUlation in plant breeding. In: Horn W, Jensen CJ, Odenbach W, Schieder O (eds) Proc Int Symp Eucarpia, Sept 8 -13, Berlin (West), Germany. de Gruyter, Berlin, pp 347 - 349Google Scholar
  72. Smith JD, Kinman ML (1965) The use of parent-offspring regression as an estimator of heritability. Crop Sci 5: 595 - 596CrossRefGoogle Scholar
  73. Snape JW, Simpson E (1981) The genetic expectations of doubled haploid lines derived from different filial generations. Theor Appl Genet 60: 123 -128Google Scholar
  74. Snape JW, Buyser de J, Henry Y, Simpson E (1986) A comparison of methods of haploid production in a cross of wheat, Triticum aestivum L. Z Pflanzenzllcht 96: 320 - 330Google Scholar
  75. Wenzel G, Uhrig H (1981) Breeding for nematode and virus resistance in potato via anther culture. Theor Appl Genet 59:333–340Google Scholar
  76. Wenzel G, Schieder 0, Przewozny T, Sopory SK, Melchers G (1979) Comparison of single cell culture derived Solanum tuberosum L. plants and model for their application in breeding programs. Theor Appl Genet 55: 49 - 55CrossRefGoogle Scholar
  77. Zhang LJ, Anceau C, Lepoivre P, Seilleur P, Semal J (1987) An efficient method for the regeneration of wheat (Triticum aestivum) from anther culture. Bull Rech Agron Gembloux 22 (4): 301 - 314Google Scholar
  78. Zhang YL, Li DS (1984) Anther culture of monosomics in Triticum aestivum. Hereditas, China 6 (3): 7 - 1Google Scholar
  79. Zhuang JJ, Jia X (1983) Increasing differentiation frequencies in wheat pollen callus. Cell and tissue culture techniques for cereal crop improvement. Sci Press, Beijing, China, pp 431 - 432Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1990

Authors and Affiliations

  • E. Picard
    • 1
  • A. Rode
    • 2
  • G. Doussinault
    • 3
  • M. Rousset
    • 4
  • M. Rives
    • 5
  1. 1.Laboratoire Blé, G. S. MoulonCNRS-INRA-Université Paris SudGif sur YvetteFrance
  2. 2.Laboratoire B.M. V., Bât 430Université Paris SudOrsay CedexFrance
  3. 3.INRA-SAPLe RheuFrance
  4. 4.INRA-SAPClermont FerrandFrance
  5. 5.INRA-SAPMMontfavetFrance

Personalised recommendations