, Volume 102, Issue 1, pp 101–108 | Cite as

In vitro chromosome doubling with colchicine during microspore culture in wheat (Triticum aestivum L.)

  • N.J.P. Hansen
  • S.B. Andersen


Isolated microspores of two DH lines of wheat were treated with 8 different colchicine concentrations up to 3 mM for either 24 h or 48 h during microspore culture. Untreated control cultures produced on average 220 embryos per spike (100,000 microspores), 68% of the regenerated plantlets were green, and 15% of the flowering plants were fertile. The colchicine treatments had a significant effect on chromosome doubling as measured by the percentage of fertile regenerants. Using colchicine concentrations around 1 mM the percentage of fertile plants among the regenerants was increased up to 53%. The highest number of embryos and regeneration rates were observed after 24 h colchicine treatment, while the highest frequencies of green plants and fertile plants were obtained with 48 h colchicine treatments. The highest number of DH plants per spike was found after treatment with colchicine concentrations of 300 to 1000 μM. Such treatments resulted in an estimated average between the two genotypes of 23 doubled haploid plants per spike.

albinism chromosome doubling colchicine treatment green plants haploids microspore embryogenesis T. aestivum 


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  1. Amssa, M., J. De Buyser & Y. Henry, 1980. Origine des plantes diploides obtenues par culture in vitrod'antheres de blé tendre (Triticum aestivumL.): influence du pretraitement au froid et de la culture in vitro sur le doublement. C R Acad Sci Paris 290: 1095-1097.Google Scholar
  2. Barnabas, B., P.L. Phaler & G. Kovacs, 1991. Direct effect of colchicine on the microspore embryogenesis to produce dihaploid plants in wheat (Triticum aestivumL.). Theor Appl Genet 81: 675-678.Google Scholar
  3. Chen, Z.Z., S. Snyder, Z.G. Fan & W.H. Loh, 1994. Efficient production of doubled haploid plants through chromosome doubling of isolated microspores in Brassica napus. Plant Breeding 113: 217-221.CrossRefGoogle Scholar
  4. Chu, C.C. & R.D. Hill, 1988. An improved anther culture method for obtaining higher frequency of pollen embryoids in Triticum aestivum L. Plant Science 55: 175-181.CrossRefGoogle Scholar
  5. Chu, C.C., R.D. Hill & A.L. Brule-Babel, 1990. High frequency of pollen embryoid formation and plant regeneration in Triticum aestivumL. on monosaccharide containing media. Plant Science 66: 255-262.CrossRefGoogle Scholar
  6. Chuong, P.V. & W.D. Beverdorf, 1985. High frequency embryogenesis through isolated microspore culture in Brassica napusL. and B. carinataBraun. Plant Science 39: 219-226.CrossRefGoogle Scholar
  7. Custers, J.B.M., J.H.G. Cordewener, Y. Noellen, H.J.M. Dons & M.M. Van Lookeren-Campagne, 1994. Temperature controls both gametophytic and sporophytic development in microspore cultures of Brassica napus. Plant Cell Reports 13: 267-271.CrossRefGoogle Scholar
  8. Datta, S.K. & G. Wenzel, 1987. Isolated microspore derived plant formation via embryogenesis in Triticum aestivumL. Plant Science 48: 49-54.CrossRefGoogle Scholar
  9. De Buyser, J. & Y. Henry, 1979. Androgenese sur des bles tendres en cours de selection. 1. L'obtention des plantes in vitro. Z Pflanzenzuchtung 83: 49-56.Google Scholar
  10. De Buyser, J., Y. Henry, P. Lonnet, R. Hertzog & A. Hespel, 1987.'Florin': a doubled haploid wheat variety developed by the anther culture method. Plant Breeding 98: 53-56.CrossRefGoogle Scholar
  11. Hansen, A.L., A. Gertz, M. Joersbo & S.B. Andersen, 1998. Antimicrotubule herbicides for in vitrochromosome doubling in Beta vulgarisL. ovule culture. Euphytica (in press).Google Scholar
  12. Hansen, F.L., S.B. Andersen, I.K. Due & A. Olesen, 1988. Nitrous oxide as a possible alternative agent for chromosome doubling of wheat haploids. Plant Science 54: 219-222.CrossRefGoogle Scholar
  13. Hansen, N.J.P. & S.B. Andersen, 1996. In vitrochromosome doubling potential of colchicine, oryzalin, trifluralin and APM in Brassica napusmicrospore culture. Euphytica 88: 159-164.CrossRefGoogle Scholar
  14. Henry, Y. & J. De Buyser, 1980. Androgenese sur des bles tendres en cours de selection. 2. L'obtention des grains. Z Pflanzenzuchtung 84: 9-17.Google Scholar
  15. Henry, Y. & J. De Buyser, 1990. Wheat anther culture: Agronomic performance of doubled haploid lines and the release of a new variety 'Florin'. In: Y. Bajaj (Ed.) Biotechnology in agriculture and forestry, vol 13 Wheat, pp. 286-352.Google Scholar
  16. Hoekstra, S., M.H. Van Ziderfeld, J.D. Louwerse, F. Heidekamp & F. Van der Mark, 1992. Anther and microspore culture of Hordeum vulgareL., cv Igri. Plant Science 86: 89-96.CrossRefGoogle Scholar
  17. Inagaki, M., 1985. Chromosome doubling of the wheat haploids obtained from crosses with Hordeum bulbosomL. Japan J Breed 35: 193-195.Google Scholar
  18. Mejza, S.J., V. Morgant, D.E. Dibona & J.R. Wong, 1993. Plant regeneration from isolated microspores of Triticum aestivum. Plant Cell Reports 12: 149-153.CrossRefGoogle Scholar
  19. Möllers, C., M.C.M. Iqbal & G. Röbbelen, 1994. Efficient production of doubled haploid Brassica napusplants by colchicine treatment of microspores. Euphytica 75: 95-104.CrossRefGoogle Scholar
  20. Navarro Alvarez, W., P.S. Baenziger, K.M. Eskridge, M. Hugo & V.D. Gustafson, 1994. Addition of colchicine to wheat anther culture media to increase doubled haploid plant production. Plant Breeding 112: 192-198.CrossRefGoogle Scholar
  21. Olsen, F.L. 1991. Isolation and cultivation of embryonic microspores from barley (Hordeum vulgareL.). Hereditas 115: 255-266.PubMedGoogle Scholar
  22. Ouyang, J.-W., H. Liang, S.-E. Jia, C. Zhang, T.-H. Zhao, L.-Z. He, X. Jia, 1994. Studies on the chromosome doubling of wheat pollen plants. Plant Science 98: 209-214.CrossRefGoogle Scholar
  23. Pechan, P.M. & W.A. Keller, 1988. Identification of potentially embryogenic microspores in Brassica napus. Phys Plant 74: 377- 384.CrossRefGoogle Scholar
  24. Puolimatka, M., S. Laine & J. Pauk, 1996. Effect of ovary co cultivation and culture medium on embryogenesis of directly isolated microspores of wheat. Cereal Research Comm 24: 393-400.Google Scholar
  25. SAS Institute Inc., 1996. SAS/STAT software, changes and enhancements through release 6.11, SAS Institute Inc., Cary, NC, USA.Google Scholar
  26. Taira, T., Z.Z. Shao, H. Hamawaki & E.N. Larter, 1991. The effect of colchicine as a chromosome doubling agent for wheat-rye hybrids as influenced by pH, method of application, and post-treatment environment. Plant Breeding 106: 329-333.CrossRefGoogle Scholar
  27. Touraev, A., A. Indrianto, I. Wratschko, O. Vicente & E. Heberle-Bors, 1996. Efficient microspore embryogenesis in wheat (Triticum aestivumL.) induced by starvation at high temperature. Sex Plant Reprod 9: 209-215.CrossRefGoogle Scholar
  28. Wan, Y., D.R. Duncan, A.L. Rayburn, J.F. Petolino & J.H. Widholm, 1991. The use of antimicrotubule herbicides for the production of doubledhaploid plants from antherderived maize callus. Theor Appl Genet 81: 205-211.CrossRefGoogle Scholar
  29. Wang, X. & H. Hu, 1984. The effect of potato IImedium for Triticale anther culture. Plant Sci Lett 36: 237-239.CrossRefGoogle Scholar
  30. Zaki, M.M.M. & H. Dickinson, 1991. Microspore-derived embryos in Brassica: The significance of division symmetry in pollen mitosis I to embryogenic development. Sex Plant Repro 4: 49- 55.Google Scholar
  31. Zaki, M.M.M. & H. Dickinson, 1995. Modification of cell development in vitro: the effect of colchicine on anther and isolated microspore culture in Brassica napus. Plant Cell Tissue Organ Culture 40: 255-270.CrossRefGoogle Scholar
  32. Zhao, J., D.H. Simmonds & W. Newcomb, 1996a. High frequency production of doubled haploid plants of Brassica napuscv. Topas derived from colchicine-induced microspore embryogenesis without heat shock. Plant Cell Reports 15: 668-671.CrossRefGoogle Scholar
  33. Zhao J, D.H. Simmonds & W. Newcomb, 1996b. Induction of embryogenesis with colchicine instead of heat in microspores of Brassica napusL. cv. Topas. Planta 198: 433-439.CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • N.J.P. Hansen
    • 1
  • S.B. Andersen
    • 1
  1. 1.Department of Agricultural Sciences, Section Plant Breeding and Crop ScienceThe Royal Veterinary and Agricultural UniversityFrederiksberg CDenmark

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