, Volume 104, Issue 2, pp 87–94 | Cite as

Non-random gametoclonal variation in rice regenerants from callus subcultured for a prolonged period under high osmotic stress

  • Shinya Yoshida
  • Kazuhiko Watanabe
  • Morihiro Fujino


Gametoclonal variations were investigated in rice plants regenerated from anther-derived callus after subculture for two different periods (4–6 and 20 weeks) in medium with two levels (292 and 438 mM) of mannitol. A high osmotic stress by mannitol in subculture medium contributed to the maintenance of high regeneration ability. The frequency of diploid regenerants (R0 plants) increased and that of haploids decreased from callus subcultured for the longer period in medium with the higher concentration of mannitol. R1 regenerant lines displayed variations in all six agronomical traits studied at the frequencies of 63% and 88% after subculture for 4–6 weeks and 20 weeks, respectively. High homozygosity was suggested in most of the R1 lines for these traits by the similar levels of variance to those in the control variety. A principal component analysis showed a skewed distribution of variants towards dwarfness and less panicle weight in the regenerants from callus subcultured for 20 weeks with 438 mM mannitol. Two short culmn variants were shown to be due to single or double recessive mutations. The results suggested that prolonged subculture under high osmotic stress causes non-random selection of homozygous gametoclonal variations in rice anther culture.

anther-derived callus Oryza sativa L. osmotic stress gametoclonal variation subculture period 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Adkins, S.W., R. Kunanuvatchaidach & I.D. Godwin, 1995. Somaclonal variation in rice - drought tolerance and other agronomic characters. Aust J Bot 43: 201-209.CrossRefGoogle Scholar
  2. Chen, C.C. & C.M. Chen, 1980. Changes in chromosome number in microspore callus of rice during successive subcultures. Can J Genet Cytol 22: 607-614.Google Scholar
  3. Chu, C.C., 1975. Establishment of an efficient medium for anther culture of rice through comparative experiments on nitrogen sources. Sci Sin 18: 659-668.Google Scholar
  4. Demarly, Y., 1986. Experimental and theoretical approach of in vitrovariation. In: J. Semal (Ed.), Somaclonal Variations and Crop Improvement, pp 84-99. Gembloux, Belgium.Google Scholar
  5. Devaux, P., A. Kilian & A. Kleinhofs, 1993. Anther culture and Hordeum bulbosum-derived barley doubled haploids: mutations and methylation. Mol Gen Genet 241: 674-679.PubMedCrossRefGoogle Scholar
  6. Evans, D.A. & W.R. Sharp, 1983. Single gene mutations in tomato plants regenerated from tissue culture. Science 221: 949-951.Google Scholar
  7. Fukui, K., 1983. Sequential occurrence of mutations in a growing rice callus. Theor Appl Genet 65: 225-230.CrossRefGoogle Scholar
  8. Gamborg, O.L., R.A. Miller & K. Ojima, 1968. Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50: 151-158.PubMedCrossRefGoogle Scholar
  9. Hirochika, H., 1993. Activation of tobacco retrotransposons during tissue culture. EMBO J 12: 2521-2528.PubMedGoogle Scholar
  10. Hirochika, H., K. Sugimoto, Y. Otsuki, H. Tsugawa & M. Kanda, 1996. Retrotransposons of rice involved in mutations induced by tissue culture. Proc Natl Acad Sci 93: 7783-7788.PubMedCrossRefGoogle Scholar
  11. Hoffman, F., E. Thomas & G. Wenzel, 1982. Anther culture as a breeding tool in rape. Theor Appl Genet 61: 225-232.Google Scholar
  12. Kanda, M., S. Kikuchi, F. Takaiwa & K. Oono, 1988. Regeneration of variant plants from rice (Oryza sativaL.) protoplasts derived from long term culture. Jpn J Genet 63: 127-136.Google Scholar
  13. Kawata, M., S. Harada, B. Antonio & K. Oono, 1992. Protoclonal variation of plant regeneration in rice. Plant Cell Tissue Organ Culture 28: 1-10.CrossRefGoogle Scholar
  14. Larkin, P.J. & W.R. Scowcroft, 1981. Somaclonal variation - a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60: 443-445.CrossRefGoogle Scholar
  15. McCoy, T.J., R.L. Phillips & H.W. Rines, 1982. Cytogenetic analysis of plants regenerated from oat tissue. High frequency of partial chromosome loss. Can J Genet Cytol 24: 37-50.Google Scholar
  16. Müller, E., P.T.H. Brown, S. Hartke & H. Lörz, 1990. DNAvariation in tissue-culture-derived rice plants. Theor Appl Genet 80: 673- 679.CrossRefGoogle Scholar
  17. Murashige, T. & F. Skoog, 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15: 473- 493.CrossRefGoogle Scholar
  18. Nakamura, K., H. Suzuki, K. Hattori & Y. Futuhara, 1994. Identification of ploidy level of the regenerated plants by anther culture in rice. Breed Sci 44: 19-22.Google Scholar
  19. Ogura, H., J. Kyozuka, Y. Hayashi, T. Koba & K. Shimamoto, 1987. Field performance and cytology of protoplast-derived rice (Oryza sativa): high yield and low degree of variation of four japonica cultivars. Theor Appl Genet 74: 670-676.CrossRefGoogle Scholar
  20. Oono, K., 1978. Test tube breeding of rice by tissue culture. Trop Agric Res Series 11: 109-123.Google Scholar
  21. Oono, K., 1983. Genetic variability in rice plants regenerated from cell culture. In: Cell and Tissue Culture Techniques for Cereal Crop Improvement Proceedings, pp 339-358. Gordon and Breach, Science Publishers, New York.Google Scholar
  22. Oono, K., 1985. Putative homozygous mutation in regenerated plants of rice. Mol Gen Genet 198: 377-384.CrossRefGoogle Scholar
  23. Sathish, P., O.L. Gamborg & M.W. Nabors, 1995. Rice anther culture: callus initiation and androclonal variation in progenies of regenerated plants. Plant Cell Rep 14: 432-436.CrossRefGoogle Scholar
  24. Sibi, M., 1986. Non-menderian heredity. Genetic analysis of variant plants regenerated from in vitroculture. In: J. Semal (Ed.), Somaclonal Variations and Crop Improvement, pp 55-83. Gembloux, Belgium.Google Scholar
  25. Yamamoto, T., Y. Soeda, A. Nishikawa & H. Hirohara, 1994. A study of somaclonal variation for rice improvement induced by three kinds of anther-derived cell culture techniques. Plant Tiss Cult Lett 11: 116-121.Google Scholar
  26. Yoshida, S., M. Iwai, K. Watanabe & M. Fujino, 1998. Stimulatory effect of sugar-alcohol during subculture on green plant regeneration in anther-derived callus of rice. Plant Cell Rep (in press).Google Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Shinya Yoshida
    • 1
  • Kazuhiko Watanabe
    • 1
  • Morihiro Fujino
    • 1
  1. 1.Hyogo Prefectural Institute of Agriculture, KasaiHyogoJapan

Personalised recommendations