, Volume 167, Issue 4, pp 473–481 | Cite as

Plant regeneration from callus cultures of several soybean genotypes via embryogenesis and organogenesis

  • U. B. Barwale
  • H. R. Kerns
  • J. M. Widholm


Using callus derived from immature embryos, regeneration of viable plants was obtained in soybean (Glycine max (L.) Merr.). Depending on the composition of the medium, regeneration occurred via embryogenesis or via organogenesis. Embryogenesis resulted when embryos were plated on Murashige and Skoog (MS) medium containing 43 μM α-naphthaleneacetic acid. In work with the cultivar Williams 82, the addition of 5.0 μM thiamine HCl increased embryogenesis from 33% to 58% of the embryos plated. Addition of 30 μM nicotinic acid to the MS medium enhanced embryogenesis further to 76%. Organogenesis was obtained when medium containing 13.3 μM 6-benzylaminopurine, 0.2 μM and α-naphthaleneacetic acid and four times the normal concentration of MS minor salts was used. Histological studies of these cultures confirmed the organogenic and embryogenic nature of the cultures, by demonstrating the formation of shoot buds and somatic embryos, respectively. Similar responses were obtained in all 54 genotypes tested in this manner. The cultures retained the ability to regenerate complete plants for at least 12 months and 12–15 subcultures. Seeds have been obtained from several regenerated plants and when grown in the field these produced normal-appearing fertile plants.

Key words

Embryogenesis in callus culture Glycine (plant regeneration) Organogenesis in callus culture Plant regeneration Tissue culture (regeneration) 





2,4-dichlorophenoxyacetio acid


gibberellic acid


indole-3-acetic acid


indole-3-butyric acid


Murashige and Shoog (1962) medium


α-naphthaleneacetic acid


4-amino-3,5,6-trichloropicolinic acid


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Armstrong, C.L., Green, C.E. (1985) Establishment and maintenance of friable, embryogenic maize callus and the involvement of l-proline. Planta 164, 207–214Google Scholar
  2. Barwale, U.B., Meyer, M.M., Widholm, J.M. (1986) Theor. Appl. Genet. (in press)Google Scholar
  3. Beversdorf, W.D., Bingham, E.T. (1977) Degrees of differentiation obtained in tissue cultures of Glycine species. Corp Sci. 17, 307–311Google Scholar
  4. Cheng, T.Y., Saka, T., Voqui-Dinh, T.H. (1980) Plant regeneration from soybean cotyledonary node segments in culture. Plant Sci. Lett. 19, 91–99Google Scholar
  5. Christianson, M.L., Warnick, D.A., Carlson, P.S. (1983) A morphogenetically competent soybean suspension culture. Science 222, 632–634Google Scholar
  6. Dos Santos, A.V.P., Cutter, E.G., Davey, M.R. (1983) Origin and development of somatic embryos in Medicago sativa L. (alfalfa). Protoplasma 117, 107–115Google Scholar
  7. Gamborg, O.L., Miller, R.A., Ojima, K. (1968) Exp. Cell Res. 50, 148–151Google Scholar
  8. Gamborg, O.L., Davis, B.P., Stahlhut, R.W. (1983) Somatic embryogenesis in cell cultures of Glycine species. Plant Cell Rep. 2, 209–212Google Scholar
  9. Grant, J. (1984) Plant regeneration from cotyledonary tissue of Glycine canescens, a perennial wild relative of soybean. Plant Cell Tissue Organ Cult 3, 169–173Google Scholar
  10. Hoagland, D.R., Arnon, D.I. (1950) The water-culture method for growing plants without soil. California Agric. Exp. Sta. Bull. No. 347Google Scholar
  11. Johansen, D.A. (1940) Plant microtechnique. McGraw-Hill Publ., New York LondonGoogle Scholar
  12. Kameya, T., Widholm, J.M. (1981) Plant regeneration from hypocotyl sections of Glycine species. Plant Sci. Lett. 21, 289–294Google Scholar
  13. Kimball, S.L., Bingham, E.T. (1973) Adventitious bud development of soybean hypocotyl sections in culture. Crop Sci. 13, 758–760Google Scholar
  14. Lippmann, B., Lippmann, G. (1984) Induction of somatic embryos in cotyledonary tissue of soybean Glycine max L. Merr. Plant Cell Rep. 3, 215–218Google Scholar
  15. Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant 15, 473–497Google Scholar
  16. Newell, C.A., Luu, H.T. (1985) Protoplast culture and plant regeneration in Glycine canescens F. J. Herm. Plant Cell Tissue Organ Cult. 4, 145–149Google Scholar
  17. Oswald, T.H., Smith, R.A., Phillips, D.V. (1977) Physiol. Plant. 39, 129–134Google Scholar
  18. Phillips, G.C., Collins, G.B. (1981) Induction and development of somatic embryos from cell suspension cultures of soybean. Plant Cell Tissue Organ Cult 1, 123–129Google Scholar
  19. Saka H.T., Voqui-Dinh, T.H., Cheng, T.Y. (1980) Stimulation of multiple shoot formation on soybean stem nodes in cultures. Plant Sci. Lett. 19, 193–201Google Scholar
  20. Widholm, J.M., Rick, S. (1983) Shoot regeneration from Glycine canescens tissue cultures. Plant Cell Rep. 2, 19–20Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • U. B. Barwale
    • 1
  • H. R. Kerns
    • 2
  • J. M. Widholm
    • 1
  1. 1.Department of AgronomyUniversity of Illinois at Urbana-ChampaignUSA
  2. 2.Department of HorticultureUniversity of Illinois at Urbana-ChampaignUSA

Personalised recommendations