Molecular and General Genetics MGG

, Volume 215, Issue 1, pp 58–64 | Cite as

Plants regenerated from auxin-auxotrophic variants are inviable

  • Anne D. Blonstein
  • Tiina Vahala
  • Maarten Koornneef
  • Patrick J. King


Shoots regenerated from auxin-auxotrophic variants of Nicotiana plumbaginifolia were inviable when cultured in vitro in the absence of auxin. Variant shoots survived longer when grafted to wild-type stocks but eventually died after a characteristic pattern of degeneration. The auxin auxotrophs were isolated after mutagen treatment by a total isolation method as infrequent variants amongst haploid protoplast-derived cell colonies. The variants responded to several active auxins but, unlike the wild type, not to cytokinin. Plant regeneration from variant cultures ceased at early stages of shoot formation after complete withdrawal of auxin from the medium.

Key words

Auxin-auxotroph Nicotiana plumbaginifolia-Haploid Protoplast Variant 







2,4-dichlorophenoxyacetic acid


dimethyl sulfoxide


indole-3-acetic acid


indole-3-butyric acid




1-naphthaleneacetic acid


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Blonstein AD (1986) Auxotroph isolation in vitro. In: Blonstein AD, King PJ (eds) A genetic approach to plant biochemistry, Plant Gene Research, vol 3. Springer, Vienna, pp 259–279Google Scholar
  2. Blonstein AD, Vahala T, Fracheboud Y, King PJ (1988) Temperature-sensitive plant mutants isolated in vitro. Mol Gen Genet 211:252–259Google Scholar
  3. Bourgin J-P, Chupeau Y, Missioner C (1979) Plant regeneration from mesophyll protoplasts of several Nicotiana species. Physiol Plant 45:288–292Google Scholar
  4. Cohen JD, Bialek K (1984) The biosynthesis of indole-3-acetic acid in higher plants. In: Crozier A, Hillman JR (eds) The biosynthesis and metabolism of plant hormones. Cambridge University Press, pp 165–182Google Scholar
  5. Eriksson TR (1985) Protoplast isolation and culture. In: Fowke LC, Constabel F (eds) Plant protoplasts. CRC Press, Boca Raton, pp 1–20Google Scholar
  6. Gebhardt Ch, Schnebli V, King PJ (1981) Isolation of biochemical mutants using haploid mesophyll protoplasts of Hyoscyamus muticus. II. Auxotrophic and temperature-sensitive clones. Planta 153:81–89Google Scholar
  7. Green CE, Phillips RL (1975) Plant regeneration from tissue cultures of maize. Crop Sci 15:417–421Google Scholar
  8. Kao KN, Constable NF, Michayluk MR, Gamborg OL (1974) Plant protoplast fusion and growth of intergeneric hybrids. Planta 120:215–227Google Scholar
  9. King PJ (1976) Studies on the growth in culture of plant cells. XX. Utilisation of 2,4-dichlorophenoxyacetic acid by steadysate cultures of Acer pseudoplatanus L. J Exp Bot 27:1053–1072Google Scholar
  10. King PJ (1988) Plant hormone mutants. Trends Genet 4:157–162Google Scholar
  11. Larkin PJ, Scowcroft WR, Geissler AE, Katekar GF (1982) Phytotropins. IV. Effects of phytotropins on cultured plant cells and protoplasts. Aust J Plant Physiol 9:297–308Google Scholar
  12. Leguay JJ, Guern J (1977) Quantitative effects of 2,4-D on growth of suspension cultured Acer pseudoplatanus cells. II. Influence of 2,4-D metabolism and intracellular pH on the control of cell division by intracellular 2,4-D concentration. Plant Physiol 60:265–270Google Scholar
  13. Leguay JJ, Piecoup M, Puckett J, Jouanneau JP (1988) Common responses of cultured soybean cells to 2,4-D starvation and fungal elicitor treatment. Plant Cell Rep 7:19–22Google Scholar
  14. Maliga P (1982) Cell culture procedures for Nicotiana plumbaginifolia. Plant Mol Biol Newslett 3:88–94Google Scholar
  15. Oetiker J, King PJ (1986) Characterisation of a temperature-sensitive, auxin auxotrophic mutant of Hyoscyamus muticus. J Cell Biol 10B:30Google Scholar
  16. Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Soc Exp Biol Symp 11:118–131Google Scholar
  17. Suter M, Schnebli V, King PJ (1988) The development of a negative selection system for the isolation of plant temperature-sensitive auxin auxotrophs. Theor Appl Genet 75:869–874Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Anne D. Blonstein
    • 1
  • Tiina Vahala
    • 1
  • Maarten Koornneef
    • 1
  • Patrick J. King
    • 1
  1. 1.Friedrich Miescher-InstitutBaselSwitzerland

Personalised recommendations