Plant Cell, Tissue and Organ Culture

, Volume 22, Issue 3, pp 191–196 | Cite as

Selection for resistance to filtrates of Fusarium spp. in embryogenic cell suspension culture of Medicago sativa L.

  • Pavla Binarová
  • J. Nedělník
  • M. Fellner
  • Božena Nedbálková


A highly embryogenic cell suspension of alfalfa derived from a genotype sensitive to Fusarium oxysporum was successfully used for selection in vitro for resistance to culture filtrates of F. oxysporum, F. solani and F. avenaceum. Fifty two stable resistant cell lines were obtained and 500 plants regenerated from them. Among the 167 regenerants tested under glass there were 12–20% more plants with increased resistance to pathogens than in the group of plants regenerated from a control cell line. It was also found that the cell suspension cultures derived from genotypes of alfalfa with increased resistance to Fusarium spp. better tolerated filtrates of the pathogen. The results of a comparison of virulence of individual isolates of several species of Fusarium with toxicity of their filtrates to plants in vivo and in cell cultures were not unequivocal.

Key words

culture filtrate disease resistance embryogenic cell suspension culture Fusarium spp. Medicago sativa L. 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alicchio R, Antonioli C & Palenzona D (1984) Karyotypic variability in plants of Solanum melongena regenerated from callus grown, in presence of culture filtrate of Verticillium dahliae. Theor. Appl. Genet. 67: 267–271Google Scholar
  2. Arcioni S, Pezzotti M & Damiani F (1987) In vitro selection of alfalfa plants resistant to Fusarium oxysporum f. sp. medicaginis. Theor. Appl. Genet. 74: 700–705Google Scholar
  3. Behnke M (1980) General resistance to late blight of Solanum tuberosum, plants regenerated from callus resistant to culture filtrates of Phytophtora infestans. Theor. Appl. Genet. 56: 151–152Google Scholar
  4. Bilay VI (1977) Fusarii. Kijev, p442Google Scholar
  5. Binarová P & Novák FJ (1985) A study on embryogenesis in two different types of cell culture of Medicago sativa. Acta Univ. Agric. Brno XXXIII(3): 293–297Google Scholar
  6. Binarová P & Doležel J (1988) Alfalfa embryogenic cell suspension culture: growth and ploidy level stability. J. Plant Physiol. 133: 561–566Google Scholar
  7. Binarová P, Novotný F & Nedbálková B (1990) Selection and characterization of alfalfa cell lines resistant to lysine plus threonine and/or ethionine. Biochem. Physiol. Pflanz. 185: 1–2 (in press)Google Scholar
  8. Blaydes JF (1966) Interation of kinetin and various inhibtors of the growth of soybean tissue. Physiol. Plant. 19: 748–753Google Scholar
  9. Booth (1971) The genus Fusarium. Kew Commonwealth Mycol Inst. p 237Google Scholar
  10. Carr DH & Walker JE (1961) Carbol fuchsin as a stain for human chromosomes. Stain Technol. 36: 233–236Google Scholar
  11. Daubt ME (1986) Tissue culture and the selection of resistance to pathogens. Annu. Rev. Phytopathol. 24: 159–186Google Scholar
  12. Harman-Mitchell C, Knows TR & McCoy TJ (1983) Toxic components produced by Fusarium oxysporum f. sp. medicaginis and their use in alfalfa cell culture selection techniques. Phytopathology 73: 829Google Scholar
  13. Hartman CL, McCoy TJ & Knows TR (1984) Selection of alfalfa (Medicago sativa) cell lines and regeneration of plants resistant to the toxins, produced by Fusarium oxysporum f. sp. medicaginis. Plant Sci. Lett. 34: 183–194Google Scholar
  14. Hijano EH, Barnes DK & Frosheiser FI (1983) Inheritance of resistance to Fusarium wilt in alfalfa. Crop Sci. 23: 31–34Google Scholar
  15. Hughes K (1984) Selection for herbicide resistance. In: Evans DJ, Sharp WR, Amirato PV & Yamada Y (Eds) Handbook of Plant Cell Culture, Vol 1 (pp 441–459). MacMillan Publ. Co., New YorkGoogle Scholar
  16. Latunde Dada AO & Lucas JA (1983) Somaclonal variation and reaction to Verticilium wilt in Medicago sativa L. plants regenerated from protoplast. Plant Sci. Lett. 32: 205–211Google Scholar
  17. Mirocha TJ & Christensen TM (1986) Mycotoxins and the fungi that produce them. Proc. Am. Phytopathol. Soc. 3: 110–125Google Scholar
  18. Manka M & Chelkowski J (1985) Phytotoxicity and pathogenicity of Fusarium nivale towards cereal seedlings. Phytopathol. Z. 114: 1–5Google Scholar
  19. Nedělník J (1986) The Sensitivity of some cultivars of red clovers to selected species of Fusarium spp. and the effect of fungicides on the growth of Fusarium spp. in vitro. Sb. UVTI. Ochr. Rostl. 22: 285–293Google Scholar
  20. Nedělník J (1988a) The resistance of some varieties of Medicago sativa L. to Fusarium oxysporum Schlecht. Sb UVTIZ. Ochr. Rostl. 24: 191–197Google Scholar
  21. Nedělník J (1988b) Pathogenicity and culture filtrate phytotoxicity in species of genus Fusarium Link. Ex Fr. Proc. XI Czechoslovak Plant Prot. Conf., Nitra 1988: 65–66Google Scholar
  22. Sacristan MD (1982) Resistance responses to Phoma lingam of plants regenerated from selected cell and embryogenic cultures of haploid Brassica napus. Theor. Appl. Genet. 61: 193–200Google Scholar
  23. Scheffer RP (1983) Toxins as chemical determinants of plant disease. In: Daly M & Deverall BJ (Eds) (pp 1–40). New York, Academic PressGoogle Scholar
  24. Widholm (1972) The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technol. 47: 189–195Google Scholar

Copyright information

© Kluwer Academic Publishers 1990

Authors and Affiliations

  • Pavla Binarová
    • 1
  • J. Nedělník
    • 2
  • M. Fellner
    • 1
  • Božena Nedbálková
    • 2
  1. 1.Institute of Experimental BotanyCzechoslovak Academy of SciencesOlomoucCzechoslovakia
  2. 2.Research and Breeding Institute of Forage CropsTroubsko u BrnaCzechoslovakia

Personalised recommendations