Plant Cell, Tissue and Organ Culture

, Volume 29, Issue 3, pp 257–268 | Cite as

The influence of plant growth regulator concentrations and callus age on somaclonal variation in callus culture regenerants of strawberry

  • Narender S. Nehra
  • Kutty K. Kartha
  • Cecil Stushnott
  • Kenneth L. Giles


The effect of plant growth regulator concentrations and ageing of callus on the extent and nature of variation among callus culture regenerants of strawberry (Fragaria × ananassa) cv. Redcoat was examined. Plants regenerated from callus culture had reduced plant vigour, shorter petiole length and smaller leaf size, but more leaves and runners under greenhouse conditions. These responses appeared to be due to a physiological influence of plant growth regulators. No distinct phenotypic variants were observed at plant growth regulator concentrations in the range of 1–10 μM each of BA and 2,4-d combination, but the highest concentration (20 μM each) of this combination produced a high frequency (10%) of dwarf type variants. The dwarf nature of these variants was maintained in the runner plants produced by the primary regenerants. The plants regenerated from 8-week-old calli did not show any distinct morphological variants. However, a significant proportion of deformed leaf shape (6–13%) and yellow leaf (21–29%) variants was obtained among plants regenerated from 16 and 24-week-old calli. The primary regenerants of the leaf shape variants were established as chimeras. The chimeric plants produced runner progeny with normal plants and plants with completely distorted leaf morphology. Both leaf shape and yellow leaf variants remained stable through runner propagation. Isozyme analysis failed to distinguish any of the variants from the standard runner plants. Flow cytometric analysis indicated the aneuploid nature of leaf shape variants but it could not distinguish dwarf and yellow leaf variants from standard runner plants.

Key words

callus culture flow cytometry Fragaria × ananassa Duch. isozyme somaclones 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Armstrong CL & Phillips RL (1988) Genetic and cytogenic variation in plants regenerated from organogenic and friable embryogenic tissue cultures of maize. Crop. Sci. 28: 363–369Google Scholar
  2. Arulsekar S & Parfitt DE (1986) Isozyme analysis procedure for stone fruits, almonds, grape, walnut, pistachio and fig. HortScience 21: 928–932Google Scholar
  3. Bayliss MW (1980) Chromosomal variation in plant tissues in culture. Int. Rev. Cytol. Suppl. 11A: 113–144Google Scholar
  4. Cameron JS & Hancock JF (1986) Enhanced vigor in vegetative progeny of micropropagated strawberry plants. HortScience 21: 1225–1226Google Scholar
  5. Cassels AC & Morrish FM (1987) Variation in adventitious regenerants of Begonia rex Putz ‘Lucille Closon’ as a consequence of cell ontogeny, callus ageing and frequency of callus subculture. Scientia Hort. 32: 135–143Google Scholar
  6. Chen THH, Lazar MD, Scoles GJ, Gusta LV & Kartha KK (1987) Somaclonal variation in population of winter wheat. J. Plant Physiol. 130: 27–36Google Scholar
  7. Damaini F, Pezzotti M & Arcioni S (1990) Somaclonal variation in Lotus corniculatus L. in relation to plant breeding purposes. Euphytica 46: 35–41Google Scholar
  8. Darrow GM (1966) The Strawberry, History, Breeding and Physiology. Holt, Rinehart and Winston, New YorkGoogle Scholar
  9. Evans DA & Bravo JE (1986) Phenotypic and genotypic stability of tissue culture plants. In: Zimmerman RH, Griesbach RJ, Hammerschlag FA & Lawson RH (Eds) Tissue Culture as a Plant Production System for Horticultural Crops (pp 73–94). Martinus Nijhoff Publishers, DordrechtGoogle Scholar
  10. Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP & Firoozabady E (1983) Rapid flow cytometric analysis of cell cycle in intact plant tissues. Science 220: 1049–1051Google Scholar
  11. Gamborg OL, Miller RA & Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Expt. Cell Res. 50: 151–158Google Scholar
  12. Gould AR (1986) Factors controlling variability in vitro. In: Vasil IK (Ed) Cell Culture and Somatic Cell Genetics of Plants, Vol 3 (pp 549–567). Academic press, New YorkGoogle Scholar
  13. Grabosch RA, Edge ME & Dalannay X (1987) Somaclonal variation in soybean plants regenerated from cotyledonary node tissue culture. Crop Sci. 27: 803–806Google Scholar
  14. Hemphill DD (1980) Weed control in strawberries. In: Childers NF (Ed) The Strawberry (pp 309–317). Horticultural Publication, Gainesville, FloridaGoogle Scholar
  15. Hughes JDA (1989) Strawberry June yellows — a review. Plant Pathology 38: 146–160Google Scholar
  16. Jones OP, Waller BJ & Beech MG (1988) The production of strawberry plants from callus culrures. Plant Cell Tiss. Org. Cult. 12: 235–241Google Scholar
  17. Karp A (1989) Can genetic instability be controlled in plant tissue cultures? IAPTC Newsletter 58: 2–11Google Scholar
  18. Karp A & Bright SWJ (1985) On the causes and orgins of somaclonal variation. In: Miffin BJ (Ed) Oxford Survey of slant Molecular and Cell Biology, Vol 2 (pp 199–234). Oxford University Press, OxfordGoogle Scholar
  19. Kartha KK, Leung NL & Pahl K (1980) Cryopreservation of strawberry meristems and mass propagation of plantlets. J. Amer. Soc. Hort. Sci. 105: 481–484Google Scholar
  20. Larkin PJ & Scowcroft WR (1981) Somaclonal variation — a novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet. 60: 197–214Google Scholar
  21. Lazar MD, Chen THH, Gusta LV & Kartha KK (1988) Somaclonal variation for freezing tolerance in a population derived from Norstar winter wheat. Theor. Appl. Genet. 75: 480–484Google Scholar
  22. Liu ZR & Sanford JC (1988) Plant regeneration by organogenesis from strawberry leaf and runner tissue. HortScience 23: 1057–1059Google Scholar
  23. Miller AR & Chandler CK (1990) Plant regeneration from excised cotyledons of mature strawberry achenes. HortScience 25: 569–571Google Scholar
  24. Murashige T & Nakano R (1966) Tissue culture as a potential tool in obtaining polyploid plants. J. Hered. 57: 114–118Google Scholar
  25. Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497Google Scholar
  26. Nehra NS, Kartha KK & Stushnoff C (1991) Nuclear DNA content and isozyme variation in relation to morphogenic potential of strawberry (Fragaria × ananassa Duch.) callus cultures. Can. J. Bot. 69: 239–244Google Scholar
  27. Nehra NS, Stushnoff C & Kartha KK (1989) Direct shoot regeneration from strawberry leaf disks. J. Amer. Soc. Hort. Sci. 114: 1014–1018Google Scholar
  28. Nehra NS, Stushnoff C & Kartha KK (1990) Regeneration of plants from immature leaf-derived callus of strawberry (Fragaria × ananassa). Plant Sci. 66: 119–126Google Scholar
  29. Nyman M & Wallin A (1988) Plant regeneration from strawberry (Fragaria × ananassa) mesophyll protoplasts. J. Plant Physiol. 133: 375–377Google Scholar
  30. Orton TJ (1983) Experimental approaches to the study of somaclonal variation. Plant Mol. Biol. Rep. 1: 67–76Google Scholar
  31. Reisch B (1983) Genetic variability in regenerated plants. In: Evans DA, Sharp WR, Ammirato PV & Yamnada Y (Eds) Handbook of Plant Cell Cultures, Vol 1 (pp 749–769). Macmillan, New YorkGoogle Scholar
  32. Skirvin RM (1978) Natural and induced variation in tissue culture. Euphytica 27: 241–266Google Scholar
  33. Swartz HJ, Galletta GJ & Zimmermann RH (1981) Field performance and phenotypic stability of tissue culturepropagated strawberries. J. Amer. Soc. Hort. Sci. 106: 667–673Google Scholar
  34. Varga A, Thoma LH & Bruinsma J (1988) Effects of auxins and cytokinins on epigenetic instability of callus-propagated Kalanchoe blossfeldiana pollen. Plant Cell Tiss. Org. Cult. 15: 223–231Google Scholar
  35. Wetter LR & Dyck J (1983) Isozyme analysis of cultured cells and somatic hybrids. In: Evans DA, Sharp WR, Ammirato PV & Yamada Y (Eds) Handbook of Plant Cell Cultures, Vol 1 (pp 607–628). Macmillan, New YorkGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • Narender S. Nehra
    • 1
  • Kutty K. Kartha
    • 2
  • Cecil Stushnott
    • 1
  • Kenneth L. Giles
    • 1
  1. 1.Department of Horticulture ScienceUniversity of SaskatchewanSaskatoonCanada
  2. 2.Plant Biotechnology InstituteNational Research CouncilSaskatoonCanada

Personalised recommendations