Plant Cell, Tissue and Organ Culture

, Volume 16, Issue 2, pp 75–87 | Cite as

Adventitions shoot formation on excised leaves of in vitro grown shoots of apple cultivars

  • Francesca Fasolo
  • Richard H. Zimmerman
  • Ingrid Fordham
General Paper


Leaves taken from micropropagated shoots of several apple (Malus domestica Borkh.) cultivars were cultured in vitro on Linsmaier & Skoog (LS) medium or the rice anther culture medium of Chu et al. (N6) containing various concentrations of either benzyladenine (BA) or thidiazuron (TDZ) plus naphthaleneacetic acid (NAA). Of the TDZ concentrations tested, 10 μM was most effective and it was equivalent to, or better than, 22 μM BA for both the percentage of leaves regenerating shoots and number of shoots formed per regenerating leaf in almost every experiment. Lower concentrations of NAA (1.1 and 5.4 μM) gave best results with both BA and TDZ. N6 medium gave consistently better results than LS. Lowering total salt concentration or total N concentration of LS to that of N6 did not improve the response nor did changing the NO3:NH4 ratio. The 3–4 leaves on the most distal part of the shoot were most responsive and tended to form the most adventitious shoots. Placing the leaf cultures in the dark for the first 2–3 weeks of the culture period produced the best results. Optimum results were obtained by culturing leaves from the distal part of the shoot in the dark for 2 weeks on N6 medium containing 10 μM TDZ and 1.1 or 5.4 μM NAA, then moving the cultures to 16 h daylight at a photon flux of 60 μmol s-1m-2.

Key words

Malus domestica organogenesis tissue culture thidiazuron 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Chu CC, Wang CC, Sun CS, Hsu C, Yin KC, Chu CY, Pi FY (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sinica 18:659–688Google Scholar
  2. 2.
    Dufour M, Zimmerman RH (1986) In vitro callogenesis and regeneration of apple cultivars. In: Moet-Hennessy Conference Fruit Trees Biotechnology (p 50) Moet-Hennessy, Paris (abstract)Google Scholar
  3. 3.
    James DJ, Passey AJ, Malhotra SB (1984) Organogenesis in callus derived from stem and leaf tissues of apple and cherry rootstocks. Plant Cell Tiss Org Cult 3:333–341Google Scholar
  4. 4.
    James DJ, Passey AJ, Rugini E (1986) In vitro regeneration in apple leaf tissues and its use for transformation via Agrobacterium tumefaciens and A. rhizogenes. In: Moet-Hennessy Conference Fruit Tree Biotechnology (p 53) Moet-Hennessy, Paris (abstract)Google Scholar
  5. 5.
    Jones OP, Gayner JA, Watkins R (1984) Plant regeneration from callus tissue cultures of cherry rootstock Colt (Prunus avium x P. pseudocerasus) and the apple rootstock M25 (Malus pumila). J Hort Sci 59:463–467Google Scholar
  6. 6.
    Korban SS, Skirvin RM (1985) In vitro shoot regeneration from an intact and a sectioned embryo-axis of apple seeds. Plant Sci 39:61–66Google Scholar
  7. 7.
    Kouider M, Korban SS, Skirvin RM, Chu MC (1984a) Influence of embryonic dominance and polarity on adventious shoot formation from apple cotyledons in vitro. J Amer Soc Hort Sci 109:381–385Google Scholar
  8. 8.
    Kouider M, Skirvin RM, Korban SS, Widholm JM, Hauptmann R (1984b) Adventitious shoot formation from Red Delicious apple cotyledons in vitro. J Hort Sci 59:295–302Google Scholar
  9. 9.
    Kouider M, Korban SS, Skirvin RM, Joung H (1985) The relationship of apple embryos and their cotyledons to maturity, dormancy, and the potential to form shoots in vitro. J Amer Soc Hort Sci 110:93–96Google Scholar
  10. 10.
    Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18:100–127Google Scholar
  11. 11.
    Liu JR, Sink KC, Dennis FG (1983) Plant regeneration from apple seedling explants and callus cultures. Plant Cell Tiss Org Cult 2:293–304Google Scholar
  12. 12.
    Liu ZR (1987) Regeneration and Genetic Transformation of Apple and Strawberry. MSc Thesis, Cornell University, Ithaca, NY (106 pp)Google Scholar
  13. 13.
    Rubos AC, Pryke JA (1984) Morphogenesis in embryonic tissues of apple. J Hort Sci 59:469–475Google Scholar
  14. 14.
    SAS Institute Inc (1982) SAS User's Guide: Statistics (1982 ed) SAS Institute Inc. Cary, NCGoogle Scholar
  15. 15.
    Snedecor GW (1956) Statistical Methods. 5th edn. (pp 316–320) The Iowa State College Press, Ames, IAGoogle Scholar
  16. 16.
    Tapia y Figueroa M, Viseur J (1986) Regeneration of apple (rootstock M26 and cultivars ‘Jonagold’, ‘JubileR Delgollune’, ‘Golden Delicious’) from calluses induced on leaves originated from plants grown in vitro. In: Moet-Hennessy Conference Fruit Tree Biotechnology (p 65) Moet-Hennessy, Paris (abstract)Google Scholar
  17. 17.
    Van Nieuwkerk JP, Zimmerman RH, Fordham I (1986) Thidiazuron stimulation of apple shoot proliferation in vitro. HortScience 21:516–518Google Scholar
  18. 18.
    Welander M (1986a) Plant regeneration in leaf segments of shoots raised in vitro from mature fruit trees. In: Somers DA, Gengenbach BG, Biesboer DD, Hackett WP, Green CE (Eds) 6th International Congress of Plant Tissue and Cell Culture, Abstracts (p 267) University of Minnesota, Minneapolis (abstract)Google Scholar
  19. 19.
    Welander M (1986b) Plant regeneration in leaf segments of shoots raised in vitro from mature apple trees. In: Moet-Hennessy Conference Fruit Tree Biotechnology (p 69) Moet-Hennessy, Paris (abstract)Google Scholar

Copyright information

© Kluwer Academic Publishers 1989

Authors and Affiliations

  • Francesca Fasolo
    • 1
  • Richard H. Zimmerman
    • 1
  • Ingrid Fordham
    • 1
  1. 1.Agricultural Research Service, Fruit LaboratoryU.S. Department of AgricultureBeltsvilleUSA

Personalised recommendations