Skip to main content

Micropropagation of Telopea speciosissima R. Br. (Proteaceae). 2: Rhizogenesis and acclimatisation to ex vitro conditions


Conditions affecting rhizogenesis in vitro and ex vitro and subsequent acclimatisation of Telopea speciosissima (waratah) were investigated. Clonal selections were successfully rooted in vitro in agar, on filter paper bridges or using crushed quartz-sand, the last substrate resulting in superior growth of roots. The in vitro substrates were impregnated with half-strength MS, 7.5 gl-1 sucrose and various concentrations of IBA. For the quartz-sand, an IBA concentration of 50 μM was optimal, 70% of microcuttings were rooted. No plantlets rooted in vitro were acclimatised to ex vitro conditions (using mist, fog or humidity tent regimes). Microcuttings (25–45 mm in length) were rooted ex vitro in a fog humidity regime (droplet size <10 μm) using an IBA powder dip (3 g IBA kg-1). Neither a mist nor a humidity-tent regime was suitable for rooting of waratah microshoots ex vitro. A peat and perlite mixture was superior to crushed quartz-sand or potting mix for the rooting of microshoots; this appeared to be related to the air-filled porosity (>20%) of the mixture, measured after the medium was saturated and then drained for 24h. Plantlets must be left under the high humidity regime until shoot growth resumes (four to eight weeks) otherwise plant mortality increase significantly. In vitro-produced leaves abscised between eight and 12 weeks after transfer to ex vitro conditions, indicating that these structures did not acclimatise ex vitro.

This is a preview of subscription content, access via your institution.




GA3 :

gibberellic acid


indole-3-butyric acid


least significant difference


Murashige and Skoog medium


  • Aitken R (1985) Propagation by cuttings. In: Lamont BB & Watkins PA (Eds) Horticulture of Australian Plants (pp 32–37). Western Australian Dept. Agriculture, Perth

    Google Scholar 

  • Aldrufeu A, Pages M, Messeguer J & Mele E (1983) In vitro rhizogenesis in different substrates. Acta Hort. 150: 315–323

    Google Scholar 

  • Bhojwani SS, Mullins K & Cohen D (1987) Micropropagation of Feijoa sellowiana Berg. Acta Hort. 212: 69–76

    Google Scholar 

  • Brand MH & Lineberger RD (1986) In vitro propagation of Halesia carolina L. and the influence of explantation of initial shoot proliferation. Plant Cell Tiss. Org. Cult. 7: 103–113

    Google Scholar 

  • Briggs B (1987) Commercial production of ericaceous plants by tissue culture. Acta Hort. 212: 644

    Google Scholar 

  • Desjardins Y, Laforge F, Lussier C & Gosselin A (1988) Effect of CO2 enrichment and high photosynthetic photon flux on the development of autotrophy and growth of tissue cultured strawberry, raspberry and asparagus plants. Acta Hort. 230: 45–53

    Google Scholar 

  • Donnelly DJ & Vidaver WE (1984) Pigment content and gas exchange of red raspberry in vitro and ex vitro. HortScience 109: 177–181

    Google Scholar 

  • Donnelly DJ, Vidaver WE & Colbow K (1984) Fixation of CO2 in tissue cultured red raspberry prior to and after transfer to soil. Plant Cell Tiss. Org. Cult. 3: 313–317

    Google Scholar 

  • Fujiwara K, Kozai T & Watanabe I (1988) Development of a photoautotrophic tissue culture system for shoots and/or plantlets at rooting and acclimatisation stages. Acta Hort. 230: 153–158

    Google Scholar 

  • Grout BWW & Millam S (1985) Photosynthetic development of micropropagated strawberry plantlets following transplanting. Ann. Bot. 55: 129–131

    Google Scholar 

  • Handreck KA & Black ND (1989) Growing media for ornamental plants and turf. Second edition, University of New South Wales Press, Sydney

    Google Scholar 

  • McClelland MT & Smith MAL (1988) Response of woody plant microcuttings to in vitro and ex vitro rooting methods. Comb. Proc. Intl. Plant Prop. Soc. 38: 593–599

    Google Scholar 

  • Macdonald AB (1985) Propagation facilities-past and present. Comb. Proc. Intl. Plant Prop. Soc. 36: 170–175

    Google Scholar 

  • Maene LM & Debergh PC (1983) Rooting of tissue cultured plants under ex vitro conditions. Acta Hort. 131: 201–208

    Google Scholar 

  • Marin JA & Gella R (1987) Acclimatisation of the micropropagated cherry rootstock ‘Masto de Montanana’ (Prunus cerasus L.). Acta Hort. 212: 603–606

    Google Scholar 

  • Murashige T (1984) Plant propagation through tissue culture. Annu. Rev. Plant Physiol. 25: 135–166

    Google Scholar 

  • Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497

    Google Scholar 

  • Noiton D (1988) Adventitious root formation in the apple. PhD thesis, University of Sydney, Sydney

    Google Scholar 

  • Offord CA, Campbell LC & Mullins MG (1992) Micropropagation of Telopea speciosissima R. Br. (Proteaceae). 1. Explant establishment and proliferation. Plant Cell Tiss. Org. Cult. (in press)

  • Press TF (1983) Propagation: fog not mist. Comb. Proc. Intl. Plant Prop. Soc. 33: 100–109

    Google Scholar 

  • Purnell HM (1960) Studies of the family Proteaceae. 1. Anatomy and morphology of the roots of some Victorian species. Austral. J. Bot. 8: 38–50

    Google Scholar 

  • Simmonds J (1983) Direct rooting of micropropagated M26 apple rootstock. Scientia Hort. 21: 233–241

    Google Scholar 

  • Sutter E (1983) Micropropagation of mature ortets of Liquidambar styraciflua. HortScience 18: 103

    Google Scholar 

  • Sutter E, Novello V & Shakel K (1988) Physiological and anatomical aspects of water stress of cultured plants. Acta Hort. 230: 113–119

    Google Scholar 

  • Walker N, Jaques R & Miginiac E (1987) Action of light on rooting in vitro and acclimatisation of Sequoia sempervirens to soil. Acta Hort. 212: 289–301

    Google Scholar 

Download references

Author information

Authors and Affiliations


Rights and permissions

Reprints and Permissions

About this article

Cite this article

Offord, C.A., Campbell, L.C. Micropropagation of Telopea speciosissima R. Br. (Proteaceae). 2: Rhizogenesis and acclimatisation to ex vitro conditions. Plant Cell Tiss Organ Cult 29, 223–230 (1992).

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI:

Key words

  • in vitro propagation
  • waratah