In vitro propagation of Corymbia torelliana × C. citriodora (Myrtaceae) via cytokinin-free node culture
S. J. Trueman A B and D. M. Richardson AA Department of Primary Industries and Fisheries, LB 16, Fraser Road, Gympie, Qld 4570, Australia.
B Corresponding author. Email: stephen.trueman@dpi.qld.gov.au
Australian Journal of Botany 55(4) 471-481 https://doi.org/10.1071/BT06163
Submitted: 21 August 2006 Accepted: 21 December 2006 Published: 20 June 2007
Abstract
Hybrids between Corymbia torelliana (F.Muell.) K.D.Hill & L.A.S.Johnson and C. citriodora subsp. variegata (F.Muell.) A.R.Bean & M.W.McDonald are used extensively to establish forestry plantations in subtropical Australia. Methods were developed for in vitro seed germination, shoot multiplication and plantlet formation that could be used to establish in vitro and ex vitro clone banks of juvenile Corymbia hybrids. Effects of sodium hypochlorite concentration and exposure time on seed contamination and germination, and effects of cytokinin and auxin concentrations on shoot multiplication and subsequent rooting, were assessed. A two-step surface sterilisation procedure, involving 70% ethanol followed by 1% sodium hypochlorite, provided almost no contamination and at least 88% germination. A novel method of cytokinin-free node culture proved most effective for in vitro propagation. Lateral bud break of primary shoots was difficult to induce by using cytokinin, but primary shoots rooted prolifically, elongated rapidly and produced multiple nodes in the absence of exogenous cytokinin. Further multiplication was obtained either by elongating lateral shoots of nodal explants in cytokinin-free medium or by inducing organogenic callus and axillary shoot proliferation with 2.2 µm benzyladenine. Plantlets were produced using an in vitro soil-less method that provided extensive rooting in sterile propagation mixture. These methods provide a means for simultaneous laboratory storage and field-testing of clones before selection and multiplication of desired genotypes.
Acknowledgements
We thank Tracey Shears, Glenn Comrie, Dan Foster, Debra Cook and Jaimie Cook for assistance, Chris Newell (Department of Agriculture, Western Australia) for advice on the IVS propagation system, and Dr David Lee for providing Corymbia seeds and helpful comments on the article.
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