Callus Induction

Callus induction takes place after two weeks (on a medium with ascorbic acid) or five weeks (on a medium without ascorbic acid). Callus induction and proliferation is affected by medium composition, explant source and the collection period of the donor plants. Fukui et al. (1991) used a half-strength MS (Murashige and Skoog, 1962) nutrient medium supplemented with the growth regulators a-napthylacetic acid (NAA), an auxin and kinetin, a cytokinin, for callus induction from V. album var. lutescens. In our study, all types of culture media consisted of full strength MS basal medium solidified with 0.8% agar and supplemented with 3% sucrose and either 2,4-dichlorophenoxyacetic acid (2,4-D) (an auxin), NAA, kinetin and benzyladenine (BA) (a cytokinin), alone or in combination, as plant growth regulators (PGRs), at various concentrations. Cultures were incubated under a photosynthetic photon flux density (PPFD) of 150 ^mol nr-2 s-1 (16/8, cool white fluorescent).

It is possible that use of culture media, such as MS, containing various salts at high concentrations is preferable for the tissue culture of mistletoe, the parasitism of which affords special adaptation to mineral nutrition (Becker, 1986; Grieve 1994).

Cultures could be established exclusively from "winter" accessions only. In a general sense, higher induction rates were observed from stem explants, reaching a maximum value of 45% on a medium supplemented with 2 mg/l 2,4-D. On the other hand, callus proliferation from leaf explants was definitely better, reaching a maximum average fresh weight of 1.78 g and a dry weight of 0.17 g five weeks after inoculation on a medium supplemented with 1 mg/1 kinetin.

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