The Anthurium genus comprises about 1500 tropical species, must of them important ornamental plants, which are normally propagated by seed (Dufour and Guerin, 2003). Vegetative propagation methods applied to Anthurium have not shown good results and tissue culture techniques appear as an alternative to increase the plant production (Pierik et al. 1974, Chen et al. 1997). The design of an efficient micropropagation system for this species could be important for its commercial applicability. This paper describes the results of the successful establishment of an alternative method for regeneration of Anthurium andreanum cv. Rubrun plants, from callus tissue through organogenesis. Micropropagation of plantlets originated from germinated seed is also described.

To reach these objectives, explants were obtained from plants of A. andreanum cv Rubrun germinated from seeds. The fruits were separated from spadixes and sterilized for 15 min in 3% NaOCl and then rinsed three times with sterile water for 30 min. Seeds were cultured on Murashige and Skoog, (1962) media with or without hormones and were incubated either in light or dark conditions at 25ºC. After one week, 74% of the seeds cultivated under continuous light conditions were germinated in comparison with 30% of germination showed by seeds cultivated on Murashige and Skoog, (1962) medium with BA incubated in darkness. The germinated seeds were all transferred to a medium without hormones under light conditions. Two weeks later, plantlets developed under light conditions were 2 cm high with 3 leaves and showed 2-3 pubescent roots (Figure 1). The plantlets obtained from seeds germinated under dark conditions showed callus proliferation at the base of the stem (Figure 2), and they were 1 cm lower than plantlets cultivated under light conditions.

The use of plants from germinated seeds permitted to eliminate the contamination problems, frequently found when taking explants from plants grown in the greenhouse. After 4 weeks, we took one of the plants germinated from seeds as source of explants. Four micro-cuttings were inoculated on Murashige and Skoog, (1962) medium supplemented with 4.4 mM BA and 0.05 mM NAA and incubated under continuous fluorescent light (50 mE. m^-2.s^-1) at 25ºC. Eight weeks later, 20 micro-cuttings were obtained for micropropagation experiments. These micro-cuttings were cultured in the conditions described above and six weeks later an average of 3.6 shoots were produced per explant, these shoots were 1.5 cm high and showed two to three leaves (Figure 3). We also observed the proliferation of callus tissue at the base of these shoots, which produced on average 6.6 small plantlets per 1 x 1 cm callus fragment.

Twenty segments of approximately 1 x 1 cm of these calli were sub cultivated on the same Murashige and Skoog, (1962) basal medium supplemented with 8.9 mM BA and 2.7 mM NAA and were incubated under continuous fluorescent light (50 mmol m-2 s-1) at 25ºC.We obtained an average of 43.8 plants per callus fragment (Figure 4). The sub-culture of these calli on suitable media, generates a higher number of shoots per callus fragment than the number of shoots originated from micro-cuttings culture.

Plantlets originated from callus tissue were maintained on the same medium during 4 months and then were transferred to Murashige and Skoog, (1962) medium without hormones for three months. Later, these 7 months old plants were potted on a mixture of soil and organic humus (1:1) and were successfully acclimated (Figure 5).

CHEN, F.C.; KUEHNLE, A.R. and SUGII, N. Anthurium roots for micropropagation and Agrobacterium tumefaciens-mediated gene transfer. Plant Cell Tissue and Organ Culture, 1997, vol. 49, no. 1, p. 71-74.

DUFOUR, L. and GUERIN, V. Growth, developmental features and flower production of Anthurium andreanum Lind. in tropical conditions. Scientia Horticulturae, 2003, vol. 98, no. 1, p. 25-35.

MURASHIGE, T. and SKOOG, F. A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiologia Plantarum, 1962, vol. 15, p. 473-497.

PIERIK, R.L.M.; STEEGMANS, H.H.M. and VAN DER MEYS, J.A.J. Plantlet formation in callus tissues of Anthurium andreanum Lind. Scientia Horticulturae, 1974, vol. 2, p. 193-198.

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