Introduction Deschampsia antarctica Desv. (Poaceae) is the only native Gramineae found in the Antarctic, where it is restricted to the Antarctic Peninsula and its offshore islands. Its ability to survive the harsh climate has attracted the interest of scientists searching for genes associated with freezing tolerance (Alberdi et al. 2002). For continuing research purposes it would be better if plants did not have to be collected from the field, but could be propagated effectively to provide the necessary experimental material D. antarctica normally reproduces both by seed produced by self- fertilization and also vegetatively from tillers (Holderegger et al. 2003). Vegetative propagation is slow and unable to generate enough plant material for laboratory requirements. This difficulty prompted us to develop a rapid micro- propagation method using tissue-culture methods for the production of large numbers of plants in relatively short periods. We describe here a method for the rapid micropropagation of D. antarctica that can solve the problem of availability, and prevent excessive collection from its natural habitat. Materials and methods Deschampsia antarctica was collected in the Coppermine Peninsula on Robert Island, South Shetland Islands during January 2001 and propagated vegetatively in soil:peat mixture (3:1) at 13°C ± 2°C in a growth chamber (Forma Scientific Inc, Marietta, OH, USA) with photon flux density of 180 umol m -2 s -1 at the top of canopy and 21/3 h light/dark period (start material). After four months the plants were thoroughly washed to eliminate all substrate, and surface sterilized for 20 min in Erlenmeyer flasks with 300 ml of an aqueous suspension of 1.5 g l -1 Captan 80W (BASF Chile-SA) and 0.5 gl -1 Benomyl 500WP (CYANAMID Chile-SA), under constant stirring, and then rinsed three times with sterile distilled water. The plants were transferred to 300 ml of 15% sodium hypochlorite and 0.01% Tween 20 for 20 min, and then rinsed three times with sterile distilled water. The explants (5 mm) with a portion of crown tissue and roots were excised from the sterile plants (Fig. 1a & b), placed horizontally in Petri dishes (15 x 90 mm; 20 explants per plate), and incubated in the dark on MS basal medium (pH 5.8 ± 1, 25 ± 2°C) containing 3% sucrose (w/v), 0.9% agar (w/v), and one of five different concentrations of 2,4-D and BAP growth regulators (Table I). When shoots were larger than 0.5 cm (after 30–60 days of incubation, depending on the hormone concentrations used), the calluses were subdivided. The shoots were incubated in culture bottles in the same MS medium without hormones under 16/8 h light/dark. After as little as 15 days the multiplication of plants was begun, being repeated three times, each time with 15 replicates for each hormone concentration. After this, the plants were planted in soil: peat mixture in the same conditions as the donor plants. To determine if regenerated plants were genetically different, we used the Amplification Fragments Length Polymorphism (AFLP) assay. DNA was extracted with the high molecular weigh DNA extraction kit "GenElute Plant Genomic DNA Kit" (SIGMA), according to the manufacturer's instructions. The AFLP reaction was performed according to the manufacturer's instructions (GIBCO-BRL, Life Technologies). The AFLP products were separated by electrophoresis in polyacrilamide/urea (6%/8M) gels in 0.5% TBE buffer and 1800 v by 2 h. For non-radioactive visualization, we used the Silver Sequence TM DNA Sequencing System Kit (Promega). Results and discussion After two weeks of culture under different hormonal treatments (Table I), cicatricial callus was observed around Antarctic Science 17 (1): 69–70 (2005) © Antarctic Science Ltd Printed in the UK DOI: 10.1017/S0954102005002440 69 Short Note Micropropagation of Deschampsia antarctica - a frost-resistant Antarctic plant MARELY CUBA, ANA GUTIÉRREZ-MORAGA, BARBARA BUTENDIECK and MANUEL GIDEKEL* Laboratorio de Fisiología y Biología Molecular Vegetal, Instituto de Agroindustria, Facultad de Ciencias Agropecuarias y Forestales, Universidad de La Frontera, Casilla 54-D, Temuco, Chile *corresponding author: mgidekel@ufro.cl Received 28 April 2004, accepted 18 August 2004 Table I. Callus and shoot response of D. antarctica cultured in MS medium containing different 2,4-D and BAP growth regulator combinations. 2,4D BAP Callus Shooting Shoots/Callus Treatment* (μM) (μM) formation (%) response (%) (mean ± SD) 1 2.2 - 100 98 22.7 ± 1.3 2 2.2 0.2 100 99 25.4 ± 1.9 3 4.5 2.0 90 85 10.3 ± 1.0 4 4.5 4.0 84 80 7.8 ± 1.1 5 9.0 4.0 58 35 5.3 ± 2.8 Control - - 0 65 15.6 ± 0.9 *All hormonal combinations were in MS medium.