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Methods in molecular biology (Clifton, N.J.)

Micropropagation of Phalaenopsis Blume.


PMID 20099092

Abstract

For high frequency regeneration of Phalaenopsis, young leaf segments are cultured on gelrite-gelled (1/2) MS medium supplemented with 2% sucrose, 2.0 mg/L BA, 0.5 mg/L NAA, 10% coconut water (CW), 2 g/L peptone and 1 g/L activated charcoal. Cultures are incubated at 24 +/- 2 degrees C under fluorescence light 50 micromol/m(2)/s for 16 h photoperiod per day. The PLBs (protocorm like bodies) are induced within 12 weeks of culture and are subcultured to proliferate on the fresh nutrient culture medium for 8 weeks. The PLBs clumps are dissected and cultured on (1/2) MS medium containing 2% sucrose, 10% coconut water (CW), 2 g/L peptone, 150 mg/L L-glutamine and 1 g/L activated charcoal. The PLB sections elongate to form shoots and new PLBs are induced from the base within 8 weeks of culture. For plantlets formation the shoots are cultured on (1/2) MS medium supplemented with 2% sucrose, 10% CW, 2 g/L peptone, 1 g/L activated charcoal and 50 g/L banana pulp. The regenerated plants are acclimatized and cultivated in the nursery, where they bloom within 2 years.