In the present study, effects of medium formulation and PGRs on seed germination and PLB and shoot formation of Paphiopedilum graxtrixianum were examined. The results showed that the best medium formulation for Paphiopedilum graxtrixianum seed germination (77.50 percent) was SH medium supplemented with 0.5 mg/l NAA, 0.1 mg/l BA, 9 g/l agar, 20 g/l sucrose and 1 g/l activated charcoal. Explants were cultured on media containing 1 mg/l 2,4-0 after 60 days of culture on media with 9 g/l agar, 20 g/1 sucrose, 1 g/l activated charcoal and Img/l 2,4-0 in combination with 2 mg/l KN after 90 qays of culture resulted in the high efficiency in PLB induction (3.5 PLBs/explant) and PLB proliferation (11 PLBs/explants) respectively. Moreover, PLBs derived from cultures on different media led to considerable differences in shoot formation. The best shoot formation (9.5 shoots/clump) was achieved when PLBs forming on media supplemented with the combination of 1 mg/l 2.4-D and 2 mg/l KN were transferred into SH media containing 20 percent (v/v) coconut water, 9 g/l agar, 20 g/l sucrose and 1 g/l activated charcoal. The successful plant regeneration was obseved when single shoots were cultured on media with 0,5 mg/l BA, 0,5 mg/l NAA, 9 g/l agar, 30 g/l sucrose and 1 g/l activated charcoal.
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