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In the present study, four nutrient media viz., Murashige and Skoog (MS), Phytamax (PM), Mitra et al. (M) and Knudson ‘C’ (KC), were evaluated for seed germination and early protocorm development in Cymbidium aloifolium (L.) Sw., a medicinally important orchid of Bangladesh. Besides, effect of peptone, activated charcoal, plant growth regulators viz., 6‐benzylaminopurine (1.0–2.0 mg/L) and 2,4‐dichlorophenoxy acetic acid (1.0–2.0 mg/L), and light and dark conditions were also studied. Varied response in terms of per cent seed germination was observed in different media. Mitra et al. medium supplemented with 2.0 g/L activated charcoal (AC) showed 100% seed germination and effective for induction of significantly large size protocorms (1.64 mm in dia.). Protocorms turned necrotic under dark condition. Within two weeks of culture spherules emerged out due to cracking of the seed coat. The spherules developed into protocorms with a leaf primordium at apical portion after 3‐4 weeks and gradually produced complete seedlings. A strong and stout root system was induced in in vitro seedlings upon transfer to half strength PM and M media fortified with 0.5 mg/L indole‐3‐acetic acid (IAA). Well‐rooted seedlings were transferred to a green house with 95% survival. Based upon cost analysis it was suggested that Mitra et al. medium supplemented with AC (2.0 g/L) was more effective and economic for high frequency germination of seeds and seedlings development. IAA (0.5 mg/L) in Mitra et al. medium was suitable for development of stout root system which helped in high survival of seedlings under greenhouse condition.

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