Abstract: The culture of protoplast derived from cotyledon and hypocotyl of Cassia obtusifolia seedlings was studied.About 1.5mm long sections of cotyledons and hypocotyls were digested in any enzyme solution for a certain time.The protoplasts were cultured in a liquid medium at a density of 5.1×10 4 No./ml and transferred onto a solid medium for callus culture and shoot regeneration.Some factors affecting protoplast isolation and division were studied.The results showed that 15 day age seedlings’ hypocotyls and cotyledons used as donor tissues were suitable for their protoplast isolation and the yield of their protoplasts amounted to 1.6Ⅹ10 4NO /g.FW;Pectolyase Y 23 was indispensible to their protoplast isolation;KM8P medium supplemented with 0.4mg/L 2,4 D,1.0mg/L NAA and 0.1mg/L KT as well as floating culture fitted in the division of protoplast derived cells.After a culture of 3 days ,the first division was observed.The plating frequency was about 29.2% in 15 days.Complete plantlets were regenerated 14 days after the shoots,3 cm or so,were transferred onto 1/2 MS medium containing 0.2mg/L IBA.

Key words: Cassia obtusifolia, Cotyledon and hypocotyl protoplast, Plant regeneration

CLC Number: 

• S567