Cell-suspension cultures of ononis-natrix L subspecies ramosissima Establishment and culture conditions

Miguel, Maria Graça; Barroso, J. G.

http://hdl.handle.net/10400.1/6290

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Authors

Miguel, Maria Graça

Barroso, J. G.

Abstract(s)

Explants from petioles, folioles or hypocotyls of Ononis natrix have been used for calli initiation. Hypocotyls inoculated on MS medium supplemented with 2% sucrose and 0.5 mg.l-1 2,4-D/1 mg.l-1 Kin showed to be the best primary explant. Cell suspension cultures were established in MS basal medium supplemented with 2% sucrose, 0.5 mg.l-1 NAA or 2,4-D and 1 mg.l-1 Kin. Different subculturing periods, inoculum density, hormonal supplementation and sucrose concentration were assayed in order to obtain the best culture growth conditions. The optimal conditions were achieved with cultures initiated with 40 g.l-1 of initial inoculum, growing in MS basal medium supplemented with 4% sucrose, 0.5 mg.l-1 NAA and 1 mg.1-1 Kin subcultured every twelve days. Under these experimental conditions, the cultures showed a doubling time of 36.3 hours.