Obtention of callus and cell suspension of different plant species

Abstract

Fifteen-day old seedlings of lucerne (Medicago sativa L.), pueraria (Pueraria phaseoloides (Roxb.) Benth) and tomato (Lycopersicum esculentum Mill), and sweet potato (Ipomoea batatas (L.) Lam) were transfered to tubes containing 20 ml of Murashige & Skoog (MS) medium, ammended with 5 mg/l of 2,4-D and 1 mg/l of cinetin per litre of medium, for the obtention of callus. The obtained calluses were transplanted to Erlenmeyer flasks containing MS medium with 2 g/l of casem, and were used for the obtention of cell suspensions, in Erlenmeyer flasks containing 25 ml of liquid MS medium, ammended with 5 mg of 2,4-D, 2 mg of biotin, 1 mg of cinetin and 1 g of casein per litre of medium. The cell suspensions were incubated, under 90 rpm shaking at 27 ºC temperature and 16 h of light a day (1,000 lux). The establishment of the growth curves and fresh weight of cells was determined, every three days, during 24 days with periodical monitoring of cell viability, through INT (iodonitrotetrazolium) monitoring technique. Suspensions from calluses of lucerne, sweet potato and pueraria hypocotyl exhibited more abundant and faster growth rate and greater viability than those from potato and tomato hypocotyl, and pueraria radicles.