In vitro regeneracija pupoljaka iz segmenata kotiledona, hipokotila i korena kod Brassica oleraceae ssp.

Abstract

Kod šest kultivara podvrsta kupusa, brokolija, kelja i karfiola vrste Brassica oleracea istražena je sposobnost in vitro regeneracije pupoljaka. Segmenti kotiledona, hipokotila i korena klijanaca starih 7 dana korišćeni su kao eksplantati. Inkubiran su na Murashige i Skoog (MS) podlogu sa različitim kombinacijama auksina i citokinina (BAP + IBA, BAP + NAA, KIN + IBA). Najviši procenat regenerisanih pupoljaka je dobijen na hipokotilima, dok su kotiledoni uglavnom produkovali korenove. Sposobnost regeneracije široko je varirala zavisno od kultivara, tipa eksplantata i koncentracije primenjene hormonalne kombinacije. Za kupuse najbolja regeneracija pupoljaka bila je na MS podlozi sa 1,0 mg/l BAP i 0,5 mg/l IBA, dok je kod brokolija i kelja to MS medijum sa 1.0 mg/l KIN i 0.2 mg/l IBA. Najviše kalusa i vitrifikovanih pupoljaka razvijalo se na podlozi sa 1,0 mg/l BAP + 0,2 mg/l NAA kod svih 6 ispitivanih kultivara.

Six cultivars of Brassica oleracea subspecies (cabbage, broccoli, kale and cauliflower) were tested for their ability to regenerate shoots in vitro. Cotyledon, hypocotyl and root segments of 7-day-old seedlings were used as explants. They were incubated on Murashige & Skoog (MS) medium supplemented with different combinations of cytokinins and auxines (BAP + IBA, BAP + NAA, KIN + IBA). Hypocotyls were the most responsive for shoot regeneration and cotyledons for root regeneration. Regeneration ability widely varied depending on cultivar, explant type and concentration of growth regulators. For cabbage the best shoot regeneration was observed on MS medium supplemented with 1.0 mgl -1 BA and 0.5 mgl-1 IBA, while for broccoli and kale the highest shoot regeneration was on MS medium supplemented with 1.0 mgl -1 KIN and 0.2 mgl-1 IBA. Callus formation in all investigated cultivars was most pronounced on MS medium with 1.0 mg/l BA + 0.2 mg/l NAA.