Callus induction from protoplasts of V. unguiculata, V. sublobata and V. mungo.

摘要: Protoplasts were isolated from hypocotyl of V. mungo (L.) Hepper or hypocotyl-derived callus sublobata (Phaseolus Roxb.) and unguiculata Walp (syn. sinensis Saviex Hassk) using an enzyme solution comprising Cellulase 2.5%, Macerozyme, Hemicellulase Driselase each at a 0.5% level in 0.5 M sorbitol. Isolated protoplasts cultured Murashige Skoog's (1962) basal liquid medium supplemented with BA, NAA, 2,4-D (1 mg/l each) sucrose (14%). After four weeks, protoplast colonies transferred to the same reduced (7%). Colonies proliferated into actively growing calli. Further attempts regenerate plants such calli not successful. However, protoclones differentiated roots on auxin/cytokinin media. Alternative methods for shoot differentiation protoplastderived cultures tried by use Agrobacterium tumefaciens “shooter” strains pGV 2215 2298 wild type strain B6S3.