Factors affecting isolation and culture of protoplasts of Somkhag (Garcinia atroviridis Griff.)
2000
Moosikapala, L. | Te-chato, S. (Prince of Songkla Univ., Songkhla (Thailand). Faculty of Natural Resources. Dept. of Plant Science)
Isolation of protoplasts was carried out from vitro-grown leaves of Somkhag at different ages of culture. Various concentrations of cellulase Onozuka R-10 and macerozyme R-10 were tested. A sample of leaves was incubated in a mixture solution of the two enzymes on a gyratory shaker at 40 rpm under darkness for 12 hours. Various densities of released protoplasts were adjusted and cultured with different kinds and concentrations of growth regulator. Leaves at 3 months of culture incubated in 2 percent cellulase Onozuka R-10 and 1 percent macerozyme R-10 gave released protoplasts at 1.25*10*[7)/g.fr.wt. Viability of protoplasts was the highest (91.19 percent). Culturing of the protoplasts embedded in MS with 0.2 percent Phytagel, 3 percent sucrose, 0.7 M manitol, 0.5 mg/l dicamba and 1 mg/l benzyladenine (BA) promoted the best result in division of protoplasts. Age of leaves and culture density play important roles in development of the protoplasts. Protoplasts from leaves after one month of culture at density of 1.5*10*[5)/ml underwent microcolony formation after 4 weeks of culture.
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