Effect of cytokinins on cell suspension culture, isolation and culture of protoplast of rubber
1999
Suraninpong, S. | Te-chato, S. (Prince of Songkla Univ., Songkhla (Thailand). Faculty of Natural Resources. Dept. of Plant Science)
Growth of cells of rubber in suspension culture in Murashige and Skoog (MS) medium supplemented with 3 percent sucrose, 0.5-1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg/l benzyladenine (BA) or 0.25 mg/l thidiazuron (TDZ) was studied during 30 days of culture. Various ages of suspension cells were exposed to a mixture of enzyme solution consisting of 1 percent cellulase Onosuka RS and 1 percent macerozyme R10. The mixture of enzyme solution was dissolved in 0.4 M mannitol in the presence of 3mM 2-N-morpholinoethylsulfonic acid (MES). The results showed that TDZ containing medium provided the better growth of cells at all times of culture/subculture. Time required for exponent growth (log phase) of the cells in TDZ supplemented medium was 16-24 days after culture while BA gave no clear situation for this stage of growth. In the case of isolation of protoplasts, the suspension maintained in TDZ-containing medium yielded a far greater number of protoplasts than those of BA-containing medium. A 10 day-old suspension provided the greater number of protoplasts of 2.56*10*[7)/ml. When protoplasts isolated from 7-10 day-old suspension were embeded in Phytagel-MS medium osmotically adjusted by mannitol to 0.4 M and supplemented with 0.1 mg/l TDZ and 1.0 mg/l 2,4-D division to microcolony (8-10 cells) could be promoted.
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