Establishment and Characterisation of Hevea Cell Suspension Cultures

A simple and rapid protocol for establishment of Hevea suspension cultures is presented. H. brasiliensis X H. nitida suspension cultures with uniformly dispersed cells and cell aggregates were obtained after one to two months of culture. The growth kinetics study showed a typical sigmoidal profile including lag phase, log phase or exponential phase, linear phase, P and deceleration phase, while the doubling time was 2-5 days. Sedimented cell volume (SCV), fresh weight (FW), growth index (GI) of FW the decrease of electrical conductivity (EC) in the medium had a very good linear correlation with dry weight (DW) or GI of DW The optimum maintenance medium for Hevea suspension cultures was the modified MS medium supplemented with 0.2 mg/L 2,4-D, 2 mg/L NAA, 1 mg/L kinetin, 50 g/L sucrose, 0.2 g/L L-Asparagine and 0.4 g/L casein hydrolysate and coconut water 5% (v/v), in which suspension cultures were still well grown with multiplication after incubating in optimised suspension medium (OSM) for more than one year This cell suspension cultures would be used for protoplast isolation, large-scale clonal propagation, preservation of genetic resources, genetic transformation and establishment of bioreactors in Hevea.