| Summary: | In this research, protoplasts were isolated from cell suspension derived from inflorescence of vetiver (Vetiveria zizanioides Nash) Surat Thani germplasm. The optimum condition for protoplast isolation was established by using 2% cellulase Onozuka R10, 2% macerozyme R10, 0.5% pectinase in 0.4 M mannitol and 7 mM CaCl2.2H2O at pH 5.8 and incubated for 10 hours in the dark on the rotary shaker at 50 rpm. Maximum protoplast yields were 8.4 × 104 protoplasts/ml PCV. Division of protoplasts was observed only in liquid medium. The first cell division was observed after 3 days of culture initiation, and the average division was 5.0% in the N6 medium supplemented with 1.0 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid) and 0.5 mg/l BA (Benzyladenine). An optimal density for culture division was 1 × 105 protoplasts/ml.
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