Somporn Prasertsongskun

pp. 411 - 416

Abstract

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.