To understand effect of inoculum size, cell density, sucrose concentration and concentrations of MS basal salts on al suspension culture and protoplast isolation of wild viola (Viola patrinii DC.) callus from petiole segments this experi-
ment was conducted. a}
In the lot of 30 mesh inoculum size, two observations were obtained ; One was that a considerable increase in
the fresh and dry weight of callus was determined. Another was that the callus mass was relatively compact compa- 104
red with others. A recommendable cell density was 0.4g for 20m1 culture medium and the higher sucrose concentra- 1~
tion, the higher fresh and dry weight were obtained. The dilution of MS basal salt was differently affected on fresh A
and dry weight ; the highest fresh weight was found in lx MS salt, while the higest dry weight was in 1/3x dilution. 2% The addition of casein hydrolysate(3g/1) was more effective to increase of both fresh and dry weight. The contents, of-protein was great in the inoculum lots with larger inoculum size and higher concentration of MS basal salts conte-
nting 3g/L of casein hydrolysate and higher sucrose compared with others. ,
The greatest protoplasts were isolated from the lot of 10 mesh size treated with 1 % pectinase SE150 and 2 % ot:
cellulase YC. In general, for optimal protoplast isolation the following conditions were recommended ; 1) Use of sma- A
L
1ler cell sizes cultured for 25 weeks, and 2) more than 5 hours incubation using the combined mixture of the q1
enzymes with proper concentrations.
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