The effect of Ca^2+ and polyamines on the activity of β-glucan synthetase Ⅱ(GSⅡ) related to cell wall synthesis was studied in carrot suspension cultured cells. The activity of GS Ⅱ is four times higher than that of β-glucan synthetase Ⅰ in carrot suspension cultured cells and in vitro experiment, the activity of GSⅡ was increased in response to increase in concentration of Ca^2+ and polyamines. When carrot suspension cultured cells were incubated together with Ca^2+ and polyamines. When carrot suspension cultured cells were incubated together with Ca^2+ and polyamines, the GSⅡ activity was high at 0.1mM of Ca^2+ and 1 mM of putrescine. Also, polycationic poly-L-lysine and poly-L-ornithine increased about 50% the GSⅡ activity than that of the control, respectively. These results may imply that Ca^2+ and polyamines were related to the enzyme activity as a polycationic nature. In addition, verapamil as the calcium channel blocker and flunarizine as an antagonist of calcium mechanism in cytoplasm decreased GSⅡ activity ramarkably, Ca^2+ and calmodulin stimulated GSⅡ activity as Ca^2+ of free ion rather than Ca^2+ calmodulin complex. The effect of 2,4-D on the GSⅡ activity in culture medum is shown to be low at 0.1㎎ per liter and GSⅡ activity increased about 30% more than that of the 0.1㎎/l at the range of 0.3-1.0㎎ per litere. Cummulative results suggest that Ca^2+ and polyfamines stimulate the cell wall synthesis by means of the enhancement of GSⅡ activity responsible for synthesizing the cell wall components.