This study was carried out to establish an improved bioassay system, whole-plant bioassay which is more effective in developing natural herbicides for foliar treatment such as herbicidal essential oils. Two bioassay systems using four weed species (Echinochloa crus-galli, Digitaria sanguinalis, Aeschynomene indica, and Abutilon theophrasti), spraying method and spotting method, were established. Spraying method is applicable if the amount of test compounds is enough, while spotting method is useful for the small amount of test compounds. The initial application rate was desirable at $2,500{sim}5,000;{mu}g;mL^{-1}$. Herbicidal activities were higher in the NOP treatment when compared to the Tween 20 treatment. To efficiently evaluate volatile compounds such as essential oils, if the compound-treated pots were incubated in dew chamber for about 10hrs, better results were obtained in the degree and stability of herbicidal responses. When the efficiency of bioassay systems established in this study was compared, the spraying method was minimized four times to the conventional method that has beed used for screening of synthetic compounds in KRICT. On the other hand, in the spotting method, screening for development of a natural herbicides was possible even in level of 1/100 test volume and 1/200 amounts of test compound compared to the spraying method.