The purpose of this study is to investigate the positive or negative effects of 5-aminolevulinic acid(ALA) on the growth of several crops and weeds, by applying a seed soaking treatment, foliar treatment, and application timing, while comparing biological activity between ALA produced by chemical synthesis (Synthetic-ALA) and extracellularly-accumulated ALA by overexpressing the hemeA gene isolated from Bradyrhizobium japonicum(Bio-ALA). Seed soaking treatment of ALA in barley (five cultivars) and wheat (five cultivars) had not shown positive effects at lower concentrations, 0.05 to 0.5 mM as well as negative effects at higher concentrations, 1 to 30 mM. In rice, there also was no positive effect by seed soaking treatment of ALA at lower concentrations, although the rice became damaged by an application of 5 and 10 mM ALA. Growth in barley cultivars, Ganghossalbori, Naehanssalbori, Songhakbori, Saessalbori, and Daehossalbori were increased up to 14%, 19-51 %, 17-64%, 18-23%, and 22-38% by ALA foliar application at lower concentrations, 0.05 to 0.5 mM, respectively. On the other hand, the growth in barley cultivars was inhibited by ALA foliar application at higher concentrations. Barley responded more positively to ALA foliar application than wheat and rice. The growth stimulation caused by ALA seed soaking treatment was less than by ALA foliar treatment. ALA treatment at the 1.5-leaf stage increased growth of barley by 19-58%, while pretreatment to seeds, post-emergence treatment at 3 days after seeding, 3-leaf stages, and 5-leaf stages had not shown positive effects. Thus, the positive effects of ALA on barley were dependent greatly upon the timing of application and its concentration. Monocots weeds were more sensitive to ALA foliar treatment than dicotyledonous weeds. A monocot weed, Setaria viridis L. was the most susceptible plant to ALA while a dicotyledonous weed, Plantago asiatica L. was the most tolerant. No significant difference in biological activity between bio-ALA and synthetic ALA on barley, wheat, rice, and weed, Ixeris dentate tested was observed. Thus, ALA produced by microorganisms would be a potent substance to be used effectively in agricultural production.