The increase in legume production in sustainable agriculture depends not only on the effectiveness of the selected nitrogen-fixing inoculants but also on their competitiveness in a soil environment containing an indigenous rhizobial population. In this study, we investigated the effect of pretreatment of red clover seeds with specific Nod factor (LCOs) on germination, growth, and nodulation of clover growing under sterile conditions and in the soil. We demonstrated that, although the symbiotic ability and competitiveness of the inoculant strain RtKO17 was not improved under competitive soil conditions, LCOs treatment of clover seeds significantly enhanced clover nodulation and growth of plants.