The allelochemicals 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and 6-methoxy-benzoxazolin-2-one (MBOA) in wheat are considered to have a role in plant defense against weeds. This study explored the effect of proximity to two weeds, wild oat and flixweed, on DIMBOA/MBOA production in wheat seedlings under hydroponic culture to identify whether the breeding of modern wheat varieties with higher concentrations of these compounds could ensure plant-mediated weed control. MBOA was detected and was noted to exert a significant response; its exudation by some wheat seedlings was significantly increased irrespective of whether the roots were in contact with or separate from those of the weeds. The weeds were a source of biotic stress to wheat when grown in proximity to it, and the stress resulted in production of higher levels of MBOA in wheat seedlings, although the concentration varied with the wheat cultivar. Therefore, the synthesis and exudation of DIMBOA/MBOA in wheat seedlings appears to be an active metabolic process influenced by the environment, particularly the presence of weeds.