A biotype of rigid ryegrass (Lolium rigidum Gaudin biotype VLR69) resistant to some ALS inhibitors was characterized to determine the mechanisms of resistance to the sulfonylurea herbicides. The biotype had a high level of resistance to chlorsulfuron (20×) and triasulfuron (25×), and an intermediate level of resistance to imazaquin (7×) and sulfometuron (7.5×) but exhibited a low level of resistance to imazapyr (2.5×). At 60 to 90 g ai ha-1 sulfometuron, 4% of the population survived without apparent herbicidal effect The same response to sulfometuron was also observed when seeds of the resistant biotype VLR69 were germinated on agar containing sulfometuron. At 27 nM sulfometuron, 4% of the seeds germinated and grew normally while the growth of the bulk of the population was retarded. This differential response to sulfometuron at the seedling stage allowed selection of sulfometuron-resistant individuals from the population. Activity of ALS extracted from these sulfometuron-resistant plants was less sensitive to inhibition by chlorsulfuron than ALS extracted from plants considered sulfometuron susceptible in the same system. Unselected plants from the VLR69 population were able to detoxify chlorsulfuron more rapidly than susceptible VLR1 plants. It is apparent that there are at least two mechanisms of sulfonylurea resistance in the resistant biotype VLR69 which occur at different frequencies within the population. These data show that more than one mechanism or resistance may develop in response to herbicide selection pressure and that the resulting populations are not necessarily homogeneous.