Resistance of Rapistrum rugosum to tribenuron and imazamox due to Trp574 or Pro197 substitution in the acetolactate synthase.

Ten putative resistant and two susceptible Rapistrum rugosum populations originating from Greece were studied for resistance to acetolactate synthase (ALS)-inhibiting herbicides, using dose-response assays, sequencing of als gene and in vitro ALS activity assays. The dose-response assays showed that one (P1) out of ten putative resistant populations was cross-resistant to tribenuron and imazamox, while another population (P4) was resistant to tribenuron only. All populations were susceptible to MCPA at the recommended rate. Gene sequencing of als revealed that the P4 population had a point mutation at Pro197 by His providing resistance to tribenuron, whereas the P1 had a Trp574 by Leu point mutation conferring cross-resistance to tribenuron and imazamox. The in vitro activity of the ALS enzyme indicated I50 values (tribenuron concentration required for 50% reduction of the ALS activity) ranging from 66.68 to 137.01 μM, whereas the respective value for the S populations ranged from 0.29 to 0.54 μM. These results strongly support that two R. rugosum populations evolved resistance to ALS-inhibiting herbicides due different point mutations in the als gene.