Papaver rhoeas biotypes displaying multiple herbicide resistance to ALS inhibitors and synthetic auxin herbicides (SAH) are spreading across Europe. In Spain, enhanced metabolism to imazamox was confirmed in one population, while cytochrome-P450 (P450) based metabolism to 2,4-D in another two. The objectives of this research were to further confirm the presence of P450 mediated enhanced metabolism and, if so, to confirm whether a putative common P450 is responsible of metabolizing both 2,4-D and imazamox. Metabolism studies were undertaken in five P. rhoeas populations with contrasted HR profiles (herbicide susceptible, only HR to ALS inhibitors, only HR to SAH, or multiple HR to both), and moreover, three different P450 inhibitors were used. The presence of enhanced metabolism to these SoA was confirmed in three more HR P. rhoeas populations. This study provides the first direct evidence that imazamox metabolism in these biotypes is P450-mediated, also in one population without an altered target site. Additionally, it was further confirmed that enhanced metabolism of 2,4-D in biotypes only HR to SAH or multiple HR to ALS inhibitors and SAH involves P450 as well. No metabolism was detected using the three inhibitors in all the herbicide-metabolizing P. rhoeas biotypes, suggesting that a common metabolic system involving P450s is responsible of degrading herbicides affecting both SoAs. Thus, selection pressure with either SAH or imidazolinone ALS inhibitors can select not only for resistance to each of them, but it can also confer cross-resistance between them in P. rhoeas.

Six Johnsongrass populations suspected of being glyphosate resistant were collected from railways and freeways near Cordoba (SW Spain), where glyphosate is the main weed control tool. The 50% reduction in shoot fresh weight (GR₅₀) values obtained for these six populations ranged from 550.4 to 1169 g ae ha⁻¹, which were 4.2 to 9 times greater than the value obtained for the susceptible population. Glyphosate was equally metabolized to the same extent in both resistant and susceptible populations, with no significant differences in either 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) inhibition or basal activity. No amino acid substitutions were observed in any of the resistant populations. Slight but significant differences in glyphosate penetration were observed among some but not all of the resistant populations and for the times of incubation assayed, although these differences were not considered further. The proposed primary mechanism of resistance in these six glyphosate-resistant Johnsongrass populations is reduced herbicide translocation, because the amount of glyphosate that translocated from treated leaves to shoots and roots in the susceptible population was double that observed in the resistant populations. As glyphosate multiple resistance due to more than one mechanism is not uncommon, this is the first time that glyphosate-resistant Johnsongrass populations have been fully described for all known mechanisms.

Glyphosate is a broad-spectrum systemic herbicide used to kill weeds, especially annual broadleaf weeds and grasses known to compete with commercial crops grown around the globe. However, weeds evolve and develop resistance to glyphosate. Until recently, no case of glyphosate resistance had been detected in France. Glyphosate resistance was indeed recently recorded in a Lolium rigidum weed population from a vineyard in the South of France. Here, we studied the mechanisms of this resistance case. Seed samples of L. rigidum were collected from the vineyard where resistance had been detected, as well as from a nearby area that had no known history of exposure to glyphosate. We studied the effect of retention of glyphosate spray, shikimic acid accumulation, glyphosate absorption and translocation, glyphosate metabolism, and the sequence of the enzyme that glyphosate targets in plants, 5-enolpyruvylshikimate-3-phosphate synthase. Our results show that glyphosate absorption decreased by 30 % in the resistant L. rigidum weed. In addition, glyphosate translocation out of the treated leaves was reduced by 52 %. Finally, the resistant biotype had a serine amino acid substitution at position 106 of the predicted protein, instead of the proline amino acid present in the susceptible population. Our results suggest that the resistant population of L. rigidum presents three different mechanisms of resistance to glyphosate, namely reduced absorption, reduced mobility in the plants, and a mutation in the gene coding for the enzyme targeted by glyphosate.

A downy brome population in a winter wheat field at Córdoba, Spain, survived use rates of chlortoluron (2.5 to 3.5 kg ai ha⁻¹) over 2 consecutive yr, where wheat monoculture and multiple annual chlortoluron applications had been carried out. The resistant (CR) biotype showed a higher ED₅₀value (7.4 kg ai ha⁻¹; the concentration required for 50% reduction of fresh weight) than the susceptible (S) control (2.2 kg ai ha⁻¹), with a 3.4-fold increase in chlortoluron tolerance. Chlortoluron resistance in the CR downy brome biotype was not caused by altered absorption, translocation, or modification of the herbicide target site but by enhanced detoxification. The inhibition of both the recovery of photosynthetic electron transport and chlortoluron metabolism in the CR biotype due to the presence of the Cyt P450 inhibitor 1-aminobenzotriazole (ABT) indicates that herbicide metabolism catalyzed by Cyt P450 monooxygenases is related to chlortoluron resistance in CR plants. Although both biotypes degraded chlortoluron byN-dealkylation and ring-methyl hydroxylation and seem to share the same ability to form polar conjugates, degradation in the resistant biotype is more efficacious as this biotype metabolizes the parent herbicide faster and to a greater extent than its susceptible counterpart. The ability of the susceptible biotype to ring-hydroxylate chlortoluron, albeit at much slower rate, probably explains its moderate tolerance to chlortoluron observed in the growth assays and its minor photosynthetic electron transport recovery observed in fluorescence measurements.