Three perennial ryegrass (Lolium perenne) populations (R1, R2, and R3) with suspected resistance (R) to acetolactate synthase (ALS) or acetyl-CoA carboxylase (ACCase) herbicides were collected from wheat (Triticum aestivum) fields in northwestern Greece to study the underlying mechanisms of resistance and their impact on growth rate and competitive ability against wheat. Preemergence and postemergence plant dose–response assays showed that the R1 population was cross-resistant to the ALS inhibitors chlorsulfuron, mesosulfuron + iodosulfuron, and pyroxsulam, but susceptible (S) to imazamox. However, all populations were susceptible to the ACCase inhibitors clodinafop-propargyl, clethodim, diclofop-methyl, and pinoxaden. The analysis of the ALS gene sequence revealed a substitution of Pro197 by His or Leu in the ALS enzyme in L. perenne, which is reported for the first time in this weed and indicates a potential mechanism of target site-mediated resistance. The R1 population grown in the absence or presence of wheat competition displayed similar aboveground biomass and tiller number trends, and therefore similar estimated growth rates. In addition, the aboveground biomass of wheat was similarly reduced by both the R1 and S populations, supporting the evidence of their similar competitive ability against wheat. In general, these findings indicate that there is no clear evidence for the fitness advantage of R1 over the S population.

Three suspected resistant (R1, R2, and R3) corn marigold populations collected from winter cereal fields located in central Greece were studied to confirm and elucidate the mechanisms of resistance to acetolactate synthase (ALS) inhibitors and their competitive ability against barley. Whole-plant dose–response assays proved that the three suspected R populations were highly cross-resistant to the ALS inhibitors tribenuron, pyroxsulam + florasulam, and imazamox, whereas their control with synthetic auxin plus ALS inhibitors co-formulated mixtures was increased in the order of tritosulfuron + dicamba < florasulam + clopyralid < tribenuron + mecoprop-P < florasulam + aminopyralid. The ALS gene sequence revealed a point mutation in 11 plants of the R1, R2, and R3 populations, which resulted in the substitution of Pro-197-Thr or Trp-574-Leu. By contrast, all three sequenced plants of the susceptible (S) population were found with the wild-type allele encoding Pro-197 and Trp-574. This is the first report of ALS-inhibitor resistance in corn marigold. The competition study between barley and four densities of the S, R2, or R3 populations indicated similar biomass rates for all three populations, suggesting lack of association between the competitive ability of the R populations and the target-site resistance mechanism, which was also confirmed by the similar biomass reduction rates of barley grown in competition with S or R populations.

Five johnsongrass populations collected from corn grown in northern Greece were studied to elucidate the levels and mechanisms of resistance to acetolactate synthase (ALS)- and acetyl-CoA carboxylase (ACCase)-inhibiting herbicides. Whole-plant response assays indicated that two populations were highly cross-resistant to all ALS inhibitors tested (foramsulfuron, nicosulfuron, rimsulfuron, and imazamox) but were effectively controlled by the recommended rate of the ACCase-inhibiting herbicides propaquizafop and clethodim. TheALSgene sequence revealed a point mutation that resulted in the substitution of Trp574 by Leu in the ALS enzyme, suggesting that the resistance mechanism is target-site mediated. These findings highlight a serious threat against the sustainable use of the ALS-inhibiting herbicides in controlling johnsongrass and other grass weeds in cornfields, suggesting rotational use of herbicides with different modes of action, along with the use of nonchemical methods, for viable Johnsongrass management.

Seventeen putative resistant late watergrass populations [Echinochloa phyllopogon (Stapf.) Koso-Pol.; syn.: Echinochloa oryzicola (Vasinger) Vasinger] originating from rice (Oryza sativa L.) monoculture fields in northern Greece were examined for possible evolution of multiple resistance to acetolactate synthase (ALS) and acetyl-CoA carboxylase (ACCase) inhibitors and auxin herbicides in rate–response pot assays. Most of the populations were highly cross-resistant to the ALS-inhibiting herbicides bispyribac-Na, imazamox, penoxsulam, and nicosulfuron + rimsulfuron, whereas three of them were also multiple resistant to both ALS and the auxin mimic quinclorac. In addition, two E. phyllopogon populations were found to be multiple resistant to the ALS and ACCase inhibitors cycloxydim, cyhalofop-butyl, profoxydim, and quizalofop-P-ethyl. Amplification and sequencing of the ACCase gene fragment from eight surviving profoxydim-treated plants of the two multiple-resistant E. phyllopogon populations to ALS- and ACCase-inhibiting herbicides, revealed an Ile to Leu substitution at codon 1781 of the ACCase enzyme. However, amplification and sequencing of the ALS gene fragment in the same E. phyllopogon plants sequenced for ACCase revealed a Trp to Leu substitution at codon 574 of the ALS enzyme in three out of the eight sequenced plants. These results strongly support the evidence of coexisting E. phyllopogon multiple target-site resistance to ALS and ACCase inhibitors, which is reported for the first time.

Field and pot experiments were conducted in Greece to study the occurrence of resistance in silky windgrass to acetolactate synthase (ALS)- and acetyl-CoA synthase (ACCase)-inhibiting herbicides. Twenty-four populations of silky windgrass were examined in whole-plant response experiments. High levels of field-evolved resistance to chlorsulfuron (0% to 28% control in terms of fresh weight reduction) with the recommended field rates were confirmed in most silky windgrass populations. However, other ALS inhibitors, such as pyroxsulam and a premix of mesosulfuron-methyl and iodosulfuron, provided adequate control (76% to 100% in terms of fresh weight reduction) of most populations, except eight silky windgrass populations that were found to be cross-resistant to all ALS-inhibiting herbicides tested (i.e., chlorsulfuron, commercial mixture of mesosulfuron-methyl plus iodosulfuron, and pyroxsulam). Conversely, most silky windgrass populations were controlled effectively (90% to 100% in terms of fresh weight reduction) with the recommended field rates of ACCase inhibitors cycloxydim, clethodim, and pinoxaden, but five populations were also found to be resistant to clodinafop-propargyl (10% to 68% control in terms of fresh weight reduction). The ALS gene sequencing of the eight silky windgrass populations, with cross-resistance to ALS inhibitors, revealed a point mutation at the Pro-197 position, causing amino acid substitution by Ser or Thr in the ALS enzyme. Overall, chlorsulfuron and clodinafop-propargyl were selecting agents of field-evolved multiple resistance to ALS- and ACCase-inhibiting herbicides in five silky windgrass populations. As the available postemergence-applied chemistries/modes of action registered for grass weed control in cereals are rather limited, adopting integrated management practices and implementing proactive and reactive measures to delay the evolution of resistant populations is essential.

Background: Weed herbicide resistance is a serious problem in crop protection globally. Giant foxtail (Setaria faberi R.A.N. Herrm.) populations cannot be controlled by acetolactate synthase (ALS)-inhibiting herbicides in a few corn (Zea mays L.) monoculture fields. Methods: Five putative resistant giant foxtail populations, originating from corn monoculture fields in northeastern Greece, were evaluated for possible evolution of ALS-inhibitor resistance (nicosulfuron, rimsulfuron). The resistance ratio, the underlying resistance mechanism, and its impact on competitive ability against corn were studied. Results: The whole-plant rate-response assays showed that these populations were resistant (R) to the sulfonylureas nicosulfuron and rimsulfuron, but susceptible (S) to imidazolinone imazamox, triketone 4-hydroxyphenylpyruvate dioxygenase inhibitor tembotrione, and acetyl-CoA carboxylase inhibitor cycloxydim. The sequencing of the ALS gene did not reveal the presence of resistance-associated point mutations, indicating that the resistance was probably not target-site mediated. This was confirmed by the application of piperonyl butoxide two hours before nicosulfuron application, which reversed the resistance in all R giant foxtail populations, supporting the evidence of enhanced metabolism-mediated resistance. The competition study between corn and R or S giant foxtail populations indicated no stable trend reduction in corn traits, suggesting that the resistance mechanism was not associated with the competitive ability of the R populations. The novel ALS genotype in S. faberi, characterized for the first time and submitted to the GenBank database with accession number PV016837, indicated a closer genetic relationship with the S. viridis ALS gene than with S. italica. Conclusions: Five giant foxtail populations have evolved metabolism-based resistance to the ALS-inhibiting herbicides nicosulfuron and rimsulfuron.