Resistance to acetolactate synthase (ALS)-inhibitor herbicides due to continuous and repeated selection is widespread in many troublesome weed species, including Palmer amaranth, throughout the United States. The objective of this research was to investigate the physiological and molecular basis of resistance to ALS inhibitors in a chlorsulfuron-resistant Palmer amaranth population (KSR). Our results indicate that the KSR population exhibits a high level of resistance to chlorsulfuron compared with two known susceptible populations, MSS and KSS from Mississippi and Kansas, respectively. MSS is highly susceptible to chlorsulfuron, whereas KSS is moderately sensitive. Dose—response analysis revealed that KSR was more than 275-fold more resistant compared with KSS. Nucleotide sequence analysis of the ALS gene from the plants that survived chlorsulfuron treatment revealed the possibility of evolution of both target site—based and non—target site based resistance to ALS inhibitors in the KSR population. The most common mutation (Pro-197-Ser) in the ALS gene associated with resistance to the sulfonylureas in many weed species was found only in 30% of the KSR population. A preliminary malathion study showed that the remaining 70% of resistant plants might have cytochrome P450—mediated non—target site resistance. This is the first report elucidating the mechanism of resistance to ALS inhibitors in Palmer amaranth from Kansas. Presence of both target site— and non—target site based mechanisms of resistance limits the herbicide options to manage Palmer amaranth in cropping systems.

Nomenclature: Chlorsulfuron; Palmer amaranth, Amaranthus palmeri S. Wats. AMAPA

Five rigid ryegrass populations suspected of being resistant to both glyphosate and oxyfluorfen were collected in southern Spain and tested under laboratory-controlled conditions. Four populations (Depuradora, Condado, AlamoRasilla, and Portichuelo) were treated with glyphosate for at least 15 consecutive years, and treatments during the last 5 yr were mixed with oxyfluorfen. The fifth population (4alamos) followed the same glyphosate treatment, although oxyfluorfen was never used to control it. Dose—response assays confirmed glyphosate resistance in all populations, with resistance indexes ranging from 11.7 to 37.5 (GR₉₀). Shikimate accumulation assays consistently supported these data, as the most glyphosate-resistant populations (Depuradora and Condado) displayed the lowest shikimate levels. Surprisingly, four populations (Depuradora, Condado, AlamoRasilla, and Portichuelo) displayed 7.93- to 70.18-fold more resistance (GR₉₀) to oxyfluorfen, despite limited selection pressure, showing a similar resistance pattern as that for glyphosate. The 4alamos population displayed oxyfluorfen GR₉₀ values that were similar to those observed in susceptible plants; however, this population was significantly more resistant in terms of plant survival (LD₉₀). Protoporphyrin IX accumulation assays supported the results of dose—response assays, in that the most oxyfluorfen-resistant populations accumulated less protoporphyrin IX. Although more studies are needed, it seems that these five glyphosate-resistant weed populations display a natural tendency to easily develop resistance to oxyfluorfen, with the populations that have higher resistance to glyphosate also having higher resistance to oxyfluorfen.

Nomenclature: Glyphosate; oxyfluorfen; rigid ryegrass, Lolium rigidum Gaudin LOLRI.

Giant ragweed is a highly competitive weed that continually threatens crop production systems due to evolved resistance to acetolactate synthase—inhibiting herbicides (ALS-R) and glyphosate (GR). Two biotypes of GR giant ragweed exist and are differentiated by their response to glyphosate, termed here as rapid response (RR) and non—rapid response (NRR). A comparison of data from surveys of Indiana crop fields done in 2006 and 2014 showed that GR giant ragweed has spread from 15% to 39% of Indiana counties and the NRR biotype is the most prevalent. A TaqMan® single-nucleotide polymorphism genotyping assay was developed to identify ALS-R populations and revealed 47% of GR populations to be ALS-R as well. The magnitude of glyphosate resistance for NRR populations was 4.6 and 5.9 based on GR₅₀ and LD₅₀ estimates, respectively. For RR populations, these values were 7.8 to 9.2 for GR₅₀ estimates and 19.3 to 22.3 for LD₅₀ estimates. A novel use of the Imaging-PAM fluorometer was developed to discriminate RR plants by assessing photosystem II quantum yield across the entire leaf surface. H₂O₂ generation in leaves of glyphosate-treated plants was also measured by 3,3′-diaminobenzidine staining and quantified using imagery analysis software. Results show photo-oxidative stress of mature leaves is far greater and occurs more rapidly following glyphosate treatment in RR plants compared with NRR and glyphosate-susceptible plants and is positively associated with glyphosate dose. These results suggest that under continued glyphosate selection pressure, the RR biotype may surpass the NRR biotype as the predominant form of GR giant ragweed in Indiana due to a higher level of glyphosate resistance. Moreover, the differential photo-oxidative stress patterns in response to glyphosate provide evidence of different mechanisms of resistance present in RR and NRR biotypes.

Nomenclature: Glyphosate; giant ragweed, Ambrosia trifida L. AMBTR.

Italian ryegrass has invaded wheat fields in China and is becoming a predominant, troublesome weed. Fenoxaprop-P-ethyl has been widely used for weed control on Chinese farms since the 1990s. However, overuse has led to fenoxaprop-P-ethyl resistance in Italian ryegrass in Chinese wheat fields. In this study, we identified a putative fenoxaprop-P-ethyl—resistant population of Italian ryegrass, HZYC-6, from Henan province, China. Mutations involving Asp-2078-Gly and Ile-1781-Leu substitutions were identified in the carboxyl-transferase domain of acetyl-coenzyme A carboxylase in this population, and these mutations are the likely cause of the target site—based resistance to fenoxaprop-P-ethyl. In addition, we identified cytochrome P450—mediated metabolism of herbicides (non—target site based resistance) in the HZYC-6 population, indicating that multiple mechanisms of resistance may be segregating in this population. Furthermore, HZYC-6 was also highly resistant to haloxyfop-P-methyl and quizalofop-P-ethyl, moderately resistant to clodinafop-propargyl and sethoxydim, and had low resistance to clethodim and pinoxaden.

Nomenclature: Clethodim; clodinafop-propargyl; fenoxaprop-P-ethyl; haloxyfop-P-methyl; pinoxaden; quizalofop-P-ethyl; sethoxydim; Italian ryegrass, Lolium perenne ssp. multiflorum (Lam.) Husnot; wheat, Triticum aestivum L.

The widespread occurrence of Palmer amaranth resistant to acetolactate synthase inhibitors and/or glyphosate led to the increased use of protoporphyrinogen oxidase (PPO)-inhibiting herbicides. This research aimed to: (1) evaluate the efficacy of foliar-applied fomesafen to Palmer amaranth, (2) evaluate cross-resistance to foliar PPO inhibitors and efficacy of foliar herbicides with different mechanisms of action, (3) survey the occurrence of the PPO Gly-210 deletion mutation among PPO inhibitor-resistant Palmer amaranth, (4) identify other PPO target-site mutations in resistant individuals, and (5) determine the resistance level in resistant accessions with or without the PPO Gly-210 deletion. Seedlings were sprayed with fomesafen (263 g ai ha^-1), dicamba (280 g ai ha^-1), glyphosate (870 g ai ha^-1), glufosinate (549 g ai ha^-1), and trifloxysulfuron (7.84 g ai ha^-1). Selected fomesafen-resistant accessions were sprayed with other foliar-applied PPO herbicides. Mortality and injury were evaluated 21 d after treatment (DAT). The PPX2L gene of resistant and susceptible plants from a selected accession was sequenced. The majority (70%) of samples from putative PPO-resistant populations in 2015 were confirmed resistant to foliar-applied fomesafen. The efficacy of other foliar PPO herbicides on fomesafen-resistant accessions was saflufenacil > acifluorfen = flumioxazin > carfentrazone = lactofen > pyraflufen-ethyl > fomesafen > fluthiacet-methyl. With small seedlings, cross-resistance occurred with all foliar-applied PPO herbicides except saflufenacil (i.e., 25% with acifluorfen, 42% with flumioxazin). Thirty-two percent of PPO-resistant accessions were multiple resistant to glyphosate and trifloxysulfuron. Resistance to PPO herbicides in Palmer amaranth occurred in at least 13 counties in Arkansas. Of 316 fomesafen survivors tested, 55% carried the PPO Gly-210 deletion reported previously in common waterhemp. The PPO gene (PPX2L) in one accession (15CRI-B), which did not encode the Gly-210 deletion, encoded an Arg-128-Gly substitution. The 50% growth reduction values for fomesafen in accessions with Gly-210 deletion were 8- to 15-fold higher than that of a susceptible population, and 3- to 10-fold higher in accessions without the Gly-210 deletion.

Nomenclature: Acifluorfen; carfentrazone; dicamba; flumioxazin; fluthiacet-methyl; fomesafen; lactofen; glufosinate; glyphosate; pyraflufen-ethyl; saflufenacil; trifloxysulfuron; common waterhemp, Amaranthus rudis Sauer, AMATA; Palmer amaranth, Amaranthus palmeri S. Wats, AMAPA.

Seedling emergence traits of susceptible (S) and resistant (R) blackgrass subpopulations isolated from a single non—target-site resistant (NTSR) population were studied in controlled conditions. The seedling emergence of the R subpopulation was lower and slower than that of the S subpopulation, especially at low temperature and deep burial. The burial depth inhibiting final emergence by 50% for the R subpopulation was significantly lower than that of the S subpopulation at low temperature. The present study revealed that under suboptimal conditions the NTSR loci conferring herbicide resistance were correlated with a fitness cost in relation to seedling emergence traits. The results suggest that deep soil cultivation and delayed sowing of autumn-sown crops can hamper germination of the R more than of the S subpopulation and thus potentially reduce the prevalence of the R subpopulation in the blackgrass population.

Nomenclature: Blackgrass, Alopecurus myosuroides Huds. ALOMY.

A 2,4-D-resistant tall waterhemp population (FS) from Nebraska was evaluated for resistance to other TIR1 auxin receptor herbicides and to herbicides having alternative mechanisms of action using greenhouse bioassays and genetic markers. Atrazine, imazethapyr, lactofen, mesotrione, glufosinate, and glyphosate were applied in a single-dose bioassay, and tissue was collected from marked plants for genetic analysis. The FS population was not injured by atrazine or by imazethapyr. Approximately 50% of the plants survived lactofen and were actively growing 28 d after treatment. The population was susceptible to mesotrione, glufosinate, and glyphosate. Ametryn, chlorimuron-ethyl, 2,4-D, aminocyclopyraclor, aminopyralid, and picloram were applied in dose—response studies. The FS population was sensitive to ametryn, and the Ser-264-Gly substitution in the D1 protein was not detected, suggesting the lack of response to atrazine is not due to a target-site mutation. The FS population exhibited less than 50% injury to chlorimuron-ethyl at application rates 20 times the labeled use rate. The Ser-653-Asn acetolactate synthase (ALS) substitution, which confers resistance to imidazolinone herbicides, was present in the FS population. However, this does not explain the lack of response to the sulfonylurea herbicide, chlorimuron-ethyl. Sequencing of a portion of the PPX2L gene did not show the ΔG210 mutation that confers resistance to protoporphyrinogen oxidase—inhibiting herbicides, suggesting that other factors were responsible for waterhemp survival after lactofen application. The FS population was confirmed to be at least 30-fold resistant to 2,4-D relative to the susceptible populations. In addition, it was at least 3-fold less sensitive to aminopyralid and picloram, two other TIR1 auxin receptor herbicides, than the 2,4-D-susceptible populations were. These data indicated that the FS population contains both target and non—target site mechanisms conferring resistance to herbicides spanning at least three mechanisms of action: TIR1 auxin receptors, ALS inhibitors, and photosystem II inhibitors.

Nomenclature: 2,4-D; ametryn; aminocyclopyrachlor; aminopyralid; atrazine; chlorimuron-ethyl; glufosinate; glyphosate; imazethapyr; lactofen; mesotrione; picloram; tall waterhemp, Amaranthus tuberculatus (Moq.) Sauer. AMATU.

Palmer amaranth is the most economically damaging glyphosate-resistant (GR) weed in the southern United States. An understanding of the basic biology, including relative growth and competitiveness of GR and glyphosate-susceptible (GS) Palmer amaranth phenotypes from a segregating population collected from the same geographical location, may yield information helpful in the management of resistant populations. A segregating population of Palmer amaranth collected in North Carolina during 2010 was used as a plant source for both GR and GS traits. Research was conducted in the greenhouse to compare the following: level of resistance and shikimate accumulation in GR and GS phenotypes following glyphosate application; interference from GR and GS phenotypes on early-season vegetative growth of corn, cotton, and peanut; effect of various durations of imposed drought stress on GR and GS phenotypes; and response of GR and GS phenotypes to POST-applied herbicides. The GR₅₀ (glyphosate rate providing 50% reduction in shoot dry biomass) was 17 times greater with the GR phenotype compared with the GS phenotype. Shikimate accumulated in both GR and GS phenotypes following glyphosate application, but greater concentrations were found in GS plants. The GR and GS phenotypes responded similarly when subjected to drought stress; grown with corn, cotton, and peanut; or treated with 2,4-D, atrazine, dicamba, fomesafen, glufosinate, paraquat, tembotrione, and thifensulfuron. These results indicate that in the absence of glyphosate selection pressure, resistance to glyphosate does not influence the growth and competitiveness of GR and GS Palmer amaranth phenotypes collected from the same geographical location.

Nomenclature: 2,4-D; atrazine; dicamba; fomesafen; glufosinate; glyphosate; paraquat; tembotrione; thifensulfuron; Palmer amaranth, Amaranthus palmeri S. Wats.; corn, Zea mays L.; cotton, Gossypium hirsutum L.; peanut, Arachis hypogaea L.

As chemical management options for weeds become increasingly limited due to selection for herbicide resistance, investigation of additional nonchemical tools becomes necessary. Harvest weed seed control (HWSC) is a methodology of weed management that targets and destroys weed seeds that are otherwise dispersed by harvesters following threshing. It is not known whether problem weeds in western Canada retain their seeds in sufficient quantities until harvest at a height suitable for collection. A study was conducted at three sites over 2 yr to determine whether retention and height criteria were met by wild oat, false cleavers, and volunteer canola. Wild oat consistently shed seeds early, but seed retention was variable, averaging 56% at the time of wheat swathing, with continued losses until direct harvest of wheat and fababean. The majority of retained seeds were >45 cm above ground level, suitable for collection. Cleavers seed retention was highly variable by site-year, but generally greater than wild oat. The majority of seed was retained >15 cm above ground level and would be considered collectable. Canola seed typically had >95% retention, with the majority of seed retained >15 cm above ground level. The suitability ranking of the species for management with HWSC was canola > cleavers > wild oat. Efficacy of HWSC systems in western Canada will depend on the target species and site- and year-specific environmental conditions.

Nomenclature: False cleavers, Galium spurium L. GALSP; volunteer canola, Brassica napus L. BRSNN; wild oat, Avena fatua L. AVEFA; fababean, Vicia faba L.; wheat, Triticum aestivum L.

The escalating evolution of weed species resistant to acetolactase synthase (ALS)-inhibitor herbicides makes alternative weed control strategies necessary for field crops that are dependent on this herbicide group. A fully integrated strategy that combined increased crop seeding rates (2X or 4X recommended), mechanical weed control with a minimum-tillage rotary hoe, and reduced-rate non—ALS inhibitor herbicides was compared with herbicides, rotary hoe, and seeding rates alone as a method of controlling ALS inhibitor—tolerant Indian mustard as a model weed. The full-rate herbicide treatment had the lowest weed biomass (98% reduction) and the highest yield of all treatments in 3 of 4 site-years, regardless of seeding rate. The fully integrated treatment at the 4X seeding rate had weed suppression rates equal to the full herbicide treatment at the recommended seeding rate. The fully integrated and reduced-rate herbicide treatments at the 4X seeding rate reduced weed biomass by 89% and 83%, respectively, compared with the control at the recommended seeding rate. The rotary hoe treatment alone resulted in poor weed control (≤38%), even at the highest seeding rate. Fully integrated and reduced-rate herbicide treatments at 2X and 4X seeding rates had yields equal to those of the full herbicide treatment at the recommended seeding rate. Partially or fully integrated weed control strategies that combine increased crop seeding rates and reduced-rate non—ALS inhibitor herbicides, with or without the use of a rotary hoe, can control weeds resistant to ALS-inhibitor herbicides, while maintaining crop yields similar to those achieved with full-rate herbicides. However, combining increased seeding rate, reduced-rate herbicides, and mechanical rotary hoe treatment into a fully integrated strategy maximized weed control, while reducing reliance on and selection pressure against any single weed control tactic.

Nomenclature: Indian mustard, Brassica juncea (L.) Czern.

Adoption of soybean that is resistant to 2,4-D will result in more use of glyphosate plus 2,4-D premixes and tank mixtures. Preliminary whole-plant greenhouse assays confirm most Palmer amaranth populations found in Indiana are glyphosate-resistant (GR), and some biotypes exhibit tolerance to 2,4-D amine. Dose—response experiments were conducted to determine the level of glyphosate resistance and 2,4-D amine tolerance in four Palmer amaranth biotypes. A premix formulation of glyphosate plus 2,4-D choline was also evaluated. The R1, R2, and R3 biotypes were 31- to 66-fold more resistant to glyphosate (R:S ratio) than the S1 biotype based on the herbicide dose to cause 90% mortality (LD₉₀). The maximum POST rate of the premix formulation of Enlist Duo® labeled in ‘Enlist®’ soybean is 2,195 g ae ha^-1 When separated by active ingredient, the maximum POST rate of Enlist Duo® is equivalent to 1,141 and 1,054 g ae ha^-1 of glyphosate and 2,4-D choline, respectively. In the absence of glyphosate, the maximum rate of 2,4-D (1,054 g ae ha^-1) in the premix formulation of Enlist Duo® controlled S1, R2, and R3 biotypes, but failed to control all plants from the R1 biotype. Estimates for LD₉₀ showed the R1 biotype was 3-fold more tolerant than the S1 biotype to 2,4-D amine. However, no plants survived the 1,155 g ae ha^-1 (600 g ae ha^-1 of glyphosate plus 555 g ae ha^-1 2,4-D) treatment with the premix formulation of glyphosate plus 2,4-D choline. Overall, results from this experiment suggest GR Palmer amaranth that also exhibits increased tolerance to 2,4-D amine will be difficult to control with glyphosate or 2,4-D alone, but can be controlled POST with Enlist Duo® at lower than labeled field rates (1,618 to 2,195 g ae ha^-1).

Nomenclature: 2,4-D amine; 2,4-D choline; Enlist Duo®; glyphosate; Palmer amaranth, Amaranthus palmeri S. Wats.; soybean, Glycine max (L.) Merr.

Four early-generation backcross populations (BC₁F₂) derived from one common recipient parental background, Weed Tolerant Rice 1 (‘WTR1’), and four different donor parents (‘Y134’, ‘Zhong 143’, ‘Khazar’, and ‘Cheng Hui-448’) were tested to identify suitable donor and recipient parents for weed competitiveness and to standardize evaluation of the weed-competitive ability in rice. ‘GSR IR2-6’ (G-6) derived from a backcross of WTR1/Y134//WTR1 was selected as the best population and was advanced for phenotypic experiments in the 2014 dry season. The introgression lines (ILs) derived from the G-6 population were evaluated for seed germination and seedling vigor in greenhouse conditions and for weed-competitive ability under field conditions (upland weed-free, upland weedy, and lowland weedy). Parents and checks were included for comparison. Selection pressure for weed competitiveness was relatively stronger in upland conditions than in lowland conditions. After three rounds of selection and based on their relative grain yield performances across conditions, a total of 21 most-promising introgression fixed lines showing superior traits and weed-competitive ability were identified. G-6-L2-WL-3, G-6-RF6-WL-3, G-6-L15-WU-1,G-6-Y16-WL-2, and G-6-L6-WU-3 were the top ILs in lowland weedy conditions, whereas G-6-Y7-WL-3, G-6-Y6-WU-3, G-6-Y3-WL-3, and G-6-Y8-WU-1 were the highest yielding in upland weedy conditions. The use of weed-competitive rice cultivars in African and Asian countries will be a highly effective strategy to reduce production costs and provide alternative solutions to the unavailability of herbicides. Competitive rice varieties will also significantly improve grain yields in aerobic rice systems and can become an important strategy for successful upland rice production.

Nomenclature: Rice, Oryza sativa L.