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A comprehensive, Wisconsin state-wide assessment of waterhemp response to a diverse group of herbicide sites of action has not been conducted. Our objective was to characterize the response of a state-wide collection of waterhemp accessions to postemergence (POST) and preemergence (PRE) herbicides commonly used in corn and soybean in Wisconsin. Greenhouse experiments were conducted with more than 80 accessions from 27 counties. POST treatments included 2,4-D, atrazine, dicamba, fomesafen, glufosinate, glyphosate, imazethapyr, and mesotrione at 1× and 3× label rates. PRE treatments included atrazine, fomesafen, mesotrione, metribuzin, and S-metolachlor at 0.5×, 1×, and 3× label rates. Ninety-eight percent and 88% of the accessions exhibited ≥50% plant survival after exposure to imazethapyr and glyphosate POST 3× rate, respectively. Seventeen percent, 16%, and 3% of the accessions exhibited ≥50% plant survival after exposure to 2,4-D, atrazine, and dicamba, respectively, applied POST at the 1× rate. Survival of all accessions was ≤25% after exposure to 2,4-D or dicamba applied POST at the 3× rate, or glufosinate, fomesafen, and mesotrione applied POST at either rate evaluated. No plant of any accession survived exposure to glufosinate at either rate. Forty-five percent and 3% of the accessions exhibited <90% plant density reduction after exposure to atrazine applied PRE at the 3× rate and fomesafen PRE at the 1× rate, respectively. Plant density reduction of all accessions was ≥96% after exposure to fomesafen applied PRE at the 3× rate, or metribuzin, S-metolachlor, and mesotrione applied PRE at the 1× rate. Our results suggest that waterhemp resistance to imazethapyr and glyphosate applied POST is widespread in Wisconsin, whereas resistance to 2,4-D, atrazine, and dicamba applied POST is present to a lower extent. One accession (A75, Fond du Lac County) exhibited multiple resistance to imazethapyr, atrazine, glyphosate, and 2,4-D when applied POST. Overall, atrazine applied PRE was ineffective for waterhemp control in Wisconsin. Proactive resistance management and the use of effective PRE and POST herbicides are fundamental for waterhemp management in Wisconsin. Nomenclature: atrazine; dicamba; fomesafen; glufosinate; glyphosate; imazethapyr; mesotrione; metribuzin; S-metolachlor; 2,4-D; waterhemp, Amaranthus tuberculatus (Moq.) Sauer; corn, Zea mays (L.); soybean, Glycine max (L.) Merr.

Efficient natural dispersal of herbicide-resistance alleles via seed and pollen can markedly accelerate the incidence of herbicide-resistant weed populations across an agroecoregion. Studies were conducted in western Canada in 2014 and 2015 to investigate pollen- and seed-mediated gene flow in kochia. Pollen-mediated gene flow (PMGF) from glyphosate-resistant (GR) to non-GR kochia was quantified in a field trial (hub and spoke design) at Saskatoon, Saskatchewan. Seed-mediated gene flow of acetolactate synthase (ALS) inhibitor-resistant kochia as a function of tumbleweed speed and distance was estimated in cereal stubble fields at Lethbridge, Alberta and Scott, Saskatchewan. Regression analysis indicated that outcrossing from GR to adjacent non-GR kochia ranged from 5.3 to 7.5%, declining exponentially to 0.1 to 0.4% at 96 m distance. However, PMGF was significantly influenced by prevailing wind direction during pollination (maximum of 11 to 17% outcrossing down-wind). Seed dropped by tumbleweeds varied with distance and plant speed, approaching 90% or more (ca. 100,000 seeds or more) at distances of up to 1,000 m and plant speeds of up to 300 cm s−1. This study highlights the efficient proximal (pollen) and distal (seed) gene movement of this important GR weed. Nomenclature: Glyphosate; kochia, Kochia scoparia (L.) Schrad., KCHSC.

The continued dispersal of Palmer amaranth can impose detrimental impacts on cropping systems in Wisconsin. Our objective was to characterize the response of a recently introduced Palmer amaranth accession in southern Wisconsin to postemergence (POST) and preemergence (PRE) herbicides commonly used in corn and soybean. Greenhouse experiments were conducted with the Wisconsin putative herbicide-resistant accession (BRO) and two additional control accessions from Nebraska, a glyphosate-resistant (KEI2) and a glyphosate-susceptible (KEI3) accession. POST treatments were 2,4-D, atrazine, dicamba, glufosinate, glyphosate, imazethapyr, lactofen, and mesotrione at 1X and 3X label rates. PRE treatments were atrazine, mesotrione, metribuzin, S-metolachlor, and sulfentrazone at 0.5X, 1X, and 3X label rates. Plant survival of each accession was ≥63% after exposure to imazethapyr POST 3X rate. Survival of BRO and KEI2 was 44% (±13) and 50% (±13), respectively, after exposure to atrazine POST 3X rate. Survival of BRO was 69% (±12) after exposure to glyphosate POST 1X rate, whereas survival of KEI2 was 44% (±13) after exposure to glyphosate POST 3X rate. After exposure to 2,4-D POST 1X rate, KEI2 and KEI3 survival was 38% (±13) and 50% (±13), respectively. Survival of all accessions was ≤31% after exposure to 2,4-D POST 3X rate or dicamba, glufosinate, lactofen, and mesotrione POST at either rate. Plant density reduction of KEI2 was 77% (±13) after exposure to atrazine PRE 1X rate, whereas density reduction of BRO was 56% (±13) after exposure to atrazine PRE 3X rate. Plant density reduction of all accessions was ≥94% after exposure to mesotrione PRE 1X and 3X rates or metribuzin, S-metolachlor, and sulfentrazone PRE at either rate. Our results suggest that each accession is resistant (≥50% survival) to imazethapyr POST, that BRO and KEI2 are resistant to atrazine and glyphosate POST, and that KEI2 and KEI3 are resistant to 2,4-D POST. The recently introduced BRO accession exhibited multiple resistance to imazethapyr, atrazine, and glyphosate POST. In addition, atrazine PRE was ineffective for BRO control, suggesting that diversified resistance management strategies will be critical for its effective management. Nomenclature: atrazine; dicamba; glufosinate; glyphosate; imazethapyr; lactofen; mesotrione; metribuzin; sulfentrazone; S-metolachlor; 2,4-D; Palmer amaranth; Amaranthus palmeri S. Watson; corn; Zea mays L.; soybean; Glycine max (L.) Merr.

Wild oat is the second-most abundant, but most economically important, weed across the Canadian Prairies of western Canada. Despite the serious economic effects of resistance to acetyl-CoA carboxylase (ACC) or acetolactate synthase (ALS) inhibitors or both in this weed throughout the Northern Great Plains of North America, little research has examined the basis for herbicide resistance. We investigated target-site and nontarget-site mechanisms conferring ACC- and ALS-inhibitor resistance in 16 wild oat populations from across western Canada (four ACC-inhibitor resistant, four ALS-inhibitor resistant, and eight ACC- and ALS-inhibitor resistant). The ACC1 mutations were found in 8 of the 12 ACC inhibitor-resistant populations. The Ile1781Leu mutation was detected in three populations, the Trp2027Cys and Asp2078Gly mutations were in two populations each, and the Trp1999Cys, Ile2041Asn, Cys2088Arg, and Gly2096Ser substitutions were in one population each. Three populations had two ACC1 mutations. Only 2 of the 12 ALS inhibitor-resistant populations had an ALS target-site mutation—Ser653Thr and Ser653Asn substitutions. This is the first global report of ALS target-site mutations in Avena spp. and four previously undocumented ACC1 mutations in wild oat. Based on these molecular analyses, seedlings of five ACC + ALS inhibitor-resistant populations (one with an ACC1 mutation; four with no ACC or ALS mutations) were treated with malathion, a known cytochrome P450 monooxygenase inhibitor, followed by application of one of four ACC- or ALS-inhibiting herbicides. Malathion treatment often resulted in control or suppression of these populations, suggesting involvement of this enzyme system in contributing to resistance to both ACC and ALS inhibitors. Nomenclature: Wild oat, Avena fatua L. AVEFA.

In response to concerns about acetolactate synthase (ALS) inhibitor–resistant weeds in wheat production systems, we explored the efficacy of managing Bromus spp., downy and Japanese bromes, in a winter wheat system using alternative herbicide treatments applied in either fall or spring. Trials were established at Lethbridge and Kipp, Alberta, and Scott, Saskatchewan, Canada over three growing seasons (2012–2014) to compare the efficacy of pyroxasulfone (a soil-applied very-long-chain fatty acid elongase inhibitor; WSSA Group 15) and flumioxazin (a protoporphyrinogen oxidase inhibitor; WSSA Group 14) against industry-standard ALSinhibiting herbicides for downy and Japanese brome control. Winter wheat injury from herbicide application was minor, with the exception of flucarbazone application at Scott. Bromus spp. control was greatest with pyroxsulam and all herbicide treatments containing pyroxasulfone. Downy and Japanese bromes were controlled least by thiencarbazone and flumioxazin, respectively, whereas Bromus spp. had intermediate responses to the other herbicides tested. Herbicides applied in fall resulted in reduced winter wheat yield relative to the spring applications. Overall, pyroxasulfone or pyroxsulam provided the most efficacious Bromus spp. control compared with the other herbicides and consistently maintained optimal winter wheat yields. Therefore, pyroxasulfone could facilitate management of Bromus spp. resistant to ALS inhibitors in winter wheat in the southern growing regions of western Canada. Improved weed control and delayed herbicide resistance may be achieved when pyroxasulfone is applied in combination with flumioxazin. Nomenclature: Flucarbazone; flumioxazin; pyroxasulfone; pyroxsulam; thiencarbazone; downy brome, Bromus tectorum L. BROTE; Japanese brome, Bromus japonicus Thunb. BROJA; winter wheat, Triticum aestivum L.

Palmer amaranth is the latest pigweed species documented in Connecticut; it was identified there in 2019. In a single-dose experiment, the Connecticut Palmer amaranth biotype survived the field-use rates of glyphosate (840 g ae ha-1) and imazaquin (137 g ai ha-1) herbicides applied separately. Additional experiments were conducted to (1) determine the level of resistance to glyphosate and acetolactate synthase (ALS) inhibitors in the Connecticut-resistant (CT-Res) biotype using whole-plant dose-response bioassays, and (2) evaluate the response of the CT-Res biotype to POST herbicides commonly used in Connecticut cropping systems. Based on the effective dose required for 90% control (ED90), the CT-Res biotype was 10-fold resistant to glyphosate when compared with the Kansas-susceptible (KS-Sus) biotype. Furthermore, the CT-Res biotype was highly resistant to ALS-inhibitor herbicides; only 18% control was achieved with 2,196 g ai ha-1 imazaquin. The CT-Res biotype was also cross-resistant to other ALS-inhibitor herbicides, including chlorimuron-ethyl (13.1 g ai ha-1), halosulfuron-methyl (70 g ai ha-1), and sulfometuron-methyl (392 g ai ha-1). The CT-Res Palmer amaranth was controlled 75% to 100% at 21 d after treatment (DAT) with POST applications of 2,4-D (386 g ae ha-1), carfentrazone-ethyl (34 g ai ha-1), clopyralid (280 g ae ha-1), dicamba (280 g ae ha-1), glufosinate (595 g ai ha-1), lactofen (220 g ai ha-1), oxyfluorfen (1,121g ai ha-1), and mesotrione (105 g ai ha-1) herbicides. Atrazine (2,240 g ai ha-1) controlled the CT-Res biotype only 52%, suggesting the biotype is resistant to this herbicide as well. Here we report the first case of Palmer amaranth from Connecticut with multiple resistance to glyphosate and ALS inhibitors. Growers should proactively use all available weed control tactics, including the use of effective PRE and alternative POST herbicides (tested in this study), for effective control of the CT-Res biotype. Nomenclature: 2, 4-D ester; atrazine; carfentrazone-ethyl; chlorimuron-ethyl; clopyralid, dicamba; glyphosate; glufosinate; halosulfuron-methyl; imazaquin; lactofen; mesotrione; oxyfluorfen, sulfometuron-methyl; Palmer amaranth, Amaranthus palmeri S. Watson

In summer, 2011, we investigated suspected glyphosate-resistant (GR) kochia in three chem-fallow fields (designated F1, F2, F3, each farmed by a different grower) in southern Alberta. This study characterizes glyphosate resistance in those populations, based on data from dose–response experiments. In a greenhouse experiment, the three populations exhibited a resistance factor ranging from 4 to 6 based on shoot biomass response (GR50 ratios), or 5 to 7 based on survival response (LD50 ratios). Similar results were found in a field dose–response experiment at Lethbridge, AB, in spring 2012 using the F2 kochia population. In fall 2011, we surveyed 46 fields within a 20-km radius of the three chem-fallow fields for GR kochia. In the greenhouse, populations were screened with glyphosate at 900 g ae ha−1. Seven populations were confirmed as GR, the farthest site located about 13 km from the three originally confirmed populations. An additional GR population more than 100 km away was later confirmed. Populations were screened for acetolactate synthase (ALS)–inhibitor (thifensulfuron ∶ tribenuron) and dicamba resistance in the greenhouse, with molecular characterization of ALS-inhibitor resistance in the F1, F2, and F3 populations. All GR populations were resistant to the ALS-inhibiting herbicide, but susceptible to dicamba. ALS-inhibitor resistance in kochia was conferred by Pro197, Asp376, or Trp574 amino acid substitutions. Based upon a simple empirical model with a parameter for selection pressure, calculated from weed relative abundance and glyphosate efficacy, and a parameter for seedbank longevity, kochia, wild oat, and green foxtail were the top three weeds, respectively, predicted at risk of selection for glyphosate resistance in the semiarid Grassland region of the Canadian prairies; wild oat, green foxtail, and cleavers species were predicted at greatest risk in the subhumid Parkland region. This study confirms the first occurrence of a GR weed in western Canada. Future research on GR kochia will include monitoring, biology and ecology, fitness, mechanism of resistance, and best management practices. Nomenclature: Dicamba; glyphosate; thifensulfuron; tribenuron; cleavers: false cleavers, Galium spurium L. or catchweed bedstraw, Galium aparine L.; green foxtail, Setaria viridis (L.) Beauv.; kochia, Kochia scoparia (L.) Schrad. KCHSC, synonym: Bassia scoparia (L.) A.J. Scott.; wild oat, Avena fatua L.

American sloughgrass [Beckmannia syzigachne (Steud.) Fernald] is one of the most predominant and troublesome weeds in wheat (Triticum aestivum L.) fields rotated with rice (Oryza sativa L.) in China. Mesosulfuron-methyl is one of the main herbicides used to selectively control B. syzigachne in winter wheat fields in China. After many years of application, mesosulfuron-methyl failed to control B. syzigachne in Yutai County. The objectives of this study were to determine the resistance level to mesosulfuron-methyl and other acetolactate synthase (ALS) inhibitors in the B. syzigachne population collected from Yutai County (R) and identify the mechanism of resistance. The results indicated that the R population was 4.1-fold resistant to mesosulfuron-methyl and was cross-resistant to pyroxsulam (600-fold), imazethapyr (4.1-fold), flucarbazone (12-fold), and bispyribac-sodium (12-fold). In vitro assays revealed that ALS in the R population was as sensitive as that in a susceptible (S) population. Gene sequence analysis identified no known resistant mutations in the ALS gene of the R population. Furthermore, real-time quantitative reverse transcriptase PCR experiments indicated that the expression level of the ALS gene in the R population was not different from that of the S population. However, the cytochrome P450 inhibitor malathion reversed the R population's resistance to mesosulfuron-methyl. The result of ultraperformance liquid chromatography–tandem mass spectrometry (UPLC-MS-MS) spectral analysis indicated that the metabolic rates of mesosulfuron-methyl in the R population were significantly faster than in the S population. Therefore, non-target resistance to mesosulfuron-methyl has been demonstrated in the R population. The resistance was very likely caused by enhanced herbicide metabolism.

Eclipta, widespread in tropical, subtropical, and temperate regions, is one of the main malignant broadleaf weeds and thrives in moist and dryland fields. Field rates of acetolactate synthase (ALS) inhibitors have failed to control eclipta in some farmlands in China. One ALS inhibitor–resistant population (R) collected from Jiangsu province in China was confirmed in the greenhouse in our preliminary work. Whole-plant assays revealed that this R population was highly resistant to four sulfonylureas (pyrazosulfuron-ethyl, 134-fold; bensulfuron-methyl, 172-fold; metsulfuron-methyl, 30-fold; and tribenuron-methyl, 195-fold), two triazolopyrimidines (pyroxsulam, 98-fold; penoxsulam, 30-fold), and one pyrimidinylthio-benzoate (bispyribac-sodium, 166-fold) and was moderately resistant to two imidazolinones (imazethapyr, 10-fold; imazapic, 19-fold). ALS enzyme-activity assays showed insensitivity of the ALS from the R population (resistance index values ranged from 12 to 293) to all of the above ALS inhibitors in vitro. Chromatograms from ALS gene sequence analysis detected a homozygous Pro-197-Ser amino acid substitution in the R population. These results confirmed that the Pro-197-Ser substitution results in broad-spectrum cross-resistance to ALS inhibitors in the eclipta R population. To our knowledge, this study is the first to report broad cross-resistance to ALS inhibitors in eclipta and to obtain the full-length ALS gene sequence.

The rapid evolution and spread of glyphosate-resistant (GR) kochia in the Northern Great Plains is an increasing threat to GR cropping systems and conservation tillage practices common in this region. GR kochia accessions with 4.6- to 11-fold levels of resistance to glyphosate have recently been reported in Montana. Those GR kochia accessions were also suspected to be resistant to acetolactate synthase (ALS) inhibitors, i.e., multiple herbicide-resistant (MHR) kochia. In this research, the level of resistance to the ALS-inhibitor herbicides (sulfonylureas) and the molecular mechanisms conferring resistance to glyphosate and ALS-inhibitor herbicides in MHR kochia was investigated. On the basis of whole-plant dose–response assays, MHR kochia accessions (GIL01, JOP01, and CHES01) were 9.3- to 30-fold more resistant to premixed thifensulfuron methyl + tribenuron methyl + metsulfuron methyl than the susceptible (SUS) accession. In an in vivo leaf-disk shikimate assay, MHR plants accumulated less shikimate than the SUS plants at a discriminate dose of 100 μM glyphosate. Sequencing of the conserved region of EPSPS revealed no target-site mutation at Thr102 or Pro106 residue. MHR kochia accessions had increased relative EPSPS gene copies (~ 4 to 10) compared with the SUS accession (single copy). Furthermore, MHR kochia accumulated higher EPSPS protein compared with the SUS plants. Resistance to the ALS-inhibitor herbicides was conferred by Pro197 amino acid substitution (proline to glutamine). EPSPS gene amplification and a single target-site mutation at Pro197 in ALS gene confer resistance to glyphosate and ALS-inhibitor herbicides, respectively, in MHR kochia accessions from Montana. This is the first confirmation of occurrence of MHR kochia in Montana. Nomenclature: Glyphosate; metsulfuron methyl; thifensulfuron methyl; tribenuron methyl; Kochia, Kochia scoparia (L.) Schrad.