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.

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.

Following six consecutive annual applications of imazaquin in combination with trifluralin or pendimethalin to several soybean fields on the Delmarva Peninsula, unacceptable smooth pigweed control was observed. Field and greenhouse studies were conducted to determine if this population of smooth pigweed was resistant to imazaquin and other herbicides. In field research, imazaquin and imazethapyr gave complete control of the susceptible (S) population while providing no control of the resistant (R) population; pyrithiobac controlled 99 and 90% of the R and S populations, respectively. Pendimethalin, metribuzin, MON-12000, and flumiclorac gave less than 75% control of both S and R populations. Chlorimuron, primisulfuron, CGA-152005, and lactofen gave above 75% control, and thifensulfuron and nicosulfuron gave above 90% control of both S and R populations. Seeds were collected from the R and S smooth pigweed populations for research in the greenhouse. Greenhouse studies confirmed high levels of resistance to imazaquin and imazethapyr and low levels of cross-resistance to rimsulfuron and chlorimuron in the R population. Susceptibility of the R population to nicosulfuron, thifensulfuron, pyrithiobac, and pendimethalin was comparable to that of the S population.

Cleavers species (false cleavers and catchweed bedstraw) are among the top 10 most abundant weeds across the prairie region of western Canada, and are increasing in relative abundance at the fastest rate since the 1970s. In 2008, two false cleavers populations from Tisdale and Choiceland, Saskatchewan, were suspected of acetolactate synthase (ALS) –inhibitor resistance. Dose-response experiments were conducted with the use of imazethapyr and florasulam, both ALS inhibitors, as well as fluroxypyr, a synthetic auxin. Additionally, a 1,954–base-pair region of the ALS gene including sites known to confer ALS resistance were sequenced. Both populations were highly resistant to imazethapyr (resistance factors greater than 100), one population (Tisdale) was highly resistant to florasulam (Choiceland population susceptible, although a second, larger screening of 200 individuals indicated low frequency [2%] florasulam resistance), and both populations were susceptible to fluroxypyr. All sequenced Tisdale individuals screened with imazethapyr posessed the Trp574Leu mutation. In contrast, three point mutations were found for Choiceland individuals sequenced: Ser653Asn, Trp574Leu, and Asp376Glu. These ALS target-site mutations have not been documented previously in this species.

A survey of 109 fields was conducted across western Canada in spring 2007 to determine the extent of ALS-inhibitor and dicamba (synthetic auxin) resistance in kochia. Weed seedlings were collected from fields in three provinces of western Canada and transplanted into the greenhouse. Seeds were harvested from selfed plants, and the F1 progeny were screened for resistance to the ALS-inhibitor mixture thifensulfuron–tribenuron or dicamba. All kochia populations were susceptible to dicamba. ALS inhibitor–resistant kochia was found in 85% of the fields surveyed in western Canada: 80 of 95 fields in Alberta, six of seven fields in Saskatchewan, and all seven fields in Manitoba. For the 93 ALS inhibitor–resistant populations, the mean frequency (±SE) of parental plants classified as resistant was 61 ± 3%. Most of the resistant populations (87%) were heterogeneous and contained both resistant and susceptible individuals. ALS sequence data (Pro197 and Asp376 mutations) and genotyping data (Trp574 mutation) obtained for 87 kochia parental (i.e., field-collected) plants confirmed the presence of all three target-site mutations as well as two mutational combinations (Pro197 + Trp574, Asp376 + Trp574) in resistant individuals.

Glyphosate-resistant (GR) giant ragweed, horseweed, and common ragweed were confirmed in southwestern Ontario, Canada in 2008, 2010, and 2011, respectively. In the western prairie provinces of Alberta and Saskatchewan, GR (plus acetolactate synthase inhibitor-resistant) kochia was discovered in 2011. This symposium paper estimates the environmental impact (EI) of the top herbicide treatments or programs used to manage these GR weed species in the major field crops grown in each region. For each herbicide treatment, EI (per ha basis) was calculated as the environmental impact quotient (EIQ), which quantifies the relative potential risk of pesticide active ingredients on human and ecological health based on risk components to farm workers, consumers, and the environment, multiplied by the application rate (kg ai ha−1). Total EI is defined as EI (per ha basis) multiplied by the application area (i.e., land area affected by a GR weed). It was assumed that all herbicide treatments would supplement the continued usage of glyphosate because of its broad spectrum weed control. For the control of these GR weeds, most treatments contain auxinic or protoporphyrinogen oxidase (PPO)-inhibiting herbicides. The majority of auxinic herbicide treatments result in low (EI ≤ 10) to moderate (11 to 20) EI, whereas all treatments of PPO inhibitors have low EI. Total EI of GR horseweed and kochia will generally be greater than that of giant or common ragweed because of rapid seed dispersal. For recommended herbicide treatments to control GR weeds (and herbicide-resistant weeds in general), EI data should be routinely included with cost and site of action in weed control extension publications and software, so that growers have the information needed to assess the EI of their actions.

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.

The molecular basis for acetolactate synthase (ALS)–inhibitor resistance was determined for 6 susceptible (HS) and 24 resistant (HR) kochia populations from western Canada. The latter included 3 HR populations from Alberta (AB), 3 from Manitoba (MB), and 18 from Saskatchewan (SK). HR plants survived application of the ALS-inhibitor herbicide thifensulfuron–tribenuron mixture in the greenhouse. Most of the HR populations were heterogeneous and contained both HR and HS individuals. The molecular basis for resistance was determined in 273 HR individuals by sequencing the ALS gene (2,270 base pair [bp]) or by conducting a TaqMan genotyping assay developed in this study using real-time polymerase chain reaction (PCR) for single nucleotide polymorphism (SNP) 1709, where a G to T substitution resulted in a Leu for Trp substitution at amino acid position 574 (Trp574Leu mutation). A total of 16 SNPs were identified in the ALS gene sequences (0.7% polymorphism), 5 of which resulted in amino acid changes that confer resistance to ALS-inhibiting herbicides. The SNPs correspond to three target-site mutations: Pro197 (SNPs 565 and 566), Asp376 (SNP 1116), and Trp574 (SNPs 1708 and 1709). The Trp574Leu mutation was predominant (189 HR plants). The next most common mutation was the highly variable residue Pro197 (44 HR plants) with substitution by one of nine amino acids. The least-frequent were Asp376Glu (9 plants) and Trp574Arg (3 plants) substitutions. The presence of two ALS target-site mutations was found in 30 individual kochia plants, the first report from field-selected weed populations. These include combinations Pro197 + Trp574 (23 plants) and Pro197 + Asp376 (7 plants). The detection of Pro197, Asp376, and Trp574 mutations, as well as both combinations, from geographically separate regions suggests multiple origins of these mutations.

ALS inhibitor-resistant biotypes are the fastest growing class of herbicide-resistant (HR) weeds. A Canadian ALS inhibitor-resistant biotype of Russian thistle was first reported in 1989. The molecular basis for ALS-inhibitor resistance is unknown for Canadian populations of this polyploid weed species, and was determined in this study for one Alberta and two Saskatchewan HR Russian thistle populations. HR plants survived spray application of the ALS-inhibitor mixture thifensulfuron : tribenuron in the greenhouse. All three HR Russian thistle populations were heterogeneous and contained both HR and herbicide-susceptible (HS) individuals. The molecular basis for resistance was determined by sequencing the ALS gene and/or conducting a TaqMan genotyping assay for single nucleotide polymorphism (SNP) for the Trp574Leu mutation. Two target-site mutations were observed: Trp574Leu in all three biotypes (554 individuals) and Pro197Gln in one biotype (one individual), suggesting multiple-founding events for Russian thistle HR populations in western Canada. Segregation patterns among F1 and F2 progeny arrays of HR lines sprayed under greenhouse conditions varied; some segregated (i.e., had HR and HS progeny), whereas other lines were exclusively HR. In contrast, no segregation of molecular types, i.e., Trp574, Trp/Leu574 and Leu574, as would be expected with heterozygosity at a single locus Trp/Leu574, was observed. Such lack of segregation is consistent with the polyploid genome structure of Russian thistle and the presence of two copies of the ALS gene. The presence of more than one ALS gene confounded the ability of the molecular techniques to accurately identify “true” heterozygotes in this study.

Acetolactate synthase (ALS)–inhibiting herbicides are often used to control Italian ryegrass in winter wheat in Texas. An Italian ryegrass biotype near Waco, TX was evaluated for resistance to mesosulfuron in field and greenhouse experiments. Control of the biotype in the field was less than 10% with the label rate of mesosulfuron (15 g ai/ha). Greenhouse studies confirmed that the biotype was resistant to mesosulfuron; control of the biotype was less than 35% at 120 g ai/ha mesosulfuron. The herbicide dose required to reduce plant biomass of a susceptible and the Waco biotype by 50% (GR50) was 1.3 and 31 g ai/ha, respectively, indicating a resistance level of 24-fold in the Waco biotype. However, the Waco biotype was controlled with the acetyl-CoA carboxylase inhibitors diclofop and pinoxaden.

Acetolactate synthase (ALS) –inhibitor resistance has been recently documented in a yellow nutsedge biotype in Arkansas rice production, with a target-site mutation resulting in an amino acid substitution from Trp574 to Leu. Preliminary observations have indicated that the resistant biotype showed distinct phenotypic characteristics. Two greenhouse experiments were conducted on the resistant biotype in comparison with three susceptible standards (1) to understand differential growth habit and spatial distribution, and (2) to characterize shoot emergence pattern and seedling vigor. The resistant biotype exhibited a drastically different growth habit with secondary and tertiary basal bulbs emerging away from the parent shoot, resulting in a wider spatial distribution and ground coverage compared to the very compact growth habit of susceptible biotypes. Unlike the susceptible biotypes, the rhizomes developing into tubers were not often connected to the primary basal bulb, but were originating randomly from daughter shoots. The resistant biotype produced an extensive subterranean network of rhizomes and basal bulbs, with wider root spread and distribution compared to the susceptible biotypes. The growth habit of the resistant biotype appeared to be intermediate between yellow and purple nutsedges. Further, the resistant biotype showed a considerably delayed emergence pattern with relatively high levels of tuber dormancy. Although the resistant plants exhibited low early-growth seedling vigor and biomass production compared to the susceptible biotypes (perhaps because of smaller tubers), final aboveground biomass production was greater than that of susceptible biotypes. The overall growth habit and phenotype of the resistant biotype may provide a competitive advantage over adjacent species through the ability to occupy niches and gain improved access to critical resources. The distinct growth pattern may also mean that tillage should not be relied upon for control because it can assist further spread by disconnecting and displacing the chains of rhizomes.

Wild buckwheat is the most abundant broadleaf weed across the Prairie region of western Canada. Acetolactate synthase (ALS)-inhibiting herbicides are commonly used to control this species and other broadleaf weeds in cereal crops. A field survey in Alberta in 2007 identified a single population that was putatively resistant to ALS-inhibiting herbicides. In herbicide resistance screening in the greenhouse, all F1 progeny tested were resistant to the ALS-inhibiting herbicides thifensulfuron/tribenuron, a sulfonylurea herbicide, or florasulam, a triazolopyrimidine herbicide; dose response of shoot biomass indicated the population was 10- and 20-fold less sensitive to thifensulfuron/tribenuron and florasulam, respectively, than a susceptible control population. ALS gene sequencing of 24 F1 progeny indicated that the Trp574Leu target-site mutation was responsible for conferring ALS-inhibitor resistance in this biotype, the first global report of ALS-inhibitor resistance for this species. Because this mutation typically endows high-level resistance across all five ALS-inhibitor classes, this wild buckwheat biotype may only be controlled by a different site-of-action herbicide.

This greenhouse experiment examined the response of homozygous susceptible and acetolactate synthase (ALS) inhibitor–resistant plants from six Canadian kochia accessions with the Pro197 or Trp574 mutation to six alternative herbicides of different sites of action. The null hypothesis was ALS-inhibitor–resistant and –susceptible plants from within and across accessions would respond similarly to herbicides of different sites of action. This hypothesis was accepted for all accessions except that of MBK2 with the Trp574 mutation. Resistant plants of that accession were 80, 60, and 50% more sensitive than susceptible plants to pyrasulfotole, mesotrione (hydroxyphenylpyruvate dioxygenase [HPPD] inhibitors), and carfentrazone (protoporphyrinogen oxidase [PPO] inhibitor), respectively. However, no differential dose response between resistant and susceptible plants of this kochia accession to bromoxynil, fluroxypyr, or glyphosate was observed. A previous study had found marked differences in growth and development between resistant and susceptible plants of this accession, but not of the other accessions examined in this experiment. Negative cross-resistance exhibited by resistant plants of accession MBK2 to PPO and HPPD inhibitors in this experiment may be a pleiotropic effect related to the Trp574 mutation.

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.

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.

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.