Multiple herbicide-resistant (MHR) kochia is a serious concern in the U.S. Great Plains and warrants alternative herbicide mixtures for its control. Greenhouse and field experiments were conducted at Kansas State University research and extension centers near Hays and Garden City, KS, to investigate the interactions of 2,4-D, dichlorprop-p, dicamba, and halauxifen/fluroxypyr premix in various combinations for MHR kochia control. Two previously confirmed MHR (resistant to glyphosate, dicamba, and fluroxypyr) populations and a susceptible population were tested in a greenhouse study. Kochia at the Hays field site was resistant to glyphosate and chlorsulfuron, whereas the population at Garden City was resistant to glyphosate, dicamba, and fluroxypyr. Results from a greenhouse study indicated that 2,4-D, dicamba, dichlorprop-p, and a halauxifen/fluroxypyr premix provided 26% to 69% control of both MHR populations at 28 d after treatment (DAT). However, the control increased to 85% to 97% when these herbicides were applied in three-way mixtures. Synergistic interactions were observed when dicamba was mixed with dichlorprop-p, 2,4-D, dichlorprop-p + 2,4-D, and halauxifen/fluroxypyr + 2,4-D for shoot dry weight reductions (86% to 92%) of both MHR populations. Results from a field study also indicated synergistic interactions when dicamba was mixed with dichlorprop-p + 2,4-D, halauxifen/fluroxypyr + dichlorprop-p, and halauxifen/fluroxypyr + 2,4-D, resulting in 84% to 95% control of MHR kochia at 28 DAT across both sites. These results indicate that synergistic effects of mixing dicamba with other auxinic herbicides in two- or three-way mixtures can help control MHR kochia.

The ability of weed populations to evolve resistance to herbicides affects management strategies and the profitability of crop production. The objective of this research was to screen Palmer amaranth accessions from Arkansas for glufosinate resistance. Additional efforts focused on the effectiveness of various herbicides, across multiple sites of action (SOAs), on each putative-resistant accession. The three putative accessions were selected from 60 Palmer amaranth accessions collected in 2019 and 2020 and screened with to 0.5× and 1× rates of glufosinate. A dose-response experiment was conducted for glufosinate on accessions A2019, A2020, and B2020. The effectiveness of various preemergence- and postemergence-applied herbicides were evaluated on each accession. Resistance ratios of A2019, A2020, and B2020 to glufosinate ranged from 5.1 to 27.4 when comparing LD50 values to two susceptible accessions, thus all three accessions were resistant to glufosinate. All three accessions (A2019, A2020, and B2020) were found to have a reduction equal to or greater than 20 percentage points in mortality to at least one herbicide from five different SOAs equal to or greater than five sites of action. Herbicides from nine different SOAs controlled A2019 at least 20 percentage points less than the susceptible accessions, which points to a need for additional research to characterize the response of this accession.

Velvetleaf is an economically important weed in agronomic crops in Nebraska and the United States. Dicamba applied alone usually does not provide complete velvetleaf control, particularly when velvetleaf is taller than 15 cm. The objectives of this experiment were to evaluate the interaction of dicamba, fluthiacet-methyl, and glyphosate applied alone or in a mixture in two- or three-way combinations for velvetleaf control in dicamba/glyphosate–resistant (DGR) soybean and to evaluate whether velvetleaf height (≤12 cm or ≤20 cm) at the time of herbicide application influences herbicide efficacy, velvetleaf density, biomass, and soybean yield. Field experiments were conducted near Clay Center, NE in 2019 and 2020. The experiment was arranged in a split-plot with velvetleaf height (≤12 cm or ≤20 cm) as the main plot treatment and herbicides as subplot treatment. Fluthiacet provided ≥94% velvetleaf control 28 d after treatment (DAT) and ≥96% biomass reduction regardless of application rate or velvetleaf height. Velvetleaf control was 31% to 74% at 28 DAT when dicamba or glyphosate was applied alone to velvetleaf ≤20 cm tall compared with 47% to 100% control applied to ≤12-cm-tall plants. Dicamba applied alone to ≤20-cm-tall velvetleaf provided <75% control and <87% biomass reduction 28 DAT compared with ≥90% control with dicamba at 560 g ae ha−1 + fluthiacet at 7.2 g ai ha−1 or glyphosate at 1,260 g ae ha−1. Dicamba at 280 g ae ha−1 + glyphosate at 630 g ae ha−1 applied to ≤20-cm-tall velvetleaf resulted in 86% control 28 DAT compared with the expected 99% control. The interaction of dicamba + fluthiacet + glyphosate was additive for velvetleaf control and biomass reduction regardless of application rate and velvetleaf height.

Barnyardgrass [Echinochloa crus-galli (L.) P. Beauv.] is a noxious grass weed that infests rice fields and causes huge crop yield losses. In this study, we collected 12 E. crus-galli populations from rice fields of Ningxia Province in China and investigated the resistance levels to the acetolactate synthase (ALS) inhibitor penoxsulam and the acetyl-CoA carboxylase (ACCase) inhibitor cyhalofop-butyl. The results showed that eight populations exhibited resistance to penoxsulam and four populations evolved resistance to cyhalofop-butyl. Moreover, all four cyhalofop-butyl–resistant populations (NX3, NX4, NX6, and NX7) displayed multiple herbicide resistance to both penoxsulam and cyhalofop-butyl. The alternative herbicides bispyribac-sodium, metamifop, and fenoxaprop-p-ethyl cannot effectively control the multiple herbicide–resistant (MHR) plants. To characterize the molecular mechanisms of resistance, we amplified and sequenced the target site–encoding genes in resistant and susceptible populations. Partial sequences of three ALS genes and six ACCase genes were examined. A Trp-574-Leu mutation was detected in EcALS1 and EcALS3 in two high-level (65.84- and 59.30-fold) penoxsulam-resistant populations, NX2 and NX10, respectively. In addition, one copy (EcACC4) of ACCase genes encodes a truncated aberrant protein due to a frameshift mutation in E. crus-galli populations. None of the amino acid substitutions that are known to confer herbicide resistance were detected in ALS and ACCase genes of MHR populations. Our study reveals the wide spread of MHR E. crus-galli populations in Ningxia Province that exhibit resistance to several ALS and ACCase inhibitors. Non–target site based mechanisms are likely to be involved in E. crus-galli resistance to the herbicides, at least in four MHR populations.

Kochia accessions (designated as KS-4A and KS-4H) collected from a corn field near Garden City, KS, have previously shown multiple resistance to glyphosate, dicamba, and fluroxypyr. These accessions were also suspected as being resistant to photosystem II (PS II) inhibitors. The main objectives of this research were to 1) confirm the coexistence of cross-resistance to PS II inhibitors (atrazine and metribuzin) applied PRE and POST, 2) investigate the underlying mechanism of PS II-inhibitor resistance, and 3) determine the effectiveness of alternative POST herbicides for control of these multiple herbicide–resistant (MHR) kochia accessions. Results from dose-response experiments revealed that the KS-4A and KS-4H kochia accessions were 23-fold to 48-fold resistant to PRE- and POST-applied atrazine and 13-fold to 18-fold resistant to POST-applied metribuzin compared to a known susceptible kochia accession (KS-SUS). Both accessions also showed putative resistance to PRE-applied metribuzin that needs to be confirmed. Sequence analyses of the psbA gene further revealed that all samples from the KS-4A and KS-4H kochia accessions had a Ser264Gly point mutation. A pretreatment with malathion followed by a POST application of atrazine at 1,120 g ha−1 or metribuzin at 630 g ha−1 did not reverse the resistance phenotypes of these MHR accessions. In a separate greenhouse study, alternative POST herbicides, including bicyclopyrone + bromoxynil; bromoxynil + pyrasulfotole; paraquat alone or in combination with atrazine, metribuzin, 2,4-D, or saflufenacil; and saflufenacil alone or in combination with 2,4-D effectively controlled the KS-4H accession (≥97% injury). To our knowledge, this research reports the first case of kochia accessions with cross-resistance to PRE-applied atrazine and POST-applied metribuzin. Growers should adopt diversified weed control strategies, including the use of competitive crops, cover crops, targeted tillage, and harvest weed seed control along with effective alternative PRE and POST herbicides with multiple sites of action to control MHR kochia seedbanks on their production fields.

Palmer amaranth is one of the most difficult-to-control weeds in row crop systems and has evolved resistance to several herbicide sites of action (SOAs). A late-season weed-escape survey had been conducted earlier to determine the distribution of protoporphyrinogen oxidase–inhibitor resistant Palmer Amaranth in Arkansas. The objective of this study was to evaluate the susceptibility of Arkansas Palmer amaranth accessions to commonly used herbicide SOAs. The SOAs evaluated were group 2 + 9, 3, 4, 5, 10, 14, 15, and 27, and the representative herbicide from each group was imazethapyr + glyphosate (79 + 860 g ha−1), trifluralin (1,120 g ha−1), dicamba (280 and 560 g ha−1), atrazine (560 g ha−1), glufosinate (594 g ha−1), fomesafen (395 g ha−1), S-metolachlor (1,064 g ha−1), and tembotrione (92 g ha−1), respectively. Palmer amaranth mortality varied among accessions across SOAs. Averaged across accessions, the mortality rates, by treatment in order from lowest to highest, were as follows: glyphosate + imazethapyr (16%), tembotrione (51%), dicamba at 280 g ha−1 (51%), fomesafen (76%), dicamba at 560 g ha−1 (82%), atrazine (85%), trifluralin (87%), S-metolachlor (96%), and glufosinate (99.5%). This study provides evidence that Palmer amaranth accessions with low susceptibility to glyphosate + imazethapyr, fomesafen, and tembotrione are widespread throughout Arkansas. Of the remaining SOAs, most Palmer amaranth accessions were sensitive; however, within each herbicide SOA, except glufosinate, control of some accessions was less than expected and resistance is suspected.

Field surveys were conducted across the Blacklands region of Texas during 2016 and 2017 to document the distribution of herbicide-resistant Lolium spp. infesting winter wheat production fields in the region. A total of 68 populations (64 Italian ryegrass, four perennial ryegrass) were evaluated in a greenhouse for sensitivity to herbicides of three different modes of action: an acetolactate synthase (ALS) inhibitor (mesosulfuron-methyl), two acetyl-coenzyme-A carboxylase (ACCase) inhibitors (diclofop-methyl and pinoxaden), and a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) inhibitor (glyphosate). Herbicides were applied at twice the label-recommended rates for mesosulfuron-methyl (29 g ai ha−1), diclofop-methyl (750 g ai ha−1), and pinoxaden (118 g ai ha−1); and at the recommended rate for glyphosate (868 g ae ha−1). The herbicide screenings were followed by dose-response assays of the most-resistant ryegrass population for each herbicide at eight rates (0.5, 1, 2, 4, 8, 16, 32, and 64×), compared with a susceptible population at six rates (0.0625, 0.125, 0.25, 0.5, 1, and 2×). The initial screening and dose-response experiments were conducted in a completely randomized design with three replications and two experimental runs. Survivors (<80% injury) were characterized as highly resistant (0% to 20% injury) or moderately resistant (21% to 79%). Results showed that 97%, 92%, 39%, and 3% of the Italian ryegrass populations had survivors to diclofop-methyl, mesosulfuron-methyl, pinoxaden, and glyphosate treatments, respectively. Of the four perennial ryegrass populations, three were resistant to diclofop-methyl and mesosulfuron-methyl, and one was resistant to pinoxaden as well. Perennial ryegrass populations did not exhibit any resistance to glyphosate. Dose-response assays revealed 37-, 196-, and 23-fold resistance in Italian ryegrass to mesosulfuron-methyl, diclofop-methyl, and pinoxaden, respectively, compared with a susceptible standard. One Italian ryegrass population exhibited three-way multiple resistance to ACCase-, ALS-, and EPSPS-inhibitors. The proliferation of multiple herbicide–resistant ryegrass is a challenge to sustainable wheat production in Texas Blacklands and warrants diversified management strategies.

We conducted a survey in the major row-crop production regions of Texas to determine the response of waterhemp to glyphosate (5-enolpyruvylshikimate-3-phosphate synthase [EPSPS] inhibitor), atrazine (photosystem II [PSII] inhibitor), pyrithiobac (acetolactate synthase [ALS] inhibitor), tembotrione (hydroxyphenylpyruvate dioxygenase [HPPD] inhibitor), fomesafen (protoporphyrinogen oxidase [PPO] inhibitor), and dicamba (synthetic auxin). We evaluated 127 accessions for these herbicides. Resistance was confirmed on the basis of plant survival within an accession, and the injury ratings of surviving plants were used to categorize each accession as resistant (<50% injury) or less sensitive (50% to 89% injury). For glyphosate, approximately 27% of all tested accessions were resistant and 20% were less sensitive. The Gulf Coast region had the most glyphosate-resistant accessions (46% of the accessions from this region), followed by the Blacklands region (9%). A dose-response assay of the most resistant waterhemp accession (TX-25) exhibited 17-fold resistance to glyphosate when compared with a susceptible standard. Waterhemp resistance to atrazine also was common in the Gulf Coast region. The accession with the greatest atrazine resistance (TX-31) exhibited 47- and 68-fold resistance to this herbicide when applied POST and PRE, respectively. Widespread resistance to pyrithiobac was observed in waterhemp accessions throughout the Blacklands and Gulf Coast regions. The most resistant accession identified in this study was 61-fold resistant compared with a susceptible standard. No high-level resistance was detected for tembotrione, dicamba, or fomesafen, but high variability in sensitivity to tembotrione and dicamba was observed. One waterhemp accession exhibited reduced sensitivity to fomesafen; the rest were sensitive. Overall, at least two accessions exhibited resistance or reduced sensitivity to herbicides with five different sites of action. The study illustrates the prevalence of multiple herbicide resistance in waterhemp accessions in Texas and emphasizes the need to implement diversified management tactics.

Kochia [Bassia scoparia (L.) A. J. Scott] is a problematic annual broadleaf weed species in the North American Great Plains. Bassia scoparia inherits unique biological characteristics that contribute to its propensity to evolve herbicide resistance. Evolution of glyphosate resistance in B. scoparia has become a serious threat to the major cropping systems and soil conservation practices in the region. Bassia scoparia populations with resistance to four different herbicide sites of action are a concern for growers. The widespread occurrence of multiple herbicide–resistant (HR) B. scoparia across the North American Great Plains has renewed research efforts to devise integrated weed management strategies beyond herbicide use. In this review, we aim to compile and document the growing body of literature on HR B. scoparia with emphasis on herbicide-resistance evolutionary dynamics, distribution, mechanisms of evolved resistance, agronomic impacts, and current/future weed management technologies. We focused on ecologically based, non-herbicidal strategies such as diverse crop rotations comprising winter cereals and perennial forages, enhanced crop competition, cover crops, harvest weed seed control (HWSC), and tillage to manage HR B. scoparia seedbanks. Remote sensing using hyperspectral imaging and other sensor-based technologies would be valuable for early detection and rapid response and site-specific herbicide resistance management. We propose research priorities based on an improved understanding of the biology, genetic diversity, and plasticity of this weed that will aid in preserving existing herbicide resources and designing sustainable, integrated HR B. scoparia mitigation plans.

Thirteen biotypes of rigid ryegrass were screened for trifluralin resistance. From these, the two most resistant biotypes, SLR 31 and SLR 10, were chosen for further studies involving exposure to other dinitroanilines, mitosis-inhibiting herbicides and 14C-trifluralin. SLR 31, and SLR 10 exhibited an approximate 10-fold reduced sensitivity to trifluralin in comparison to susceptible biotypes. Resistance to five other dinitroaniline herbicides was observed, with reduced sensitivity varying from 32-fold for ethalfluralin to 2.5-fold for isopropalin. The resistance in rigid ryegrass to other herbicides and drugs that affect mitosis were tested. Resistance comparable to that of trifluralin was recorded for the herbicides terbutol and DCPA, while low levels of cross-resistance to amiprophosmethyl was present. Trifluralin affected mitotic indices at a much lower level in the susceptible biotypes than in the resistant biotypes. No differences in the uptake and translocation of 14C-trifluralin were observed between resistant and susceptible biotypes. Most of the 14C detected in the plant material was in the root tissue. A small level of 14C was detected in the seeds, and no substantial increases were noted in coleoptile tissue. The resistance spectra in SLR 31 and SLR 10 were phenotypically similar to those occurring in an intermediate trifluralin-resistant goosegrass and trifluralin-resistant green foxtail.

An overreliance on herbicides in several important grain- and cotton-producing regions of the world has led to the widespread evolution of herbicide-resistant weed populations. Of particular concern are weed populations that exhibit simultaneous resistance to multiple herbicides (MHR). Too often, herbicides are the only tool used for weed control. We use the term herbicide-only syndrome (HOS) for this quasi-addiction to herbicides. Growers and their advisers focus on herbicide technology, unaware of or ignoring basic evolutionary principles or the necessary diversity provided by other methods of weed control. Diversity in weed control practices disrupts resistance evolution. Significant challenges exist to implementing diversity, including how to address information so that producers choose to alter existing behaviors (HOS) and take calculated risks by attempting new and more complex strategies. Herbicide resistance management in the long term will require creativity in many sectors, including roles for growers, industry, researchers, consultants, retailers, and regulators. There can be creativity in herbicide registration and regulation, as exemplified by the recent U.S. Environmental Protection Agency program that encourages herbicide registrants to register products in minor crops. We propose one idea for a regulatory incentive to enable herbicide registrants in jurisdictions such as the United States to receive an extended data exclusivity period in exchange for not developing one new herbicide in multiple crops used together in rotation, or for implementing stewardship practices such as robust mixtures or limitations on application frequency. This incentive would provide a mechanism to register herbicides in ways that help to ensure herbicide longevity. Approaches based only on market or financial incentives have contributed to the current situation of widespread MHR. Our suggestion for regulatory creativity is one way to provide both financial and biological benefits to the registering company and to the overall stakeholder community by incentivizing good resistance management.

A population of oriental mustard from Port Broughton in South Australia was reported as not being controlled by 2,4-D. Dose response experiments determined this population was resistant to both 2,4-D and MCPA, requiring greater than 20 times more herbicide for equivalent control compared to a known susceptible population (from Roseworthy, South Australia) and a population resistant only to the acetohydroxyacid synthase (AHAS)-inhibiting herbicides (from Tumby Bay, South Australia). The Port Broughton population was also found to be resistant to three chemical groups that inhibit AHAS; however, the level of resistance was lower than the known acetolactate synthase–resistant population from Tumby Bay. Herbicides from other modes of action were able to control the Port Broughton population. Assays of isolated AHAS from the Port Broughton population showed high levels of resistance to the sulfonylurea and sulfonamide herbicide groups, but not to the imidazolinone herbicides. A single nucleotide change in the AHAS gene that predicted a Pro to Ser substitution at position 197 in the protein was identified in the Port Broughton population. This population of oriental mustard has evolved multiple resistance to AHAS-inhibiting herbicides (AHAS inhibitors) and auxinic herbicides, through a mutation in AHAS and a second nontarget-site mechanism. Whether the same mechanism provides resistance to both AHAS inhibitors and auxinic herbicides remains to be determined. Multiple resistance to auxinic herbicides and AHAS inhibitors in the Port Broughton population will make control of this population more difficult.

The inheritance and linkage of enhanced metabolism-based herbicide cross-resistance was examined in a multiple resistant population of rigid ryegrass. F1 hybrids between resistant and susceptible populations showed an intermediate response to all the four herbicides tested, with no indication of maternal inheritance. Segregation of F2 families fitted a single-gene model for resistance to simazine, chlorotoluron, and chlorsulfuron. But there was more than the expected mortality from the low dose of tralkoxydim. Segregating F2 populations were selected by high rates of each of the four herbicides to create selected F2 families. Analysis of the response of these families demonstrated that simazine resistance is linked to chlorotoluron resistance. No other herbicide resistances were linked. Therefore, in this population of rigid ryegrass, at least three separate genes are responsible for metabolism-based cross-resistance. This study shows that multiple herbicide resistance in rigid ryegrass is the result of accumulation of a number of different resistance genes.

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.

Field and greenhouse experiments were conducted to determine the level of glyphosate resistance in common waterhemp populations from Platte County (MO1) and Holt County, Missouri (MO2), and to determine the level and distribution of resistance to glyphosate, acetolactate synthase (ALS)–inhibiting herbicides, and protoporophyrinogen oxidase (PPO)–inhibiting herbicides across the MO1 site. Results from greenhouse experiments revealed that the MO1 and MO2 waterhemp populations were 19 and 9 times more resistant to glyphosate, respectively, than a susceptible waterhemp population. In field experiments, greater than 54% of waterhemp at the MO1 site survived 1.7 kg glyphosate ae ha−1 (twice the labeled rate) 6 wk after treatment. Tank-mix combinations of ALS- and PPO-inhibiting herbicides with glyphosate also failed to provide complete control of the waterhemp population at the MO1 site. Collection and screening of seed from individual female waterhemp accessions revealed multiple resistance to glyphosate, ALS-, and PPO-inhibiting herbicides across the MO1 site. All 14 waterhemp accessions collected across the MO1 site exhibited greater than 65% survival to 2× rates of glyphosate and thifensulfuron, and these accessions were spread across a 5-km2 (503-ha) area. Four waterhemp accessions collected across a 0.9-km2 (87-ha) area also exhibited 26 to 38% survival to 2× rates of lactofen. The results from these experiments provide evidence and confirmation of the first glyphosate-resistant waterhemp population in the United States and reveal that multiple resistance to glyphosate, ALS-, and PPO-inhibiting herbicides can occur in waterhemp.

Surviving rigid ryegrass plants were collected from a cropping field at Pindar, Western Australia (population WALR 50), after inadequate control by glyphosate applied at the normal field rate. Plants were grown to maturity in pots and seeds were collected. Glyphosate dose–response experiments with known susceptible and resistant control populations confirmed the resistant status of the WALR 50 population. The glyphosate rate resulting in 5% mortality (LD50) and GR50 (the glyphosate rate required to reduce mean growth of individuals to 50% of the untreated control) values for this population were 1,069 and 217 g ae ha−1, respectively, corresponding to R:S ratios of 3.4 and 1.9 for mortality and growth. In addition, a novel root growth–based assay of glyphosate resistance was developed and validated, giving a root growth GR50 R:S ratio of 3.4. A resistance profile was established by assessing population-level survival of WALR 50 after applications at recommended rates of a range of herbicides commonly used for rigid ryegrass control in Australia. High levels of resistance to the acetolactate synthase (ALS)–inhibiting sulfonylurea herbicides chlorsulfuron and sulfometuron, moderate resistance to the acetyl coenzyme A carboxylase (ACCase)–inhibiting herbicide diclofop, and low levels of resistance to the imidazilinone herbicide imazethapyr were found. More detailed dose–response experiments confirmed resistance to chlorsulfuron, sulfometuron, and diclofop. In vitro enzyme-inhibition studies demonstrated that ALS resistance in WALR 50 is due to an insensitive target enzyme and that ACCase resistance is due to a nontarget site–based mechanism. WALR 50 is the first glyphosate-resistant weed population with confirmed resistance to ACCase- and ALS-inhibiting herbicides.