Accounting for 53% of United States peanuts (Arachis hypogaea L.), Georgia is the top peanut-producing state with approximately 1.42 billion kg produced in 2023. Peanut producers often use the ALS imidazolinone herbicide imazapic but reduced yellow nutsedge (Cyperus esculentus L.) control was reported in Georgia peanuts after four-years of continuous imazapic use. This study aimed to determine the level of resistance (LD50, I50, and GR50), potential cross-resistance for the suspected resistant population, and identify the associated genetic mutations conferring resistance. A susceptible biotype was treated with 0, 0.0088, 0.0175, 0.035, 0.07, 0.14, 0.28, and 0.56 kg ai ha-1, and a resistant biotype was sprayed with 0, 0.07, 0.14, 0.28, 0.56, 1.13, 2.26, and 4.5 kg ai ha-1 of imazapic. To determine if the suspected resistant biotype was cross-resistant to halosulfuron-methyl, an ALS herbicide used to control nutsedge spp., both biotypes were treated with 0, 0.0117, 0.0233, 0.0466, 0.0933, 0.187, 0.373, and 0.746 g ai ha-1 of halosulfuron-methyl. Plants were rated for injury 7, 14, and 28 days after treatment (DAT), and above-ground biomass harvested at 28 DAT. For imazapic, LD50 was 0.041 and 1.503 kg ai ha-1 and the GR50 was estimated to be 0.0128 and 1.853 kg ai ha-1 for Sus and Res biotypes, respectively, indicating 36 and 145-fold increase in resistance of the Res biotype for I50 and GR50, respectively. Both biotypes responded similarly to applications of halosulfuron-methyl, with biomass reductionat rates greater than 0.023 kg ai ha-1. Transcriptome profiles revealed a mutation in the target-site gene of the resistant biotype, causing an amino acid substitution from Alanine to Valine at position 205 (Ala205Val). Growers should continue to rotate chemistries and implement integrated weed management approaches for control of C. esculentus as the use of imazapic over consecutive years has led to resistance in C. esculentus.

Downy brome (Bromus tectorum L.) is a difficult species to control in the dryland wheat (Triticum aestivum L.) production areas of northeastern Oregon. The selection of herbicide-resistant B. tectorum populations has further complicated B. tectorum management. A survey of wheat growers was conducted in 2021 and 2022 to understand B. tectorum management practices. The survey included four questions based on the growing seasons from 2017 to 2022 related to crop rotation, tillage versus no-tillage, irrigation versus dryland, and herbicide programs. To determine the extent of herbicide resistance, seeds were collected from 49 B. tectorum populations in wheat fields in northeastern Oregon and tested for resistance. Herbicides tested were clethodim, glyphosate, imazamox, mesosulfuron, metribuzin, propoxycarbazone, pyroxasulfone, pyroxsulam, quizalofop, and sulfosulfuron. Winter wheat–summer fallow rotation was the most predominant cropping system in the region. Most of the fields were in no-tillage systems, and none were irrigated. Pyroxasulfone applied preemergence and acetolactate synthase (ALS) inhibitors applied postemergence were the most often used herbicides for B. tectorum control in winter wheat. Glyphosate was the most frequently used herbicide for B. tectorum control in summer fallow. Resistance screenings confirmed that 46 of the 49 B. tectorum populations were resistant to ALS inhibitors with different cross-resistance patterns. Two populations were resistant to metribuzin and exhibited multiple resistance to ALS inhibitors. All populations were susceptible to clethodim, glyphosate, pyroxasulfone, and quizalofop. The widespread occurrence of ALS inhibitor–resistant B. tectorum populations limits effective postemergence herbicide options in winter wheat.

Digitaria ciliaris var. chrysoblephara (Fig. & De Not.) R.R. Stewart is an annual xeromorphic weed that severely infests direct-seeded rice fields in China. Herbicide resistance is emerging in D. ciliaris var. chrysoblephara owing to extensive and recurrent use of the acetyl-CoA carboxylase (ACCase)-inhibiting herbicide metamifop. In this study, a total of 53 D. ciliaris var. chrysoblephara populations randomly sampled from direct-seeded rice fields across Jiangsu Province were investigated for metamifop resistance and potential resistance-endowing mutations. Single-dose assays revealed that 17 (32.1%) populations evolved resistance to metamifop and 5 (9.4%) populations were in the process of developing resistance. The resistance index (RI) of metamifop-resistant populations ranged from 2.7 to 32.1. Amino acid substitutions (Ile-1781-Leu, Trp-2027-Cys/Ser, and Ile-2041-Asn) in ACCase genes were detected in resistant D. ciliaris var. chrysoblephara plants and caused various cross-resistance patterns to ACCase-inhibiting herbicides. All of four resistant populations (YC07, YZ09, SQ03, and HA06), with different ACCase mutations, exhibited cross-resistance to the aryloxyphenoxypropionate (APP) herbicides cyhalofop-butyl (RI values: 10.0 to 19.9), fenoxaprop-P-ethyl (RI values: 53.7 to 132.8), and haloxyfop-P-methyl (RI values: 6.2 to 62.6), and the phenylpyrazoline (DEN) pinoxaden (RI values: 2.3 to 5.4), but responded differently to the cyclohexanedione (CHD) herbicides clethodim and sethoxydim. It is noteworthy that four postemergence herbicides used for rice cropping, including bispyribac-sodium, pyraclonil, quinclorac, and anilofos, showed poor control effect against D. ciliaris var. chrysoblephara, suggesting few alternations for managing this weed in rice fields except ACCase inhibitors. In conclusion, this work demonstrated that the D. ciliaris var. chrysoblephara had developed resistance to ACCase-inhibiting herbicides in rice cultivation of China, and target-site amino acid substitutions in ACCase were primarily responsible for metamifop resistance.

Herbicide-resistant annual bluegrass (Poa annua L.) has become a problem in non-arable land areas. In arable fields, P. annua is frequently of lower priority in weed control program due to the variety of control options available and the relatively modest impact on crop yield compared with other species. In Ireland, postemergence herbicides are not primarily intended for P. annua control, but some herbicides, including the acetolactate synthase (ALS) inhibitor mesosulfuron-methyl + iodosulfuron-methyl, exhibit P. annua activity. In this study, a suspected P. annua population (POAAN-R) that survived mesosulfuron-methyl + iodosulfuron-methyl at 0.75 of the field recommended rate was sampled from a wheat (Triticum aestivum L.) field in County Dublin, Ireland. Single-dose testing confirmed that the suspected POAAN-R had evolved resistance to mesosulfuron-methyl + iodosulfuron-methyl and, additionally, to pyroxsulam (not registered in Ireland for P. annua control), but was sensitive to clethodim, glyphosate, pendimethalin, and flufenacet. Dose–response experiments indicated that POAAN-R was more resistant (GR50 resistance index) to both mesosulfuron-methyl + iodosulfuron-methyl (47.8 times) and pyroxsulam (38.0 times) than sensitive POAAN-S, and this was associated with the mutation at Trp-574 in the ALS protein. Malathion (a cytochrome P450 [P450] inhibitor) pretreatment did not reverse POAAN-R resistance to mesosulfuron-methyl + iodosulfuron-methyl or pyroxsulam at the field rate or above. The natural inherent mutation at Ile-1781 in acetyl-CoA carboxylase protein had no effect on both POAAN-R and POAAN-S sensitivity to clethodim. The glyphosate sensitivity of POAAN-R also corresponded with no known mutation in 5-enolpyruvylshikimate-3-phosphate synthase protein. Based on field histories, poor early-season weed control coupled with intensive use of mesosulfuron-methyl + iodosulfuron-methyl (often at reduced rates) has unintentionally selected for ALS inhibitor–resistant POAAN-R. This is the first report to characterize resistance in P. annua to ALS-inhibiting herbicides mesosulfuron-methyl + iodosulfuron-methyl and pyroxsulam in an arable setting. There is an opportunity to effectively control POAAN-R using herbicides, but this needs a wide-ranging and varied approach, coupled with cultural/nonchemical practices.

Shortawn foxtail (Alopecurus aequalis Sobol.) is an obligate wetland plant that is widely distributed throughout Europe, temperate Asia, and North America. In China, it is widespread in the middle and lower reaches of the Yangtze River as a noxious weed in winter cropping fields with a rice (Oryza sativa L.) rotation. The acetolactate synthase (ALS)-inhibiting herbicide mesosulfuron-methyl has been widely used to control annual grass and broadleaf weeds, including A. aequalis, in wheat (Triticum aestivum L.) fields, leading to the selection of herbicide-resistant weeds. In this study, an A. aequalis population, AHFT-4, that survived mesosulfuron-methyl at the field-recommended rate (9 g ai ha−1) was collected in Anhui Province. Single-dose testing confirmed that the suspected resistant AHFT-4 had evolved resistance to mesosulfuron-methyl. Target gene sequencing revealed a resistance mutation of Pro-197-Ala in ALS1 of the resistant plants, and a derived cleaved amplified polymorphic sequence marker was developed to specifically detect the mutation. A relative expression assay showed no significant difference in ALS expression between AHFT-4 and a susceptible population without or with mesosulfuron-methyl treatment. Whole-plant dose–response bioassays indicated that AHFT-4 had evolved broad-spectrum cross-resistance to ALS-inhibiting herbicides of all five chemical families tested, with GR50 resistance index (RI) values ranging from 21 to 206. However, it remained susceptible to the photosystem II inhibitor isoproturon. Pretreatment with the cytochrome P450 inhibitor malathion or the glutathione S-transferase inhibitor 4-chloro-7-nitrobenzoxadiazole had no significant effects on the resistance of AHFT-4 to mesosulfuron-methyl. To our knowledge, this study reports for the first time the ALS gene Pro-197-Ala substitution conferring broad-spectrum cross-resistance to ALS-inhibiting herbicides in A. aequalis.

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

The transition from puddled-transplanted rice (Oryza sativa L.) (PTR) to direct-seeded rice (DSR) is gaining popularity in central China. In contrast, the PTR system is the most common practice in southwest China. Weeds are a major problem in the paddy fields of the DSR systems, and herbicides are widely used for weed control. However, the increased frequency and rate of herbicide use leads to the rapid evolution of resistance. Smallflower umbrella sedge (Cyperus difformis L.) is a troublesome weed species in rice fields of China and is usually controlled by the acetolactate synthase (ALS)-inhibiting herbicide bensulfuron-methyl. Here, we collected 32 C. difformis populations from DSR systems (Hunan Province) and PTR systems (Guangxi Province) and investigated their resistance to bensulfuron-methyl. Results revealed 80% (8 out of 10) populations from Hunan Province and 14% (3 out of 22) populations from Guangxi Province had evolved resistance to bensulfuron-methyl. Five populations from Hunan Province (HN-2, HN-3, HN-5, HN-9, HN-10) possessing the Trp-574-Leu mutation had high-level resistance (ranging from 169- to >1,309-fold) based on GR50 ratios. The resistant populations from Guangxi Province had a lower level of resistance to bensulfuron-methyl due to a Pro-197-Ser mutation. The Asp-376-Glu mutation was only identified in the HN-4 population. In addition, the GX-3 population from the PTR systems was resistant to bensulfuron-methyl without ALS gene mutations, indicating non–target site resistance (NTSR). Although some resistant populations of both regions exhibited cross-resistance to multiple ALS-inhibiting herbicides, including pyrazosulfuron-ethyl, bispyribac-sodium, penoxsulam, and imazapic, sensitivity was also detected to the auxin herbicide MCPA and the photosystem II–inhibiting herbicides bentazone and propanil. These results indicate that cultivation practices affect resistance evolution in C. difformis. DSR systems exert high selection pressure by selecting the Trp-574-Leu mutation, resulting in high-level resistance. In contrast, a mutation at Pro-197 plus NTSR likely plays a significant role in ALS resistance in the PTR systems.

Herbicide-resistant (HR) kochia is a growing problem in the Great Plains region of Canada and the United States. Resistance to up to four herbicide sites of action, including photosystem II inhibitors, acetolactate synthase inhibitors, synthetic auxins, and the 5-enolpyruvylshikimate-3-phosphate synthase inhibitor glyphosate have been reported in many areas of this region. Despite being present in the United States since 1993/1994, auxinic-HR kochia is a recent and growing phenomenon in Canada. This study was designed to characterize 1) the level of resistance and 2) patterns of cross-resistance to dicamba and fluroxypyr in 12 putative auxinic-HR kochia populations from western Canada. The incidence of dicamba-resistant individuals ranged among populations from 0% to 85%, while fluroxypyr-resistant individuals ranged from 0% to 45%. In whole-plant dose-response bioassays, the populations exhibited up to 6.5-fold resistance to dicamba and up to 51.5-fold resistance to fluroxypyr based on visible injury 28 d after application. Based on plant survival estimates, the populations exhibited up to 3.7-fold resistance to dicamba and up to 72.5-fold resistance to fluroxypyr. Multiple patterns of synthetic auxin resistance were observed, in which one population from Cypress County, Alberta, was resistant to dicamba but not fluroxypyr, whereas another from Rocky View County, Alberta, was resistant to fluroxypyr but not dicamba based on single-dose population screening and dose-response bioassays. These results suggest that multiple mechanisms may confer resistance to dicamba and/or fluroxypyr in Canadian kochia populations. Further research is warranted to determine these mechanisms. Farmers are urged to adopt proactive nonchemical weed management practices in an effort to preserve efficacy of the remaining herbicide options available for control of HR kochia.

Ludwigia prostrata is a problematic weed in rice fields in China, where acetolactate synthase (ALS)-inhibiting herbicides (e.g., bensulfuron-methyl) are widely used for the management of broadleaf weeds. Recently, an L. prostrata biotype (JS-R) that failed to be controlled with ALS-inhibiting herbicides was found in Jiangsu Province, China. This study aims to determine the level and molecular mechanism of resistance to bensulfuron-methyl in this JS-R biotype and to evaluate its spectrum of cross-resistance to other ALS-inhibiting herbicides. The dose–response assays indicated that the JS-R L. prostrata biotype had evolved 21.2-fold resistance to bensulfuron-methyl compared with the susceptible biotype (JS-S). ALS gene sequencing revealed that a nucleotide mutation (CCA to TCA) at codon 197, resulting in a Pro-197-Ser mutation, was detected in the resistant plants. Moreover, while the JS-R biotype contained the Pro-197-Ser resistance mutation and showed cross-resistance to pyrazosulfuron-ethyl (12.0-fold), it was sensitive to penoxsulam, bispyribac-sodium, and imazethapyr, which may serve as alternative herbicides to control the resistant L. prostrata biotype. This is the first confirmation of an L. prostrata biotype resistant to bensulfuron-methyl due to a Pro-197-Ser resistance mutation in the ALS gene.

Italian ryegrass [Lolium perenne L. spp. multiflorum (Lam.) Husnot] is one of the most troublesome weeds worldwide. Lolium multiflorum is also a grass seed crop cultivated on 50,000 ha in Oregon, where both diploid and tetraploid cultivars are grown. For this work, we will refer to the species as L. multiflorum, since the common names annual ryegrass and Italian ryegrass both refer to the same species. A survey was conducted to understand the distribution and frequency of L. multiflorum and its susceptibility to selected herbicides used in its control. The herbicides selected were clethodim, glufosinate, glyphosate, mesosulfuron-methyl (mesosulfuron), paraquat, pinoxaden, pyroxsulam, quizalofop-P-ethyl (quizalofop), pronamide, flufenacet + metribuzin, and pyroxasulfone. The ploidy levels of the populations were also tested. A total of 150 fields were surveyed between 2017 and 2018, of which 75 (50%) had L. multiflorum present. Herbicide-resistant populations were documented in 88% of the 75 populations collected. The most frequent resistances were to acetyl-CoA carboxylase (ACCase), acetolactate synthase (ALS), 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) inhibitors, and combinations thereof. Multiple resistance and cross-resistance, found in 75% of the populations, were the most frequent patterns of resistance. Paraquat-resistant biotypes were confirmed in six orchard crop populations for the first time in Oregon. Herbicide resistance was spatially clustered, with most cases of resistance in the northern part of the surveyed area. Populations resistant to ALS and ACCase inhibitors were prevalent in wheat (Triticum aestivum L.) fields. Multiple resistance was positively correlated with plant density. Tetraploid feral populations were identified, but no cases of herbicide resistance were documented. This is the first survey of herbicide resistance and ploidy diversity in L. multiflorum in western Oregon. Resistant populations were present across the surveyed area, indicating that the problem is widespread.

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.

A non-GMO trait called Inzen™ was recently commercialized in grain sorghum to combat weedy grasses, allowing the use of nicosulfuron POST in the crop. Inzen™ grain sorghum carries a double mutation in the acetolactate synthase (ALS) gene Val560Ile and Trp574Leu, which potentially results in cross-resistance to a wide assortment of ALS-inhibiting herbicides. To evaluate the scope of cross-resistance to Weed Science Society of America Group 2 herbicides in addition to nicosulfuron, tests were conducted in 2016 and 2017 at the Lon Mann Cotton Research Station near Marianna, AR, the Arkansas Agricultural Research and Extension Center in Fayetteville, AR, and in 2016 at the Pine Tree Research Station near Colt, AR. The tests included ALS-inhibiting herbicides from all five families: sulfonylureas, imidazolinones, pyrimidinylthiobenzoics, triazolinones, and triazolopyrimidines. Treatments were made PRE or POST to grain sorghum at a 1× rate for crops in which each herbicide is labeled. Grain sorghum planted in the PRE trial were Inzen™ and a conventional cultivar. Visible estimates of injury and sorghum heights were recorded at 2 and 4 wk after herbicide application, and yield data were collected at crop maturity. In the PRE trial, no visible injury, sorghum height reduction, or yield loss were observed in plots containing the Inzen™ cultivar. Applications made POST to the Inzen™ grain sorghum caused visible injury, sorghum height reduction, and yield loss of 20%, 13%, and 35%, respectively, only in plots where bispyribac-Na was applied. There was no impact on the crop from other POST-applied ALS-inhibiting herbicides. These results demonstrate that the Inzen™ trait confers cross-resistance to most ALS-inhibiting herbicides and could offer promising new alternatives for weed control and protection from carryover of residual ALS-inhibiting herbicides in grain sorghum.

The evolution of resistance to multiple herbicides in Palmer amaranth is a major challenge for its management. In this study, a Palmer amaranth population from Hutchinson, Kansas (HMR), was characterized for resistance to inhibitors of photosystem II (PSII) (e.g., atrazine), acetolactate synthase (ALS) (e.g., chlorsulfuron), and EPSP synthase (EPSPS) (e.g., glyphosate), and this resistance was investigated. About 100 HMR plants were treated with field-recommended doses (1×) of atrazine, chlorsulfuron, and glyphosate, separately along with Hutchinson multiple-herbicide (atrazine, chlorsulfuron, and glyphosate)–susceptible (HMS) Palmer amaranth as control. The mechanism of resistance to these herbicides was investigated by sequencing or amplifying the psbA, ALS, and EPSPS genes, the molecular targets of atrazine, chlorsulfuron, and glyphosate, respectively. Fifty-two percent of plants survived a 1× (2,240 g ai ha−1) atrazine application with no known psbA gene mutation, indicating the predominance of a non–target site resistance mechanism to this herbicide. Forty-two percent of plants survived a 1× (18 g ai ha−1) dose of chlorsulfuron with proline197serine, proline197threonine, proline197alanine, and proline197asparagine, or tryptophan574leucine mutations in the ALS gene. About 40% of the plants survived a 1× (840 g ae ha−1) dose of glyphosate with no known mutations in the EPSPS gene. Quantitative PCR results revealed increased EPSPS copy number (50 to 140) as the mechanism of glyphosate resistance in the survivors. The most important finding of this study was the evolution of resistance to at least two sites of action (SOAs) (~50% of plants) and to all three herbicides due to target site as well as non–target site mechanisms. The high incidence of individual plants with resistance to multiple SOAs poses a challenge for effective management of this weed.

Organophosphate insecticides, which have the capacity to inhibit specific herbicide-degrading (cytochrome P450) enzymes, have been used to explore metabolic herbicide-resistance mechanisms in weeds. This study investigates the response of seven field-selected rigid ryegrass (Lolium rigidum Gaudin) populations to herbicides from three different sites of action in the presence or absence of the P450 inhibitor phorate. Phorate antagonized the thiocarbamate herbicides triallate and prosulfocarb (8-fold increase in LD50) in multiple resistant L. rigidum populations with resistance to three different site-of-action herbicides. In contrast, phorate synergized trifluralin and propyzamide in some populations, reducing the LD50 by 50%. Conversely, treatment with phorate had no significant effect on the LD50 for S-metolachlor or pyroxasulfone (inhibitors of very-long-chain fatty-acid synthesis). Phorate has diverse effects that are herbicide and population dependant in field-selected L. rigidum, suggesting P450 involvement in the metabolism of trifluralin and failure to activate thiocarbamate herbicides in these populations. This research highlights the need for implementation of diverse approaches other than herbicide alone as part of a long-term integrated strategy to reduce the likelihood of metabolism-based resistance to PPI herbicides in L. rigidum.

Diversity is key for sustainable weed management and can be achieved via both chemical and nonchemical control tactics. Genetically modified crops with two-way or three-way stacked herbicide-tolerant traits allow use of herbicide mixtures that would otherwise be phytotoxic to the crop. Early weed management (EWM) strategies promote the use of PRE herbicides with residual activity to keep the field free of weeds early in the season for successful crop establishment. To evaluate the respective sustainability and practicality of the two chemical-based management tactics (i.e., stacked traits and EWM), we used a population model of waterhemp, Amaranthus tuberculatus (Moq.) Sauer (syn. rudis), to simulate the evolution of resistance in this key weed species in midwestern U.S. soybean [Glycine max (L.) Merr.] agroecosystems. The model tested scenarios with a varying number of herbicide sites of action (SOAs), application timings (PRE and POST), and preexisting levels of resistance. Results showed that both tactics provided opportunity for controlling resistant A. tuberculatus populations. In general, each pass over the field should include at least two effective herbicide SOAs. Nevertheless, the potential evolution of cross-resistance may void the weed control programs embraced by stacked traits and diverse herbicide SOAs. Economic calculations suggested that the diversified programs could double long-term profitability when compared to the conventional system, because of improved yield and grain quality. Ultimately, the essence of a sustainable herbicide resistance management strategy is to be proactive. Although a herbicide-dominated approach to diversifying weed management has been prevalent, the increasing presence of weed populations with multiple resistance means that finding herbicides to which weed populations are still susceptible is becoming increasingly difficult, and thus the importance of reintroducing cultural and mechanical practices to support herbicides must be recognized.

The widespread, rapid evolution of herbicide-resistant weeds is a serious and escalating agronomic problem worldwide. During China’s economic boom, the country became one of the most important herbicide producers and consumers in the world, and herbicide resistance has dramatically increased in the past decade and has become a serious threat to agriculture. Here, following an evidence-based PRISMA (preferred reporting items for systematic reviews and meta-analyses) approach, we carried out a systematic review to quantitatively assess herbicide resistance in China. Multiple weed species, including 26, 18, 11, 9, 5, 5, 4, and 3 species in rice (Oryza sativa L.), wheat (Triticum aestivum L.), soybean [Glycine max (L.) Merr.], corn (Zea mays L.), canola (Brassica napus L.), cotton (Gossypium hirsutum L.)., orchards, and peanut (Arachis hypogaea L.) fields, respectively, have developed herbicide resistance. Acetolactate synthase inhibitors, acetyl-CoA carboxylase inhibitors, and synthetic auxin herbicides are the most resistance-prone herbicides and are the most frequently used mechanisms of action, followed by 5-enolpyruvylshikimate-3-phosphate synthase inhibitors and protoporphyrinogen oxidase inhibitors. The lack of alternative herbicides to manage weeds that exhibit cross-resistance or multiple resistance (or both) is an emerging issue and poses one of the greatest threats challenging the crop production and food safety both in China and globally.

Bearded sprangletop [Diplachne fusca (L.) P. Beauv. ex Roem. & Schult. ssp. fascicularis (Lam.) P. M. Peterson & N. Snow] is a noxious annual grass weed of paddy fields, distributed in coastal regions of the Jiangsu and Hebei provinces in China. Cyhalofop-butyl has been widely used to control grass weeds since 2006 in China. Overreliance on cyhalofop-butyl has led to the evolution of resistant weeds. In this study, the resistance level and cyhalofop-butyl resistance mechanisms were investigated in the putative resistant (JSHH) population. The dose–response experiments showed that the JSHH D. fusca population had evolved 8.9-fold resistance to cyhalofop-butyl. Acetyl-CoA carboxylase (ACCase) sequencing revealed a point mutation (GGC to GCC) at amino acid position 2096, resulting in a Gly-2096-Ala substitution in the resistant population. To our knowledge, this is the first case of cyhalofop-butyl resistance in D. fusca and the first report of a target-site mutation conferring resistance to ACCase-inhibiting herbicides in D. fusca. In addition, the resistant D. fusca population (JSHH) with the Gly-2096-Ala mutation was cross-resistant to the aryloxyphenoxypropionate herbicide metamifop, the cyclohexanedione herbicide sethoxydim, and the phenylpyrazolin herbicide pinoxaden.

Late watergrass [Echinochloa phyllopogon (Stapf) Koso-Pol.] is one of the most persistent weeds in rice fields and shows resistance to some acetolactate synthase (ALS)-inhibiting herbicides, such as penoxsulam. Previous studies of E. phyllopogon’s herbicide resistance have focused on non–target site resistance mechanisms. In this study, E. phyllopogon populations from Heilong Jiang Province, China, that were possibly resistant to penoxsulam were used to identify the target site–based mechanisms of resistance. Population HSRH-520 showed a 25.4-fold higher resistance to penoxsulam than the sensitive population, HSRH-538. HSRH-520 was resistant to other ALS inhibitors, with resistance indexes ranging from 17.1 to 166. Target-gene sequence analysis revealed two different ALS genes in E. phyllopogon; a Pro-197-Ser substitution occurred in the ALS-2 gene of HSRH-520. In vitro activity assays revealed that the penoxsulam concentrations required to inhibit 50% of the ALS activity were 13.7 times higher in HSRH-520 than in HSRH-538. Molecular-docking tests showed that the Pro-197-Ser mutation reduced the binding affinity between ALS and ALS inhibitors belonging to the triazolopyrimidine, sulfonylaminocarbonyltriazolinone, and sulfonylurea families, and there were almost no effects on binding affinity when the ALS inhibitors were of the pyrimidinylthiobenzoate and imidazolinone families. Overall, the results indicated and verified that the Pro-197-Ser mutation leads to increased ALS activity by reducing the binding affinity of the inhibitor and ALS. This is the first report on the Pro-197-Ser mutation in the complete ALS gene of E. phyllopogon and will aid future research of target site–based resistance mechanisms of E. phyllopogon to ALS inhibitors.

Palmer amaranth is one of the most problematic weeds in cropping systems of North America, especially in midsouthern United States, because of its competitive ability and propensity to evolve resistance to several herbicide sites of action. Previously, we confirmed and characterized the first case of nontarget site resistance (NTSR) to fomesafen in a Palmer amaranth accession from Randolph County, AR (RCA). The primary basis of the present study was to evaluate the cross- and multiple-resistance profile of the RCA accession. The fomesafen dose-response assay in the presence of malathion revealed a lower level of RCA resistance when compared with fomesafen alone. The resistance index of the RCA accession, based on 50% biomass reduction, ranged from 63-fold (fomesafen alone) to 22-fold (malathion plus fomesafen), when compared with a 2007 susceptible, and 476-fold and 167-fold, respectively, relative to a 1986 susceptible check. The RCA accession was resistant to other protoporphyrinogen oxidase (PPO) inhibitors (i.e., flumioxazin, acifluorfen, saflufenacil) as well as the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor tembotrione and acetolactate synthase (ALS) inhibitor pyrithiobac sodium. Sequencing of the ALS gene revealed no point mutations, indicating that a target-site mechanism is not involved in conferring ALS-inhibitor resistance in the RCA accession. Of the three PPO-inhibiting herbicides tested in combination with the malathion, saflufenacil resulted in the greatest biomass reduction (80%; P < 0.05) and lowest survival rate (23%; P < 0.05) relative to nontreated plants. The application of cytochrome P450 or glutathione S-transferase inhibitors with fomesafen did not lead to any adverse effects on soybean, suggesting a possible role for these compounds for management of NTSR under field conditions. These results shed light on the relative unpredictability of NTSR in conferring herbicide cross- and multiple resistance in Palmer amaranth.