The aim of this paper is to bring attention to weed ecology research that is taking place in an unexpected discipline: archaeology. While archaeobotanists (archaeologists or botanists who specialize in archaeological plant remains) have been accessing literature in weed ecology for decades and applying the findings to their own studies, their results are almost exclusively published in archaeological journals such as the Journal of Archaeological Science or Vegetation History and Archaeobotany. For this reason, their work is underutilized by weed ecologists, especially those who have an interest in historical weed ecology. Archaeobotanical research could help weed scientists understand the long-term effects of agricultural practices on weed communities and predict the potential impacts of climate change. This paper begins with a brief review of the history of archaeobotany as a discipline, then describes ways in which weed ecology is applied in archaeobotany, including Functional Interpretation of Botanical Surveys (FIBS). Finally, we present opportunities for future collaboration between archaeobotanists and weed scientists.

Herbicide resistance has been studied extensively in agronomic crops across North America but is rarely examined in vegetables. It is widely assumed that the limited number of registered herbicides combined with the adoption of diverse weed management strategies in most vegetable crops effectively inhibits the development of resistance. It is difficult to determine whether resistance is truly less common in vegetable crops or whether the lack of reported cases is due to the lack of resources focused on detection. This review highlights incidences of resistance that are thought to have arisen within vegetable crops. It also includes situations in which herbicide-resistant weeds were likely selected for within agronomic crops but became a problem when vegetables were grown in sequence or in adjacent fields. Occurrence of herbicide resistance can have severe consequences for vegetable growers, and resistance management plans should be adopted to limit selection pressure. This review also highlights resistance management techniques that should slow the development and spread of herbicide resistance in vegetable crops.

Germination and emergence assays represent the most notable examples of time-to-event data in agriculture and related disciplines. In spite of the peculiar characteristics of this type of data, there has been little effort to establish a specific and comprehensive framework for their analyses. Indeed, a brief survey of the literature shows that germination and emergence data, along with other phenological measurements such as flowering time, have been analyzed through myriad approaches, giving rise to confusion and uncertainty among scientists and practitioners as to what may represent the best statistical practice. This lack of coherence in statistical approach may reduce the efficiency of research, while making the communication of results and the cross-study comparisons extremely challenging. Here, we attempt to provide a coherent framework and protocol for the analyses of germination/emergence and other time-to-event data in weed science and related disciplines, together with a software implementation in the form of a new R package. We propose a similar approach to biological assays in ecotoxicology, based on: (1) fitting a time-to-event model to describe the whole time course of events; (2) comparing time-to-event curves across experimental treatments, and (3) deriving further information from the fitted model to better focus on some traits of interest. The most appropriate methods to accomplish this procedure were carefully selected from the framework of survival analysis and related sources and were modified to comply with the specific needs of weed, seed, and plant sciences. Finally, they were implemented in the new R package drcte. In this article, we describe the procedure and its limitations by way of providing examples of several types of germination/emergence assays. We highlight that our proposed procedure can also serve as the first step of data analyses, with its output subsequently submitted to traditional or meta-analytic approaches.

Weeds compete with crops for soil moisture, along with other resources, which can impact the germination, growth, and seed production of weeds; however, this impact has not been systematically recorded and synthesized across diverse studies. To address this knowledge gap, a global meta-analysis was conducted using 1,196 paired observations from 86 published articles assessing the effect of water stress on weed germination, growth characteristics, and seed production. These studies were conducted and published during 1970 through 2020 across four continents (Asia, Australia, Europe, and North America). Imposed water stress was expressed as solution osmotic potential (ψsolution), soil water potential (ψsoil), or soil moisture as percent field capacity. Meta-analysis revealed that water stress inhibits weed germination, growth, and seed production, and the quantitative response intensified with increasing water stress. A ψsolution greater than −0.8 MPa completely inhibits germination of both grass and broadleaf weeds. A ψsolution from −0.09 to −0.32 MPa reduces weed germination by 50% compared with the unstressed condition. Moderate soil water stress, equivalent to 30% to 60% field capacity, inhibits growth characteristics (branches or tillers per plant, leaf area, leaves per plant, plant height, root, and shoot biomass) by 33% and weed seed production by 50%. Severe soil water stress, below 30% field capacity, inhibits weed growth by 51% and seed production by 88%. Although water stress inhibits weed growth, it does not entirely suppress the ability to germinate, grow, and produce seeds, resulting in weed seedbank accumulation. This creates management challenges for producers, because weed seeds can survive in the soil for many years, depending on weed species and environmental conditions. Quantitative information compiled in this meta-analysis can be instrumental to model the weeds’ multidimensional responses to water stress and designing integrated weed management strategies for reducing the weed seedbank.

Recent innovations in 3D imaging technology have created unprecedented potential for better understanding weed responses to management tactics. Although traditional 2D imaging methods for mapping weed populations can be limited in the field by factors such as shadows and tissue overlap, 3D imaging mitigates these challenges by using depth data to create accurate plant models. Three-dimensional imaging can be used to generate spatiotemporal maps of weed populations in the field and target weeds for site-specific weed management, including automated precision weed control. This technology will also help growers monitor cover crop performance for weed suppression and detect late-season weed escapes for timely control, thereby reducing seedbank persistence and slowing the evolution of herbicide resistance. In addition to its many applications in weed management, 3D imaging offers weed researchers new tools for understanding spatial and temporal heterogeneity in weed responses to integrated weed management tactics, including weed–crop competition and weed community dynamics. This technology will provide simple and low-cost tools for growers and researchers alike to better understand weed responses in diverse agronomic contexts, which will aid in reducing herbicide use, mitigating herbicide-resistance evolution, and improving environmental health.

Glufosinate resistance in Palmer amaranth (Amaranthus palmeri S. Watson) was recently detected in three accessions from Arkansas, USA. Amaranthus palmeri is the first and only broadleaf weed species resistant to this herbicide, and the resistance mechanism is still unclear. A previous study characterized the glufosinate resistance level in the accessions from Arkansas. A highly glufosinate-resistant accession was further used to investigate the mechanism conferring glufosinate resistance in A. palmeri. Experiments were designed to sequence the herbicide target enzyme cytosolic and chloroplastic glutamine synthetase isoforms (GS1 and GS2, respectively) and quantify copy number and expression. Absorption, translocation, and metabolism of glufosinate using the 14C-labeled herbicide were also evaluated in the resistant and susceptible accessions. The glufosinate-resistant accession had an increase in copy number and expression of GS2 compared with susceptible plants. All accessions showed only one GS1 copy and no differences in expression. No mutations were identified in GS1 or GS2. Absorption (54% to 60%) and metabolism (13% to 21%) were not different between the glufosinate-resistant and glufosinate-susceptible accessions. Most residues of glufosinate (94% to 98%) were present in the treated leaf. Glufosinate translocation to tissues above the treated leaf and in the roots was not different among accessions. However, glufosinate translocation to tissues below the treated leaf (not including roots) was greater in the resistant A. palmeri (2%) compared with the susceptible (less than 1%) accessions. The findings of this paper strongly indicate that gene amplification and increased expression of the chloroplastic glutamine synthetase enzyme are the mechanisms conferring glufosinate resistance in the A. palmeri accession investigated. Thus far, no additional resistance mechanism was observed, but further investigations are ongoing.

The full spectrum of herbicide resistance in a weed can vary according to the mechanistic basis and cannot be implied from the selective pressure. Common ragweed (Ambrosia artemisiifolia L.) is an important weed species of horticultural crops that has developed resistance to linuron based on either target site– or non–target site resistance mechanisms. The objective of the study is to characterize the cross-resistance to metribuzin of linuron-selected biotypes of A. artemisiifolia with target site– and non–target site resistance and determine its genetic basis. Crosses were made between two types of linuron-resistant biotype and a linuron-susceptible biotype, and the progeny were further backcrossed with susceptible plants to the third backcross (BC3) generation to determine their responses to both herbicides compared with parental lines. The target site–based linuron-resistant biotype was cross-resistant to metribuzin, and resistance to both herbicides was maintained at the same level in the BC3 line. In contrast, the linuron-selected biotype with a non–target site resistance mechanism was not cross-resistant to metribuzin. In addition, the BC3 lines deriving from the non–target site resistant parents had very low-level resistance. While the target site–resistance trait is maintained through multiple crosses, non–target site based resistance would be lost over time when selection is absent or insufficient to retain all genes involved in resistance as a complex trait. This would imply A. artemisiifolia biotypes with different mechanisms would need to be managed differently over time.

The control of multiple-resistant wild radish (Raphanus raphanistrum L.) populations in no-till Australian wheat (Triticum aestivum L.) crops has relied upon 4-hydroxyphenylpyruvate dioxygenase (HPPD)-inhibiting herbicides over the last decade. Two R. raphanistrum populations identified as putatively resistant to pyrasulfotole + bromoxynil in an initial large-scale screening trial were characterized and confirmed to be 5- to 8-fold (comparison of LD50 values) less sensitive than the susceptible control population to the HPPD inhibitor pyrasulfotole when plants were treated at the 4-leaf stage. The two pyrasulfotole-resistant populations exhibited up to 4-fold resistance to the coformulated herbicide mixture pyrasulfotole + bromoxynil and up to 9- and 11-fold cross-resistance to mesotrione and topramezone postemergence, respectively. A small-plot trial was conducted in the field from which of one of the populations suspected of resistance was originally collected. Pyrasulfotole + bromoxynil or topramezone + bromoxynil applied postemergence delivered reduced R. raphanistrum control (79% to 87%), whereas mesotrione applied preemergence was >99% effective. We report here the first case of field resistance to HPPD-inhibiting herbicides in R. raphanistrum, caused by 12 yr of continuous reliance on that mode of action. The mitigation of herbicide resistance in continuous no-till cropping requires a constant optimization of the herbicide technology via alternation and mixtures of multiple sites of action, use of preemergence herbicides, and ensuring postemergence herbicides are applied at the most sensitive plant growth stages.

Waterhemp [Amaranthus tuberculatus (Moq.) Sauer] is one of the most troublesome agronomic weeds in the midwestern United States. The rapid evolution and selection of herbicide-resistance traits in A. tuberculatus is a major challenge in managing this species. An A. tuberculatus population, designated CHR, was identified in 2012 in Champaign County, IL, and previously characterized as resistant to herbicides from six site-of-action groups: 2,4-D (Group 4), acetolactate synthase inhibitors (Group 2), protoporphyrinogen oxidase inhibitors (Group 14), 4-hydroxyphenylpyruvate dioxygenase inhibitors (Group 27), photosystem II inhibitors (Group 5), and very-long-chain fatty-acid synthesis inhibitors (Group 15). Recently, ineffective control of CHR was observed in the field after dicamba application. Therefore, this research was initiated to confirm dicamba resistance, quantify the resistance level, and investigate its inheritance in CHR. Multiple field trials were conducted at the CHR location to confirm poor control with dicamba and compare dicamba treatments with other herbicides. Greenhouse trials were conducted to quantify the resistance level in CHR and confirm genetic inheritance of the resistance. In field trials, dicamba did not provide more than 65% control, while glyphosate and glufosinate provided at least 90% control. Multiple accessions were generated from controlled crosses and evaluated in greenhouse trials. Greenhouse dicamba dose–response experiments indicated a resistance level of 5- to 10-fold relative to a sensitive parental line. Dose–response experiments using F1 lines indicated that dicamba resistance was an incompletely dominant trait. Segregation analysis with F2 and backcross populations indicated that dicamba resistance had moderate heritability and was potentially a multigenic trait. Although dicamba resistance was predominantly inherited as a nuclear trait, minor maternal inheritance was not completely ruled out. To our knowledge, CHR is one of the first cases of dicamba resistance in A. tuberculatus. Further studies will focus on elucidating the genes involved in dicamba resistance.

Herbicide-resistant (HR) crops are widely grown throughout the United States and Canada. These crop-trait technologies can enhance weed management and therefore can be an important component of integrated weed management (IWM) programs. Concomitantly, evolution of HR weed populations has become ubiquitous in agricultural areas where HR crops are grown. Nevertheless, crop cultivars with new or combined (stacked) HR traits continue to be developed and commercialized. This review, based on a symposium held at the Western Society of Weed Science annual meeting in 2021, examines the impact of HR crops on HR weed management in the U.S. Great Plains, U.S. Pacific Northwest, and the Canadian Prairies over the past 25 yr and their past and future contributions to IWM. We also provide an industry perspective on the future of HR crop development and the role of HR crops in resistance management. Expanded options for HR traits in both major and minor crops are expected. With proper stewardship, HR crops can reduce herbicide-use intensity and help reduce selection pressure on weed populations. However, their proper deployment in cropping systems must be carefully planned by considering a diverse crop rotation sequence with multiple HR and non-HR crops and maximizing crop competition to effectively manage HR weed populations. Based on past experiences in the cultivation of HR crops and associated herbicide use in the western United States and Canada, HR crops have been important determinants of both the selection and management of HR weeds.

Research was conducted using a functional malachite green colorimetric assay to evaluate acetyl-coenzyme A carboxylase (ACCase) activity previously identified as resistant to sethoxydim and select aryloxyphenoxypropionate (FOPs) herbicides, fenoxaprop, and fluazifop. Two resistant southern crabgrass [Digitaria ciliaris (Retz.) Koeler] biotypes, R1 and R2, containing an Ile-1781-Leu amino acid substitution and previously identified as resistant to sethoxydim, pinoxaden, and fluazifop but not clethodim was utilized as the resistant chloroplastic ACCase source compared with known susceptible (S) ACCase. Dose-response studies with sethoxydim, clethodim, fluazifop-p-butyl, and pinoxaden (0.6 to 40 µM) were conducted to compare the ACCase–herbicide interactions of R1, R2, and S using the malachite green functional assay. Assay results indicated that R biotypes required more ACCase-targeting herbicides to inhibit ACCase activity compared with S. IC50 values of all four herbicides for R biotypes were consistently an order of magnitude greater than those of S. No sequencing differences in the carboxyltransferase domain was observed for R1 and R2; however, R2 IC50 values were greater across all herbicides. These results indicate the malachite green functional assay is effective in evaluating ACCase activity of R and S biotypes in the presence of ACCase-targeting herbicides, which can be used as a replacement for the 14C-based radiometric functional assays.

Site-specific weed management using open-source object detection algorithms could accurately detect weeds in cropping systems. We investigated the use of object detection algorithms to detect Palmer amaranth (Amaranthus palmeri S. Watson) in soybean [Glycine max (L.) Merr.]. The objectives were to (1) develop an annotated image database of A. palmeri and soybean to fine-tune object detection algorithms, (2) compare effectiveness of multiple open-source algorithms in detecting A. palmeri, and (3) evaluate the relationship between A. palmeri growth features and A. palmeri detection ability. Soybean field sites were established in Manhattan, KS, and Gypsum, KS, with natural populations of A. palmeri. A total of 1,108 and 392 images were taken aerially and at ground level, respectively, between May 27 and July 27, 2021. After image annotation, a total of 4,492 images were selected. Annotated images were used to fine-tune open-source faster regional convolutional (Faster R-CNN) and single-shot detector (SSD) algorithms using a Resnet backbone, as well as the “You Only Look Once” (YOLO) series algorithms. Results demonstrated that YOLO v. 5 achieved the highest mean average precision score of 0.77. For both A. palmeri and soybean detections within this algorithm, the highest F1 score was 0.72 when using a confidence threshold of 0.298. A lower confidence threshold of 0.15 increased the likelihood of species detection, but also increased the likelihood of false-positive detections. The trained YOLOv5 data set was used to identify A. palmeri in a data set paired with measured growth features. Linear regression models predicted that as A. palmeri densities increased and as A. palmeri height increased, precision, recall, and F1 scores of algorithms would decrease. We conclude that open-source algorithms such as YOLOv5 show great potential in detecting A. palmeri in soybean-cropping systems.

Acetolactate synthase (ALS) inhibitors provide postemergence control of green kyllinga (Kyllinga brevifolia Rottb.) in turfgrass and other cropping systems. A suspected resistant (R) biotype of K. brevifolia was collected from a golf course and evaluated for resistance to ALS inhibitors. In greenhouse experiments, the sulfosulfuron rates required to cause 50% shoot biomass reduction from the nontreated at 4 wk after treatment (WAT) were 10 and 792 g ai ha−1 for the susceptible (S) and R biotypes, respectively. The rates required to cause 50% injury at 4 WAT were 189 and >3,360 g ai ha−1, respectively. In other experiments, shoot mass of the R biotype was not reduced by imazaquin, trifloxysulfuron-sodium, pyrimisulfan, thiencarbazone + foramsulfuron + halosulfuron, florasulam + halauxifen-methyl, and bentazon compared with the nontreated, while sulfentrazone reduced biomass similarly for both R and S biotypes. Gene sequencing of the R biotype revealed a mutation at Asp-376-Glu that has previously conferred resistance to five families of ALS inhibitors. This is the first report of ALS-inhibitor resistance in K. brevifolia.

Palmer amaranth (Amaranthus palmeri S. Watson) is one of the most problematic weeds in many cropping systems in the midsouthern United States because of its multiple weedy traits and its propensity to evolve resistance to many herbicides with different mechanisms of action. In Arkansas, A. palmeri has evolved metabolic resistance to S-metolachlor, compromising the effectiveness of an important weed management tool. Greenhouse studies were conducted to evaluate the differential response of A. palmeri accessions from three states (Arkansas, Mississippi, and Tennessee) to (1) assess the occurrence of resistance to S-metolachlor among A. palmeri populations, (2) evaluate the resistance level in selected accessions and their resistant progeny, (3) and determine the susceptibility of most resistant accessions to other soil-applied herbicides. Seeds were collected from 168 crop fields between 2017 and 2019. One hundred seeds per accession were planted in silt loam soil without herbicide for >20 yr and sprayed with the labeled rate of S-metolachlor (1,120 g ai ha−1). Six accessions (four from Arkansas and two from Mississippi) were classified resistant to S-metolachlor. The effective doses (LD50) to control the parent accessions ranged between 73 and 443 g ha−1, and those of F1 progeny of survivors were 73 to 577 g ha−1. The resistance level was generally greater among progeny of surviving plants than among resistant field populations. The resistant field populations required 2.2 to 7.0 times more S-metolachlor to reduce seedling emergence 50%, while the F1 of survivors needed up to 9.2 times more herbicide to reduce emergence 50% compared with the susceptible standard.

Hydrilla [Hydrilla verticillata (L. f.) Royle] is often called the “perfect aquatic weed,” as it has numerous physiological adaptations that make it highly aggressive and competitive. Hydrilla verticillata has historically been managed effectively using fluridone; however, the overreliance on this single mechanism of action (MOA) resulted in evolved fluridone resistance in the late 1990s. Where fluridone-resistant H. verticillata populations evolved, endothall became widely used for H. verticillata control. In 2018, florpyrauxifen-benzyl, a highly active auxin-mimic herbicide, was registered for H. verticillata control, and its use has increased since its introduction. Endothall and florpyrauxifen-benzyl provide two effective MOAs for H. verticillata management, and combining these two MOAs would be an effective strategy to delay further resistance evolution. The objective of this research was to determine whether combining endothall and florpyrauxifen-benzyl would significantly impact the behavior of either herbicide in dioecious (DHV) or monoecious (MHV) H. verticillata compared with their behavior when applied alone. Endothall and florpyrauxifen-benzyl absorption and accumulation alone and in combination were measured over a 192-h time course. Translocation patterns were also determined. Herbicide accumulation in MHV and DHV was not impacted when these herbicides were applied in combination. Endothall translocation from shoots to roots in DHV was not impacted (alone = 18.7 ± 1.4%; combination = 23.2 ± 2.2%); however, endothall shoot-to-root translocation in MHV was reduced from 16.2 ± 1.3% applied alone to 2.2 ± 0.1% when applied in combination with florpyrauxifen-benzyl. Florpyrauxifen-benzyl shoot-to-root translocation was reduced in both MHV and DHV when applied in combination with endothall. Florpyrauxifen-benzyl translocation was reduced by 16- and 6-fold in DHV and MHV, respectively. These data do not suggest that there would be operational impacts from endothall and florpyrauxifen-benzyl mixtures. Still, there appear to be changes in herbicide behavior, primarily shoot-to-root translocation, when these two herbicides are applied in combination.

Glyphosate’s efficacy is influenced by the amount absorbed and translocated throughout the plant to inhibit 5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS). Glyphosate resistance can be due to target-site (TS) or non–target site (NTS) resistance mechanisms. TS resistance includes an altered target site and gene overexpression, while NTS resistance includes reduced absorption, reduced translocation, enhanced metabolism, and exclusion/sequestration. The goal of this research was to elucidate the mechanism(s) of glyphosate resistance in common ragweed (Ambrosia artemisiifolia L.) from Ontario, Canada. The resistance factor for this glyphosate-resistant (GR) A. artemisiifolia biotype is 5.1. No amino acid substitutions were found at positions 102 or 106 of the EPSPS enzyme in this A. artemisiifolia biotype. Based on [14C]glyphosate studies, there was no difference in glyphosate absorption or translocation between glyphosate-susceptible (GS) and GR A. artemisiifolia biotypes. Radio-labeled glyphosate metabolites were similar for GS and GR A. artemisiifolia 96 h after application. Glyphosate resistance in this A. artemisiifolia biotype is not due to an altered target site due to amino acid substitutions at positions 102 and 106 in the EPSPS and is not due to the NTS mechanisms of reduced absorption, reduced translocation, or enhanced metabolism.

A Palmer amaranth (Amaranthus palmeri S. Watson) population (KCTR: KS Conservation Tillage Resistant) collected from a conservation tillage field was confirmed with resistance to herbicides targeting at least six sites of action, including 2,4-D. The objectives of this research were using KCTR A. palmeri to investigate (1) the level of 2,4-D resistance, (2) 2,4-D absorption and translocation profiles, (3) the rate of 2,4-D metabolism compared with 2,4-D–tolerant wheat (Triticum aestivum L.), and (4) the possible role of cytochrome P450s (P450s) in mediating resistance. Dose–response experiments were conducted to assess the level of 2,4-D resistance in KCTR compared with susceptible plants, KSS (KS 2,4-D susceptible) and MSS (MS 2,4-D susceptible). KSS, MSS, and KCTR plants were treated with [14C]2,4-D to determine absorption, translocation, and metabolic patterns. Additionally, whole-plant dose–response assays were conducted by treating KCTR and KSS plants with P450 inhibitors (malathion, piperonyl butoxide [PBO]) before 2,4-D application. Dose–response experiments indicated a 6- to 11-fold 2,4-D resistance in KCTR compared with susceptible plants. No difference was found in percent [14C]2,4-D absorption among the populations. However, 10% less and 3 times slower translocation of [14C]2,4-D was found in KCTR compared with susceptible plants. Importantly, [14C]2,4-D was metabolized faster in KCTR than susceptible plants. At 24, 48, and 72 h after treatment (HAT), KCTR metabolized ∼20% to 30% more [14C]2,4-D than susceptible plants. KCTR plants and wheat generated metabolites with similar polarity. Nonetheless, at 24 HAT, ∼70% of [14C]2,4-D was metabolized in wheat, compared with only 30% in KCTR A. palmeri. Application of malathion before 2,4-D increased the sensitivity to 2,4-D in KCTR, suggesting involvement of P450s in mediating 2,4-D metabolism. However, no such impact of PBO was documented. Overall, this study confirms that enhanced metabolism is the primary mechanism of 2,4-D resistance in KCTR.

Ammannia multiflora Roxb. is a dominant broadleaf weed that is a serious problem in southern China rice fields, and acetolactate synthase (ALS)-inhibiting herbicides have been used for its control for more than 20 years. Excessive reliance on ALS-inhibiting herbicides has led to herbicide resistance in A. multiflora. In this study, 10 A. multiflora populations from the Jiangsu Province of China were collected, and the resistance levels and target site–resistance mechanisms to ALS-inhibiting herbicides bensulfuron-methyl and penoxsulam were investigated. The dose–response assays showed that eight populations evolved resistance to bensulfuron-methyl (9.1- to 90.9-fold) and penoxsulam (5.0- to 103.1-fold). Amplification of ALS genes indicated that there were three ALS genes (AmALS1, AmALS2, and AmALS3) in A. multiflora. Sequence analysis revealed amino acid mutations at Pro-197 in either AmALS1 (Pro-197-Ala, Pro-197-Ser, and Pro-197-His) or AmALS2 (Pro-197-Ser and Pro-197-Arg) in resistant populations, and no mutations were found in AmALS3. Moreover, two independent mutations (Pro-197-Ala in AmALS1 and Pro-197-Ser in AmALS2 or Pro-197-Ala in AmALS1 and Pro-197-Arg in AmALS2) coexisted in two resistant populations, respectively. In addition, the auxin mimic herbicides MCPA and florpyrauxifen-benzyl, the photosystem II inhibitor bentazon, and the protoporphyrinogen oxidase inhibitor carfentrazone-ethyl can effectively control the resistant A. multiflora populations. Our study demonstrates the wide prevalence of ALS inhibitor–resistant A. multiflora populations in Jiangsu Province and the diversity of Pro-197 mutations in ALS genes and provides alternative herbicide options for controlling resistant A. multiflora populations.