Herbicide-resistant Crops Research Articles

New Approaches to Herbicide and Bioherbicide Discovery

Abstract During the past 30 yr an impasse has developed in the discovery and commercialization of synthetic herbicides with new molecular targets and novel chemistries. Similarly, there has been little success with bioherbicides, both microbial and chemical. These bioherbicides are needed to combat fast-growing herbicide resistance and to fulfill the need for more environmentally and toxicologically safe herbicides. In response to this substantial and growing opportunity, numerous start-up companies are utilizing novel approaches to provide new tools for weed management. These diverse new tools broaden the scope of discovery, encompassing advanced computational, bioinformatic, and imaging platforms; plant genome–editing and targeted protein degradation technologies; and machine learning and artificial intelligence (AI)-based strategies. This review contains summaries of the presentations of 10 such companies that took part in a symposium held at the WSSA annual meeting in 2024. Four of the companies are developing microbial bioherbicides or natural product–based herbicides, and the other six are using advanced technologies, such as AI, to accelerate the discovery of herbicides with novel molecular target sites or to develop non-GMO, herbicide-resistant crops.

Transport of herbicides by PIN-FORMED auxin transporters.

Auxins are a group of phytohormones that control plant growth and development 1. Their crucial role in plant physiology has inspired development of potent synthetic auxins that can be used as herbicides 2. Phenoxyacetic acid derivatives are a widely used group of auxin herbicides in agriculture and research. Despite their prevalence, the identity of the transporters required for distribution of these herbicides in plants is both poorly understood and the subject of controversial debate 3,4. Here we show that PIN-FORMED auxin transporters transport a range of phenoxyacetic acid herbicides across the membrane and we characterize the molecular determinants of this process using a variety of different substrates as well as protein mutagenesis to control substrate specificity. Finally, we present Cryo-EM structures of Arabidopsis thaliana PIN8 with 2,4-dichlorophenoxyacetic acid (2,4-D) or 4-chlorophenoxyacetic acid (4-CPA) bound. These structures represent five key states from the transport cycle, allowing us to describe conformational changes associated with substrate binding and transport across the membrane. Overall, our results reveal that phenoxyacetic acid herbicides use the same export machinery as endogenous auxins and exemplify how transporter binding sites undergo transformations that dictate substrate specificity. These results enable development of novel synthetic auxins and for guiding precision breeding of herbicide resistant crop plants.

A natural glutathione S-transferase gene GSTU23 confers metabolic resistance to metamifop in Echinochloa crus-galli

Herbicides are essential for farmers to control weed. However, prolonged use of herbicides has caused the development of herbicide resistance in weeds. Here, the resistant Echinochloa crus-galli (RL5) was obtained by continuous treatment with metamifop for five generations in paddy fields. RL5 plants showed a 13.7-fold higher resistance to metamifop compared to susceptible E. crus-galli (SL5) plants. Pre-treatment with GST inhibitor (NBD-Cl) significantly increased the susceptibility of RL5 plants to metamifop. Faster metamifop metabolism and higher GST activity in RL5 plants than in SL5 plants were also confirmed, highlighting the role of GST in metabolic resistance. RNA-Seq analysis identified EcGSTU23 as a candidate gene, and this gene was up-regulated in RL5 and field-resistant E. crus-galli plants. Furthermore, the EcGSTU23 gene was overexpressed in the transgenic EcGSTU23-Maize, and the EcGSTU23-Maize showed resistance to metamifop. In vitro metabolic studies also revealed that the purified EcGSTU23 displayed catalytic activity in glutathione (GSH) conjugation, and metamifop was rapidly metabolized in the co-incubation system containing EcGSTU23 protein. These results provide direct experimental evidence of EcGSTU23's involvement in the metabolic resistance of E. crus-galli to metamifop. Understanding the resistance mechanism can help in devising effective strategies to combat herbicide resistance and breeding of genetically modified herbicide resistant crops.

Cytochrome P450 CYP81A104 in Eleusine indica confers resistance to multiherbicide with different modes of action.

Developing herbicide-resistant (HR) crop cultivars is an efficient way to control weeds and minimize crop yield losses. However, widespread and long-term herbicide application has led to the evolution of resistant weeds. Here, we established a resistant (R) E. indica population, collected from imidazolinone-resistant rice cultivar fields. The R population evolved 4.5-fold resistance to imazamox. Acetolactate synthase (ALS) gene sequencing and ALS activity assays excluded the effect of target-site resistance in this population. P450 inhibitor malathion pretreatment significantly reversed resistance to imazamox. RNA sequencing showed that a P450 gene CYP81A104 was expressed higher in R versus susceptible (S) plants. Arabidopsis overexpressing CYP81A104 showed resistance to ALS inhibitors (imazamox, tribenuron-methyl, penoxsulam and flucarbazone-sodium), PSII inhibitor (bentazone), hydroxyphenyl pyruvate dioxygenase inhibitor (mesotrione) and auxin mimics (MCPA), which was generally consistent with the results presented in the R population. This study confirmed that the CYP81A104 gene endowed resistance to multiherbicides with different modes-of-action. Our findings provide an insight into the molecular characteristics of resistance and contribute to formulating an appropriate strategy for weed management in HR crops. © 2024 Society of Chemical Industry.

Nanobody Mediated Atrazine Resistance in Plants.

In planta expression of recombinant antibodies has been proposed as a strategy for herbicide resistance but is not well advanced yet. Here, an atrazine nanobody gene fused with a green fluorescent protein tag was transformed to Arabidopsis thaliana, which was confirmed with PCR, ELISA, and immunoblotting. High levels of nanobody accumulation were observed in the nucleus, cytoderm, and cytosol. The nanobody expressed in the plant had similar affinity, sensitivity, and selectivity as that expressed in Escherichia coli. The T3 homozygous line showed resistance in a dose-dependent manner up to 380 g ai/ha of atrazine, which is approximately one-third of the recommended field application rate. This is the first report of utilizing a nanobody in plants against herbicides. The results suggest that utilizing a high-affinity herbicide nanobody gene rather than increasing the expression of nanobodies in plants may be a technically viable approach to acquire commercial herbicide-resistant crops and could be a useful tool to study plant physiology.

The impact of herbicide-resistant canola systems on the weed community dynamics in the Canadian Prairies

Increased adoption of no-till and extensive cultivation of herbicide-resistant (HR) crops in low diversity rotations are some transformations that occurred during the 1970s–2000s in western Canada. Historical data analysis of weed survey data (Saskatchewan, Manitoba, and Alberta) in canola crops and a canola rotation field experiment were used to understand how tillage, herbicide-resistant cropping systems, and the temporal frequency of canola in the rotation can impact weed abundance, composition, and diversity in the Canadian prairies. Weed survey data revealed that the spatial frequency of many weed species declined in years after 1995 compared with before 1995, the year HR cultivars were introduced. False cleavers ( Galeum spurium L .) and volunteer wheat ( Triticum aestivum L.) were the only species showing a continuous increase in frequency throughout the surveyed years. Species richness showed no trajectories but varied depending on climatic conditions. Roundup-Ready (RR) systems were associated with green foxtail ( Setaria viridis L.) irrespective of their frequency in the rotation. Continuous canola systems had an obvious association with chickweed ( Stellaria media L.) regardless of herbicide systems. Wild oats ( Avena fatua L.) were predominant in the longer rotations with glufosinate and imidazolinone-resistant canola. In the field study, weeds associated before in crop herbicide application were relatively controlled after herbicides were applied, except for foxtail barley ( Hordeum Jubatum L.), which was not controlled in most HR systems. Overall, both weed survey data and field experiment data revealed greater control of most weed species due to the adoption of HR canola cultivars and the associated cultural practices.

Induced mutation in the SiALS gene offers new weed management opportunities for sesame crop

Weeds are the primary biotic constraint affecting sesame growth and production. Here, we applied EMS mutagenesis to an elite sesame cultivar and discovered a novel point mutation in the sesame SiALS gene conferring resistance to imidazolinone, a group of acetolactate-synthase (ALS)-inhibitors. The mutant line exhibited high resistance to imazamox, an ALS-inhibitor, with hybrid plants displaying an intermediate response. Field-based validation confirmed the mutant line’s substantial resistance, leading to a significantly higher yield under imazamox treatment. Under pre-emergence application of imazapic, the mutant plants sustained growth, whereas wild-type and weed were effectively controlled. Field trials using s-metolachlor and imazapic combined resulted in weed-free plots compared to untreated controls. Consequently, this treatment showed a significantly greater yield (2280 vs. 880 Kg ha−1) than the commercial practice (s-metolachlor). Overall, our study unveils the potential of utilizing this point mutation in sesame breeding programs, offering new opportunities for integrated weed management strategies for sesame cultivation. Developing herbicide-resistant crop plants holds promise for supporting sustainable production and addressing the challenges of weed infestations in sesame farming.

"Peanut (Arachis hypogaea) Response to Fungicides and Herbicides Applied using Different Spray Nozzles"

Late leaf spot [caused by Nothopassalora personata (Berk. & M.A. Curtis) U. Braun, C. Nakash, Videira & Crous] is an economically important disease in peanut (Arachis hypogaea L.) and fungicides are used routinely to protect peanut from infection and the resulting yield loss. Herbicides are an important component of effective weed management in peanut. Concern over particle drift of pesticides exists in the farming community, especially after stewardship issues associated with synthetic auxin application in herbicide resistant crops in the United States. Spray nozzles that deliver larger droplets that are less prone to drift is a possible solution to this issue. However, fungicides may be less effective in penetrating the crop canopy and covering foliage for protection from pathogens with these nozzles compared with nozzles delivering smaller droplets. Research was conducted to compare suppression of late leaf spot disease when chlorothalonil, prothioconazole plus tebuconazole, azoxystrobin, pyraclostrobin, and chlorothalonil were applied sequentially every 14 days using hollow cone nozzles (fine droplet size), regular flat fan nozzles (medium droplet size), air induction flat fan nozzles (coarse droplet size), and turbo teejet induction nozzles (ultra-coarse droplet size). Regular flat fan and hollow cone nozzles were slightly more effective in delivering fungicides based on canopy defoliation at harvest compared with turbo teejet induction nozzles and in some cases air induction nozzles. However, suppression of pathogens by fungicides applied using all four nozzle types prevented canopy defoliation adequately to protect peanut from yield loss. In a separate experiment, peanut yield and control of common ragweed (Ambrosia artemisiifolia L.) and late leaf spot did not differ at harvest when herbicides and fungicides were applied with air induction or turbo teejet induction nozzles throughout the cropping cycle. These results indicate that farmers can apply fungicides and herbicides to peanut using nozzles that limit potential for particle drift with minimal concern over reduced pesticide efficacy and protection of yield from pests.

A Critical Review of Conventional and Modern Approaches to Develop Herbicide-Resistance in Rice.

Together with rice, weeds strive for nutrients and space in farmland, resulting in reduced rice yield and quality. Planting herbicide-resistant rice varieties is one of the effective ways to control weeds. In recent years, a series of breakthroughs have been made to generate herbicide-resistant germplasm, especially the emergence of biotechnological tools such as gene editing, which provides an inherent advantage for the knock-out or knock-in of the desired genes. In order to develop herbicide-resistant rice germplasm resources, gene manipulation has been conducted to enhance the herbicide tolerance of rice varieties through the utilization of techniques such as physical and chemical mutagenesis, as well as genome editing. Based on the current research and persisting problems in rice paddy fields, research on the generation of herbicide-resistant rice still needs to explore genetic mechanisms, stacking multiple resistant genes in a single genotype, and transgene-free genome editing using the CRISPR system. Current rapidly developing gene editing technologies can be used to mutate herbicide target genes, enabling targeted genes to maintain their biological functions, and reducing the binding ability of target gene encoded proteins to corresponding herbicides, ultimately resulting in herbicide-resistant crops. In this review article, we have summarized the utilization of conventional and modern approaches to develop herbicide-resistant cultivars in rice as an effective strategy for weed control in paddy fields, and discussed the technology and research directions for creating herbicide-resistant rice in the future.

Gene flow and spontaneous mutations are responsible for imidazolinone herbicide-resistant weedy rice (Oryza sativa L.)

For more than two decades, weedy rice (Oryza sativa L.) has been controlled in rice fields by using imidazolinone (IMI) herbicide-resistant rice technology (Clearfield®). Outcrossing in weedy rice populations and spontaneous mutations are potential problems with herbicide-resistant crop management technologies, such as the IMI-resistant rice. The aim of this study was to characterize the mechanism of IMI herbicide resistance in weedy rice through dose-response bioassay study and evaluating amino acid substitutions in acetolactate synthase (ALS) protein. A total of 118 suspected IMI-resistant weedy rice samples, which survived in the field after an IMI herbicide application, were collected at harvest time from Türkiye in 2020 and 2021. Single-dose imazamox application experiment revealed that 38 plants survived herbicide treatment. The imazamox resistance of the surviving plants was confirmed by dose-response experiment. ALS gene region underwent a sanger DNA partial sequencing. No substitution was found in 10 samples, however, amino acid substitutions were found in 26 samples with S563N, one sample with S653T, and one sample with E630D. The S653N point is the same substitution point that serves as the origin of resistance for the Clearfield® rice varieties that are commonly cultivated in the region. It has been hypothesized that the gene flow from IMI-resistant rice may be the cause of resistance in the IMI resistant weedy rice samples with S653N. The other substitution, S653T, were considered spontaneous mutation to IMI resistance. Interestingly, the S653T mutation was detected for the first time in weedy rice. The mechanism of resistance of 10 resistant weedy rice was not confirmed in this study, however, it may be a non-target resistance or another mutation point in target site, but evidently, they did not acquire resistance by gene flow from IMI-resistant rice. It has been concluded that the effectiveness of IMI-resistant rice technology in controlling weedy rice has drastically decreased due to possible gene flow, spontaneous mutation and non-target resistance. In addition to cultural controls like clean seed, clean machinery and crop rotation, other herbicide-tolerant rice systems such as Provisia® and Roxy-RPS® rice are needed to create a diverse weedy rice management ensemble available for rice production and move towards sustainable rice farming.

Multi-omics analysis identifies EcCS4 is negatively regulated in response to phytotoxin isovaleric acid stress in Echinochloa crus-galli.

Knowledge of herbicidal targets is critical for weed management and food safety. The phytotoxin isovaleric acid (ISA) is effective against weeds with a broad spectrum, carries low environmental risks, and is thus an excellent herbicide lead. However, the biochemical and molecular mechanisms underlying the action of ISA remain unclear. Multi-omics data showed that acetyl coenzyme A (acetyl-CoA) was the key affected metabolite, and that citrate synthase (CS) 4 was substantially down-regulated under ISA treatment in Echinochloa crus-galli leaves. In particular, the transcript level of EcCS4 was the most significantly regulated among the six genes involved in the top 10 different pathways. The EcCS4 encodes a protein of 472 amino acids and is localized to the cell membrane and mitochondria, similar to the CS4s of other plants. The protein content of EcCS4 was down-regulated after ISA treatment at 0.5 h. ISA markedly inhibited the CS4 activity in vitro in a concentration-dependent manner (IC50 = 41.35 μM). In addition, the transgenic rice plants overexpressing EcCS4 (IC50 = 111.8 mM for OECS4-8 line) were more sensitive, whereas loss-of-function rice mutant lines (IC50 = 746.5 mM for oscs4-19) were more resistant to ISA, compared to wild type (WT) plants (IC50 = 355.6 mM). CS4 was first reported as a negative regulator of plant responses to ISA. These results highlight that CS4 is a candidate target gene for the development of novel herbicides and for breeding herbicide-resistant crops. © 2023 Society of Chemical Industry.

Screening and Selection for Herbicide Tolerance among Diverse Tomato Germplasms

Solanum lycopersicum, the domesticated species of tomato, is produced and consumed globally. It is one of the most economically important vegetable crops worldwide. In the commercial production of tomatoes, tomatoes are extremely sensitive to herbicide drifts from row crops in the vicinity. Injury to tomatoes from auxin herbicides and glyphosate can occur at rates as low as 0.01×. This results in a substantial yield reduction, and at high drift rates, plants may not show signs of recovery. With the new herbicide-resistant crop technologies on the market, which include 2,4-D and dicamba-resistant crops, there is an increase in the usage of these herbicides, causing more serious drift problems. There is a diverse germplasm of tomatoes that includes wild relatives which are tolerant to numerous biotic and abiotic stresses. Herbicide/chemical stress is an abiotic stress, and wild tomato accessions may have a natural tolerance to herbicides and other abiotic stresses. In the current study, diverse tomato genotypes consisting of 110 accessions representing numerous species, Solanum habrochaites, S. cheesmaniae, S. pimpinellifolium, S. chilense, S. lycopersicum, S. pimpinellifolium, S. galapagense, S. chimelewskii, S. corneliomulleri, S. neorickii, and S. lycopersicoides, were used for screening drift rate herbicide tolerance. The herbicides tested included simulated drift rates of 2,4-D, dicamba, glyphosate, quinclorac, aminopyralid, aminocyclopyrachlor, and picloram. The visual injury rating of each accession for each herbicide treatment was taken 7, 14, 21, and 28 days after treatment (DAT) on a scale of 0–100%. Numerous accessions were found to have minimal injury (less than 20%) for each of the herbicides tested; nine accessions were found for both 2,4-D and glyphosate, eleven for dicamba, five for quinclorac, eight for aminocyclopyrachlor and two for both aminopyralid and picloram at 28 DAT. The identification of genotypes with a higher herbicide tolerance will provide valuable genetic resources for the development of elite tomato varieties that can resist herbicide injury and produce competitive yields.

Exploring the effect of region on diversity and composition of weed seedbanks in herbicide-resistant crop systems in the United States.

Soil seedbanks have been recognized as one of the crucial components of agricultural ecosystems. However, studies on the shift in structure and biodiversity of soil seedbanks in herbicide-resistant crop systems are limited, and a functional trait perspective of the soil seedbank is often overlooked. A 6 years experiment was conducted to investigate the roles of region, crop system, and weed management strategy on species richness, functional trait diversity, and composition of the weed seedbank. Species richness was different across the interaction of region and crop system, while functional trait diversity only showed difference across regions. Species and functional trait compositions were affected by the interaction of region and crop system. Specifically, the compositional difference among crop systems was mainly determined by the significant heterogeneity of group dispersion. Growers and practitioners should consider weed functional traits in developing lasting agricultural management strategies. Long-term weed research should draw attention to the impact of transgenic crop systems and specific management tactics on weed dispersal, functional composition, and resistance evolution of weed species in such agroecosystems. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A perspective on how glyphosate and 2,4-D in wetlands may impact climate change

An increase in herbicide use is occurring due to a growing population and herbicide-resistant crops in agriculture, which has resulted in more herbicide tolerant target species. Glyphosate and 2,4-Dichlorophenoxyacetic acid (2,4-D) are two of the most commonly used herbicides worldwide and are more recently being used in combination in pre-mixed commercial formulas. Subsequently, herbicide contamination of wetlands will increase exposure of microorganisms to multiple chemical stressors. Methane is a potent greenhouse gas naturally emitted from wetlands, but herbicides may disrupt biogeochemical processes leading to an unbalanced methane cycle. We review the impacts of these herbicides on aquatic microbial communities from glyphosate-derived nutrient enrichment and 2,4-D inhibition of methane oxidation, and examine how these altered metabolic processes may lead to increased methane production in wetlands. The response of wetland ecosystems to herbicide contamination will vary across regions, in part due to the complexity of microbial communities, however, this perspective gives a glimpse into the potential global implications of continuing herbicide use on wetlands and demonstrates the importance for research on ecosystem-level co-stressors.

Functional characterization of novel mutations in the conserved region of EPSPS for herbicide resistance in pigeonpea: structure-based coherent design

Prospectively, agroecosystems for the growth of crops provide the potential fertile, productive, and tropical environment which attracts infestation by weedy plant species that compete with the primary crop plants. Infestation by weed is a major biotic stress factor faced by pigeonpea that hampers the productivity of the crop. In the modern era with the development of chemicals the problem of weed infestation is dealt with armours called herbicides. The most widely utilized, post-emergent, broad-spectrum herbicide has an essential active ingredient called glyphosate. Glyphosate mechanistically inhibits a chloroplastic enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) by competitively interacting with the PEP binding site which hinders the shikimate pathway and the production of essential aromatic amino acids (Phe, Tyr, Trp) and other secondary metabolites in plants. Moreover, herbicide spray for weed management is lethal to both the primary crop and the weeds. Therefore, it is critical to develop herbicide-resistant crops for field purposes to reduce the associated yield and economic losses. In this study, the in-silico analysis drove the selection and validation of the point mutations in the conserved region of the EPSPS gene, which confers efficient herbicide resistance to mutated-CcEPSPS enzyme along with the retention of the normal enzyme function. An optimized in-silico validation of the target mutation before the development of the genome-edited resistant plant lines is a prerequisite for testing their efficacy as a proof of concept. We validated the combination of GATIPS mutation for its no-cost effect at the enzyme level via molecular dynamic (MD) simulation. Communicated by Ramaswamy H. Sarma

Effects of Pesticides on the Arbuscular Mycorrhizal Symbiosis

Substantial amounts of pesticides, used in agricultural production to control pests, diseases, and weeds, and thereby attain high product quantities and quality, can severely affect the ecosystem and human health. The amounts of pesticides used depend on the specifics of the current production system but also exhibit large effects of past practices. Pesticides do not act only on the target organisms but also on organisms for which the chemicals were not specifically formulated, constituting hazardous molecules for humans and the environment. Pesticides, therefore, also influence soil microbial communities including organisms that engage in mutualistic plant symbioses that play a crucial role in its mineral nutrition, such as arbuscular mycorrhizal fungi. In this review, we summarize the current knowledge on the effects of synthetic and natural (‘green’) pesticides (fungicides, herbicides, and insecticides) on arbuscular mycorrhizal symbiosis. We deal with both the direct effects (spore germination and extraradical and intraradical growth of the mycelium) and indirect effects on the agroecosystem level. Such indirect effects include effects through the spread of herbicide-resistant crops and weeds to neighboring ecosystems, thereby modifying the mycorrhizal inoculum potential and altering the plant–plant interactions. We also briefly discuss the possibility that mycorrhizal plants can be used to enhance the phytoremediation of organic pesticides.

Potential risks of future herbicide-resistant weeds in New Zealand revealed through machine learning

ABSTRACT New Zealand has fewer numbers of herbicide-resistant crop weeds than many other highly developed economies, yet these numbers are likely to increase in the future. A clear indication of the scale of this risk can be derived from the predictable structure in the global occurrence of herbicide-resistant weeds that results from similarities in agronomic and environmental conditions. Hierarchical cluster analysis was used to identify groups of countries with similar herbicide-resistant weed assemblages to New Zealand. A distinct cluster of 11 European countries with strong climatic and agronomic affinities to New Zealand was identified. The combined assemblage of herbicide-resistant weeds within this cluster consisted of 27 species and the potential risk of a species evolving herbicide resistance was calculated as its frequency among these European countries. Species with potential to become herbicide resistant in New Zealand included established crop weeds (e.g. Senecio vulgaris, Tripleurospermum inodorum) as well as species only encountered as contaminants of seed imports (e.g. Alopecurus myosuroides, Apera spica-venti). All eight species already known to be herbicide-resistant in New Zealand were found in the high-risk assemblage and this indicates that the analysis provided a realistic measure of future risk.

Comparative proteomic and physiological analyses reveal tribenuron-methyl phytotoxicity and nontarget-site resistance mechanisms in Brassicanapus.

The application of herbicides is the most effective strategy for weed control and the development of herbicide-resistant crops will facilitate the weed management. The acetolactate synthase-inhibiting herbicide, tribenuron-methyl (TBM), is broadly used for weed control. However, its application in rapeseed field is restricted since rapeseed is sensitive to TBM. Herein, an integrated study of cytological, physiological and proteomic analysis of the TBM-resistant rapeseed mutant M342 and its wild-type (WT) plants was conducted. After TBM spraying, M342 showed improved tolerance to TBM, and proteins implicated in non-target-site resistance (NTSR) to herbicides had a significantly higher level in M342 as compared with the WT. Differentially accumulated proteins (DAPs) between these two genotypes were enriched in glutathione metabolism and oxidoreduction coenzyme metabolic process, which protected the mutant from oxidative stress triggered by TBM. Important DAPs related to stress or defence response were up-accumulated in M342regardless of the TBM treatment, which might serve as the constitutive part of NTSR to TBM. These results provide new clues for further exploration of the NTSR mechanism in plants and establish a theoretical basis for the development of herbicide-resistant crops.

Enlist™ corn tolerance to preemergence and postemergence applications of synthetic auxin and ACCase-inhibiting herbicides

AbstractAllowing the use of two additional modes of action (MOAs), Enlist™ corn is a novelty in the continuum of herbicide-resistant crop development efforts that have occurred since the 1990s. Knowledge of Enlist corn tolerance to labeled herbicides and other herbicides within the same MOA for various use and/or exposure scenarios is not well established. Four site-year field experiments for preemergence (PRE) and postemergence (POST) applications were conducted at sites in Fayetteville (2021 and 2022) and Tillar (2020 and 2021), Arkansas, to evaluate Enlist corn response following PRE or POST applications of synthetic auxin herbicides or those that inhibit acetyl-CoA carboxylase (ACCase). A non-Enlist and an Enlist corn hybrid were used for each herbicide treatment to establish differential tolerance. Injury response to PRE application varied among site-years; clethodim was the only herbicide that occasionally caused significant (7% to 17%) injury to Enlist corn. None of the PRE treatments affected plant height, stand, or yield of Enlist corn; these responses were generally similar or better for Enlist corn compared to non-Enlist corn. Enlist corn showed significant injury to POST applications of florpyrauxifen-benzyl (>10%), fluazifop-P-butyl and quizalofop-P-ethyl (>5%), and clethodim and sethoxydim (>75%) 1 wk after application (WAA). These initial injury responses to clethodim and sethoxydim were generally reflected in Enlist corn yield; however, the minimal injury from fluazifop-P-butyl and quizalofop-P-ethyl did not affect yield. Injury to non-Enlist corn with POST-applied ACCase-inhibiting herbicides 2 WAA was >80%, resulting in a proportionate yield reduction. Even though florpyrauxifen-benzyl caused more initial injury to non-Enlist corn, yield reduction in non-Enlist corn was occasionally less than of Enlist corn, with both hybrids experiencing >75% yield reduction. In summary, Enlist corn may occasionally show transient injury even to labeled herbicides when applied POST, and even though the injury from florpyrauxifen-benzyl is initially mild, it nonetheless results in substantial yield loss.

A LuALS Mutation with High Sulfonylurea Herbicide Resistance in Linum usitatissimum L.

The cultivation of herbicide-resistant crops is an effective tool for weed management in agriculture. Weed control in flax (Linum usitatissimum L.) remains challenging due to the lack of available herbicide-resistant cultivars. In this study, a mutant resistant to acetolactate synthase (ALS)-inhibiting herbicides was obtained by ethyl methanesulphonate (EMS) mutagenesis using an elite cultivar, Longya10. Whole-plant dose-response assays revealed that, compared to Longya10, the mutant was 11.57-fold more resistant to tribenuron-methyl (TBM) and slightly resistant to imazethapyr (resistance index (mutant/Longya10) < 3). In vitro acetolactate synthase assays showed that the relative resistance of the mutant was 12.63 times more than that of Longya10. A biochemical analysis indicated that there was a Pro197Ser (relative to the Arabidopsis thaliana ALS sequence) substitution within the LuALS1, conferring high resistance to sulfonylurea herbicides in the mutant. Additionally, two cleaved amplified polymorphic sequence (CAPS) markers, BsaI-LuALS1 and EcoO109I-LuALS1, were developed based on the mutation site for marker assistant selection in breeding. Moreover, the mutant did not cause losses in natural field conditions. We find a mutant with ALS-inhibiting herbicide resistance chemically induced by EMS mutagenesis, providing a valuable germplasm for breeding herbicide-resistant flax varieties.