A weed survey was conducted on 134 Palmer amaranth (Amaranthus palmeri S. Watson) populations from Mississippi and Arkansas in 2017 to investigate the spread of resistance to protoporphyrinogen oxidase (PPO) inhibitors using fomesafen as a proxy. Fomesafen resistance was found in 42% of the A. palmeri populations. To investigate the resistance basis of different PPO inhibitors, we further characterized 10 representative populations by in planta bioassay in a controlled environment and molecular characterizations (DNA sequencing and TaqMan® gene expression assay). A total of 160 plants were sprayed with a labeled field rate (1X) of fomesafen or salfufenacil and screened for the presence of three known resistance-endowing mutations in the mitochondrial PPX2 gene (ΔGly-210, Arg-128-Gly, Gly-399-Ala). To compare the potencies of fomesafen and saflufenacil, dose–response studies were conducted on two highly resistant and one sensitive populations. The interaction of the two herbicides with the target protein harboring known PPX2 mutations was also analyzed. Our results showed that: (1) 90% of the fomesafen- or saflufenacil-resistant plants have at least one of the three known PPX2 mutations, with ΔGly-210 being the most prevalent; (2) saflufenacil is more potent than fomesafen, with five to nine times lower resistance/susceptible (R/S) ratios; (3) fomesafen selects for more diverse mutations, and computational inhibitor/target modeling of fomesafen suggest a weaker binding affinity in addition to a smaller interaction volume and volume overlap with the substrate protoporphyrinogen IX than saflufenacil. As a result, saflufenacil shows reduced sensitivity to PPX2 target-site mutations. Results from current study can help pave the way for designing weed management strategies to delay resistance development and maintain the efficacy of PPO inhibitors.

Broomrape species (Orobanche spp. and Phelipanche spp.) pose a tremendous threat to economically important crops in Iran; however, there is little information on the country’s major broomrape species, their distribution and host ranges, and the subsequent crop losses they cause. The literature from Iran describes Egyptian broomrape (Orobanche aegyptiaca Pers.), hemp broomrape (Orobanche ramosa L.), Phelipanche nana (Reut.) Beck, and Orobanche cumana Wallr. as the major species of broomrape infesting agricultural fields. Solanaceae crops are among the most susceptible crops to infestation by broomrape in the country, and farmers are either shifting production to crops with lesser value or abandoning crop production entirely. Among infested crops, tomato (Solanum lycopersicum L.) is the most susceptible and widely infested, resulting in annual crop yield reductions of up to 40%. Chick pea (Cicer arietinum L.) and canola (Brassica napus L.) are also highly susceptible to broomrape infestation. Current control methods (physical, agronomical, biological, and chemical) have not proven to be effective or economic. The microscopic size of broomrape seeds and the presence of a wide range of host plants (weeds and crops) have promoted the spread of broomrape to new areas of Iran. Hence, the familiarization of farmers with the biology of broomrape would be highly effective in finding suitable control measures and in preventing its further distribution.

The widespread evolution of herbicide resistance in weed populations has become an increasing concern for no-tillage (NT) growers in semiarid regions of the U.S. Great Plains. Lack of cost-effective and alternative new herbicide sites of action further exacerbates the problem of herbicide-resistant (HR) weeds and threatens the long-term sustainability of prevailing cropping systems in the region. A recent decline in commodity prices and increasing herbicide costs to manage HR weeds has spurred research efforts to build a strong rationale for developing ecologically based integrated weed management (IWM) strategies in the U.S. Great Plains. Integration of cover crops (CCs) in NT dryland production systems potentially offers several ecosystem services, including weed control, soil health improvement, decline in selective pest pressure, and overall reduction in pest management inputs. This review article aims to document the role of CCs for IWM, with emphasis on exploring emerging weed issues; ecological, economic, and agronomic benefits of growing CCs; and constraints preventing adoption of CCs in NT cropping systems in the semiarid Great Plains. We attempt to focus on changes in weed management practices, their long-term impacts on weed seedbanks, weed shifts, and herbicide-resistance evolution in the most common weed species in the region. We also highlight current knowledge gaps and propose new research priorities based on an improved understanding of CC management strategies that will ultimately aid in achieving sustainable weed management goals and preserving natural resources in water-limited environments.

Wild poinsettia (Euphorbia heterophylla L.) is a troublesome broadleaf weed in grain production areas in South America. Herbicide resistance to multiple sites of action has been documented in this species, including protoporphyrinogen oxidase (PPO) inhibitors. We investigated the physiological and molecular bases for PPO-inhibitor resistance in a E. heterophylla population (RPPO) from Southern Brazil. Whole-plant dose–response experiments revealed a cross-resistance profile to three different chemical groups of PPO inhibitors. Based on dose–response parameters, RPPO was resistant to lactofen (47.7-fold), saflufenacil (8.6-fold), and pyraflufen-ethyl (3.5-fold). Twenty-four hours after lactofen treatment (120 g ha−1) POST, RPPO accumulated 27 times less protoporphyrin than the susceptible population (SPPO). In addition, RPPO generated 5 and 4.5 times less hydrogen peroxide and superoxide than SPPO, respectively. The chloroplast PPO (PPO1) sequences were identical between the two populations, whereas 35 single-nucleotide polymorphisms were found for the mitochondrial PPO (PPO2). Based on protein homology modeling, the Arg-128-Leu (homologous to Arg-98-Leu in common ragweed [Ambrosia artemisiifolia L.] was the only one located near the catalytic site, also in a conserved region of PPO2. The cytochrome P450 monooxygenase inhibitor malathion did not reverse resistance to lactofen in RPPO, and both populations showed similar levels of PPO1 and PPO2 expression, suggesting that metabolic resistance and PPO overexpression are unlikely. This is the first report of an Arg-128-Leu mutation in PPO2 conferring cross-resistance to PPO inhibitors in E. heterophylla.

Protoporphyrinogen oxidase (PPO) is an important target for discovering new herbicides that interfere with the synthesis of porphyrin. To discover new PPO inhibitors with improved biological activity, a series of new diphenyl ethers containing tetrahydrophthalimide were designed and synthesized. Among them, J6.1 (IC50 = 4.7 nM) and J6.3 (IC50 = 30.0 nM) show higher maize (Zea mays L.) PPO inhibitory activity than the commercial herbicides oxyfluorfen (IC50 = 117.9 nM) and flumioxazin (IC50 = 157.1 nM). The greenhouse herbicidal activity of J6.3 is comparable to that of oxyfluorfen, and it is greater than that of flumioxazin. Even at a dose of 300 g ai ha−1, cotton (Gossypium hirsutum L.) and peanut (Arachis hypogaea L.) show greater tolerance to J6.3, suggesting that J6.3 could be used for further development of new herbicide candidates in those fields. In addition, molecular docking has been used to further study the mechanism of action of J6.3. The results show that the introduction of a nitro group and tetrahydrophthalimide into the diphenyl ether structure is beneficial to biological activity.

Glufosinate inhibits glutamine synthetase (GS), a key enzyme for amino acid metabolism and photorespiration. Protoporphyrinogen oxidase (PPO) inhibitors block chlorophyll biosynthesis and cause protoporphyrin accumulation, a highly photodynamic intermediate. Both herbicides ultimately lead to plant death by a massive accumulation of reactive oxygen species (ROS) through different mechanisms. We investigated a potential synergistic effect by the mixture of the two herbicide mechanisms of action (MoAs). The tank mix between a low rate of glufosinate (280 g ai ha−1) with an ultra-low dose of saflufenacil (1 g ha−1) provided enhanced herbicidal activity compared with the products applied individually on Palmer amaranth (Amaranthus palmeri S. Watson). The synergism between the two herbicides was also confirmed by isobole analysis and field trials. The herbicide combination provided high levels of efficacy when applied at low temperature and low humidity. Mechanistically, glufosinate caused a transient accumulation of glutamate, the building block for chlorophyll biosynthesis. Consequently, inhibition of both GS and PPO resulted in greater accumulation of protoporphyrin and ROS, forming the physiological basis for the synergism between glufosinate and PPO inhibitors. While the synergy between the two herbicide MoAs provided excellent efficacy on weeds, it caused low injury to PPO-resistant waterhemp [Amaranthus tuberculatus (Moq.) Sauer] and high injury to both glufosinate-resistant and glufosinate-susceptible soybean [Glycine max (L.) Merr.]. Glufosinate enhances the activity of PPO inhibitors through glutamate and protoporphyrin accumulation, leading to increased levels of ROS and lipid peroxidation. The synergism between the two herbicide MoAs can help to overcome environmental effects limiting the efficacy of glufosinate. Future research is needed to optimize the uses for this herbicidal composition across different cropping systems.

Plants of Sumatran fleabane [Conyza sumatrensis (Retz.) E. Walker] were identified in a field with an unusual rapid leaf-injury herbicide symptoms after application of 2,4-D in mixture with glyphosate. The objectives of this study were to confirm the occurrence of resistance to 2,4-D herbicide and to characterize the occurrence of rapid necrosis as the mechanism associated with the herbicide resistance in C. sumatrensis. The studies performed were an initial screening, effect of 2,4-D alone and associated with glyphosate, cross- and multiple-resistance evaluation, effect of commercial formulation and analytical product, and rate of H2O2 evolution. The Marpr9-rn accession was identified with rapid necrosis symptoms and survival to 804 g ae ha−1 of 2,4-D. The resistance factor to the herbicide 2,4-D was 18.6 at 49 d after spraying. The analytical product 2,4-D and the commercial formulation resulted in similar symptoms of rapid necrosis. This symptom did not occur for the six other auxinic herbicides (dicamba, florpyrauxifen-benzyl, fluroxypyr, halauxifen-methyl, picloram, and triclopyr), indicating absence of cross-resistance. Multiple resistance to the herbicides paraquat, saflufenacil, and ammonium glufosinate was not identified in the Marpr9-rn population. However, survival following treatment with the herbicides glyphosate and chlorimuron-ethyl occurred. The evolution of H2O2 began at 15 min after application and was less pronounced in low light. These results indicate the first case of resistance to 2,4-D and occurrence of rapid necrosis in C. sumatrensis.

In Argentina, Lolium spp. occur in 40% of winter cereal crops from the Pampas. Several years ago, cases of glyphosate-resistant perennial ryegrass (Lolium perenne L.) were detected, and the use of acetyl-CoA carboxylase (ACCase)-inhibiting herbicides to eradicate these plants has been considered. The aim of this study was to evaluate the sensitivity of a putative pinoxaden-resistant L. perenne population to ACCase-inhibiting herbicides. Around 80% of plants from the putative resistant population survived at a recommended dose of pinoxaden, and they produced viable seeds. The resistance indices (RIs) to pinoxaden were 5.1 and 2.8 for plant survival and seed production, respectively. A single point mutation that conferred a Asp-2078-Gly substitution in ACCase was the source of the resistance. To match the plant control achieved in the susceptible population, the resistant population required 5.4- and 10.4-fold greater doses of clethodim and quizalofop, respectively. RIs for viable seed production when treated with clethodim and quizalofop were 3.3 and 6.6, respectively. The Asp-2078-Gly mutation endowed significant levels of resistance to pinoxaden, clethodim, and quizalofop. For three herbicides, the level of resistance of a pinoxaden-resistant L. perenne population to ACCase inhibitors was evaluated, based on an evaluation of dose response for plant survival and seed production. The RIs were higher for plant survival than for seed production. In Argentina, the selection pressure associated with clethodim and haloxifop preplant application and pinoxaden use on wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) crops, would have favored the propagation of the Asp-2078-Gly mutation with its associated resistance.

Cowpea witchweed [Striga gesnerioides (Willd.) Vatke] is a primary constraint of cowpea [Vigna unguiculata (L.) Walp.] production in West Africa. Previously, seven S. gesnerioides races were classified based upon host specificity and genotypic profiling. Because race number and distribution are dynamic systems influenced by gene flow, genetic drift, and natural selection, a thorough investigation of S. gesnerioides diversity and the effectiveness of known sources of resistance in cowpea is needed to develop varieties with durable and broad-spectrum Striga resistance. In this study, we screened seven cowpea lines against 58 unique S. gesnerioides populations collected from across nine West African countries. Individuals from 10 S. gesnerioides populations were genotyped with simple sequence repeat (SSR) markers. We identified six races of S. gesnerioides based on their parasitism of the seven cowpea lines with known differential resistance genotypes. No cowpea line was resistant to all 58 Striga populations and none of the Striga populations were able to overcome the resistance of all seven lines. A novel race, SG6, of the parasite collected from Kudu, Nigeria, was found to overcome more cowpea resistance genes than any previously reported race. SSR analysis indicates that Striga populations are highly differentiated and genetic relatedness generally corresponds with geographic proximity rather than their host compatibility. Due to the dearth of broad-spectrum resistance found among Striga-resistant cowpea lines, there exists a need to stack multiple Striga resistance genes in order to confer broad-spectrum and durable resistance.

The developmental status of perennial weeds such as rush skeletonweed (Chondrilla juncea L.) can influence herbicide absorption and translocation. Differential efficacy between fall and spring applications suggests vernalization impacts herbicide absorption and translocation in other perennial asters. Clopyralid and aminopyralid absorption and translocation were quantified in nonvernalized and vernalized plants following application of 14C-labeled herbicides 2, 4, 8, 24, and 72 h after treatment. Less 14C clopyralid was absorbed, and at a slower rate, in vernalized plants. Movement out of the treated leaf was slower, with 14C clopyralid translocating more rapidly than 14C aminopyralid. More 14C moved to the roots in nonvernalized plants compared with vernalized plants, regardless of herbicide. Increased translocation to belowground survival structures is needed for effective control of C. juncea.

The growth response of annual sowthistle (Sonchus oleraceus L.) to anticipated future climate conditions is currently unknown, and thus two parallel studies were conducted dealing with glyphosate-resistant (GR) and glyphosate-susceptible (GS) biotypes of S. oleraceus. The glyphosate efficacy study was conducted using different doses of glyphosate (0 [control], 180, 360, 720 [recommended dose], and 1,440 g ae ha−1) at two different moisture levels (well-watered and water-stressed conditions). In the second study, the growth and seed production of these biotypes were studied under different atmospheric carbon dioxide (CO2) concentrations (450 and 750 ppm) and under well-watered (100% field capacity) and water-stressed (50% field capacity) conditions. Results showed that the GR biotype survived (>68%) at 1,440 g ha−1, but for the GS biotype, no plant survived, and both biotypes were slightly (<10%) affected by moisture regimes. In the elevated CO2 condition, the GS biotype plants were >38% taller and produced >44%, >18%, and >21% more leaves, buds, and seeds, respectively, compared with the ambient CO2 concentration under both moisture regimes. The biomass also increased by 27% in comparison with the ambient CO2 concentration. For the GR biotype, plants at the elevated CO2 level, while they also grew 38% taller in comparison with the ambient CO2 concentration, the numbers of leaves, buds, and seeds and biomass were not affected by this increase in CO2. Results showed that there were minimal changes in response to glyphosate for GR and GS biotypes of S. oleraceus with or without moisture stress. Our study suggests that future climate change with elevated CO2 levels can affect the response of S. oleraceus to glyphosate, and such knowledge will be helpful for weed management in the future.

The dicamba-resistant cropping system was developed to be used as a tool to control multiple-resistant weed species, particularly Palmer amaranth (Amaranthus palmeri S. Watson). However, dicamba applications have resulted in off-target movement of the herbicide to susceptible neighboring vegetation, with frequent damage to non–dicamba resistant soybean [Glycine max (L.) Merr.]. Pod malformation and subsequent auxin-like injury to progeny is common when parent soybean plants are exposed to the herbicide post-flowering. Yet no publication to date has conveyed the presence of dicamba in seed. The objective of this study was to determine whether dicamba exists and at what quantities inside soybean seed following a low-dose exposure in the pod-filling stage using radiolabeled herbicide as a tracer. Non–dicamba resistant soybean plants were grown in the greenhouse until the pod-filling growth stage and then treated with 2.8 g ae ha−1 of dicamba (1/200 of the recommended rate of 560 g ae ha−1). Immediately afterward, [14C]dicamba (approximately 6.4 kBq per plant) was applied to the adaxial surface of one trifoliate leaf located in the midportion of each plant. The greatest amount of [14C]dicamba recovered was in seeds and in pods, and these plant parts accumulated 44% and 38% of the total absorbed, respectively. Chromatography results showed that the totality of the [14C]dicamba present in the soybean seeds was in the phytotoxic form, except for a single sample, in which one metabolite was detected (possibly 5-hydroxy dicamba). Precautions should be taken to avoid dicamba exposure to sensitive soybean fields, especially those dedicated to seed production, as this may result in low seed quality and symptomology on progeny plants.

Glyphosate is the most used herbicide worldwide, which has contributed to concerns about its environmental impact. Compared with most other herbicides, glyphosate has a half-life in soil and water that is relatively short (averaging about 30 d in temperate climates), mostly due to microbial degradation. Its primary microbial product, aminomethylphosphonic acid, is slightly more persistent than glyphosate. In soil, glyphosate is virtually biologically inactive due to its strong binding to soil components. Glyphosate does not bioaccumulate in organisms, largely due to its high water solubility. Glyphosate-resistant crops have greatly facilitated reduced-tillage agriculture, thereby reducing soil loss, soil compaction, carbon dioxide emissions, and fossil fuel use. Agricultural economists have projected that loss of glyphosate would result in increased cropping area, some gained by deforestation, and an increase in environmental impact quotient of weed management. Some drift doses of glyphosate to non-target plants can cause increased plant growth (hormesis) and/or increased susceptibility to plant pathogens, although these non-target effects are not well documented. The preponderance of evidence confirms that glyphosate does not harm plants by interfering with mineral nutrition and that it has no agriculturally significant effects on soil microbiota. Glyphosate has a lower environmental impact quotient than most synthetic herbicide alternatives.

A redroot pigweed (Amaranthus retroflexus L.) population (HN-02) collected from Nenjiang County, Heilongjiang Province, exhibited multiple resistance to fomesafen and nicosulfuron. The purposes of this study were to characterize the herbicide resistance status of an HN-02 population for both acetolactate synthase (ALS) and protoporphyrinogen oxidase (PPO) inhibitors and the response to other herbicides and to investigate the target site-based mechanism governing fomesafen and nicosulfuron resistance. Three mutations, Ala-205-Val and Trp-574-Leu mutations in the ALS gene and an Arg-128-Gly mutation in the PPX2 gene, were identified in individual resistant plants. An HN-02F1-1 subpopulation homozygous for the Ala-205-Val and Arg-128-Gly mutations was generated, and whole-plant experiments confirmed multiple resistance to PPO inhibitors (fomesafen, fluoroglycofen-ethyl, and acifluorfen) and ALS inhibitors (imidazolinones [IMI], sulfonylureas [SU], and triazolopyrimidines [TP]) in the HN-02F1-1 plants, which presented resistance index values ranging from 8.3 to 110; however, these plants were sensitive to flumioxazin, fluroxypyr-meptyl, and 2,4-D butylate. In vitro ALS enzyme activity assays revealed that, compared with ALS from susceptible plants, ALS from the HN-02F1-1 plants was 15-, 28- and 320-fold resistant to flumetsulam, nicosulfuron, and imazethapyr, respectively. This study confirms the first case of multiple resistance to PPO and ALS inhibitors in A. retroflexus and determines that the target-site resistance mechanism was produced by Ala-205-Val and Arg-128-Gly mutations in the ALS gene and PPX2 gene, respectively. In particular, the Ala-205-Val mutation was found to endow resistance to three classes of ALS inhibitors: TP, SU, and IMI.

The WSSA Group 15 (HRAC Group K3) herbicide flufenacet is a key compound in weed resistance management, primarily used for PRE control of grass weeds in winter cereal–based crop rotations in Europe. Although resistance to compounds of its mechanism of action (inhibition of the synthesis of very-long-chain fatty acids) generally evolves slowly, reduced flufenacet efficacy due to enhanced glutathione transferase (GST) activity has been described in several blackgrass (Alopecurus myosuroides Huds.) populations. The present study aimed to better understand of the mechanism of flufenacet detoxification in A. myosuroides. Therefore, we characterized four A. myosuroides populations with different levels of flufenacet sensitivity. Flufenacet degradation was significantly slowed down in a sensitive population and a population with reduced flufenacet sensitivity by the use of the GST inhibitors tridiphane and ethacrynic acid at sublethal rates. Finally, an RNA sequencing (RNA-seq) study with the four A. myosuroides populations was conducted. In total, six differentially expressed GSTs and nine transcription factors as well as a keto-acyl-CoA reductase involved in the biosynthesis of very-long-chain fatty acids were identified as candidate genes among a set of 319 significantly more highly expressed gene-associated contigs. Among a set of 218 contigs with significantly lower expression levels, receptor kinase activity was the most frequent annotation. In summary, the likely GST-mediated reduction in sensitivity evolves in A. myosuroides at a slow rate and can partially be reversed by an interaction between flufenacet and the GST inhibitors tridiphane and ethacrynic acid. This provides further evidence for enhanced GST activity as a key mechanism in flufenacet resistance in A. myosuroides and supports the hypothesis that the six differentially expressed GSTs detected in the present RNA-seq study are potentially involved in flufenacet resistance.

Palmer amaranth (Amaranthus palmeri S. Watson) populations resistant to acetolactate synthase (ALS)-inhibiting herbicides and glyphosate are fairly common throughout the state of North Carolina (NC). This has led farm managers to rely more heavily on herbicides with other sites of action (SOA) for A. palmeri control, especially protoporphyrinogen oxidase and glutamine synthetase inhibitors. In the fall of 2016, seeds from A. palmeri populations were collected from the NC Coastal Plain, the state’s most prominent agricultural region. In separate experiments, plants with 2 to 4 leaves from the 110 populations were treated with field use rates of glyphosate, glufosinate-ammonium, fomesafen, mesotrione, or thifensulfuron-methyl. Percent visible control and survival were evaluated 3 wk after treatment. Survival frequencies were highest following glyphosate (99%) or thifensulfuron-methyl (96%) treatment. Known mutations conferring resistance to ALS inhibitors were found in populations surviving thifensulfuron-methyl application (Ala-122-Ser, Pro-197-Ser, Trp-574-Leu, and/or Ser-653-Asn), in addition to a new mutation (Ala-282-Asp) that requires further investigation. Forty-two populations had survivors after mesotrione application, with one population having 17% survival. Four populations survived fomesafen treatment, while none survived glufosinate. Dose–response studies showed an increase in fomesafen needed to kill 50% of two populations (LD50); however, these rates were far below the field use rate (less than 5 g ha−1). In two populations following mesotrione dose–response studies, a 2.4- to 3.3-fold increase was noted, with LD90 values approaching the field use rate (72.8 and 89.8 g ha−1). Screening of the progeny of individuals surviving mesotrione confirmed the presence of resistance alleles, as there were a higher number of survivors at the 1X rate compared with the parent population, confirming resistance to mesotrione. These data suggest A. palmeri resistant to chemistries other than glyphosate and thifensulfuron-methyl are present in NC, which highlights the need for weed management approaches to mitigate the evolution and spread of herbicide-resistant populations.