Waterhemp Populations Research Articles

Environmental influences on common waterhemp germination and tolerance to salinity and herbicides

AbstractThis study investigated the potential of common waterhemp [Amaranthus tuberculatus (Moq.) Sauer] invading the semiarid, salinity‐prone areas of the Central Valley (CV) of California, and its response to commonly used herbicides. Despite its status as one of the worst weeds in the US agriculture, common waterhemp is not very prevalent in California. Limited information exists on its germination ecology and potential to invade the CV, or its response to herbicides. Seeds were collected from a local common waterhemp population, and germination was tested under a range of water potential (0 to −4.17 MPa), salinity (0–25 dS m−1), and pH (5–9) levels. Plant tolerance to salinity at different electrical conductivity (EC) levels (0–20 EC) and mortality under applications of label rates of postemergence herbicides (glyphosate, glufosinate, rimsulfuron, or caprylic + capric acid) at the 4‐ to 6‐leaf or 8‐ to 10‐leaf stage were evaluated. Results showed that the seeds were moderately tolerant to drought and salinity as 50% and 55% of them germinated at −0.5 MPa and 10 EC, respectively. Germination was greater in neutral to alkaline than in acidic conditions. Plants were not very tolerant to salinity as 96% of them died at 10 EC. Most of the plants were controlled by herbicide applications at the 4‐ to 6‐leaf stage, but escapes occurred at the later stage. Therefore, the local population has the potential to germinate and establish in the CV but can be controlled by tested herbicides when applied at early stages.

Norflurazon and metribuzin as components of postemergence tank mixtures with HPPD-inhibiting herbicides for multiple-resistant waterhemp control

Field and greenhouse studies investigated the phytoene desaturase (PDS) and photosystem II (PSII)-inhibiting herbicides, norflurazon and metribuzin, as components of two- and three-way postemergence (POST) tank mixtures for controlling multiple herbicide-resistant (MHR) waterhemp (Amaranthus tuberculatus). Studies were conducted at two locations (Champaign County, Illinois, U.S.A (CHR) and McLean County, Illinois, U.S.A. (MCR)) during 2019 and 2020. One of three commercial HPPD-inhibiting herbicides (mesotrione, tembotrione or topramezone) was added to a tank mixture with norflurazon, a reduced rate of metribuzin, or both to determine if greater MHR waterhemp control with tank mixtures could be achieved compared to individual herbicides and assess the potential benefit of norflurazon. Field results indicated the addition of norflurazon to HPPD-inhibiting herbicides resulted in greater waterhemp control (14% increase on average) compared to individual herbicides applied alone. Additionally, metribuzin in combination with HPPD-inhibiting herbicides consistently increased control of MHR waterhemp populations compared to the herbicides applied alone. However, only the three-way tank mixture containing tembotrione at the MCR site reduced biomass relative to two-way treatments containing tembotrione, metribuzin or norflurazon. Greenhouse experiments characterized mixtures of norflurazon with metribuzin and the HPPD-inhibiting herbicides, mesotrione and topramezone. Waterhemp biomass was significantly reduced when metribuzin and HPPD inhibitors were combined in comparison to individual components. Two-way tank mixtures containing norflurazon with mesotrione or metribuzin also resulted in significant biomass reductions compared to herbicides applied alone in MHR waterhemp. However, lower control levels were noted in MHR waterhemp when norflurazon was mixed with either topramezone or topramezone plus metribuzin compared to topramezone alone. In summary, norflurazon and metribuzin can be effective tank mixture components with HPPD-inhibiting herbicides for MHR waterhemp control. However, more research on these tank mixtures is warranted to optimize weed control and crop selectivity, which includes investigating active ingredient ratios, application timings and droplet spectrum effects on coverage.

Confirmation of 4-Hydroxyphenylpyruvate Dioxygenase Inhibitor-Resistant and 5-Way Multiple-Herbicide-Resistant Waterhemp in Ontario, Canada

This is the first confirmation of 4-hydroxyphenylpyruvate dioxygenase-(HPPD)-inhibitor resistant waterhemp and the first report of 5-way multiple-herbicide-resistant (MHR) waterhemp in Ontario to Group 2, 5, 9, 14, and 27 herbicides. Seed was collected across southern Ontario in 2021 and tested for resistance to imazethapyr, atrazine, metribuzin, glyphosate, lactofen, mesotrione, and mesotrione + atrazine representing Group 2, 5, 5, 9, 14, 27, and 27 + 5 herbicides, respectively. All of the waterhemp populations collected in 2021 had 4-way resistance to imazethapyr, atrazine, glyphosate, and mesotrione. In seven Ontario counties, 5-way resistant waterhemp was confirmed which includes resistance to the HPPD-inhibiting herbicides.

Control of waterhemp (Amaranthus tuberculatus) at multiple locations in Illinois with single preemergence applications of VLCFA-inhibiting herbicides

AbstractHerbicides that inhibit very-long-chain fatty acids (VLCFAs) have been widely used for preemergence control of annual monocot and small-seeded dicot weed species, such as waterhemp, since their discovery in the 1950s. VLCFA-inhibiting herbicides are often applied in combination with active ingredients that possess residual activity on small-seeded broadleaf weeds, which can make their contribution to preemergence waterhemp control difficult to quantify. Bare-ground field experiments were designed to investigate the efficacy of eight VLCFA-inhibiting herbicides applied at their minimum and maximum labeled rates for control of Illinois waterhemp populations. Four different locations were selected, two of which contained previously characterized VLCFA inhibitor–resistant waterhemp populations in Champaign County (CHR) and McLean County (MCR). Two locations with VLCFA inhibitor–sensitive waterhemp populations included the University of Illinois South Farm in Urbana, IL, and the Orr Research Center in Perry, IL. Soils at the CHR, MCR, and Urbana locations contained greater than 3% organic matter, but less than 3% organic matter at Perry. Non-encapsulated acetochlor and alachlor controlled CHR and MCR waterhemp populations 28 d after treatment (DAT), whereas other VLCFA-inhibiting herbicides resulted in 61% and 76% control of the CHR and MCR populations, respectively. In contrast, all VLCFA-inhibiting herbicides resulted in 81% and 88% control of the Perry and Urbana waterhemp populations, respectively, 28 DAT. Waterhemp control decreased by 42 DAT, especially for the VLCFA inhibitor–resistant CHR and MCR populations. Overall, VLCFA-inhibiting herbicides remain effective for controlling sensitive waterhemp, but most are not effective for controlling VLCFA inhibitor–resistant waterhemp populations. Proper herbicide stewardship and integrated weed management practices should be implemented to maintain VLCFA-inhibiting herbicide efficacy for waterhemp management in the future.

Resistance to a nonselective 4-hydroxyphenylpyruvate dioxygenase-inhibiting herbicide via novel reduction-dehydration-glutathione conjugation in Amaranthus tuberculatus.

Summary Metabolic resistance to 4‐hydroxyphenylpyruvate dioxygenase (HPPD)‐inhibiting herbicides is a threat in controlling waterhemp (Amaranthus tuberculatus) in the USA. We investigated resistance mechanisms to syncarpic acid‐3 (SA3), a nonselective, noncommercial HPPD‐inhibiting herbicide metabolically robust to Phase I oxidation, in multiple‐herbicide‐resistant (MHR) waterhemp populations (SIR and NEB) and HPPD inhibitor‐sensitive populations (ACR and SEN).Dose–response experiments with SA3 provided ED50‐based resistant : sensitive ratios of at least 18‐fold. Metabolism experiments quantifying parent SA3 remaining in excised leaves during a time course indicated MHR populations displayed faster rates of SA3 metabolism compared to HPPD inhibitor‐sensitive populations.SA3 metabolites were identified via LC‐MS‐based untargeted metabolomics in whole plants. A Phase I metabolite, likely generated by cytochrome P450‐mediated alkyl hydroxylation, was detected but was not associated with resistance. A Phase I metabolite consistent with ketone reduction followed by water elimination was detected, creating a putative α,β‐unsaturated carbonyl resembling a Michael acceptor site. A Phase II glutathione–SA3 conjugate was associated with resistance.Our results revealed a novel reduction–dehydration–GSH conjugation detoxification mechanism. SA3 metabolism in MHR waterhemp is thus atypical compared to commercial HPPD‐inhibiting herbicides. This previously uncharacterized detoxification mechanism presents a unique opportunity for future biorational design by blocking known sites of herbicide metabolism in weeds.

Investigating the Efficacy of Selected Very-Long-Chain Fatty Acid-Inhibiting Herbicides on Iowa Waterhemp (Amaranthus tuberculatus) Populations with Evolved Multiple Herbicide Resistances

Very long chain fatty acid (VLCFA)-inhibiting herbicides (Herbicide group (HG) 15) have been applied to corn and soybean fields in Iowa since the 1960s. The VLCFA-inhibiting herbicides are now applied more frequently to control multiple herbicide-resistant (MHR) waterhemp (Amaranthus tuberculatus Moq. J.D. Sauer) populations that are ubiquitous across the Midwest United States as resistance to the VLCFA-inhibiting herbicides is not widespread. Waterhemp has evolved multiple resistances to herbicides from seven sites of action (HG 2, 4, 5, 9, 14, 15, and 27), and six-way herbicide-resistant populations have been confirmed. Thus, the objective of this study was to determine if selected Iowa waterhemp populations are less sensitive to VLCFA-inhibiting herbicides when additional herbicide resistance traits have evolved within the selected population. Dose–response assays were conducted in a germination chamber to determine the efficacy of three selected VLCFA-inhibiting herbicides (acetochlor, S-metolachlor, and flufenacet) on selected Iowa MHR waterhemp populations. An herbicide-susceptible, three-way, four-way, and five-way herbicide-resistant waterhemp population responded to the herbicide treatments differently; however, several of the four-way and five-way herbicide-resistant populations exhibited resistance ratios greater than 1 when treated with acetochlor and S-metolachlor. Selected four-way herbicide-resistant waterhemp populations from Iowa were subjected to a dose–response assay in the field using the same VLCFA-inhibiting herbicides, and all herbicides achieved control greater than 80% at the maximum labeled rate. The results of the experiments provide evidence that some MHR waterhemp populations may exhibit decreased susceptibility the VLCFA-inhibiting herbicides, but generally, these herbicides remain efficacious on Iowa MHR waterhemp populations.

Molecular confirmation of resistance to PPO inhibitors in Amaranthus tuberculatus and Amaranthus palmeri, and isolation of the G399A PPO2 substitution in A. palmeri

AbstractDuring the 2017 to 2019 growing seasons, samples of waterhemp and Palmer amaranth that had reportedly survived field-rate applications of protoporphyrinogen oxidase (PPO)–inhibiting herbicides were collected from the American Midwest and tested for target-site mutations known at the time to confer resistance. Target-site resistance was identified in nearly all (135 of 145) tested common waterhemp populations but in only 8 of 13 Palmer amaranth populations. Follow-up research on one population of Palmer amaranth (W-8), which tested negative for all such mutations, confirmed it was resistant to lactofen, with a magnitude of resistance comparable to that conferred by the ΔG210 PPO2 mutation. Gene sequences from both isoforms of PPO (PPO1 and PPO2) were compared between W-8 and known PPO inhibitor–sensitive sequence. A glycine-to-alanine substitution at the 399th amino acid position (G399A) of PPO2, recently identified to reduce target-site herbicide sensitivity, was observed in a subset of resistant W-8 plants. Because no missense mutation completely delimited resistant and sensitive sequences, we initially suspected the presence of a secondary, non-target-site resistance mechanism in this population. To isolate G399A, a segregating F2 population was produced and screened with a delimiting rate of lactofen. χ2 goodness-of-fit analysis of dead/alive ratings indicated single-locus inheritance of resistance in the F2 population, and molecular markers for the W-8 parental PPO2 coding region co-segregated tightly, but not perfectly, with resistance. More research is needed to fully characterize Palmer amaranth PPO inhibitor–resistance mechanisms, which appear to be more diverse than those found in common waterhemp.

Metabolic resistance to S-metolachlor in two waterhemp (Amaranthus tuberculatus) populations from Illinois, USA.

Two waterhemp (Amaranthus tuberculatus) populations from Illinois demonstrating multiple-resistance to acetolactate synthase (ALS)-, 4-hydroxyphenylpyruvate dioxygenase, and photosystem II (PSII)-inhibiting herbicides (designated CHR and SIR) also displayed reduced sensitivity to very-long-chain fatty acid-inhibiting herbicides, including S-metolachlor. We hypothesized that a physiological mechanism, such as enhanced metabolism, could be responsible for the reduced efficacy of S-metolachlor. Metabolism experiments indicated that resistant populations degraded S-metolachlor more rapidly than sensitive populations and equally as rapidly as corn 2-24 h after treatment (HAT). Resistant waterhemp and corn metabolized 90% (DT90 ) of absorbed S-metolachlor in less than 3.2 h whereas DT90 values for sensitive waterhemp exceeded 6 h. The glutathione S-transferase inhibitor 4-chloro-7-nitrobenzofurazon and cytochrome P450-inhibitor malathion decreased the amount of S-metolachlor metabolized in resistant waterhemp at 4 HAT but not in sensitive waterhemp or corn, and altered the abundance of certain metabolites in resistant waterhemp. Results from this research demonstrate that resistance to S-metolachlor in these waterhemp populations is due to enhanced herbicide metabolism relative to sensitive populations. In addition, our results indicate that resistant waterhemp might utilize metabolic pathway(s) more intricate than either sensitive waterhemp or corn based on their metabolite profiles. © 2020 Society of Chemical Industry.

Herbicide drift exposure leads to reduced herbicide sensitivity in Amaranthus spp.

While the introduction of herbicide tolerant crops provided growers new options to manage weeds, the widespread adoption of these herbicides increased the risk for herbicide spray drift to surrounding vegetation. The impact of herbicide drift in sensitive crops is extensively investigated, whereas scarce information is available on the consequences of herbicide drift in non-target plants. Weeds are often abundant in field margins and ditches surrounding agricultural landscapes. Repeated herbicide drift exposure to weeds could be detrimental to long-term management as numerous weeds evolved herbicide resistance following recurrent-selection with low herbicide rates. The objective of this study was to evaluate if glyphosate, 2,4-D, and dicamba spray drift could select Amaranthus spp. biotypes with reduced herbicide sensitivity. Palmer amaranth and waterhemp populations were recurrently exposed to herbicide drift in a wind tunnel study over two generations. Seeds from survival plants were used for the subsequent rounds of herbicide drift exposure. Progenies were subjected to herbicide dose-response studies following drift selection. Herbicide drift exposure rapidly selected for Amaranthus spp. biotypes with reduced herbicide sensitivity over two generations. Weed management programs should consider strategies to mitigate near-field spray drift and suppress the establishment of resistance-prone weeds on field borders and ditches in agricultural landscapes.

Comparing responses of sensitive and resistant populations of Palmer amaranth (Amaranthus palmeri) and waterhemp (Amaranthus tuberculatus var. rudis) to PPO inhibitors

AbstractResistance to protoporphyrinogen oxidase (PPO) inhibitors was first observed in waterhemp in 2001 and was conferred by the deletion of a glycine residue at the 210th position (ΔGly-210) of the PPO enzyme. PPO-inhibitor resistance in Palmer amaranth was first observed in 2011, 10 years later. The objectives of this study were to directly compare PPO inhibitor responses in plants of both species with or without the ΔGly-210 mutation. Using greenhouse experiments, early (EPOST) and late (LPOST) postemergence dose responses using lactofen and fomesafen, and preemergence (PRE) dose responses using fomesafen and flumioxazin, were obtained for a sensitive and resistant population each of waterhemp and Palmer amaranth. An additional spray study confirmed each sensitive population used in the dose responses was representative of its respective species, with regards to PPO-inhibitor sensitivity. When treated at either POST timing, Palmer amaranth was more tolerant than waterhemp, and the ΔGly-210 mutation provided greater resistance in Palmer amaranth (48-fold to >3,440-fold, depending on timing and herbicide) than in waterhemp (31-fold to 123-fold). The level of tolerance in Palmer amaranth was striking; the sensitive Palmer amaranth population treated LPOST survived as well or better than the resistant waterhemp population treated EPOST. With PRE applications, response differences both between species and between resistant and sensitive populations generally were less pronounced, relative to POST applications. Collectively, this research indicates Palmer amaranth tolerance to POST-applied PPO inhibitors could have initially slowed (relative to waterhemp) evolution of resistance to these herbicides, and resistant and sensitive populations of both species are more likely to be effectively controlled with PRE rather than POST applications.

Characterization of a waterhemp (Amaranthus tuberculatus) population from Illinois resistant to herbicides from five site-of-action groups

AbstractExperiments were initiated to characterize a waterhemp population (CHR) discovered in a central Illinois corn field after it was not controlled by the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor topramezone. Field experiments conducted during 2014–2015 indicated that acetolactate synthase (ALS)-, protoporphyrinogen oxidase (PPO)-, photosystem II (PSII)-, and HPPD-inhibiting herbicides and the synthetic auxin 2,4-D did not control the CHR population. Laboratory experiments confirmed target site–based resistance mechanisms to ALS- and PPO-inhibiting herbicides. Herbicide doses required to reduce dry biomass 50% (GR50) were determined in greenhouse dose–response experiments, and indicated 16-fold resistance to the HPPD inhibitor mesotrione, 9.5-fold resistance to the synthetic auxin 2,4-D, and 252-fold resistance to the PSII inhibitor atrazine. Complementary results from field, laboratory, and greenhouse investigations indicate that the CHR population has evolved resistance to herbicides from five sites of action (SOAs): ALS-, PPO-, PSII-, and HPPD-inhibiting herbicides and 2,4-D. Herbicide use history for the field in which CHR was discovered indicates no previous use of 2,4-D.

Investigating target-site resistance mechanism to the PPO-inhibiting herbicide fomesafen in waterhemp and interspecific hybridization of Amaranthus species using next generation sequencing.

Waterhemp (Amaranthus tuberculatus (Moq.) J. D. Sauer) is one of the most pernicious weeds in cropping systems of the USA due to its evolved resistance against several herbicide sites-of-action, including protoporphyrinogen oxidase inhibitors (PPO-R). Currently, the only source of PPO-R documented in waterhemp is ΔG210 of PPX2. Gene flow may not only lead to a transfer of herbicide-resistant alleles, but also produce a hybrid genotype more competitively fit than one or both parents. However, investigating gene flow of Amaranthus species has been of interest in the past two decades with limited evidence. Here, a high-throughput MiSeq amplicon sequencing method was used to investigate alterations of the PPX2 gene in 146 PPO-R waterhemp populations across five Midwest states of the USA. Five R128 codons of PPX2, novel to waterhemp, were found including AGG (R), GGA (G), GGG (G), AAA (K) and ATA (I). R128G, R128I, and R128K were found in 11, 3, and 2 populations, respectively. R128G and R128I, but not R128K, conferred fomesafen resistance in a bacterial system. Sequence alignment of the R128 region of PPX2 identified a tumble pigweed (Amaranthus albus)-type and Palmer amaranth (Amaranthus palmeri)-type PPX2 allele to be present and widespread in the surveyed waterhemp populations, thus providing strong evidence of gene flow between Amaranthus species. Using a next-generation sequencing method, we identified two PPO target-site mutations R128G/I novel to waterhemp and provided evidence of gene flow of Amaranthus species in a large group of screened waterhemp populations from five Midwest states of the USA. © 2019 Society of Chemical Industry.

Using RNA-seq to characterize responses to 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor herbicide resistance in waterhemp (Amaranthus tuberculatus)

BackgroundWaterhemp (Amaranthus tuberculatus (Moq.) J.D. Sauer) is a problem weed commonly found in the Midwestern United States that can cause crippling yield losses for both maize (Zea mays L.) and soybean (Glycine max L. Merr). In 2011, 4-hydroxyphenylpyruvate-dioxygenase (HPPD, EC 1.13.11.27) inhibitor herbicide resistance was first reported in two waterhemp populations. Since the discovery of HPPD-herbicide resistance, studies have identified the mechanism of resistance and described the inheritance of the herbicide resistance. However, no studies have examined genome-wide gene expression changes in response to herbicide treatment in herbicide resistant and susceptible waterhemp.ResultsWe conducted RNA-sequencing (RNA-seq) analyses of two waterhemp populations (HPPD-herbicide resistant and susceptible), from herbicide-treated and mock-treated leaf samples at three, six, twelve, and twenty-four hours after treatment (HAT). We performed a de novo transcriptome assembly using all sample sequences. Following assessments of our assembly, individual samples were mapped to the de novo transcriptome allowing us to identify transcripts specific to a genotype, herbicide treatment, or time point. Our results indicate that the response of HPPD-herbicide resistant and susceptible waterhemp genotypes to HPPD-inhibiting herbicide is rapid, established as soon as 3 hours after herbicide treatment. Further, there was little overlap in gene expression between resistant and susceptible genotypes, highlighting dynamic differences in response to herbicide treatment. In addition, we used stringent analytical methods to identify candidate single nucleotide polymorphisms (SNPs) that distinguish the resistant and susceptible genotypes.ConclusionsThe waterhemp transcriptome, herbicide-responsive genes, and SNPs generated in this study provide valuable tools for future studies by numerous plant science communities. This collection of resources is essential to study and understand herbicide effects on gene expression in resistant and susceptible weeds. Understanding how herbicides impact gene expression could allow us to develop novel approaches for future herbicide development. Additionally, an increased understanding of the prolific traits intrinsic in weed success could lead to crop improvement.

Carfentrazone-ethyl resistance in an Amaranthus tuberculatus population is not mediated by amino acid alterations in the PPO2 protein.

To date, the only known mechanism conferring protoporphyrinogen IX oxidase (PPO)-inhibitor resistance in waterhemp (Amaranthus tuberculatus) is a glycine deletion in PPO2 (ΔG210), which results in cross-resistance to foliar PPO-inhibiting herbicides. However, a metabolism-based, HPPD-inhibitor resistant waterhemp population from Illinois (named SIR) was suspected of having a non-target site resistance (NTSR) mechanism due to its resistance to carfentrazone-ethyl (CE) but sensitivity to diphenylethers (DPEs). In greenhouse experiments, SIR sustained less injury than two PPO inhibitor-sensitive populations (WCS and SEN) after applying a field-use rate of CE, and after initial rapid necrosis, regrowth of SIR plants was comparable to a known PPO inhibitor-resistant population (ACR) possessing the ΔG210 mutation. Dose-response analysis determined 50% growth reduction rates in CE-resistant (SIR and ACR) and sensitive (SEN) waterhemp populations, which showed SIR was 30-fold resistant compared to SEN and two-fold more resistant than ACR. Deduced amino acid sequences derived from SIR PPX2 partial cDNAs did not contain the ΔG210 mutation found in ACR or other target-site mutations that confer PPO-inhibitor resistance previously reported in Palmer amaranth (Amaranthus palmeri). Although several SIR cDNAs contained amino acid substitutions, none were uniform among samples. Additionally, SIR plants treated with malathion and CE showed a significant reduction in biomass accumulation compared to CE alone. These results indicate robust CE resistance in SIR is not mediated by amino acid changes in the PPO2 protein, but instead resistance may be conferred through a NTSR mechanism such as enhanced herbicide metabolism.

Metabolic Pathway of Topramezone in Multiple-Resistant Waterhemp (Amaranthus tuberculatus) Differs From Naturally Tolerant Maize.

Waterhemp [Amaranthus tuberculatus (Moq.) Sauer] is a problematic dicot weed in maize, soybean, and cotton production in the United States. Waterhemp has evolved resistance to several commercial herbicides that inhibit the 4-hydroxyphenylpyruvate-dioxygenase (HPPD) enzyme in sensitive dicots, and research to date has shown that HPPD-inhibitor resistance is conferred by rapid oxidative metabolism of the parent compound in resistant populations. Mesotrione and tembotrione (both triketones) have been used exclusively to study HPPD-inhibitor resistance mechanisms in waterhemp and a related species, A. palmeri (S. Wats.), but the commercial HPPD inhibitor topramezone (a pyrazolone) has not been investigated from a mechanistic standpoint despite numerous reports of cross-resistance in the field and greenhouse. The first objective of our research was to determine if two multiple herbicide-resistant (MHR) waterhemp populations (named NEB and SIR) metabolize topramezone more rapidly than two HPPD inhibitor-sensitive waterhemp populations (named SEN and ACR). Our second objective was to determine if initial topramezone metabolite(s) detected in MHR waterhemp are qualitatively different than those formed in maize. An excised leaf assay and whole-plant study investigated initial rates of topramezone metabolism (<24 h) and identified topramezone metabolites at 48 hours after treatment (HAT), respectively, in the four waterhemp populations and maize. Results indicated both MHR waterhemp populations metabolized more topramezone than the sensitive (SEN) population at 6 HAT, while only the SIR population metabolized more topramezone than SEN at 24 HAT. Maize metabolized more topramezone than any waterhemp population at each time point examined. LC-MS analysis of topramezone metabolites at 48 HAT showed maize primarily formed desmethyl and benzoic acid metabolites, as expected based on published reports, whereas SIR formed two putative hydroxylated metabolites. Subsequent LC-MS/MS analyses identified both hydroxytopramezone metabolites in SIR as different hydroxylation products of the isoxazole ring, which were also present in maize 48 HAT but at very low levels. These results indicate that SIR initially metabolizes and detoxifies topramezone in a different manner than tolerant maize.

Inheritance of 4-hydroxyphenylpyruvate dioxygenase inhibitor herbicide resistance in an Amaranthus tuberculatus population from Iowa, USA

Waterhemp (Amaranthus tuberculatus (Moq.) J.D. Sauer) is a weed prevalent in the Midwest United States and can cause yield losses up to 74% in maize (Zea mays L.) and 56% in soybean (Glycine max (L.) Merr.). An important adaptive trait commonly found in waterhemp is the ability to evolve herbicide resistance and waterhemp populations have evolved resistance to six herbicide sites of action. In 2011, two waterhemp populations were discovered resistant to p-hydroxyphenylpyruvate-dioxygenase (HPPD, EC 1.13.11.27) inhibitor herbicides. We reciprocally crossed a known HPPD-resistant waterhemp population with a known HPPD-susceptible waterhemp population and then intermated the F1 families to established a pseudo-F2 generation. We challenged the parent, F1 and pseudo-F2 generations against four HPPD-inhibiting herbicide rates (mesotrione). Our results suggest the HPPD-resistance trait is polygenic. Furthermore, the number of genes involved with the herbicide resistance increase at higher herbicide rates. These data indicated at least one dominant allele at each major locus is required to confer HPPD herbicide resistance in waterhemp. Using different waterhemp populations and methodologies, this study confirms the reported “complex” HPPD resistance inheritance while providing new information in the response of HPPD-resistant waterhemp to HPPD herbicides.

Metabolism of 2,4-dichlorophenoxyacetic acid contributes to resistance in a common waterhemp (Amaranthus tuberculatus) population.

Synthetic auxins such as 2,4-dichlorophenoxyacetic acid (2,4-D) have been widely used for selective control of broadleaf weeds since the mid-1940s. In 2009, an Amaranthus tuberculatus (common waterhemp) population with 10-fold resistance to 2,4-D was found in Nebraska, USA. The 2,4-D resistance mechanism was examined by conducting [14 C] 2,4-D absorption, translocation and metabolism experiments. No differences were found in 2,4-D absorption or translocation between resistant and susceptible A. tuberculatus plants. Resistant plants metabolized [14 C] 2,4-D more rapidly than did susceptible plants. The half-life of [14 C] 2,4-D in susceptible plants was 105 h, compared with 22 h in resistant plants. Pretreatment with the cytochrome P450 inhibitor malathion inhibited [14 C] 2,4-D metabolism in resistant plants and reduced the 2,4-D dose required for 50% growth inhibition (GR50 ) of resistant plants by 7-fold to 27 g ha-1 , similar to the GR50 for susceptible plants in the absence of malathion. Our results demonstrate that rapid 2,4-D metabolism is a contributing factor to resistance in A. tuberculatus, potentially mediated by cytochrome P450. Metabolism-based resistance to 2,4-D could pose a serious challenge for A. tuberculatus control because of the potential for cross-resistance to other herbicides. © 2017 Society of Chemical Industry.

Multiple-Herbicide Resistance in a 2,4-D–Resistant Waterhemp (Amaranthus tuberculatus) Population from Nebraska

A 2,4-D-resistant tall waterhemp population (FS) from Nebraska was evaluated for resistance to other TIR1 auxin receptor herbicides and to herbicides having alternative mechanisms of action using greenhouse bioassays and genetic markers. Atrazine, imazethapyr, lactofen, mesotrione, glufosinate, and glyphosate were applied in a single-dose bioassay, and tissue was collected from marked plants for genetic analysis. The FS population was not injured by atrazine or by imazethapyr. Approximately 50% of the plants survived lactofen and were actively growing 28 d after treatment. The population was susceptible to mesotrione, glufosinate, and glyphosate. Ametryn, chlorimuron-ethyl, 2,4-D, aminocyclopyraclor, aminopyralid, and picloram were applied in dose–response studies. The FS population was sensitive to ametryn, and the Ser-264-Gly substitution in the D1 protein was not detected, suggesting the lack of response to atrazine is not due to a target-site mutation. The FS population exhibited less than 50% injury to chlorimuron-ethyl at application rates 20 times the labeled use rate. The Ser-653-Asn acetolactate synthase (ALS) substitution, which confers resistance to imidazolinone herbicides, was present in the FS population. However, this does not explain the lack of response to the sulfonylurea herbicide, chlorimuron-ethyl. Sequencing of a portion of thePPX2Lgene did not show the ΔG210 mutation that confers resistance to protoporphyrinogen oxidase–inhibiting herbicides, suggesting that other factors were responsible for waterhemp survival after lactofen application. The FS population was confirmed to be at least 30-fold resistant to 2,4-D relative to the susceptible populations. In addition, it was at least 3-fold less sensitive to aminopyralid and picloram, two other TIR1 auxin receptor herbicides, than the 2,4-D-susceptible populations were. These data indicated that the FS population contains both target and non–target site mechanisms conferring resistance to herbicides spanning at least three mechanisms of action: TIR1 auxin receptors, ALS inhibitors, and photosystem II inhibitors.

Glyphosate- and multiple-resistant waterhemp (Amaranthus tuberculatus var. rudis) in Ontario, Canada

Waterhemp is one of the most troublesome weeds in the US and is spreading into Ontario. In 2014, a waterhemp population was not controlled with glyphosate in a field in Lambton County, ON. This population was the first confirmed glyphosate-resistant (GR) waterhemp in Canada. In 2015, waterhemp seeds were collected from 48 fields in Lambton (32), Chatham-Kent (2), and Essex (14) counties to determine the occurrence and distribution of GR waterhemp in Ontario. Waterhemp plants were grown in a greenhouse and sprayed when 10 cm in height. In addition to glyphosate (a group 9 herbicide), collected populations were screened for resistance to imazethapyr and atrazine, representing herbicide groups 2 and 5, respectively. Visual control estimates for biomass reduction were completed at 1, 3, and 5 wk after application. Glyphosate-resistant waterhemp was confirmed in 40 fields, representing 82% of all sampled fields from the three Ontario counties. Of the 49 populations collected, all were resistant to imazethapyr ...

Biochemical characterization of metabolism-based atrazine resistance in Amaranthus tuberculatus and identification of an expressed GST associated with resistance.

SummaryRapid detoxification of atrazine in naturally tolerant crops such as maize (Zea mays) and grain sorghum (Sorghum bicolor) results from glutathione S‐transferase (GST) activity. In previous research, two atrazine‐resistant waterhemp (Amaranthus tuberculatus) populations from Illinois, U.S.A. (designated ACR and MCR), displayed rapid formation of atrazine‐glutathione (GSH) conjugates, implicating elevated rates of metabolism as the resistance mechanism. Our main objective was to utilize protein purification combined with qualitative proteomics to investigate the hypothesis that enhanced atrazine detoxification, catalysed by distinct GSTs, confers resistance in ACR and MCR. Additionally, candidate AtuGST expression was analysed in an F2 population segregating for atrazine resistance. ACR and MCR showed higher specific activities towards atrazine in partially purified ammonium sulphate and GSH affinity‐purified fractions compared to an atrazine‐sensitive population (WCS). One‐dimensional electrophoresis of these fractions displayed an approximate 26‐kDa band, typical of GST subunits. Several phi‐ and tau‐class GSTs were identified by LC‐MS/MS from each population, based on peptide similarity with GSTs from Arabidopsis. Elevated constitutive expression of one phi‐class GST, named AtuGSTF2, correlated strongly with atrazine resistance in ACR and MCR and segregating F2 population. These results indicate that AtuGSTF2 may be linked to a metabolic mechanism that confers atrazine resistance in ACR and MCR.