Resistance to HPPD-inhibiting herbicides in a population of waterhemp (Amaranthus tuberculatus) from Illinois, United States

BACKGROUND: A population of waterhemp in a seed maize production field in central Illinois, United States, was not adequately controlled after post‐emergence applications of herbicides that inhibit 4‐hydroxyphenylpyruvate dioxygenase (HPPD). RESULTS: Progeny from the field population survived following treatment with mesotrione, tembotrione or topramezone applied to the foliage either alone or in combination with atrazine in greenhouse experiments. Dose–response experiments indicated that the level of resistance to the HPPD inhibitor mesotrione is at least tenfold relative to sensitive biotypes. CONCLUSION: These studies confirm that waterhemp has evolved resistance to HPPD‐inhibiting herbicides. Copyright © 2011 Society of Chemical Industry     

Influence of multiple herbicide resistance on growth in Amaranthus tuberculatus

Plant defence traits, such as herbicide resistance mutations, may incur a fitness cost to plants that become evident when the trait is not needed. However, individuals with multiple herbicide resistance traits may decrease fitness beyond that of plants with a single herbicide resistance mutation. Multiple herbicide‐resistant (MHR) Amaranthus tuberculatus populations are becoming more prevalent in Midwest United States agroecosystems. The objective was to determine whether selected MHR A. tuberculatus populations express differential development when grown in a herbicide‐free environment. The hypothesis was that MHR A. tuberculatus populations become increasingly less fit when additional herbicide resistances evolve. Multiple herbicide‐resistant and herbicide‐susceptible A. tuberculatus populations were grown in a herbicide‐free field for 20 weeks for two seasons. Differences (P < 0.001) in apical growth were detected 5 and 7 weeks after transplanting for all populations in 2016 and 2017 respectively. Gender and population influenced (P < 0.001) flowering date, with males flowering up to 1.5 weeks earlier than females, but did not cause pollination asynchrony. Shoot biomass was not different (P = 0.84) across A. tuberculatus populations, but there were differences (P < 0.001) for gender and year. Seed production was different amongst A. tuberculatus populations (P = 0.001), but was not influenced by the number of MHR traits. Conversely, a negative quadratic relationship between seed mass and the number of MHR traits was observed (r² = 0.32; P < 0.001). The experiment results demonstrate that MHR in A. tuberculatus populations is not incurring a fitness penalty that will remove the populations in the immediate future.     

Waterhemp (Amaranthus tuberculatus) control under drought stress with 2,4‐dichlorophenoxyacetic acid and glyphosate

Waterhemp (Amaranthus tuberculatus) is a common and troublesome weed in cropping systems throughout the United States. With the potential for future periods of low rainfall or drought, the need for improved weed control under drought stress is necessary. Drought stress typically reduces herbicide efficacy by reducing the foliar uptake of herbicides and their translocation. The objectives of this research were to determine the efficacy of 2,4‐dichlorophenoxyacetic acid (2,4‐D) and glyphosate, applied alone or when tank‐mixed, on waterhemp under varying levels of drought stress, the effect of the timing of drought stress in relation to herbicide application and the absorption and translocation of each herbicide in drought‐stressed waterhemp. At reduced herbicide rates, 2,4‐D had a greater level of control of waterhemp under drought stress, compared to glyphosate. The level of herbicide efficacy was lower when the amount of water that was applied to the plants was reduced. The level of waterhemp control was greatest when drought stress occurred before the herbicide application and when the plants were watered to saturation after the application, compared to when drought stress occurred after the herbicide application or restricted watering levels occurred throughout the entire study. Glyphosate absorption and translocation were reduced in the drought‐stressed plants, but 2,4‐D absorption and translocation were not altered. The absence of a reduction in 2,4‐D translocation in the drought‐stressed weeds has not been previously reported. Applying herbicides prior to a rainfall event could increase the weed control level, even if the weed is stressed. Determining how and why 2,4‐D absorption and translocation levels, compared to those of glyphosate, are unaffected by drought stress in waterhemp can aid in improving the control of drought‐stressed weeds with other postemergence herbicides.     

Triazine resistance in Amaranthus tuberculatus (Moq) Sauer that is not site-of-action mediated

While surveying Illinois Amaranthus tuberculatus (Moq) Sauer (tall waterhemp) half-sib populations for herbicide response variability, several were observed to segregate for resistance to atrazine. Studies were conducted on greenhouse-grown A tuberculatus plants to compare atrazine responses among populations that were segregating for resistance (SegR), uniformly sensitive (UniS) or uniformly resistant (UniR). In chlorophyll fluorescence assays, leaves of plants from the SegR and UniS populations displayed changes in fluorescence after treatment with atrazine, indicating that atrazine was inhibiting electron transport of photosystem II in chloroplasts. Sequencing of a fragment of psbA, which encodes the D1 protein, revealed that the SegR population did not contain the amino acid substitution that is typically found in triazine-resistant plants. Whole-plant herbicide dose-response experiments revealed that, relative to the UniS population, atrazine resistances in the UniR and SegR populations were > 770-fold and 16-fold, respectively. The SegR population was also resistant to cyanazine (59-fold), but not to metribuzin, linuron or pyridate. Triazine resistance in the SegR population was shown to be a nuclear inherited trait, unlike maternal inheritance of site-of-action mediated triazine resistance found in the UniR population. Taken collectively, these findings confirm the existence of two distinct triazine resistance mechanisms in A tuberculatus.     

Growth and fitness of triazine-susceptible and triazine-resistant common waterhemp (Amaranthus tuberculatus var. rudis)

Two greenhouse trials were conducted over a 2 year period (2004 and 2005) to compare the relative growth potential and fitness of triazine-susceptible (TS) and triazine-resistant (TR) waterhemp biotypes. Waterhemp plants from each biotype were grown in 6 L pots in the greenhouse in a completely randomized design. There was no difference in leaf number, plant height, plant biomass, time to bud or time to flower between the two biotypes. The TR female waterhemp plants produced 30% less reproductive biomass and 39% less seed than the TS biotype. The fitness penalty associated with triazine resistance was related to reproductive capability, not to vegetative growth.     

Herbicidal activity on acetolactate synthase‐resistant barnyardgrass (Echinochloa crus‐galli) in Arkansas,USA

The intensive use of the acetolactate synthase (ALS)‐inhibiting herbicides, imazethapyr, penoxsulam and bispyribac‐sodium, in imidazolinone‐resistant (Clearfield) rice increases the risk of the evolution of ALS‐resistant barnyardgrass. In 2009, imazethapyr failed to control barnyardgrass that was collected from a field in Arkansas, USA, following the failure of the herbicide in 2008. A greenhouse experiment was conducted to confirm and document the level of resistance of the biotype against three ALS‐inhibiting herbicides that currently are labeled in rice. The level of control of the resistant biotype at the labeled rate of bispyribac‐sodium of 35 g ai ha^?1 was 10%, penoxsulam at 22 g ai ha^?1 was 0% and imazethapyr at 70 g ai ha^?1 was 25%. The level of mortality of the susceptible biotype was 100% with all the herbicides at the labeled rate. The dose needed to kill 50% of the resistant plants was 49 g ha^?1 of bispyribac‐sodium, 254 g ha^?1 of penoxsulam and 170 g ha^?1 of imazethapyr. For the susceptible biotype, bispyribac‐sodium at 6 g ha^?1, penoxsulam at 10 g ha^?1 and imazethapyr at 12 g ha^?1 killed 50% of the treated plants. Based on these findings, it was confirmed that a barnyardgrass population has evolved cross‐resistance to three ALS‐inhibiting herbicides in rice culture in Arkansas. Furthermore, an experiment was conducted to determine if the ALS‐resistant biotype could be controlled using other mechanisms of action. The results indicated that propanil, a photosystem II inhibitor, and quinclorac, a synthetic auxin, failed to control the resistant biotype at the labeled rates, whereas all the other evaluated herbicides provided effective control of both biotypes.     

Differential response of Amaranthus tuberculatus (Moq ex DC) JD Sauer to glyphosate

Midwest USA farmers have reported inconsistent control of Amaranthus tuberculatus (= rudis) (Moq ex DC) JD Sauer by glyphosate in glyphosate-resistant crops. The potential of selection for decreased A tuberculatus sensitivity to glyphosate was therefore investigated in a reportedly resistant Everly, IA population (P0-EV). Evaluation of six A tuberculatus populations from the Midwest USA estimated a seedling baseline sensitivity of 2.15 mM glyphosate. Based on these results, three generations of divergent recurrent selection were implemented on P0-EV to isolate resistant and susceptible populations. A seedling assay was developed to screen large amounts of seeds and thus expedite the selection process. Whole-plant and seedling rate responses of P0-EV and a known pristine A tuberculatus population from Paint Creek, OH (P0-WT) identified no significant difference in response to glyphosate; however, greater phenotypic variance was ostensibly evident in P0-EV. The first recurrent generation selected for resistance at 3.2 mM glyphosate (RS1-R) had a 5.9- and 1.7-fold resistance increase at the seedling and whole-plant levels, respectively, compared with the susceptible generation selected at 32 microM glyphosate. After three cycles of recurrent selection, 14.6-fold difference in resistance at the seedling level and 3.1-fold difference at the whole-plant level were observed when comparing the populations selected for resistance (RS3-R) and susceptibility (RS3-S). Overall, recurrent selection increased the frequency of resistant individuals and decreased the variability to glyphosate at the population level. Nevertheless, variability for glyphosate resistance was still evident in RS3-R. Results herein suggested that A tuberculatus is inherently variable to glyphosate and that selection decreased the sensitivity to glyphosate. We purport that evolved glyphosate resistance in A tuberculatus may require multiple cycles of selection under field conditions. Historic estimated use of glyphosate alludes to the evolution of tolerant weed populations.     

Occurrence of sulfonylurea resistance in Sagittaria trifolia,a basal monocot species,based on target‐site and non‐target‐site resistance

Sagittaria trifolia L. is one of the most serious weeds in paddy fields in Japan. Since the late 1990s, severe infestations of S. trifolia have occurred following applications of sulfonylurea herbicides in Akita prefecture. In this study, two accessions of S. trifolia, R1 and R2, were collected from paddy fields with severe infestations and their resistance profiles were determined in comparison to a susceptible accession, S1. R1 and R2 were highly resistant to bensulfuron‐methyl. R1 was also highly resistant to pyrazosulfuron‐ethyl, but R2 was susceptible. Relative to S1, R1 had an amino acid substitution at the Pro₁₉₇ residue of acetolactate synthase (ALS), a well‐known mutation that confers sulfonylurea resistance, suggesting that R1 has a target‐site‐based resistance (TSR) mechanism. The sequence of the ALS gene in R2 was identical to that in S1. A Southern blot analysis indicated that there was only one copy of the ALS gene in S1 and R2. These results suggest that R2 has a non‐target‐site‐based resistance (NTSR) mechanism. R2 was moderately resistant to imazosulfuron but susceptible to thifensulfuron‐methyl. R2 and S1 were susceptible to pretilachlor, benfuresate, MCPA‐ethyl and bentazon. The results reveal the occurrence of two sulfonylurea‐resistant biotypes of S. trifolia that show different mechanisms of cross‐resistance to sulfonylureas related to TSR in R1 and NTSR in R2.     

Mechanism of resistance to bensulfuron-methyl in Alisma plantago-aquatica biotypes from Portuguese rice paddy fields

Two Alisma plantago‐aquatica biotypes resistant to bensulfuron‐methyl were detected in rice paddy fields in Portugal’s Mondego (biotype T) and Tagus and Sorraia (biotype Q) River valleys. The fields had been treated with bensulfuron‐methyl‐based herbicide mixtures for 4–6 years. In order to characterize the resistant (R) biotypes, dose–response experiments, absorption and translocation assays, metabolism studies and acetolactate synthase (ALS) activity assays were performed. There were marked differences between R and susceptible (S) biotypes, with a resistance index (ED₅₀R/S) of 500 and 6.25 for biotypes Q and T respectively. Cross‐resistance to azimsulfuron, cinosulfuron and ethoxysulfuron, but not to metsulfuron‐methyl, imazethapyr, bentazone, propanil and MCPA was demonstrated. No differences in the absorption and translocation of ¹⁴C‐bensulfuron‐methyl were found between the biotypes studied. Maximum absorption attained 1.12, 2.02 and 2.56 nmol g⁻¹ dry weight after 96 h incubation with herbicide, for S, Q and T biotypes respectively. Most of the radioactivity taken up by the roots was translocated to shoots. Bensulfuron‐methyl metabolism in shoots was similar in all biotypes. The R biotypes displayed a higher level of ALS activity than the S biotype, both in the presence and absence of herbicide and the resistance indices (IC₅₀R/S) were 20 197 and 10 for biotypes Q and T respectively. These data confirm for the first time that resistance to bensulfuron‐methyl in A. plantago‐aquatica is target‐site‐based. In practice, to control target site R biotypes, it would be preferable to use mixtures of ALS inhibitors with herbicides with other modes of action.