Status of black grass (Alopecurus myosuroides) resistance to acetyl-coenzyme A carboxylase inhibitors in France

We assessed the contributions of target site‐ and non‐target site‐based resistance to herbicides inhibiting acetyl‐coenzyme A carboxylase (ACC) in Alopecurus myosuroides (black grass). A total of 243 A. myosuroides populations collected across France were analysed using herbicide sensitivity bioassay (24 300 seedlings analysed) and ACC genotyping (13 188 seedlings analysed). Seedlings resistant to at least one ACC‐inhibiting herbicide were detected in 99.2% of the populations. Mutant, resistant ACC allele(s) were detected in 56.8% of the populations. Among the five resistant ACC alleles known in A. myosuroides, alleles containing an isoleucine‐to‐leucine substitution at codon 1781 were predominant (59.5% of the plants containing resistant ACC alleles). Comparison of the results from herbicide sensitivity bioassays with genotyping indicated that more than 75% of the plants resistant to ACC‐inhibiting herbicides in France would be resistant via increased herbicide metabolism. Analysis of herbicide application records suggested that in 15.9% of the populations studied, metabolism‐based resistance to ACC‐inhibiting herbicides was mostly selected for by herbicides with other modes of action. Our study revealed the importance of non‐target site‐based resistance in A. myosuroides. Using herbicides with alternative modes of action to control populations resistant to ACC‐inhibiting herbicides, the recommended management approach, may thus be jeopardised by the widespread occurrence of metabolism‐based resistance mechanisms conferring broad‐spectrum cross‐resistance.     

Simulating evolution of glyphosate resistance in Lolium rigidum I: population biology of a rare resistance trait

Despite frequent use for the past 25 years, resistance to glyphosate has evolved in few weed biotypes. The propensity for evolution of resistance is not the same for all herbicides, and glyphosate has a relatively low resistance risk. The reasons for these differences are not entirely understood. A previously published two‐herbicide resistance model has been modified to explore biological and management factors that account for observed rates of evolution of glyphosate resistance. Resistance to a post‐emergence herbicide was predicted to evolve more rapidly than it did to glyphosate, even when both were applied every year and had the same control efficacy. Glyphosate is applied earlier in the growing season when fewer weeds have emerged and hence exerts less selection pressure on populations. The evolution of glyphosate resistance was predicted to arise more rapidly when glyphosate applications were later in the growing season. In simulations that assumed resistance to the post‐emergence herbicide did not evolve, the evolution of glyphosate resistance was less rapid, because post‐emergence herbicides were effectively controlling rare glyphosate‐resistant individuals. On their own, these management‐related factors could not entirely account for rates of evolution of resistance to glyphosate observed in the field. In subsequent analyses, population genetic parameter values (initial allele frequency, dominance and fitness) were selected on the basis of empirical data from a glyphosate‐resistant Lolium rigidum population. Predicted rates of evolution of resistance were similar to those observed in the field. Together, the timing of glyphosate applications, the rarity of glyphosate‐resistant mutants, the incomplete dominance of glyphosate‐resistant alleles and pleiotropic fitness costs associated with glyphosate resistance, all contribute to its relatively slow evolution in the field.     

Glyphosate resistance in four different populations of Lolium rigidum is associated with reduced translocation of glyphosate to meristematic zones

Weed populations with resistance to glyphosate have evolved over the last 7 years, since the discovery of the first glyphosate‐resistant populations of Lolium rigidum in Australia. Four populations of L. rigidum from cropping, horticultural and viticultural areas in New South Wales and South Australia were tested for resistance to glyphosate by dose–response experiments. All populations required considerably more glyphosate to achieve 50% control compared with a known susceptible population, indicating they were resistant to glyphosate. Translocation of glyphosate within these resistant populations was examined by following the movement of radiolabelled glyphosate applied to a mature leaf. All resistant plants translocated significantly more herbicide to the tip of the treated leaf than did susceptible plants. Susceptible plants translocated twice as much herbicide to the stem meristematic portion of the plant compared with resistant plants. These different translocation patterns suggest an association between glyphosate resistance in L. rigidum and the ability of glyphosate to accumulate in the shoot meristem.     

EPSPS gene amplification in glyphosate-resistant Italian ryegrass (Lolium perenne ssp. multiflorum) from Arkansas

BACKGROUND: Resistance to glyphosate in weed species is a major challenge for the sustainability of glyphosate use in crop and non‐crop systems. A glyphosate‐resistant Italian ryegrass population has been identified in Arkansas. This research was conducted to elucidate its resistance mechanism. RESULTS: The investigation was conducted on resistant and susceptible plants from a population in Desha County, Arkansas (Des03). The amounts of glyphosate that caused 50% overall visual injury were 7 to 13 times greater than those for susceptible plants from the same population. The EPSPS gene did not contain any point mutation that has previously been associated with resistance to glyphosate, nor were there any other mutations on the EPSPS gene unique to the Des03 resistant plants. The resistant plants had 6‐fold higher basal EPSPS enzyme activities than the susceptible plants, but their I₅₀ values in response to glyphosate were similar. The resistant plants contained up to 25 more copies of EPSPS gene than the susceptible plants. The level of resistance to glyphosate correlated with increases in EPSPS enzyme activity and EPSPS copy number. CONCLUSION: Increased EPSPS gene amplification and EPSPS enzyme activity confer resistance to glyphosate in the Des03 population. This is the first report of EPSPS gene amplification in glyphosate‐resistant Italian ryegrass. Other resistance mechanism(s) may also be involved. Copyright © 2012 Society of Chemical Industry     

Intraregional and inter‐regional variability of herbicide sensitivity in common arable weed populations

The question on intraregional versus inter‐regional variability in herbicide sensitivity for weed populations is of major importance, both in extrapolation of model parameters and in herbicide zonal approval procedures. We hypothesised that inter‐regional variability in herbicide sensitivity for field populations would be the same as intraregional variability for regions with similar climatic conditions. Seeds of field weed populations were collected in a Danish, German and Polish region. Herbicide sensitivity was tested in dose–response experiments in the glasshouse with flufenacet and iodosulfuron (Apera spica‐venti), florasulam and tribenuron (Tripleurospermum inodorum), diflufenican, diflufenican + flurtamone and pendimethalin (Viola arvensis). ED₅₀ values and variance components of the ED₅₀ values were estimated to describe the influence of region, year and population. The regions accounted for a maximum of 26% of the variance and always less than the variance accounted for by individual populations. Sensitivity indices (SI₅₀) were calculated as the ratio between ED₅₀ of the test population and a reference population. There was considerable intraregional variability in SI₅₀ values and SI₅₀ values from a single region did not consistently differ from other regions. The large intraregional variability in herbicide sensitivity between populations, with no evidence of resistance, is of interest both for zonal evaluation of herbicides and resistance research. For practical weed management, we conclude that dose–response functions can be transferred between the study regions, for example for the common use in decision support systems with proper insurance for the control of less sensitive populations.     

A target-site mutation is present in a glyphosate-resistant Lolium rigidum population

Annual ryegrass (Lolium rigidum) is the only weed species to have evolved resistance to the broad‐spectrum herbicide glyphosate in Australia. A population that had failed to be controlled by glyphosate was collected from a vineyard in the Adelaide Hills region of South Australia. Dose–response experiments on this population (SLR 77) showed that it was glyphosate resistant, with an LD₅₀ that was 1.9–3.4 times higher than that of a susceptible population (VLR 1). The movement of radiolabelled glyphosate within SLR 77 plants showed that this population did not have the differential glyphosate translocation mechanism of resistance common to several other Australian glyphosate‐resistant populations. Subsequent analysis of shikimic acid accumulation within the plant after glyphosate treatment showed that this population accumulated significantly less shikimic acid than a susceptible population, but more than a glyphosate‐resistant population with the translocation mechanism, indicating the possible involvement of another mechanism of resistance. Sequencing of a portion of the SLR 77 5‐enolpyruvylshikimate‐3‐phosphate synthase gene was carried out and a mutation causing an amino acid change at position 106 from proline to threonine was identified. This mutation is likely to be responsible for glyphosate resistance in this population, as mutations in this position have been found to be responsible for glyphosate resistance in goosegrass (Eleusine indica) from Malaysia. This paper represents the first report of target‐site glyphosate resistance in L. rigidum and provides evidence that this species has at least two mechanisms of glyphosate resistance present in Australia.     

First glyphosate‐resistant Lolium spp. biotypes found in a European annual arable cropping system also affected by ACCase and ALS resistance

In Europe, glyphosate‐resistant weeds have so far only been reported in perennial crops. Following farmers' complaints of poor herbicide efficacy, resistance to glyphosate as well as to ACCase and ALS inhibitors was investigated in 11 populations of Lolium spp. collected from annual arable cropping systems in central Italy. Field histories highlighted that farmers had relied heavily on glyphosate, often at low rates, as well as in a non‐registered crop. The research aimed at elucidating the resistance status, including multiple resistance, of Lolium spp. populations through glasshouse screenings and an outdoor dose–response experiment. Target‐site resistance mechanism was also investigated for the substitutions already reported for EPSPs, ALS and ACCase genes. Three different resistant patterns were identified: glyphosate resistant only, multiple resistant to glyphosate and ACCase inhibitors and multiple resistant to glyphosate and ALS inhibitors. Amino acid substitutions were found at position 106 of the EPSPs gene, at position 1781, 2088 and 2096 of the ACCase gene and at position 197 and 574 of the ALS gene. Not all populations displayed amino acid substitutions, suggesting the presence of non‐target‐site‐mediated resistance mechanisms. After 39 years of commercial availability of glyphosate, this is the first report of multiple resistance involving glyphosate selected in annual arable crops in Europe. Management implications and options are discussed.     

The effect of temperature on glutathione S-transferase activity and glutathione content in Alopecurus myosuroides (black grass) biotypes susceptible and resistant to herbicides

Herbicide‐resistant populations of Alopecurus myosuroides (black grass) have become widespread throughout the UK since the early 1980s. Previous observations in this laboratory have demonstrated that natural climatic fluctuations caused increases in endogenous glutathione S‐transferase (GST) enzyme activity in A. myosuroides plants as they mature, which is thought to be linked to herbicide resistance in this species. The present study has investigated the effects of plant growth at 10°C and 25°C, and reports GST specific activity and glutathione (GSH) pool size in resistant and susceptible A. myosuroides biotypes. Findings demonstrate differences in GST activity between resistant and susceptible populations, which are transient at lower growth temperatures. The GSH pool size was elevated at lower growth temperature in both biotypes. We speculate that these endogenous responses are part of a natural mechanism of acclimation to environmental change in this species and suggest that resistant plants are more able to adapt to environmental stress, as indicated in this instance by temperature change. These observations imply that the control of resistant A. myosuroides by graminicides may be more effective when applied at lower temperatures and at earlier growth stages.     

Control by glyphosate and its alternatives of glyphosate‐susceptible and glyphosate‐resistant Echinochloa colona in the fallow phase of crop rotations in subtropical Australia

Echinochloa colona is the most common grass weed of summer fallows in the grain‐cropping systems of the subtropical region of Australia. Glyphosate is the most commonly used herbicide for summer grass control in fallows in this region. The world's first population of glyphosate‐resistant E. colona was confirmed in Australia in 2007 and, since then, >70 populations have been confirmed to be resistant in the subtropical region. The efficacy of alternative herbicides on glyphosate‐susceptible populations was evaluated in three field experiments and on both glyphosate‐susceptible and glyphosate‐resistant populations in two pot experiments. The treatments were knockdown and pre‐emergence herbicides that were applied as a single application (alone or in a mixture) or as part of a sequential application to weeds at different growth stages. Glyphosate at 720 g ai ha^?1 provided good control of small glyphosate‐susceptible plants (pre‐ to early tillering), but was not always effective on larger susceptible plants. Paraquat was effective and the most reliable when applied at 500 g ai ha^?1 on small plants, irrespective of the glyphosate resistance status. The sequential application of glyphosate followed by paraquat provided 96–100% control across all experiments, irrespective of the growth stage, and the addition of metolachlor and metolachlor + atrazine to glyphosate or paraquat significantly reduced subsequent emergence. Herbicide treatments have been identified that provide excellent control of small E. colona plants, irrespective of their glyphosate resistance status. These tactics of knockdown herbicides, sequential applications and pre‐emergence herbicides should be incorporated into an integrated weed management strategy in order to greatly improve E. colona control, reduce seed production by the sprayed survivors and to minimize the risk of the further development of glyphosate resistance.     

An approach to investigate the costs of herbicide‐resistant Alopecurus myosuroides

Herbicide resistance in Alopecurus myosuroides causes severe problems in Western European cropping systems. Costs of herbicide resistance were investigated in this study by analysing variable production costs and sales revenues. Three farms were selected for this study, with winter wheat as the dominating crop in all farms. Resistance in A. myosuroides populations was verified at all locations. Four farming approaches were simulated over a period of 20 years: (i) continuing the actual cropping system without increase of resistance, (ii) continuing the actual cropping system with increase of resistance, (iii) changing cropping practice to overcome resistance and (iv) changing cropping practice to prevent resistance. Contribution margins representing the proportion of sales revenue that is not consumed by variable costs were calculated for all approaches. Comparative static simulations showed that average contribution margins in a cropping system with more than 60% winter cereals and reduced tillage practice dropped from 807 € ha^?1 a^?1 without herbicide resistance to 307 € ha^?1 a^?1 with herbicide resistance. Alopecurus myosuroides population densities increased to more than 1000 plants m^?2. Diverse crop rotations, including spring crops, clover–grass leys and intensive tillage, suppressed A. myosuroides populations, and average contribution margin was 630 € ha^?1 a^?1. Preventive methods with rotations of winter cereals and spring crops with less clover–grass leys resulted in an average contribution margin of 691 € ha^?1 a^?1. In conclusion, rotations of winter cereals and spring crops combined with inversion tillage and herbicides provide stable yields and can prevent weed population increase.     

Chlorophyll fluorescence imaging: a new method for rapid detection of herbicide resistance in Alopecurus myosuroides

Due to the steadily increasing number of putative herbicide‐resistant weed populations, the demand for rapid in‐season tests is rising. In this study, we introduce a new quantitative herbicide‐resistance test system based on chlorophyll fluorescence imaging analysis of photosynthesis‐related parameters. Susceptible and herbicide‐resistant populations of Alopecurus myosuroides (black‐grass) were cultivated in multiwell tissue culture plates containing nutrient agar and different dosages of fenoxaprop‐P‐ethyl and mesosulfuron+iodosulfuron. The maximum quantum efficiency of the PSII was measured 3 h after transplanting (HAT) and then for seven days every 24 h. Data of maximum quantum efficiency of the PSII were compared with standard whole‐plant pot tests and molecular tests for target‐site mutations. It was possible to fit dose‐response curves and calculate corresponding resistance factors for ED90 for all populations tested using the chlorophyll fluorescence imaging. It was possible to distinguish between resistant and susceptible populations. The results of the chlorophyll fluorescence imaging corresponded well with the standard whole‐plant pot tests in the glasshouse. However, populations with proved target‐site mutations did not differ from other herbicide‐resistant populations in the maximum quantum efficiency values of the PSII. We conclude that the chlorophyll fluorescence imaging provides reliable data on herbicide resistance for both modes of action tested in a shorter time and using less space, compared with standard whole‐plant pot tests in the glasshouse.     

Multiple herbicide‐resistant Lolium rigidum (annual ryegrass) now dominates across the Western Australian grain belt

Lolium rigidum (annual or rigid ryegrass) is a widespread annual weed in cropping systems of southern Australia, and herbicide resistance in L. rigidum is a common problem in this region. In 2010, a random survey was conducted across the grain belt of Western Australia to determine the frequency of herbicide‐resistant L. rigidum populations and to compare this with the results of previous surveys in 1998 and 2003. During the survey, 466 cropping fields were visited, with a total of 362 L. rigidum populations collected. Screening of these populations with the herbicides commonly used for control of L. rigidum revealed that resistance to the ACCase‐ and ALS‐inhibiting herbicides was common, with 96% of populations having plants resistant to the ACCase herbicide diclofop‐methyl and 98% having plants resistant to the ALS herbicide sulfometuron. Resistance to another ACCase herbicide, clethodim, is increasing, with 65% of populations now containing resistant plants. Resistance to other herbicide modes of action was significantly lower, with 27% of populations containing plants with resistance to the pre‐emergent herbicide trifluralin, and glyphosate, atrazine and paraquat providing good control of most of the populations screened in this survey. Ninety five per cent of L. rigidum populations contained plants with resistance to at least two herbicide modes of action. These results demonstrate that resistance levels have increased dramatically for the ACCase‐ and ALS‐inhibiting herbicides since the last survey in 2003 (>95% vs. 70–90%); therefore, the use of a wide range of integrated weed management options are required to sustain these cropping systems in the future.     

Identification of resistance to either paraquat or ALS-inhibiting herbicides in two Western Australian Hordeum leporinum biotypes

BACKGROUND: Hordeum populations are becoming increasingly difficult to control in cropping fields. Two herbicide‐resistant H. leporinum populations were identified during a random crop survey after herbicides were applied. The study aimed to determine the herbicide resistance profile of these H. leporinum biotypes to a range of herbicides used for their control. RESULTS: Based on dose–response studies, one H. leporinum population was very highly resistant to sulfosulfuron and sulfometuron (both sulfonylurea herbicides) and also displayed low‐level resistance to imazamox (an imidazolinone herbicide). Reduced sensitivity of the ALS enzyme was identified with in vitro activity assays. Gene sequence analysis revealed a proline‐to‐threonine substitution at amino acid position 197 of ALS, which is likely to be the molecular basis for resistance in this population. Herbicide screening also revealed a different H. leporinum population with resistance to the bipyridyl herbicide paraquat. CONCLUSION: This study established the first cases of (1) sulfonylurea‐to‐imidazolinone cross‐resistance and (2) field‐evolved paraquat resistance in a Hordeum species in Western Australia. Copyright © 2012 Society of Chemical Industry     

Frequency and distribution of herbicide resistance in Raphanus raphanistrum populations randomly collected across the Western Australian wheatbelt

In 2003, a random survey was conducted across the Western Australian wheatbelt to establish the extent and frequency of herbicide resistance in Raphanus raphanistrum populations infesting crop fields. Five hundred cropping fields were visited, with 90 R. raphanistrum populations collected, representative of populations present in crop fields throughout the Western Australian wheatbelt. Collected populations were screened with four herbicides of various modes of action that are commonly used for the control of this weed. The majority of Western Australian R. raphanistrum populations were found to contain plants resistant to the acetolactate synthase (ALS)‐inhibiting herbicide chlorsulfuron (54%) and auxin analogue herbicide, 2,4‐D amine (60%). This survey also determined that over half (58%) of these populations were multiple resistant across at least two of the four herbicide modes of action used in the screening. Only 17% of R. raphanistrum populations have retained their initial status of susceptibility to all four herbicides. The distribution patterns of the herbicide‐resistant populations identified that there were higher frequencies of resistant and developing resistance populations occurring in the intensively cropped northern regions of the wheatbelt. These results clearly indicate that the reliance on herbicidal weed control in cropping systems based on reduced tillage and stubble retention will lead to higher frequencies of herbicide‐resistant weed populations. Therefore, within intensive crop production systems, there is a need to diversify weed management strategies and not rely entirely on too few herbicide control options.     

Reduced translocation is involved in resistance to glyphosate and paraquat in Conyza bonariensis and Conyza canadensis from California

Resistance to glyphosate and paraquat has evolved in some populations of Conyza spp. from California, USA. This study evaluated whether herbicide absorption and translocation were involved in the mechanism of resistance to both herbicides. Three lines of each species were used: glyphosate‐paraquat‐susceptible (GPS), glyphosate‐resistant (GR) and glyphosate‐paraquat‐resistant (GPR). Radiolabelled herbicide was applied to a fully expanded leaf, and absorption and movement out of the treated leaf were monitored for up to 24 h for paraquat and 72 h for glyphosate. Plants treated with paraquat were incubated in darkness for the first 16 h and then subjected to light conditions. More glyphosate was absorbed in C. bonariensis (52.9–58.3%) compared with C. canadensis (28.5–37.6%), but no differences in absorption were observed among lines within a species. However, in both species, the GR and GPR lines translocated less glyphosate out of the treated leaf when compared with their respective GPS lines. Paraquat absorption was similar among lines and across species (71.3–77.6%). Only a fraction of paraquat was translocated in the GPR lines (3% or less) when compared with their respective GPS or GR lines (20% or more) in both species. Taken together, these results indicate that reduced translocation is involved in the mechanism of resistance to glyphosate and paraquat in C. bonariensis and C. canadensis.     

Identification of a target‐site mutation conferring resistance to triazine herbicides in oriental mustard (Sisymbrium orientale L.) from Australia

In southern Australia, oriental mustard (Sisymbrium orientale) has been controlled successfully by triazine herbicides for several decades. The screening of 40 populations that were collected from the southern grain belt of Australia during 2010 and 2013 for resistance to six different herbicides (glyphosate, diflufenican, imazamox, chlorsulfuron, atrazine and 2,4‐dichlorophenoxyacetic acid) identified two oriental mustard populations as highly resistant to atrazine. Compared to the known oriental mustard‐susceptible populations (S1 and S2), these two resistant populations (P17 and P18) from near Horsham, Victoria, Australia, were 311‐ and 315‐fold resistant to atrazine, as determined by a comparison of the LD₅₀ values. However, there was no resistance to diuron detected in these populations. Sequencing of the chloroplast psbA gene identified a missense mutation of serine 264 to glycine in both herbicide‐resistant oriental mustard populations, which is known to confer high‐level atrazine resistance in other species.     

Glyphosate-resistant horseweed made sensitive to glyphosate: low-temperature suppression of glyphosate vacuolar sequestration revealed by 31P NMR

BACKGROUND: Horseweed has been the most invasive glyphosate‐resistant (GR) weed, spreading to 16 states in the United States and found on five continents. The authors have previously reported that GR horseweed employs rapid vacuolar sequestration of glyphosate, presumably via a tonoplast transporter, substantively to reduce cytosolic glyphosate concentrations. 1 It was hypothesized that glyphosate sequestration was the herbicide resistance mechanism. If resistance is indeed endowed by glyphosate sequestration, suppression of sequestration offers the potential for controlling GR horseweed at normal herbicide field‐use rates. RESULTS: Low‐temperature ³¹P NMR experiments performed in vivo with GR cold‐acclimated horseweed showed markedly suppressed vacuolar accumulation of glyphosate even 3 days after glyphosate treatment. [In stark contrast, 85% of the visible glyphosate was sequestered 24 h after spraying warm‐acclimated GR horseweed.] Cold‐acclimated GR horseweed treated at normal use rates and maintained at low temperature succumbed to the lethal effects of glyphosate over a 40 day period. Treatment of GR horseweed in the field when temperatures were cooler showed the predicted positive herbicidal response. CONCLUSIONS: Low temperature markedly diminishes vacuolar sequestration of glyphosate in the GR horseweed biotype, yielding a herbicide response equivalent to that of the sensitive biotype. This supports the recent hypothesis 1 that glyphosate sequestration is the resistance mechanism employed by GR horseweed. Copyright © 2011 Society of Chemical Industry     

Are herbicide‐resistant crops the answer to controlling Cuscuta?

BACKGROUND: Herbicide‐resistant crop technology could provide new management strategies for the control of parasitic plants. Three herbicide‐resistant oilseed rape (Brassica napus L.) genotypes were used to examine the response of attached Cuscuta campestris Yuncker to glyphosate, imazamox and glufosinate. Cuscata campestris was allowed to establish on all oilseed rape genotypes before herbicides were applied. RESULTS: Unattached seedlings of C. campestris, C. subinclusa Durand & Hilg. and C. gronovii Willd. were resistant to imazamox and glyphosate and sensitive to glufosinate, indicating that resistance initially discovered in C. campestris is universal to all Cuscuta species. Glufosinate applied to C. campestris attached to glufosinate‐resistant oilseed rape had little impact on the parasite, while imazamox completely inhibited C. campestris growth on the imidazolinone‐resistant host. The growth of C. campestris on glyphosate‐resistant host was initially inhibited by glyphosate, but the parasite recovered and resumed growth within 3–4 weeks. CONCLUSION: The ability of C. campestris to recover was related to the quality of interaction between the host and parasite and to the resistance mechanism of the host. The parasite was less likely to recover when it had low compatibility with the host, indicating that parasite‐resistant crops coupled with herbicide resistance could be highly effective in controlling Cuscuta. Published 2009 by John Wiley & Sons, Ltd.