A novel mutation in IAA16 is associated with dicamba resistance in Chenopodium album.

BACKGROUND: Resistance to dicamba in Chenopodium album was first documented over a decade ago, however, the molecular basis of dicamba resistance in this species has not been elucidated. In this research, the resistance mechanism in a dicamba-resistant C. album phenotype was investigated using a transcriptomics (RNA-sequence) approach. RESULTS: The dose-response assay showed that the resistant (R) phenotype was nearly 25-fold more resistant to dicamba than a susceptible (S) phenotype of C. album. Also, dicamba treatment significantly induced transcription of the known auxin-responsive genes, Gretchen Hagen 3 (GH3), small auxin-up RNAs (SAURs), and 1-aminocyclopropane-1-carboxylate synthase (ACS) genes in the susceptible phenotype. Comparing the transcripts of auxin TIR/AFB receptors and auxin/indole-3-acetic acid (AUX/IAA) proteins identified from C. album transcriptomic analysis revealed that the R phenotype contained a novel mutation at the first codon of the GWPPV degron motif of IAA16, resulting in an amino acid substitution of glycine (G) with aspartic acid (D). Sequencing the IAA16 gene in other R and S individuals further confirmed that all the R individuals contained the mutation. CONCLUSION: In this research, we describe the dicamba resistance mechanism in the only case of dicamba-resistant C. album reported to date. Prior work has shown that the dicamba resistance allele confers significant growth defects to the R phenotype investigated here, suggesting that dicamba-resistant C. album carrying this novel mutation in the IAA16 gene may not persist at high frequencies upon removal of dicamba application. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Inheritance of dicamba-resistance in allotetraploid Chenopodium album.

BACKGROUND: Chenopodium album L. is a troublesome weed in spring-planted crops, and different levels of ploidy have been documented for this weed species. A population of C. album has evolved resistance to dicamba. The level of ploidy and inheritance of dicamba resistance was studied in this population. RESULTS: The resistant and susceptible individuals of C. album were confirmed as tetraploid by flow cytometry. Pair-crosses were made between ten resistant and susceptible individuals. Eight F1 individuals from five crosses were confirmed resistant after treating with dicamba at 400 g a.e. ha-1 . These individuals were selfed, and the response of their progenies to dicamba was assessed in dose-response experiments, and the results confirmed the resistance trait was dominant. Furthermore, an analysis of the segregation patterns revealed that the segregation response of all F2 progenies fitted a 3:1 (resistant/susceptible) ratio when treated with dicamba at 200, 400 and 800 g a.e. ha-1 , suggesting a single gene was responsible for dicamba resistance. CONCLUSIONS: Dicamba resistance in the studied tetraploid population of C. album is governed by a single dominant gene. This type of inheritance suggests that selection for dicamba resistance can occur readily. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Pinoxaden resistance in Lolium perenne L. is due to both target-site and non-target-site mechanisms.

Application of herbicides inhibiting acetyl CoA carboxylase (ACCase) has been one of the main strategies for selectively controlling grass weed species such as perennial ryegrass (Lolium perenne L.) in wheat and barley crops in New Zealand. In this study, we have confirmed and characterized resistance to pinoxaden, an ACCase-inhibiting herbicide, in a population of L. perenne. Dose-response experiments were conducted to assess the level of pinoxaden resistance, and based on the LD50 values, the studied population was 41.4-times more resistant to pinoxaden than a susceptible population. Application of malathion, an inhibitor of the cytochrome P450s, preceding pinoxaden treatment reduced the level of resistance to 9.7-fold. However, pre-treatment with the glutathione S-transferase (GST) inhibitor 4-chloro7- nitrobenzoxadiazole prior to pinoxaden treatment did not affect pinoxaden resistance. Partial sequencing of the ACCase gene revealed that the resistant population had an isoleucine to valine replacement at position 2041. These results suggest that both cytochrome P450-based and target-site mechanisms are jointly associated with this instance of pinoxaden resistance in L. perenne. The pinoxaden-resistant L. perenne individuals were also resistant to quizalofop-p-ethyl (108.6-fold), but they were susceptible to clethodim, which can, therefore, be used to manage this pinoxaden-resistant L. perenne. This is the first report of a L. perenne population in which a rare target-site mutation works in concert with enhanced cytochrome P-450 activity to confer pinoxaden resistance. Evolution of resistance to ACCase-inhibiting herbicides in this L. perenne population indicates that integrated weed management practices are required to prevent widespread resistance developing in New Zealand cereal crop systems.

The leaching behaviour of herbicides in cropping soils amended with forestry biowastes.

Leaching of herbicides in cropping soils not only impacts the groundwater sources but also reduces their effect in controlling weeds. Leaching studies were carried out in two cropping soils and two forestry biowaste media, wood pulp and sawdust with two herbicides, atrazine and bromacil in a packed lysimeter with simulated rainfall. The hypothesis was that high organic matter forestry biowaste soil amendments reduce the leaching of herbicides through the soil profile. Results from the experimental setups varied due to the impact of the simulated rainfall on the surface structure of the media. Organic carbon content, pH and structure of the media were all factors which affected the leaching of the two herbicides. The hypothesis was true for wood pulp, but for sawdust, organic matter content had less bearing on the leaching of the herbicides than other over-riding factors, such as pH, that were media specific. In sawdust, its large particle size and related pore volume allowed preferential flow of herbicides. Overall, the data indicated that both forestry biowastes were retentive to herbicide leaching, but the effect was more pronounced with wood pulp than sawdust.

Resistance to post-emergent herbicides is becoming common for grass weeds on New Zealand wheat and barley farms.

To estimate the prevalence of herbicide-resistant weeds, 87 wheat and barley farms were randomly surveyed in the Canterbury region of New Zealand. Over 600 weed seed samples from up to 10 mother plants per taxon depending on abundance, were collected immediately prior to harvest (two fields per farm). Some samples provided by agronomists were tested on an ad-hoc basis. Over 40,000 seedlings were grown to the 2-4 leaf stage in glasshouse conditions and sprayed with high priority herbicides for grasses from the three modes-of-action acetyl-CoA carboxylase (ACCase)-inhibitors haloxyfop, fenoxaprop, clodinafop, pinoxaden, clethodim, acetolactate synthase (ALS)-inhibitors iodosulfuron, pyroxsulam, nicosulfuron, and the 5-enolpyruvyl shikimate 3-phosphate synthase (EPSPS)-inhibitor glyphosate. The highest manufacturer recommended label rates were applied for the products registered for use in New Zealand, often higher than the discriminatory rates used in studies elsewhere. Published studies of resistance were rare in New Zealand but we found weeds survived herbicide applications on 42 of the 87 (48%) randomly surveyed farms, while susceptible reference populations died. Resistance was found for ALS-inhibitors on 35 farms (40%) and to ACCase-inhibitors on 20 (23%) farms. The number of farms with resistant weeds (denominator is 87 farms) are reported for ACCase-inhibitors, ALS-inhibitors, and glyphosate respectively as: Avena fatua (9%, 1%, 0% of farms), Bromus catharticus (0%, 2%, 0%), Lolium spp. (17%, 28%, 0%), Phalaris minor (1%, 6%, 0%), and Vulpia bromoides (0%, not tested, 0%). Not all farms had the weeds present, five had no obvious weeds prior to harvest. This survey revealed New Zealand's first documented cases of resistance in P. minor (fenoxaprop, clodinafop, iodosulfuron) and B. catharticus (pyroxsulam). Twelve of the 87 randomly sampled farms (14%) had ALS-inhibitor chlorsulfuron-resistant sow thistles, mostly Sonchus asper but also S. oleraceus. Resistance was confirmed in industry-supplied samples of the grasses Digitaria sanguinalis (nicosulfuron, two maize farms), P. minor (iodosulfuron, one farm), and Lolium spp. (cases included glyphosate, haloxyfop, pinoxaden, iodosulfuron, and pyroxsulam, 9 farms). Industry also supplied Stellaria media samples that were resistant to chlorsulfuron and flumetsulam (ALS-inhibitors) sourced from clover and ryegrass fields from the North and South Island.

Characterization of clopyralid resistance in lawn burweed (Soliva sessilis).

Soliva sessilis is a troublesome annual weed species in New Zealand turfgrass. This weed has been controlled selectively in New Zealand turfgrass for many years using pyridine herbicides such as clopyralid. However, in some golf courses, the continuous application of pyridine herbicides has resulted in the selection of S. sessilis populations that are resistant to these herbicides. This study focuses on a clopyralid-resistant population of S. sessilis collected from a golf course with a long history of clopyralid applications. The resistant phenotype of S. sessilis was highly resistant to clopyralid (over 225-fold). It was also cross-resistant to dicamba, MCPA and picloram but not mecoprop. The level of resistance to dicamba was high (7-14-fold) but much lower (2-3-fold) for both MCPA and picloram. The phenotype was morphologically distinct from its susceptible counterpart. Individuals of the clopyralid-resistant phenotype had fewer lobes on their leaves and were slightly larger compared to the susceptible phenotype. Resistant individuals also had a larger leaf area and greater root dry weight than the susceptible plants. An evaluation of internal transcribed spacer (ITS) regions confirmed that clopyralid-resistant phenotypes are conspecific with S. sessilis. In summary, the cross-resistance to several auxinic herbicides in this S. sessilis phenotype greatly reduces chemical options for controlling it; thus, other integrated management practices may be needed such as using turfgrass competition to reduce weed germination. However, the morphological differences between resistant and susceptible plants make it easy to see, which will help with its management.

A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne.

The first step in managing herbicide-resistant weeds is to confirm their resistance status. It is, therefore, crucial to have a rapid, reliable and cost-effective technique to assess samples for herbicide resistance. We designed and evaluated three derived cleaved amplified polymorphic sequence (dCAPS) markers for detecting glyphosate resistance in Lolium perenne. conferred by non-synonymous mutations at codon-106 in the enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene. The dCAPS markers involve amplification of the target region, digestion of the amplified products with restriction enzymes and gel-based visualisation of the digested products. The results showed that all three dCAPS markers could successfully detect mutations at codon-106 in the target enzyme. The dCAPS markers can also inform us of the zygosity state of the resistance allele and was confirmed by sequencing the target region of the EPSPS gene. The markers described here are effective quick tests for the monitoring and evaluation of the target-enzyme mechanism of glyphosate resistance in Lolium perenne.

Degradation of atrazine and bromacil in two forestry waste products.

The persistence and degradation of two common herbicides, atrazine and bromacil in two organic media, wood pulp and sawdust were compared with two soils. The hypothesis tested was that herbicide degradation will be faster in high organic matter media compared to soil. Degradation of two herbicides was carried out in four different temperature regimes and in sterilised media. The degradation half-life (t½) was determined under above-mentioned conditions then compared to degradation in soil. The degradation as quantified by t½ of the herbicides was generally longer in both organic media. Although microbial degradation was an important factor in the mineralisation of these herbicides, overall, the pH of the media had a more profound effect on the desorption and subsequent degradation rate than the organic carbon content. The results of this study revealed that the hypothesis was only partially correct as organic matter content per se did not strongly relate to degradation rates which were mainly governed by pH and microbial activity.

Perspective: root exudation of herbicides as a novel mode of herbicide resistance in weeds.

The evolution of resistance to herbicides in weeds has become a great challenge for global agricultural production. Weeds have evolved resistance to herbicides through many different physiological mechanisms. Some weed species are known to secrete herbicide molecules from roots into the rhizosphere upon being treated. However, root exudation of herbicides as a mechanism of resistance has only recently been identified in two weed species. Root exudation pathways have been investigated in Arabidopsis, and this work suggested that ATP-binding cassette (ABC) and multidrug and toxic compound extrusion (MATE) transporters play a role in the secretion of primary and secondary plant products from roots. We hypothesize that the mechanisms involved in root exudation of herbicides that result in resistance are mediated by overactive or overexpressed transporters, probably similar to those found for the exudation of primary and secondary compounds from roots. Elucidating the molecular and physiological basis of root exudation in herbicide-resistant weeds would improve our understanding of the pathways involved in herbicide root secretion mediated by transporters in plants. © 2020 Society of Chemical Industry.

Effect on herbicide adsorption of organic forestry waste products used for soil remediation.

Organic soil amendments can be useful for improving degraded soil, but this increase in organic matter (OM) may influence adsorption of herbicides subsequently applied to the treated soil, even though the particle size of amendments and their nature differ from typical soil OM. In this study, a batch equilibrium method was used to measure adsorption of five herbicides following application to two organic media, wood pulp and sawdust, comparing these with two cropping soils. Herbicide adsorption, quantified by distribution coefficients (kd), was much higher in the two organic media than in the cropping soils. The increases in adsorption were strongly correlated to the percentage of organic carbon. When the kd was normalized to adsorption coefficients corrected for OM content (koc), variation in results between the media was greatly reduced, indicating that OM is an important factor influencing adsorption in these media. The results of this study suggest that herbicides will be less effective when applied to soils in which sawdust and wood pulp have been added. Using organic amendments to remediate soil will increase adsorption of pesticides, reducing their bio-availability and efficacy, but also reducing their tendency to leach into root zones of deep-rooted crops and into groundwater.

Fitness costs associated with multiple resistance to dicamba and atrazine in Chenopodium album.

MAIN CONCLUSION: Detrimental pleiotropic effects of resistance mutation(s) were observed for multiple-resistant phenotypes (resistant to both atrazine and dicamba). The multiple-resistant phenotypes had lower growth rates and less capacity for vegetative growth compared to the phenotypes only resistant to atrazine. The fitness costs that are conferred by herbicide resistance alleles can affect the rate of herbicide resistance evolution within populations. We evaluated the direct fitness costs involved with multiple resistance to dicamba and atrazine (R1 and R2) in Chenopodium album by comparing the performance of multiple-resistant phenotypes to those phenotypes that were only resistant to atrazine (S1 and S2). The R1 and R2 phenotypes were consistently shorter and produced less dry matter than the S1 and S2 phenotypes. The R1 and R2 phenotypes were shown to have lower relative growth rates (RGR) and net assimilation rates (NAR) than the S1 and S2 phenotypes at an early stage of growth. However, there was no significant difference in RGR between the R1 and R2 and, S1 and S2 phenotypes at a later stage of growth, though the R1 and R2 phenotypes still had a lower NAR at this later stage. Further investigations using a neighbouring crop competition approach showed that the R1 and R2 phenotypes were weaker competitors, and exhibited significantly less capacity for vegetative growth compared to the S1 and S2 phenotypes during competition. Overall, the results of this study revealed multiple- resistance to atrazine and dicamba endowed a significant fitness penalty to C. album, and it is possible that the frequency of multiple-resistant individuals would gradually decline once selection pressure from herbicides was discontinued.

Perspectives on non-target site mechanisms of herbicide resistance in weedy plant species using evolutionary physiology.

Evolutionary physiology merges the disciplines of evolution and physiology, and it is a research approach that has not received much attention for studying the development of herbicide resistance. This paper makes a case for using evolutionary physiology more frequently when studying herbicide resistance, and illustrates this using three areas where more work would be useful: (i) the interaction among major and minor alleles over many generations during the evolution of physiological responses that lead to specific mechanisms of resistance; (ii) the role of epigenetic factors, especially at an early stage of evolution, on the physiological modifications that result in phenotypes that become insensitive to herbicides; and (iii) the interaction between fitness and physiological performance over time, with emphasis on understanding mechanisms that improve the fitness of herbicide-resistant phenotypes during selection.

Mechanisms of glyphosate resistance in two perennial ryegrass (Lolium perenne) populations.

BACKGROUND: Perennial ryegrass (Lolium perenne) has developed resistance to glyphosate within New Zealand vineyards following many years of herbicide application. The objectives of this work were to confirm resistance within two populations obtained from affected vineyards, and to determine the mechanism of resistance to glyphosate. RESULTS: Population O was confirmed to have a 25-fold resistance to glyphosate, whereas population J had a sevenfold resistance. Results of genotyping assays demonstrated a single nucleotide substitution at codon 106 of 5-enolpyruvylshikimate-3-phosphate synthase in population O but not population J. Glyphosate-resistant and glyphosate-susceptible populations did not differ in glyphosate absorption. However, in both resistant populations, much more of the absorbed (14) C-glyphosate remained in the treated leaf than occurred in the susceptible population. Significantly more glyphosate was found in the pseudostem region of susceptible plants compared with resistant plants. CONCLUSION: Both target-site and non-target-site mechanisms of glyphosate resistance were found in the perennial ryegrass population with 25-fold resistance, whereas only the non-target-site mechanism of resistance was found in the population with sevenfold resistance. This is the first study of the mechanism of glyphosate resistance in perennial ryegrass.