Segregation of non‐target‐site resistance to herbicides in multiple‐resistant Alopecurus myosuroides plants

Non‐target‐site resistance (NTSR) comprises a set of mechanisms conferring resistance to multiple modes of action. Investigation of the number of loci involved in NTSR will aid in the understanding of these resistance mechanisms. Therefore, six different multiple herbicide‐resistant Alopecurus myosuroides plants with different herbicide history were crossed in two generations with a susceptible wild type. Seeds from the backcrossing generation were studied for their segregation rate for resistance to five herbicides with four different modes of action (HRAC groups C₂, A, B and K₃). Taking into account that NTSR is a set of quantitative traits, the numbers of loci controlling NTSR were estimated using a normal mixture model fitted by the NLMIXED procedure of SAS. Each herbicide was controlled by a different number of loci comparing the six plants. In most of the cases, chlorotoluron resistance was controlled by one locus, whereas resistance to fenoxaprop‐P‐ethyl needed one or two loci. Resistance to pinoxaden was in all plants conferred by two loci. Cross‐resistance of fenoxaprop‐P‐ethyl and pinoxaden was found in all backcrossings, indicating that at least one of the two loci is responsible for both resistances. Resistance to mesosulfuron + iodosulfuron was conferred by a minimum of two loci. Results indicated that a minimum of five different loci can be involved in a multiple NTSR plant. Furthermore, the plant‐specific accumulation of NTSR loci was demonstrated. Such behaviour should be taken into account when evaluating the development and further spread of herbicide resistance.     

Non‐target site resistance to flucarbazone,imazamethabenz and pinoxaden is controlled by three linked genes in Avena fatua

Extensive herbicide usage has led to the evolution of resistant weed populations that cause substantial crop yield losses and increase production costs. The multiple herbicide‐resistant (MHR) Avena fatua populations utilised in this study are resistant to members of all selective herbicide families, across five modes of action, available for A. fatua control in US small grain production, and thus pose significant agronomic and economic threats. Resistance to acetolactate synthase and acetyl‐CoA carboxylase inhibitors is not conferred by known target site mutations, indicating that non‐target site resistance (NTSR) mechanisms are involved. Understanding the inheritance of NTS MHR is of upmost importance for continued agricultural productivity in the face of the rapid increase in resistant weed populations worldwide. As few studies have examined the inheritance of NTSR in autogamous weeds, we investigated the inheritance and genetic control of NTSR in the highly autogamous, allohexaploid species A. fatua. We found that NTSR in MHRA. fatua is controlled by three separate, closely‐linked nuclear genes for flucarbazone‐sodium, imazamethabenz‐methyl and pinoxaden. The single‐gene NTSR inheritance patterns reported here contrast with other examples in allogamous species and illustrate the diversity of evolutionary responses to strong selection.     

Evolving understanding of the evolution of herbicide resistance

A greater number of, and more varied, modes of resistance have evolved in weeds than in other pests because the usage of herbicides is far more extensive than the usage of other pesticides, and because weed seed output is so great. The discovery and development of selective herbicides are more problematic than those of insecticides and fungicides, as these must only differentiate between plant and insect or pathogen. Herbicides are typically selective between plants, meaning that before deployment there are already some crops possessing natural herbicide resistance that weeds could evolve. The concepts of the evolution of resistance and the mechanisms of delaying resistance have evolved as nature has continually evolved new types of resistance. Major gene target‐site mutations were the first types to evolve, with initial consideration devoted mainly to them, but slowly ‘creeping’ resistance, gradually accruing increasing levels of resistance, has become a major force owing to an incremental accumulation of genetic changes in weed populations. Weeds have evolved mechanisms unknown even in antibiotic as well as other drug and pesticide resistances. It is even possible that cases of epigenetic ‘remembered’ resistances may have appeared. Copyright © 2009 Society of Chemical Industry     

The role of population and quantitative genetics and modern sequencing technologies to understand evolved herbicide resistance and weed fitness

Evolution of resistance to multiple herbicides with different sites of action and of nontarget site resistance (NTSR) often involves multiple genes. Thus, single‐gene analyses, typical in studies of target site resistance, are not sufficient for understanding the genetic architecture and dynamics of NTSR and multiple resistance. The genetics of weed adaptation to varied agricultural environments is also generally expected to be polygenic. Recent advances in whole‐genome sequencing as well as bioinformatic and statistical tools have made it possible to use population and quantitative genetics methods to expand our understanding of how resistance and other traits important for weed adaptation are genetically controlled at the individual and population levels, and to predict responses to selection pressure by herbicides and other environmental factors. The use of tools such as quantitative trait loci mapping, genome‐wide association studies, and genomic prediction will allow pest management scientists to better explain how pests adapt to control tools and how specific genotypes thrive and spread across agroecosystems and other human‐disturbed systems. The challenge will be to use this knowledge in developing integrated weed management systems that inhibit broad resistance to current and future weed‐control methods. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

A quantitative genetic examination of non‐target‐site resistance applied to Avena species

Quantitative genetics tools can be used to assess whether using herbicides at low doses drive selection on standing genetic variation in populations leading to non‐target‐site resistance (NTSR). These tools are particularly important for estimating the number of genes involved and the potential speed of evolution. A short cut to answering questions about the evolution of NTSR may be to measure heritability. The heritability index (H) provides a measure of the potential to develop NTSR and can be simply calculated from classical dose–response experiments. This measure and the associated experimental designs are discussed with two applied examples on Avena spp. (A. fatua and A. sterilis). In these examples, H values ranged from 0.24 to 0.73, which means that selection for NTSR is highly probable in cases with high H value. We suggest that structuring plants into genetic groups (e.g. families or populations) can contribute to our understanding of the evolutionary potential of populations and plant species to evolve resistance, without increasing experimental cost and time.     

Prevalence of herbicide‐resistant weed species in Malaysian rice fields: A review

The management of weeds in Malaysian rice fields is very much herbicide‐based. The heavy reliance on herbicide for weed control by many rice‐growers arguably eventually has led to the development and evolution of herbicide‐resistant biotypes in Malaysian rice fields over the years. The continuous use of synthetic auxin (phenoxy group) herbicides and acetohydroxyacid synthase‐inhibiting herbicides to control rice weeds was consequential in leading to the emergence and prevalence of resistant weed biotypes. This review discusses the history and confirmed cases and incidence of herbicide‐resistant weeds in Malaysian rice fields. It also reviews the Clearfield Production System and its impact on the evolution of herbicide resistance among rice weed species and biotypes. This review also emphasizes the strategies and management options for herbicide‐resistant rice field weeds within the framework of herbicide‐based integrated weed management. These include the use of optimum tillage practices, certified clean seeds, increased crop competition through high seeding rates, crop rotation, the application of multiple modes of action of herbicides in annual rotations, tank mixtures and sequential applications to enable a broad spectrum of weed control, increase the selective control of noxious weed species in a field and help to delay the resistance evolution by reducing the selection pressure that is forced on those weed populations by a specific herbicidal mode of action.     

杂草对9类常用不同作用机制除草剂的非靶标抗性机制研究进展

除草剂的应用为农业生产带来便利, 但长期、单一使用某一种或相同机制的除草剂也引发了杂草对除草剂的抗性问题。抗性杂草种类逐渐增加, 抗性形成机制复杂, 导致农田杂草的治理难度增加。杂草对除草剂的抗性机制主要分为两种, 一种是除草剂靶标位点基因的突变或过量表达导致的靶标抗性, 另一种是杂草对除草剂吸收、转运、固存和代谢等一个或多个生理过程发生变化导致的非靶标抗性。本文综述了杂草对9类不同作用方式除草剂的非靶标抗性机制的生理、生化和分子基础的研究进展, 以期为抗性杂草综合治理提供参考。     

Herbicides – a two‐edged sword*

Summary Weeds cause yield losses and reductions in crop quality. Prior to the introduction of selective herbicides, the drudgery of manual weeding forced farmers to adhere to a suit of weed management tactics by carefully combining crop rotation, appropriate tillage and fallow systems. The introduction of selective herbicides in the late 1940s and the constant flow of new herbicides in the succeeding decades provided farmers with a new tool, ‘the chemical hoe’, putting them in a position to consider weed control more independently of the crop production system than hitherto. The reliance on herbicides for weed control, however, resulted in shifts in the weed flora and the selection of herbicide‐resistant biotypes. In the 1980s, the public concern about side‐effects of herbicides on the environment and human health resulted in increasingly strict registration requirements and, in some countries, political initiatives to reduce the use of pesticides were launched. Today, the number of new herbicides being introduced has decreased significantly and integrated weed management has become the guiding concept. Farmers also have the option of growing herbicide‐resistant crops where the biology of the crop has been adapted to tolerate herbicides considered safe to humans and environmentally benign. This paper discusses some of the recent developments in herbicide discovery, technology and fate, and sketches important future developments.     

Challenges for herbicide resistance evolution and management: 50 years after Harper

Evolved resistance to herbicides is a classic example of ‘evolution in action’. This paper calls for a greater integration of ‘evolutionary‐thinking’ into herbicide resistance research. This integration, it is argued, should lead weed scientists to become less focused on simply describing resistance and more driven towards a deeper understanding of the evolutionary forces that underpin resistance evolution. I have attempted in this short paper to initiate a debate into how this might be done. In the first instance, I have highlighted the widespread misunderstanding and mis‐measurement by weed scientists of fitness and fitness costs. I have also speculated on the potential for herbicide rotations to exacerbate resistance problems by selecting for generalist (metabolic) resistance. Finally, I have discussed in greater detail the contribution of herbicide rates to resistance evolution and have reported work conducted in Australia which has shown the potential for low herbicide doses to rapidly select for very high levels of resistance in Lolium rigidum. The controversial hypotheses and suggestions put forward need to be tested by field experimentation. They may prove to be unfounded or incorrect, but if they cause us to question and expand the current resistance paradigm they will have been useful.     

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.     

The impacts of traditional and novel herbicide application methods on target plants,non‐target plants and production in intensive grasslands

Herbicides contribute significantly to agricultural intensification, but some negatively impact non‐target organisms. Much research has focused on reducing herbicide use through technological improvements in application and herbicide selectivity, but impacts on non‐target organisms are less well understood. Using experimental plots in silage systems, we investigated impacts of herbicides (both narrow spectrum targeting broad‐leaved plants and selective and non‐selective broad spectrum) applied using traditional techniques (blanket‐ and manual spot‐spraying) and a novel application technique (automated spot‐spraying) on non‐target plant richness/diversity, target weed presence (Rumex species) and production (DM yield). All herbicides reduced non‐target plant richness/diversity and sometimes target weeds (when applied using traditional methods). Automated spot‐spraying had fewer negative effects on non‐target organisms, but did not reduce target weeds. No differences in production levels among treatments were observed. The automated spot‐spraying technique requires further research and development. Our results indicate that 20–30% weed cover does not significantly alter production and so, as herbicides are expensive, their effects on non‐target organisms and the environment can be more significant than their benefits to production. We advocate more research into the relationships between weed infestation and production in grasslands, so that the propensity to overuse herbicides is reduced.     

An aspartate to glycine change in the carboxyl transferase domain of acetyl CoA carboxylase and non‐target‐site mechanism(s) confer resistance to ACCase inhibitor herbicides in a Lolium multiflorum population

BACKGROUND: The increasing use of ACCase‐inhibiting herbicides has resulted in evolved resistance in key grass weeds infesting cereal cropping systems worldwide. Here, a thorough and systematic approach is proposed to elucidate the basis of resistance to three ACCase herbicides in a Lolium multiflorum Lam. (Italian rye grass) population from the United Kingdom (UK24). RESULTS: Resistance to sethoxydim and pinoxaden was always associated with a dominant D2078G (Alopecurus myosuroides Huds. equivalent) target‐site mutation in UK24. Conversely, whole‐plant herbicide assays on predetermined ACCase genotypes showed very high levels of resistance to diclofop‐methyl for all three wild DD2078 and mutant DG2078 and GG2078 ACCase genotypes from the mixed resistant population UK24. This indicates the presence of other diclofop‐methyl‐specific resistance mechanism(s) yet to be determined in this population. The D2078G mutation could be detected using an unambiguous DNA‐based dCAPS procedure that proved very transferable to A. myosuroides, Avena fatua L., Setaria viridis (L.) Beauv. and Phalaris minor Retz. CONCLUSION: This study provides further understanding of the molecular basis of resistance to ACCase inhibitor herbicides in a Lolium population and a widely applicable PCR‐based method for monitoring the D2078G target‐site resistance mutation in five major grass weed species. Copyright © 2010 Society of Chemical Industry     

Perspectives on transgenic,herbicide‐resistant crops in the United States almost 20 years after introduction

Herbicide‐resistant crops have had a profound impact on weed management. Most of the impact has been by glyphosate‐resistant maize, cotton, soybean and canola. Significant economic savings, yield increases and more efficacious and simplified weed management have resulted in widespread adoption of the technology. Initially, glyphosate‐resistant crops enabled significantly reduced tillage and reduced the environmental impact of weed management. Continuous use of glyphosate with glyphosate‐resistant crops over broad areas facilitated the evolution of glyphosate‐resistant weeds, which have resulted in increases in the use of tillage and other herbicides with glyphosate, reducing some of the initial environmental benefits of glyphosate‐resistant crops. Transgenic crops with resistance to auxinic herbicides, as well as to herbicides that inhibit acetolactate synthase, acetyl‐CoA carboxylase and hydroxyphenylpyruvate dioxygenase, stacked with glyphosate and/or glufosinate resistance, will become available in the next few years. These technologies will provide additional weed management options for farmers, but will not have all of the positive effects (reduced cost, simplified weed management, lowered environmental impact and reduced tillage) that glyphosate‐resistant crops had initially. In the more distant future, other herbicide‐resistant crops (including non‐transgenic ones), herbicides with new modes of action and technologies that are currently in their infancy (e.g. bioherbicides, sprayable herbicidal RNAi and/or robotic weeding) may affect the role of transgenic, herbicide‐resistant crops in weed management. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.     

A SNaPshot assay for the rapid and simple detection of known point mutations conferring resistance to ACCase‐inhibiting herbicides in Lolium spp.

Evolution of resistance to herbicides in weeds is becoming an increasing problem worldwide. To develop effective strategies for weed control, a thorough knowledge of the basis of resistance is required. Although non‐target‐site‐based resistance is widespread, target site resistance, often caused by a single nucleotide change in the gene encoding the target enzyme, is also a common factor affecting the efficacies of key herbicides. Therefore, fast and relatively simple high‐throughput screening methods to detect target site resistance mutations will represent important tools for monitoring the distribution and evolution of resistant alleles within weed populations. Here, we present a simple and quick method that can be used to simultaneously screen for up to 10 mutations from several target site resistance‐associated codons in a single reaction. As a proof of concept, this SNaPshot multiplex method was successfully applied to the genotyping of nine variable nucleotide positions in the CT domain of the chloroplastic ACCase gene from Lolium multiflorum plants from 54 populations. A total of 10 nucleotide substitutions at seven of these nine positions (namely codons 1781, 1999, 2027, 2041 2078, 2088 and 2096) are known to confer resistance to ACCase‐inhibiting herbicides. This assay has several advantages when compared with other methods currently in use in weed science. It can discriminate between different nucleotide changes at a single locus, as well as screening for SNPs from different target sites by pooling multiple PCR products within a single reaction. The method is scalable, allowing reactions to be carried out in either 96‐ or 384‐well plate formats, thus reducing work time and cost.     

Herbicide‐resistant weed management: focus on glyphosate

This review focuses on proactive and reactive management of glyphosate‐resistant (GR) weeds. Glyphosate resistance in weeds has evolved under recurrent glyphosate usage, with little or no diversity in weed management practices. The main herbicide strategy for proactively or reactively managing GR weeds is to supplement glyphosate with herbicides of alternative modes of action and with soil‐residual activity. These herbicides can be applied in sequences or mixtures. Proactive or reactive GR weed management can be aided by crop cultivars with alternative single or stacked herbicide‐resistance traits, which will become increasingly available to growers in the future. Many growers with GR weeds continue to use glyphosate because of its economical broad‐spectrum weed control. Government farm policies, pesticide regulatory policies and industry actions should encourage growers to adopt a more proactive approach to GR weed management by providing the best information and training on management practices, information on the benefits of proactive management and voluntary incentives, as appropriate. Results from recent surveys in the United States indicate that such a change in grower attitudes may be occurring because of enhanced awareness of the benefits of proactive management and the relative cost of the reactive management of GR weeds. Copyright © 2011 Society of Chemical Industry     

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.     

杂草对AHAS抑制剂的抗药性分子机理研究进展

除草剂在田间的重复及不合理使用,导致了杂草抗药性的发生和发展。其中AHAS抑制剂由于靶标单一,抗性发展十分迅速。截至2009年,已有103种杂草对AHAS抑制剂产生了抗药性,占19类化学除草剂总抗药性杂草生物型的近1/3。从AHAS基因突变位点及种类与杂草抗药性水平的关系、AHAS基因突变与AHAS酶活性的关系、AHAS基因拷贝数与杂草抗药性的关系以及AHAS酶与除草剂结合前后的三维结构等方面,综述了杂草对AHAS抑制剂产生抗药性的机理,旨在为AHAS抑制剂抗性研究提供参考。并对自然种群目标基因的等位基因检测技术(ECOTILLING)和衍生型酶切扩增多态性序列(dCAPS)两种通过检测等位基因多态性的手段快速诊断抗药性杂草的新技术进行了介绍,讨论了延缓杂草抗药性发生和发展的策略。