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The introduction of harvest weed seed control (HWSC) techniques and associated machinery has enabled the routine use of an alternative weed control technology at a novel weed control timing in global grain cropping fields. Driven by the significant threat of widespread populations of annual ryegrass (Lolium rigidum) with multiple-herbicide resistance, in the 1990s Australian growers and researchers developed techniques to target, at grain harvest, the seed production of annual ryegrass and other important weed species. The HWSC approach to weed management is now routinely used by a majority of Australian grain producers as an integral component of effective weed control programs. Here we detail the development and introduction of current HWSC systems and describe their efficacy in Australian grain production systems. The use of HWSC has likely contributed to lower annual ryegrass population densities and thus mitigated the impacts of herbicide resistance as well as slowing further evolution of resistance. In addition, low weed densities enable the introduction of site-specific weed control technologies and the opportunity to target specific in-crop weeds with non-selective alternative weed control techniques. With an awareness of the evolutionary potential of weed species to adapt to all forms of weed control, there is an understanding that HWSC treatments need to be judiciously used in grain cropping systems to ensure their ongoing efficacy. The successful use of Australian developed HWSC systems has attracted global interest and there is now a considerable international research effort aimed at introducing this alternative weed control approach and timing into the world's major cropping systems. [ABSTRACT FROM AUTHOR]

Weeds are a major contributing factor to crop yield loss. Weed control is regularly practiced during the growing season, with many growers making a conscious effort to minimise weed-seed return to the soil seedbank during the cropping program. However, growers may be unintentionally introducing weed seeds through sowing of contaminated crop seed. Using samples of crop seed obtained from 29 growers across two Western Australian grain-growing regions, 81 samples were hand-cleaned to determine weed-seed contamination levels. Of those samples, 41% were weed-free, and in the remaining 59%, the main contaminant was Lolium rigidum (annual ryegrass), occurring in 49% of contaminated samples. Crop type and cleaning method had significant effects on the level of weed-seed contamination, with barley having higher levels of contamination than other crops, and professional contractors providing lower contamination than other methods of cleaning. However, any seed-cleaning method provided significantly cleaner grain samples than no seed cleaning. This study established that crop-seed contamination was evident on Western Australian farms and that growers may be unintentionally sowing weed seeds with their crops. Seed cleaning combined with judicious paddock selection and weed-seed removal during the growing season can lead to weed-free crop seed. Crop seed cleaning is an important component of reducing weed-seed contamination in crop grain. This article examines seed-cleaning operations and other factors affecting levels of weed contamination in farmer-saved crop seed, finding that seed cleaning significantly reduces the level of weed-seed contamination in grain samples. Seed cleaning combined with judicious paddock selection and weed-seed removal during the growing season can lead to weed-free crop seed. [ABSTRACT FROM AUTHOR]

Weed population simulations can be useful to predict the effects of alternative management practices on herbicide resistance (HR) evolution. Almost all previous simulations have ignored the possibility of within-field spatial structure in a weed population, instead making the implicit assumption of perfect dispersal and spatial homogeneity in population density and genetics. The effects of this simplifying assumption have not been examined, despite the fact that dispersal limitations and spatial structure within the population are likely to occur and to affect the evolution of resistance. Therefore, we developed a new spatially-explicit model called SOMER, and examined how changing the following factors affected the predicted evolution of resistance: the degree of spatial resolution used in the model; whether resistance was semi-dominant or fully-dominant; distances of pollen and natural seed dispersal; and inadvertent collection and grain harvester weed seed dispersal (GHWSD). Simulations showed that spatial resolution is important when modelling HR evolution, with the size of sub-population divisions, the pollen dispersal parameter, the level of dominance, and GHWSD all being important factors in predicting the rate and type of HR evolution. Our results show that accounting for spatial structure and dispersal does affect predictions of HR evolution, with the non-spatial model generally predicting faster resistance evolution compared to the more realistic equivalent spatial model. Most importantly, GHWSD increased the speed of HR evolution. Our spatial model also allowed us to investigate the dynamics of density and genetic structure within patches of herbicide resistant weeds, and we found that resistance genes were spread several times wider than the visible patch, and that homozygous mutations were commonly found in more centrally located weeds. We conclude that an ‘integrated spatial modelling’ approach that accounts for spatial structure should be considered when modelling HR evolution, and the evolution of resistance in general. [ABSTRACT FROM AUTHOR]

A relatively low number of weed species have evolved resistance to auxinic herbicides despite their use for almost 70 years. This inheritance study with two Raphanus raphanistrum populations multiple-resistant 2,4-D and the ALS-inhibiting herbicide chlorsulfuron determined the number of genes and genetic dominance of 2,4-D resistance and investigated the association between traits conferring resistance to the two herbicide modes of action. Levels of 2,4-D phenotypic resistance and resistance segregation patterns were assessed in parental populations, F 1 and F 2 families. [ABSTRACT FROM AUTHOR]

Avena is a problematic weed of cropping regions of southern Australia and many areas of the world. In 2010, a random survey was conducted across 14 million hectares of the Western Australian grain belt to monitor the change in herbicide resistance levels by comparing resistance frequency results with a survey conducted in 2005. Screening Avena populations with herbicides commonly used to control this weed revealed that 48% of Avena populations displayed resistance to the commonly used acetyl-Co A carboxylase-inhibiting herbicides, which was lower than that found in 2005 (71%). The broad-spectrum herbicides glyphosate and paraquat provided good control of all Avena populations. Resistance to acetolactate synthase-inhibiting herbicides and to flamprop were detected for the first time in Western Australia in this survey. Therefore, a wide range of weed management options that target all phases of the cropping program are needed to sustain these cropping systems in the future. [ABSTRACT FROM AUTHOR]

RIM, or ‘Ryegrass Integrated Management’, is a model-based decision support system (DSS) for weed management in broadacre cropping systems that was updated to continue aid the delivery of key recommendations to manage herbicide resistance. This article complements earlier publications by documenting the rationales that underpinned the re-development efforts. The objectives are to inform the next development cycle of RIM and its delivery, as well as its adaptation to other situations. Specifically, the article aims at providing developers and project managers with key aspects to be considered before and after (re-)developing this type of model-based agricultural DSS. Reviewers report a lack of similar efforts, with modelling aspects generally better documented than underpinning rationales, including those related to implementation. Yet, this type of initiative is necessary considering that agricultural DSS can become expensive projects, and that uptake by target audiences is typically low in spite of known pitfalls and limitations. The key elements that contributed to the thought process behind upgrade choices are thus provided, as well as practical consequences for modelling. Clearly re-asserting cost-effectiveness objectives and favouring human aspects led to: retaining the ‘what-if’ learning strategy rather than developing optimisation features; renouncing added modelling intricacies; enhancing the software accessibility; and anticipating future maintenance and distribution requirements. Strategies to maximise the impact of RIM are also discussed, particularly the need for qualified workshop facilitators, as well as transparency and evaluation to build user confidence. [ABSTRACT FROM AUTHOR]

Weedy plant species that have evolved resistance to herbicides due to enhanced metabolic capacity to detoxify herbicides (metabolic resistance) are a major issue. Metabolic herbicide resistance in weedy plant species first became evident in the 1980s in Australia (in Lolium rigidum) and the United Kingdom (in Alopecurus myosuroides) and is now increasingly recognized in several crop-weed species as a looming threat to herbicide sustainability and thus world crop production. Metabolic resistance often confers resistance to herbicides of different chemical groups and sites of action and can extend to new herbicide(s). Cytochrome P450 monooxygenase, glycosyl transferase, and glutathione S-transferase are often implicated in herbicide metabolic resistance. However, precise biochemical and molecular genetic elucidation of metabolic resistance had been stalled until recently. Complex cytochrome P450 superfamilies, high genetic diversity in metabolic resistant weedy plant species (especially cross-pollinated species), and the complexity of genetic control of metabolic resistance have all been barriers to advances in understanding metabolic herbicide resistance. However, next-generation sequencing technologies and transcriptome-wide gene expression profiling are now revealing the genes endowing metabolic herbicide resistance in plants. This Update presents an historical review to current understanding of metabolic herbicide resistance evolution in weedy plant species. [ABSTRACT FROM AUTHOR]

Background and Aims α-Amylase in grass caryopses (seeds) is usually expressed upon commencement of germination and is rarely seen in dry, mature seeds. A heat-stable α-amylase activity was unexpectedly selected for expression in dry annual ryegrass (Lolium rigidum) seeds during targeted selection for low primary dormancy. The aim of this study was to characterize this constitutive activity biochemically and determine if its presence conferred insensitivity to the germination inhibitors abscisic acid and benzoxazolinone. Methods α-Amylase activity in developing, mature and germinating seeds from the selected (low-dormancy) and a field-collected (dormant) population was characterized by native activity PAGE. The response of seed germination and α-amylase activity to abscisic acid and benzoxazolinone was assessed. Using an alginate affinity matrix, α-amylase was purified from dry and germinating seeds for analysis of its enzymatic properties. Key Results The constitutive α-amylase activity appeared late during seed development and was mainly localized in the aleurone; in germinating seeds, this activity was responsive to both glucose and gibberellin. It migrated differently on native PAGE compared with the major activities in germinating seeds of the dormant population, but the enzymatic properties of α-amylase purified from the low-dormancy and dormant seeds were largely indistinguishable. Seed imbibition on benzoxazolinone had little effect on the low-dormancy seeds but greatly inhibited germination and α-amylase activity in the dormant population. Conclusions The constitutive α-amylase activity in annual ryegrass seeds selected for low dormancy is electrophoretically different from that in germinating seeds and its presence confers insensitivity to benzoxazolinone. The concurrent selection of low dormancy and constitutive α-amylase activity may help to enhance seedling establishment under competitive conditions. [ABSTRACT FROM AUTHOR]

Abstract: Dormancy release in freshly matured, imbibed annual ryegrass (Lolium rigidum) seeds is inhibited by light and involves a decrease in seed sensitivity to abscisic acid. Other processes involved in dormancy release in the dark were investigated by measuring seed storage compound mobilisation and the activity of cell wall-degrading enzymes. Activities of endo-β-mannanase and total peroxidase were higher in dark-stratified compared to light-stratified seeds, indicating that weakening of the structures constraining the embryo was accelerated in the dark. A dramatic degradation of storage proteins in light-stratified seeds, accompanied by induction of a high molecular mass protease, suggests that maintenance of storage(-like) proteins is also important in dark-mediated dormancy release. α-Amylase activity was induced in dark-stratified seeds at least 48h prior to radicle emergence upon transfer to conditions permitting germination, or in light-stratified seeds supplied with exogenous gibberellin A4. This suggests that (a) α-amylase is involved in stimulation of germination of non-dormant L. rigidum seeds, and (b) dark-stratified seeds have an increased sensitivity to gibberellins which permits the rapid induction of α-amylase activity upon exposure to germination conditions. Overall, it appears that a number of processes, although possibly minor in themselves, occur in concert during dark-stratification to contribute to dormancy release. [Copyright &y& Elsevier]

Seed dormancy in wild Lolium rigidum Gaud (annual ryegrass) populations is highly variable and not well characterized at the biochemical level. To identify some of the determinants of dormancy level in these seeds, the proteomes of subpopulations selected for low and high levels of primary dormancy were compared by two-dimensional polyacrylamide gel electrophoresis of extracts from mature, dry seeds. High-dormancy seeds showed higher expression of small heat shock proteins, enolase, and glyoxalase I than the low-dormancy seeds. The functional relevance of these differences in protein expression was confirmed by the fact that high-dormancy seeds were more tolerant to high temperatures imposed at imbibition and had consistently higher glyoxalase I activity over 0–42 d dark stratification. Higher expression of a putative glutathione peroxidase in low-dormancy seeds was not accompanied by higher activity, but these seeds had a slightly more oxidized glutathione pool and higher total peroxidase activity. Overall, these biochemical and physiological differences suggest that L. rigidum seeds selected for low dormancy are more prepared for rapid germination via peroxidase-mediated cell wall weakening, whilst seeds selected for high dormancy are constitutively prepared to survive environmental stresses, even in the absence of stress during seed development. [ABSTRACT FROM PUBLISHER]

Modern herbicides make major contributions to global food production by easily removing weeds and substituting for destructive soil cultivation. However, persistent herbicide selection of huge weed numbers across vast areas can result in the rapid evolution of herbicide resistance. Herbicides target specific enzymes, and mutations are selected that confer resistance-endowing amino acid substitutions, decreasing herbicide binding. Where herbicides bind within an enzyme catalytic site very few mutations give resistance while conserving enzyme functionality. Where herbicides bind away from a catalytic site many resistance-endowing mutations may evolve. Increasingly, resistance evolves due to mechanisms limiting herbicide reaching target sites. Especially threatening are herbicide-degrading cytochrome P450 enzymes able to detoxify existing, new, and even herbicides yet to he discovered. Global weed species are accumulating resistance mechanisms, displaying multiple resistance across many herbicides and posing a great challenge to herbicide sustainability in world agriculture. Fascinating genetic issues associated with resistance evolution remain to be investigated, especially the possibility of herbicide stress unleashing epigenetic gene expression. Understanding resistance and building sustainable solutions to herbicide resistance evolution are necessary and worthy challenges. [ABSTRACT FROM AUTHOR]

• The molecular basis of an acetyl-CoA carboxylase (ACCase) target-based resistant Lolium rigidum population (WLR 96) was studied here. • The carboxyl-transferase domain of the plastidic ACCase gene from resistant individuals was amplified by PCR and sequenced. The DNA sequences were aligned and compared with a susceptible population. • Six amino acid substitutions were identified in the resistant population. The substitution Ile-2041-Asn, known to confer resistance to ACCase-inhibiting herbicides aryloxyphenoxypropionate (APP) in Alopecurus myosuroides, was identified in most resistant plants but it is always linked with other amino acid substitutions. This was confirmed by a cleaved amplified polymorphism (CAP) marker and an allele-specific PCR. The sole amino acid substitution Ile-2041-Asn was not found in this population. It is likely this mutation evolved later among individuals already possessing the other substitutions. Three haplotypes were identified from the resistant population based on the six amino acid combinations, and two are linked with herbicide resistance in this population. • The multiple amino acid substitutions including the Ile-2041-Asn form the molecular basis endowing a high degree of resistance to ACCase-inhibiting herbicides in this L. rigidum population. [ABSTRACT FROM AUTHOR]

There has been much debate regarding the potential for reduced rates of herbicide application to accelerate evolution of herbicide resistance. We report a series of experiments that demonstrate the potential for reduced rates of the acetyl-co enzyme A carboxylase (ACCase)-inhibiting herbicide diclofop-methyl to rapidly select for resistance in a susceptible biotype ofLolium rigidum. Thirty-six percent of individuals from the original VLR1 population survived application of 37.5 g diclofop-methyl ha-1 (10% of the recommended field application rate). These individuals were grown to maturity and bulk-crossed to produce the VLR1 low dose-selected line VLR1 (0.1). Subsequent comparisons of the dose-response characteristics of the original and low dose-selected VLR1 lines demonstrated increased tolerance of diclofop-methyl in the selected line. Two further rounds of selection produced VLR1 lines that were resistant to field-applied rates of diclofop-methyl. The LD50 (diclofop-methyl dose required to cause 50% mortality) of the most resistant line was 56-fold greater than that of the original unselected VLR1 population, indicating very large increases in mean population survival after three cycles of selection. In vitro ACCase inhibition by diclofop acid confirmed that resistance was not due to an insensitive herbicide target-site. Cross-resistance studies showed increases in resistance to four herbicides: fluazifop-P-butyl, haloxyfop-R-methyl, clethodim and imazethapyr. The potential genetic basis of the observed response and implications of reduced herbicide application rates for management of herbicide resistance are discussed. [ABSTRACT FROM AUTHOR]

The development of herbicide multiple‐resistance in weed species represents a major threat to current agricultural practices. The mechanistic basis for herbicide multiple‐resistance has been investigated in a population of the annual grass weed Lolium rigidum Gaud. (annual ryegrass) resistant to herbicides affecting 6 target sites. A subset of the resistant population (R2 subset) has been isolated by germination on a medium containing the acetyl‐CoA carboxylase (ACCase, EC 6.4.1.2) inhibiting herbicide, sethoxydim ((2‐[1‐(ethoxyimino)butyl]‐5‐[2‐(ethylthio)propyl]‐3‐hydroxy‐2‐cyclohexen‐1‐one)). This 12% R2 subset of the population is 600 times more resistant to sethoxydim and between 30 to 200 times more resistant to other ACCase inhibitors than the bulk of the R population. The subset has a form of ACCase which is 6 to 55 times less sensitive to inhibition by these herbicides than the enzyme present in the bulk of the resistant or in the susceptible population. There was no difference in the uptake and metabolic degradation of [4‐14C]sethoxydim between the R2 subset and the unselected R population. These results show the accumulation of different resistance mechanisms in that single population. Furthermore we propose that this accumulation of multiple resistance mechanisms is the basis for herbicide multiple‐resistance in this biotype. [ABSTRACT FROM AUTHOR]

Fully expanded leaves of 25°C grown Phaseolus vulgaris and six other species were exposed for 3 h to chilling temperatures at photon flux densities equivalent to full sunlight. In four of the species this treatment resulted in substantial inhibition of the subsequent quantum yield of CO2 uptake, indicating reduction of the photochemical efficiency of photosynthesis. The extent of inhibition was dependent on the photon flux density during chilling and no inhibition occurred when chilling occurred at a low photon flux density. No inhibition occurred at temperatures above 11.5°C, even in the presence of the equivalent of full sunlight. This interaction between chilling and light to cause inhibition of photosynthesis was promoted by the presence of oxygen at normal air partial pressures and was unaffected by the CO2 partial pressure present when chilling occured in the air. When chilling occurred at low O2 partial pressures, CO2 was effective in reducing the degree of inhibition. Apparently, when leaves of chilling-sensitive plants are exposed to chilling temperatures in air of normal composition then light is instrumental in inducing rapid damage to the photochemical efficiency of photosynthesis. [ABSTRACT FROM AUTHOR]

Summary: We reviewed the literature to understand the effects of glyphosate resistance on plant fitness at the molecular, biochemical and physiological levels. A number of correlations between enzyme characteristics and glyphosate resistance imply the existence of a plant fitness cost associated with resistance‐conferring mutations in the glyphosate target enzyme, 5‐enolpyruvylshikimate‐3‐phosphate synthase (EPSPS). These biochemical changes result in a tradeoff between the glyphosate resistance of the EPSPS enzyme and its catalytic activity. Mutations that endow the highest resistance are more likely to decrease catalytic activity by reducing the affinity of EPSPS for its natural substrate, and/or slowing the velocity of the enzyme reaction, and are thus very likely to endow a substantial plant fitness cost. Prediction of fitness costs associated with EPSPS gene amplification and overexpression can be more problematic. The validity of cost prediction based on the theory of evolution of gene expression and resource allocation has been cast into doubt by contradictory experimental evidence. Further research providing insights into the role of the EPSPS cassette in weed adaptation, and estimations of the energy budget involved in EPSPS amplification and overexpression are required to understand and predict the biochemical and physiological bases of the fitness cost of glyphosate resistance. [ABSTRACT FROM AUTHOR]

In this article the author discusses the role of gene amplification in glyphosate-resistant weed prevention that affects food production. The author states that the excessive reliance on glyphosate alongside with the prevalence of transgenic glyphosate-resistant crops resulted to the resistance of weeds on the herbicide. Moreover, he highlights the fact that gene amplification is a revolutionary response to herbicide selection pressure as well as the need to use glyphosate with more diversity.

Resistance to auxinic herbicides is increasing in a range of dicotyledonous weed species, but in most cases the biochemical mechanism of resistance is unknown. Using 14C-labelled herbicide, the mechanism of resistance to 2,4-dichlorophenoxyacetic acid (2,4-D) in two wild radish (Raphanus raphanistrum L.) populations was identified as an inability to translocate 2,4-D out of the treated leaf. Although 2,4-D was metabolized in wild radish, and in a different manner to the well-characterized crop species wheat and bean, there was no difference in metabolism between the susceptible and resistant populations. Reduced translocation of 2,4-D in the latter was also not due to sequestration of the herbicide, or to reduced uptake by the leaf epidermis or mesophyll cells. Application of auxin efflux or ABCB transporter inhibitors to 2,4-D-susceptible plants caused a mimicking of the reduced-translocation resistance phenotype, suggesting that 2,4-D resistance in the populations under investigation could be due to an alteration in the activity of a plasma membrane ABCB-type auxin transporter responsible for facilitating long-distance transport of 2,4-D. [ABSTRACT FROM AUTHOR]

Raphanus raphanistrum is a problematic weed, which has become increasingly difficult to control in Australian cropping regions. In 2010, a random survey was conducted across 14 million ha of the Western Australian grain belt to establish the frequency of herbicide resistance in R. raphanistrum and to monitor the change in resistance levels by comparing results with a previous survey in 2003. Screening R. raphanistrum populations with herbicides commonly used to control this weed revealed that most populations (84%) contained individual plants resistant to the acetolactate synthase-inhibiting herbicide chlorsulfuron, whereas 49% of populations also had plants resistant to the imidazolinone herbicides. Resistance to other mode of action herbicides (2,4-D (76%) and diflufenican (49%)) was also common. Glyphosate, atrazine and pyrasulfotole + bromoxynil remained effective on most R. raphanistrum populations. These results demonstrate that resistance to some herbicides has increased significantly since 2003 when the values were 54% for chlorsulfuron and 60% for 2,4-D; therefore, a wide range of weed management options that target all phases of the cropping program are needed to sustain these cropping systems in the future. Random surveys conducted in Western Australia have shown that herbicide resistance in major crop weeds is common and increasing. The latest survey on resistance in Raphanus raphanistrum revealed that 84% of populations were resistant to the ALS-inhibiting herbicides, 76% were resistant to synthetic auxins, and 49% to PDS inhibitors. Resistance in Raphanus has increased over the 12 year survey period and a wide range of management tools are needed to control this species. [ABSTRACT FROM AUTHOR]

Glufosinate is an important and widely used non-selective herbicide active on a wide range of plant species. Evolution of resistance to glufosinate in weedy plant species (including the global weed Eleusine indica) is underway. Here, we established the molecular basis of target site glufosinate resistance in Eleusine indica. Full-length E. indica glutamine synthetase (GS) iso-genes (EiGS1-1 , 1-2 , 1-3 , and EiGS2) were cloned, and expression of EiGS1-1 and EiGS1-2 was higher than that of EiGS2. A novel point mutation resulting in a Ser59Gly substitution in EiGS1-1 was identified in glufosinate-resistant plants. Rice calli and seedlings transformed with the mutant EiGS1-1 gene were resistant to glufosinate. Purified mutant EiGS1-1 expressed in yeast was more tolerant to glufosinate than the wild-type variant. These transgenic results correlate with a more glufosinate-resistant GS in the crude tissue extract of resistant versus susceptible E. indica plants. Structural modelling of the mutant EiGS1-1 revealed that Ser59 is not directly involved in glufosinate binding but is in contact with some important binding residues (e.g. Glu297) and especially with Asp56 that forms an intratoroidal contact interface. Importantly, the same Ser59Gly mutation was also found in geographically isolated glufosinate-resistant populations from Malaysia and China, suggesting parallel evolution of this resistance mutation. [ABSTRACT FROM AUTHOR]

The interaction between environment and genetic traits under selection is the basis of evolution. In this study, we have investigated the genetic basis of herbicide resistance in a highly characterized initially herbicide-susceptible Lolium rigidum population recurrently selected with low (below recommended label) doses of the herbicide diclofop-methyl. We report the variability in herbicide resistance levels observed in F1 families and the segregation of resistance observed in F2 and back-cross (BC) families. The selected herbicide resistance phenotypic trait(s) appear to be under complex polygenic control. The estimation of the effective minimum number of genes (NE), depending on the herbicide dose used, reveals at least three resistance genes had been enriched. A joint scaling test indicates that an additive-dominance model best explains gene interactions in parental, F1, F2 and BC families. The Mendelian study of six F2 and two BC segregating families confirmed involvement of more than one resistance gene. Cross-pollinated L. rigidum under selection at low herbicide dose can rapidly evolve polygenic broad-spectrum herbicide resistance by quantitative accumulation of additive genes of small effect. This can be minimized by using herbicides at the recommended dose which causes high mortality acting outside the normal range of phenotypic variation for herbicide susceptibility. [ABSTRACT FROM AUTHOR]

Global grain production is under threat from the escalating evolution of herbicide-resistant weed populations. Worldwide, herbicide-reliant grain crop production systems have driven the proliferation of herbicide resistant populations of major weed species. Alternatives or adjuncts to herbicides are needed for sustainable control of crop weeds in grain crops worldwide. Here we introduce and prove the ability of a new weed control tool, the Harrington Seed Destructor (HSD), to intercept and destroy weed seeds during grain crop harvest. The interception and destruction of weed seeds exiting the grain harvester in the chaff fraction during grain crop harvest is a hitherto unrealized opportunity. We have developed a cage millbased chaff processing unit that consistently destroys weed seed infesting grain crop chaff fractions. The subsequent construction of the HSD incorporating this unit had >95% weed seed destruction efficacy when used during commercial harvest of three major grain crops, wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), and lupin (Lupinus angustifolius L.). The HSD system has the potential for a dramatic impact on global grain production by providing the unique combination of effective weed control with complete retention of grain crop harvest residues. Only herbicides offer this same combination of highly effective weed control and complete residue retention in large scale grain crop production systems. [ABSTRACT FROM AUTHOR]

Abstract: In two different locations of the Western Australian “wheatbelt”, Lolium rigidum (rigid ryegrass) seeds were collected from organic fields (no herbicide use) and neighbouring conventional fields (persistent herbicide use), the latter infested with herbicide-resistant plants, to investigate the occurrence of gene flow among field populations as revealed by herbicide resistance gene transfer. Herbicides targeting acetyl-CoA carboxylase (ACCase) or acetolactate-synthase (ALS) were used to detect herbicide-resistant plants. Overall, the frequency of plants resistant to ACCase- or ALS-inhibiting herbicides was, respectively, 21% and 74% in the conventional fields and 2% and 37% in neighbouring organic fields. Mutant, herbicide-resistant ACCase and ALS alleles were detected in 16% and 38% of plants from conventional fields and in 0.53% and 3.7% of plants from organic fields. Identical mutant, herbicide-resistant ALS haplotypes were detected both in conventional and organic fields, supporting the occurrence of gene flow between L. rigidum populations in different fields. Gene flow can thus substantially increase the frequency of herbicide-resistant plants in unselected L. rigidum populations. Although gene flow cannot be prevented, it can be limited or managed. Hygiene tactics such as clean crop seed, weed seed removal at harvest and seed destruction post-harvest should be considered in order to minimize gene transfer among farms. [Copyright &y& Elsevier]

Abstract: The physiological and biochemical bases of seed dormancy in Lolium rigidum (annual ryegrass) are largely unknown, and study of this process is complicated by the outcrossing nature of the species and the strong influence of environment on seed dormancy. In order to identify heritable biochemical factors contributing to seed dormancy in L. rigidum, seeds from a field-collected population were used to select sub-populations with consistently low or high seed dormancy over four generations. Low-dormancy seeds showed constitutive α-amylase activity prior to imbibition, higher concentrations of polyphenols and cis-zeatin, and lower abscisic acid and cis-zeatin riboside concentrations than high-dormancy seeds. Selection for high dormancy was associated with a reduction in response to dark-stratification for 21d at 20°C (an effective means of releasing dormancy in the original, unselected population) over successive generations, but fluridone remained effective in breaking dormancy. Crossing of low- and high-dormancy populations indicated that dormancy level was not dependent upon the maternal genotype of the seed, and that the constitutive α-amylase activity and high seed anthocyanin concentrations characteristic of the low-dormancy populations were not correlated to high basal germination ability. [ABSTRACT FROM AUTHOR]

Abstract: Sequestration of paraquat away from its target site in the chloroplast has been proposed as a mechanism of paraquat resistance. However, no consensus has been reached as to where paraquat is sequestered. This study quantifies paraquat in leaf protoplasts of paraquat resistant (R) and susceptible (S) Lolium rigidum. Intact protoplasts were prepared from plants treated with commercial dose of paraquat for 2h. Paraquat absorbed by the leaf protoplasts was determined by light absorption of reduced paraquat following concentration and purification using a cation-exchange resin. Leaf protoplasts from treated paraquat resistant plants contained 2- to 3-fold more paraquat than leaf protoplasts isolated from susceptible plants. Since paraquat is not metabolised in L. rigidum and paraquat readily enters chloroplasts of both R and S plants, this greater amount of paraquat in leaf protoplasts of R plant must be kept away from the target site (chloroplast). This result indicates that paraquat resistance in L. rigidum is associated with a cytoplasmic mechanism, most likely a greater rate of vacuolar sequestration. [Copyright &y& Elsevier]

Contents Predictions based on evolutionary theory suggest that the adaptive value of evolved herbicide resistance alleles may be compromised by the existence of fitness costs. There have been many studies quantifying the fitness costs associated with novel herbicide resistance alleles, reflecting the importance of fitness costs in determining the evolutionary dynamics of resistance. However, many of these studies have incorrectly defined resistance or used inappropriate plant material and methods to measure fitness. This review has two major objectives. First, to propose a methodological framework that establishes experimental criteria to unequivocally evaluate fitness costs. Second, to present a comprehensive analysis of the literature on fitness costs associated with herbicide resistance alleles. This analysis reveals unquestionable evidence that some herbicide resistance alleles are associated with pleiotropic effects that result in plant fitness costs. Observed costs are evident from herbicide resistance-endowing amino acid substitutions in proteins involved in amino acid, fatty acid, auxin and cellulose biosynthesis, as well as enzymes involved in herbicide metabolism. However, these resistance fitness costs are not universal and their expression depends on particular plant alleles and mutations. The findings of this review are discussed within the context of the plant defence trade-off theory and herbicide resistance evolution. [ABSTRACT FROM AUTHOR]

1. In some cases, evaluation of resource competitive interactions between herbicide resistant vs. susceptible weed ecotypes provides evidence for the expression of fitness costs associated with evolved herbicide-resistant gene traits. Such fitness costs impact in the ecology and evolutionary trajectory of resistant populations. 2. Neighbourhood experiments were performed to quantify competitive effects and responses between herbicide-susceptible (S) and resistant (R) Lolium rigidum individuals in which resistance is due to enhanced herbicide metabolism mediated by cytochrome P450. 3. In two-way competitive interactions between the S and R phenotypes, individuals of the S phenotype were the stronger effect competitors on both a per capita and per unit-size basis. The S phenotype also exhibited a stronger competitive response to wheat plants than did the R phenotype, displaying significantly greater (30%) above-ground biomass at the vegetative stage. When subjected to competition from wheat, R individuals produced significantly fewer reproductive tillers and allocated fewer resources to reproductive traits than individuals of the S phenotype. 4. The role of potential mechanisms underlying this resistance cost driven by traits such as plant size and tolerance to low resource availability, as well as the evolutionary implications of the results are discussed. 5. Synthesis. Evolved herbicide resistance due to enhanced-herbicide metabolism mediated by cytochrome-P450 in L. rigidum has been shown to be accompanied with an impaired ability to compete for resources. These results are consistent with the resource-based theory that predicts a negative trade-off between growth and plant defence. [ABSTRACT FROM AUTHOR]

• Light plays an important role in two separate processes within the seeds of Lolium rigidum (annual ryegrass). Dormant seeds of L. rigidum remain dormant when imbibed in the light, but once seeds have lost dormancy through dark-stratification, light stimulates their germination. This study characterizes the light qualities and quantities which are effective in maintenance of dormancy. • Dormant seeds were stratified under narrow- and broad-waveband light to identify the potential photoreceptors involved in dormancy maintenance, and to determine whether dark-induced dormancy loss is reversible by light. • Blue and green light both mediated dormancy maintenance in a far-red-independent manner. Red light resulted in dormancy maintenance only when far-red wavelengths were excluded, suggesting a redundant function of phytochrome. At low fluence rates, white light was more effective than monochromatic light, suggesting the action of multiple photoreceptors in dormancy maintenance. By contrast, nondormant seeds did not germinate unless provided with red light. • These results indicate that seed dormancy maintenance is potentially mediated through the actions of blue and green light photoreceptors. Seed dormancy could thus be added to the growing list of plant responses that may be mediated by green light in a cryptochrome-independent manner. [ABSTRACT FROM AUTHOR]

Abstract: Multi-species RIM, a bio-economic model that simulates the population dynamics of two annual weed species over a 20-year period, was used to investigate the value of transgenic glyphosate-resistant canola in the management of herbicide resistant annual ryegrass and wild radish in a Western Australian dryland farming system. The perceived advantage of growing this crop is the potential to control post-emergent weeds with the broad-spectrum herbicide glyphosate, and without the yield penalty evident in triazine-resistant canola. We found that the economic value of glyphosate-resistant canola is consistently higher than that of the commonly grown triazine-resistant canola. However, the benefits of glyphosate-resistant canola would need to be weighed up against potential risks to marketability (due to consumer resistance) and risks of increased weed resistance to glyphosate (due to increased selection pressure). [Copyright &y& Elsevier]

Concurrent natural evolution of glyphosate resistance single‐ and double‐point EPSPS mutations in weed species provides an opportunity for the estimation of resistance fitness benefits and prediction of equilibrium resistance frequencies in environments under glyphosate selection. Assessment of glyphosate resistance benefit was conducted for the most commonly identified single Pro‐106‐Ser and less‐frequent double TIPS mutations in the EPSPS gene evolved in the global damaging weed Eleusine indica. Under glyphosate selection at the field dose, plants with the single Pro‐106‐Ser mutation at homozygous state (P106S‐rr) showed reduced survival and compromised vegetative growth and fecundity compared with TIPS plants. Whereas both homozygous (TIPS‐RR) and compound heterozygous (TIPS‐Rr) plants with the double TIPS resistance mutation displayed similar survival rates when exposed to glyphosate, a significantly higher fecundity in the currency of seed number was observed in TIPS‐Rr than TIPS‐RR plants. The highest plant fitness benefit was associated with the heterozygous TIPS‐Rr mutation, whereas plants with the homozygous Pro‐106‐Ser and TIPS mutations exhibited, respectively, 31% and 39% of the fitness benefit revealed by the TIPS‐Rr plants. Populations are predicted to reach stable allelic and genotypic frequencies after 20 years of glyphosate selection at which the WT allele is lost and the stable genotypic polymorphism is comprised by 2% of heterozygous TIPS‐Rr, 52% of homozygous TIPS‐RR and 46% of homozygous P106S‐rr. The high inbreeding nature of E. indica is responsible for the expected frequency decrease in the fittest TIPS‐Rr in favour of the homozygous TIPS‐RR and P106S‐rr. Mutated alleles associated with the glyphosate resistance EPSPS single EPSPS Pro‐106‐Ser and double TIPS mutations confer contrasting fitness benefits to E. indica under glyphosate treatment and therefore are expected to exhibit contrasting evolution rates in cropping systems under recurrent glyphosate selection. [ABSTRACT FROM AUTHOR]

Glyphosate is the most widely used herbicide in world agriculture and for general vegetation control in a wide range of situations. Global and often intensive glyphosate selection of very large weedy plant populations has resulted in widespread glyphosate resistance evolution in populations of many weed species. Here, working with a glyphosate-resistant (GR) Echinochloa colona population that evolved in a Western Australia agricultural field, we identified an ATP-binding cassette (ABC) transporter (EcABCC8) that is consistently up-regulated in GR plants. When expressed in transgenic rice, this EcABCC8 transporter endowed glyphosate resistance. Equally, rice, maize, and soybean overexpressing the EcABCC8 ortholog genes were made resistant to glyphosate. Conversely, CRISPR/Cas9-mediated knockout of the EcABCC8 ortholog gene OsABCC8 increased rice susceptibility to glyphosate. Subcellular localization analysis and quantification of glyphosate cellular levels in treated ABCC8 transgenic rice plants and isolated leaf protoplasts as well as structural modeling support that EcABCC8 is likely a plasma membrane-localized transporter extruding cytoplasmic glyphosate to the apoplast, lowering the cellular glyphosate level. This is a report of a membrane transporter effluxing glyphosate in a GR plant species, and its function is likely conserved in crop plant species. [ABSTRACT FROM AUTHOR]

SUMMARY: Rapid and widespread evolution of multiple herbicide resistance in global weed species endowed by increased capacity to metabolize (degrade) herbicides (metabolic resistance) is a great threat to herbicide sustainability and global food production. Metabolic resistance in the economically damaging crop weed species Lolium rigidum is well known but a molecular understanding has been lacking. We purified a metabolic resistant (R) subset from a field evolved R L. rigidum population. The R, the herbicide susceptible (S) and derived F2 populations were used for candidate herbicide resistance gene discovery by RNA sequencing. A P450 gene CYP81A10v7 was identified with higher expression in R vs. S plants. Transgenic rice overexpressing this Lolium CYP81A10v7 gene became highly resistant to acetyl‐coenzyme A carboxylase‐ and acetolactate synthase‐inhibiting herbicides (diclofop‐methyl, tralkoxydim, chlorsulfuron) and moderately resistant to hydroxyphenylpyruvate dioxygenase‐inhibiting herbicide (mesotrione), photosystem II‐inhibiting herbicides (atrazine and chlorotoluron) and the tubulin‐inhibiting herbicide trifluralin. This wide cross‐resistance profile to many dissimilar herbicides in CYP81A10v7 transgenic rice generally reflects what is evident in the R L. rigidum. This report clearly showed that a single P450 gene in a cross‐pollinated weed species L. rigidum confers resistance to herbicides of at least five modes of action across seven herbicide chemistries. [ABSTRACT FROM AUTHOR]

The combination of herbicides with different modes of action has been adopted not only to improve weed control but also to increase the environmental sustainability of plant-protection products. In this study, we showed a synergistic effect of the auxin herbicide 2,4-D amine with the PSII-inhibiting herbicide metribuzin to control the global grass weed wild oat (Avena sterilis) population and investigated the underlying mechanisms. Pretreatment with 2,4-D amine did not change the foliar absorption of metribuzin but did increase metribuzin translocation to the roots and new leaves, although enhancement of the metribuzin metabolism was also observed. Considering that the expression level of the target site psbA gene is significantly higher in leaves than in roots, increased metribuzin translocation to new leaves is likely the major cause of the observed synergism, even though enhanced metribuzin metabolism may offset the metribuzin efficacy. This is the first report on the synergistic mechanism between 2,4-D amine and metribuzin in weed control. [ABSTRACT FROM AUTHOR]

Background and Aims Resistance to the synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) in wild radish (Raphanus raphanistrum) appears to be due to a complex, multifaceted mechanism possibly involving enhanced constitutive plant defence and alterations in auxin signalling. Based on a previous gene expression analysis highlighting the plasma membrane as being important for 2,4-D resistance, this study aimed to identify the components of the leaf plasma membrane proteome that contribute to resistance. Methods Isobaric tagging of peptides was used to compare the plasma membrane proteomes of a 2,4-D-susceptible and a 2,4-D-resistant wild radish population under control and 2,4-D-treated conditions. Eight differentially abundant proteins were then targeted for quantification in the plasma membranes of 13 wild radish populations (two susceptible, 11 resistant) using multiple reaction monitoring. Key Results Two receptor-like kinases of unknown function (L-type lectin domain-containing receptor kinase IV.1-like and At1g51820-like) and the ATP-binding cassette transporter ABCB19, an auxin efflux transporter, were identified as being associated with auxinic herbicide resistance. The variability between wild radish populations suggests that the relative contributions of these candidates are different in the different populations. Conclusions To date, no receptor-like kinases have been reported to play a role in 2,4-D resistance. The lectin-domain-containing kinase may be involved in perception of 2,4-D at the plasma membrane, but its ability to bind 2,4-D and the identity of its signalling partner(s) need to be confirmed experimentally. ABCB19 is known to export auxinic compounds, but its role in 2,4-D resistance in wild radish appears to be relatively minor. [ABSTRACT FROM AUTHOR]

Glyphosate, the most commonly used herbicide in the world, controls a wide range of plant species, mainly because plants have little capacity to metabolize (detoxify) glyphosate. Massive glyphosate use has led to world-wide evolution of glyphosate-resistant (GR) weed species, including the economically damaging grass weed Echinochloa colona. An Australian population of E. colona has evolved resistance to glyphosate with unknown mechanisms that do not involve the glyphosate target enzyme 5-enolpyruvylshikimate-3-P synthase. GR and glyphosate-susceptible (S) lines were isolated from this population and used for resistance gene discovery. RNA sequencing analysis and phenotype/genotype validation experiments revealed that one aldo-keto reductase (AKR) contig had higher expression and higher resultant AKR activity in GR than S plants. Two full-length AKR (EcAKR4-1 and EcAKR4-2) complementary DNA transcripts were cloned with identical sequences between the GR and S plants but were upregulated in the GR plants. Rice (Oryza sativa) calli and seedlings overexpressing EcAKR4-1 and displaying increased AKR activity were resistant to glyphosate. EcAKR4-1 expressed in Escherichia coli can metabolize glyphosate to produce aminomethylphosphonic acid and glyoxylate. Consistent with these results, GR E. colona plants exhibited enhanced capacity for detoxifying glyphosate into aminomethylphosphonic acid and glyoxylate. Structural modeling predicted that glyphosate binds to EcAKR4-1 for oxidation, and metabolomics analysis of EcAKR4-1 transgenic rice seedlings revealed possible redox pathways involved in glyphosate metabolism. Our study provides direct experimental evidence of the evolution of a plant AKR that metabolizes glyphosate and thereby confers glyphosate resistance. [ABSTRACT FROM AUTHOR]

• Trifluralin resistance inheritance in a Lolium rigidum population is determined. • Plants in this population contain the α-tubulin mutation Val-202-Phe. • Resistance is largely inherited as a monogenic, recessive nuclear trait. • Polygenic resistance may also be involved, especially at low herbicide rates. • Recessive resistance has implications in resistance evolution and management. The increasing number of weedy species resistant to dinitroaniline herbicides warrants studies on the evolutionary factors contributing to resistance evolution, including genetic inheritance of resistance traits. In this study, we investigated the genetic control of trifluralin resistance in a well-characterised Lolium rigidum Gaud. population from Western Australia. This population was purified to contain plants homozygous for the Val-202-Phe α-tubulin mutation, and used as the resistant (R) parents and crossed with susceptible (S) parents to produce eight reciprocal F1 families. Trifluralin dose response curves of the eight F1 families indicate that trifluralin resistance in this population is inherited as an incomplete recessive nuclear trait. The F1 plants were crossed within each families to establish eight pseudo-F2 (ψ -F2) families. Segregation ratio of resistance and susceptibility in ψ -F2 families were determined using the discriminating trifluralin rates of 120 and 480 g a.i. ha−1. At 480 g a.i. ha−1 trifluralin, the segregation ratio in almost all ψ -F2 families (except one) was fit to 1:3 (resistance: susceptibility) one recessive gene control model. However, at 120 g a.i. ha−1 trifluralin, the segregation ratios in half of the families did not fit this model, indicating involvement of one or more genes in resistance at the lower rate. These results showed complexity of genetic inheritance of trifluralin resistance in this L. rigidum population possessing the Val-202-Phe α-tubulin mutation. [ABSTRACT FROM AUTHOR]

Background and Aims Resistance to synthetic auxin herbicides such as 2,4-dichlorophenoxyacetic acid (2,4-D) is increasing in weed populations worldwide, which is of concern given the recent introduction of synthetic auxin-resistant transgenic crops. Due to the complex mode of action of the auxinic herbicides, the mechanisms of evolved resistance remain largely uncharacterized. The aims of this study were to assess the level of diversity in resistance mechanisms in 11 populations of the problem weed Raphanus raphanistrum, and to use a high-throughput, whole-genome transcriptomic analysis on one resistant and one susceptible population to identify important changes in gene expression in response to 2,4-D. Methods Levels of 2,4-D and dicamba (3,6-dichloro-2-methoxybenzoic acid) resistance were quantified in a dose–response study and the populations were further screened for auxin selectivity, 2,4-D translocation and metabolism, expression of key 2,4-D-responsive genes and activation of the mitogen-activated proein kinase (MAPK) pathway. Potential links between resistance levels and mechanisms were assessed using correlation analysis. Key Results The transcriptomic study revealed early deployment of the plant defence response in the 2,4-D-treated resistant population, and there was a corresponding positive relationship between auxinic herbicide resistance and constitutive MAPK phosphorylation across all populations. Populations with shoot-wide translocation of 2,4-D had similar resistance levels to those with restricted translocation, suggesting that reduced translocation may not be as strong a resistance mechanism as originally thought. Differences in auxin selectivity between populations point to the likelihood of different resistance-conferring alterations in auxin signalling and/or perception in the different populations. Conclusions 2,4-D resistance in wild radish appears to result from subtly different auxin signalling alterations in different populations, supplemented by an enhanced defence response and, in some cases, reduced 2,4-D translocation. This study highlights the dangers of applying knowledge generated from a few populations of a weed species to the species as a whole. [ABSTRACT FROM AUTHOR]

The evolution of resistant weed populations in response to intensive herbicide selection pressure is a global issue. Resistance to post-emergence herbicides is widespread, whereas soil-applied pre-emergence herbicides can often remain effective. For example, in Australia pyroxasulfone is a new pre-emergence soil-applied herbicide which provides control of Lolium rigidum populations resistant to multiple post-emergence herbicide modes of action. A fundamental knowledge of the genetic basis of metabolic resistance in weeds is important for understanding plant evolution pathways under herbicide selection and sustaining long-term weed resistance management. In this study we define the mechanistic basis of resistance to pyroxasulfone in a L. rigidum population. TLC provides evidence that pyroxasulfone resistance is metabolism-based with approximately 88% of parental [ 14 C]-labelled pyroxasulfone metabolized in resistant plants 24 h after the herbicide treatment. HPLC-MS allowed identification of several metabolites of pyroxasulfone formed via a glutathione (GSH) conjugation pathway in pyroxasulfone-resistant L. rigidum plants . However, the initial pyroxasulfone-glutathione conjugate was not found likely due to its labile nature. The observed constitutive over-expression from six to nine-fold of two putative resistance-endowing GST genes was associated with the pyroxasulfone resistance phenotype. The most logical conclusion, based on the data thus far available, is that rapid detoxification of pyroxasulfone mediates pyroxasulfone resistance in L. rigidum plants. Future research is warranted to confirm the hypothesis advanced by this study of rapid pyroxasulfone metabolism due to GSH conjugation mediated by GST over-expressed in pyroxasulfone-resistant plants which similarly leads to the production of distinctive GSH-pyroxasulfone metabolites in L. rigidum and wheat. [ABSTRACT FROM AUTHOR]

A novel glyphosate resistance double point mutation (T102I/P106S, TIPS) in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene has been recently identified for the first time only in the weed species Eleusine indica. Quantification of plant resistance cost associated with the TIPS and the often reported glyphosate resistance single P106S mutation was performed. A significant resistance cost (50% in seed number currency) associated with the homozygous TIPS but not the homozygous P106S EPSPS variant was identified in E. indica plants. The resistance cost associated with the TIPS mutation escalated to 85% in plants under resource competition with rice crops. The resistance cost was not detected in nonhomozygous TIPS plants denoting the recessive nature of the cost associated with the TIPS allele. An excess of 11-fold more shikimate and sixfold more quinate in the shikimate pathway was detected in TIPS plants in the absence of glyphosate treatment compared to wild type, whereas no changes in these compounds were observed in P106S plants when compared to wild type. TIPS plants show altered metabolite levels in several other metabolic pathways that may account for the expression of the observed resistance cost. [ABSTRACT FROM AUTHOR]

Herbicides have been the primary tool for controlling large populations of yield depleting weeds from agro-ecosystems, resulting in the evolution of widespread herbicide resistance. In response, nonherbicidal techniques have been developed which intercept weed seeds at harvest before they enter the soil seed bank. However, the efficiency of these techniques allows an intense selection for any trait that enables weeds to evade collection, with early-flowering ecotypes considered likely to result in early seed shedding. Using a field-collected wild radish population, five recurrent generations were selected for early maturity and three generations for late maturity. Phenology associated with flowering time and growth traits were measured. Our results demonstrate the adaptive capacity of wild radish to halve its time to flowering following five generations of early-flowering selection. Early-maturing phenotypes had reduced height and biomass at maturity, leading to less competitive, more prostrate growth forms. Following three generations of late-flowering selection, wild radish doubled its time to flowering time leading to increased biomass and flowering height at maturity. This study demonstrates the potential for the rapid evolution in growth traits in response to highly effective seed collection techniques that imposed a selection on weed populations within agro-ecosystems at harvest. [ABSTRACT FROM AUTHOR]

The rate of herbicide resistance evolution in plants depends on fitness traits endowed by alleles in both the presence and absence (resistance cost) of herbicide selection. The effect of two Lolium rigidum spontaneous homozygous target-site resistance-endowing mutations (Ile-1781-Leu, Asp-2078-Gly) on both ACCase activity and various plant growth traits have been investigated here. Relative growth rate (RGR) and components (net assimilation rate, leaf area ratio), resource allocation to different organs, and growth responses in competition with a wheat crop were assessed. Unlike plants carrying the Ile-1781-Leu resistance mutation, plants homozygous for the Asp-2078-Gly mutation exhibited a significantly lower RGR (30%), which translated into lower allocation of biomass to roots, shoots, and leaves, and poor responses to plant competition. Both the negligible and significant growth reductions associated, respectively, with the Ile-1781-Leu and Asp-2078-Gly resistance mutations correlated with their impact on ACCase activity. Whereas the Ile-1781-Leu mutation showed no pleiotropic effects on ACCase kinetics, the Asp-2078-Gly mutation led to a significant reduction in ACCase activity. The impaired growth traits are discussed in the context of resistance costs and the effects of each resistance allele on ACCase activity. Similar effects of these two particular ACCase mutations on the ACCase activity of Alopecurus myosuroides were also confirmed. [ABSTRACT FROM AUTHOR]

Glyphosate is the most important and widely used herbicide in world agriculture. Intensive glyphosate selection has resulted in the widespread evolution of glyphosate-resistant weed populations, threatening the sustainability of this valuable once-in-a-century agrochemical. Field-evolved glyphosate resistance due to known resistance mechanisms is generally low to modest. Here, working with a highly glyphosate-resistant Eleusine indica population, we identified a double amino acid substitution (T102I + P106S [TIPS]) in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene in glyphosate-resistant individuals. This TIPS mutation recreates the biotechnology-engineered commercial first generation glyphosate-tolerant EPSPS in corn (Zea mays) and now in other crops. In E. indica, the naturally evolved TIPS mutants are highly (more than 180-fold) resistant to glyphosate compared with the wild type and more resistant (more than 32-fold) than the previously known P106S mutants. The E. indica TIPS EPSPS showed very high-level (2,647-fold) in vitro resistance to glyphosate relative to the wild type and is more resistant (600-fold) than the P106S variant. The evolution of the TIPS mutation in crop fields under glyphosate selection is likely a sequential event, with the P106S mutation being selected first and fixed, followed by the T102I mutation to create the highly resistant TIPS EPSPS. The sequential evolution of the TIPS mutation endowing high-level glyphosate resistance is an important mechanism by which plants adapt to intense herbicide selection and a dramatic example of evolution in action. [ABSTRACT FROM AUTHOR]

Background and Aims Dormancy in Lolium rigidum (annual ryegrass) seeds can be alleviated by warm stratification in the dark or by application of fluridone, an inhibitor of plant abscisic acid (ABA) biosynthesis via phytoene desaturase. However, germination and absolute ABA concentration are not particularly strongly correlated. The aim of this study was to determine if cytokinins of both plant and bacterial origin are involved in mediating dormancy status and in the response to fluridone. Methods Seeds with normal or greatly decreased (by dry heat pre-treatment) bacterial populations were stratified in the light or dark and in the presence or absence of fluridone in order to modify their dormancy status. Germination was assessed and seed cytokinin concentration and composition were measured in embryo-containing or embryo-free seed portions. Key Results Seeds lacking bacteria were no longer able to lose dormancy in the dark unless supplied with exogenous gibberellin or fluridone. Although these seeds showed a dramatic switch from active cytokinin free bases to O-glucosylated storage forms, the concentrations of individual cytokinin species were only weakly correlated to dormancy status. However, cytokinins of apparently bacterial origin were affected by fluridone and light treatment of the seeds. Conclusions It is probable that resident microflora contribute to dormancy status in L. rigidum seeds via a complex interaction between hormones of both plant and bacterial origin. This interaction needs to be taken into account in studies on endogenous seed hormones or the response of seeds to plant growth regulators. [ABSTRACT FROM AUTHOR]

Weed control failures due to herbicide resistance are an increasing and worldwide problem that significantly affect crop yields. Metabolism-based herbicide resistance (referred to as metabolic resistance) in weeds is not well characterized at the genetic level. An RNA-Seq transcriptome analysis was used to find candidate genes that conferred metabolic resistance to the herbicide diclofop in a diclofop-resistant population (R) of the major global weed Lolium rigidum. A reference c DNA transcriptome (19 623 contigs) was assembled and assigned putative annotations. Global gene expression was measured using Illumina reads from untreated control, adjuvant-only control, and diclofop treatment of R and susceptible (S). Contigs that showed constitutive expression differences between untreated R and untreated S were selected for further validation analysis, including 11 contigs putatively annotated as cytochrome P450 (CytP450), glutathione transferase ( GST), or glucosyltransferase ( GT), and 17 additional contigs with annotations related to metabolism or signal transduction. In a forward genetics validation experiment, nine contigs had constitutive up-regulation in R individuals from a segregating F2 population, including three CytP450, one nitronate monooxygenase ( NMO), three GST, and one GT. Principal component analysis using these nine contigs differentiated F2-R from F2-S individuals. In a physiological validation experiment in which 2,4-D pre-treatment induced diclofop protection in S individuals due to increased metabolism, seven of the nine genetically validated contigs were induced significantly. Four contigs (two CytP450, NMO, and GT) were consistently highly expressed in nine field-evolved metabolic resistant L. rigidum populations. These four contigs were strongly associated with the resistance phenotype and are major candidates for contributing to metabolic diclofop resistance. [ABSTRACT FROM AUTHOR]

Highlights: [•] Semi-dominant alleles segregate at one major locus for pyroxasulfone resistance. [•] Cross-resistance is also conferred by major locus(i). [•] A combination of herbicide options can reduce pyroxasulfone selection pressure. [•] This study will enable more accurate predictions by computational modelling. [•] Inheritance studies of novel herbicides can help sustain their longevity. [ABSTRACT FROM AUTHOR]

Synthetic herbicides have been used globally to control weeds in major field crops. This has imposed a strong selection for any trait that enables plant populations to survive and reproduce in the presence of the herbicide. Herbicide resistance in weeds must be minimized because it is a major limiting factor to food security in global agriculture. This represents a huge challenge that will require great research efforts to develop control strategies as alternatives to the dominant and almost exclusive practice of weed control by herbicides. Weed scientists, plant ecologists and evolutionary biologists should join forces and work towards an improved and more integrated understanding of resistance across all scales. This approach will likely facilitate the design of innovative solutions to the global herbicide resistance challenge. [ABSTRACT FROM AUTHOR]

Weed populations can have high genetic plasticity and rapid responses to environmental selection pressures. For example, 100-fold amplification of the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene evolved in the weed species Amaranthus palmeri to confer resistance to glyphosate, the world’s most important herbicide. However, the gene amplification mechanism is unknown. We sequenced the EPSPS gene and genomic regions flanking EPSPS loci in A. palmeri, and searched for mobile genetic elements or repetitive sequences. The EPSPS gene was 10,229 bp, containing 8 exons and 7 introns. The gene amplification likely proceeded through a DNA-mediated mechanism, as introns exist in the amplified gene copies and the entire amplified sequence is at least 30 kb in length. Our data support the presence of two EPSPS loci in susceptible (S) A. palmeri, and that only one of these was amplified in glyphosate-resistant (R) A. palmeri. The EPSPS gene amplification event likely occurred recently, as no sequence polymorphisms were found within introns of amplified EPSPS copies from R individuals. Sequences with homology to miniature inverted-repeat transposable elements (MITEs) were identified next to EPSPS gene copies only in R individuals. Additionally, a putative Activator (Ac) transposase and a repetitive sequence region were associated with amplified EPSPS genes. The mechanism controlling this DNA-mediated amplification remains unknown. Further investigation is necessary to determine if the gene amplification may have proceeded via DNA transposon-mediated replication, and/or unequal recombination between different genomic regions resulting in replication of the EPSPS gene. [ABSTRACT FROM AUTHOR]

Background and Aims Karrikinolide (KAR1) is a smoke-derived chemical that can trigger seeds to germinate. A potential application for KAR1 is for synchronizing the germination of weed seeds, thereby enhancing the efficiency of weed control efforts. Yet not all species germinate readily with KAR1, and it is not known whether seemingly non-responsive species can be induced to respond. Here a major agronomic weed family, the Brassicaceae, is used to test the hypothesis that a stimulatory response to KAR1 may be present in physiologically dormant seeds but may not be expressed under all circumstances. Methods Seeds of eight Brassicaceae weed species (Brassica tournefortii, Raphanus raphanistrum, Sisymbrium orientale, S. erysimoides, Rapistrum rugosum, Lepidium africanum, Heliophila pusilla and Carrichtera annua) were tested for their response to 1 µm KAR1 when freshly collected and following simulated and natural dormancy alleviation, which included wet–dry cycling, dry after-ripening, cold and warm stratification and a 2 year seed burial trial. Key Results Seven of the eight Brassicaceae species tested were stimulated to germinate with KAR1 when the seeds were fresh, and the remaining species became responsive to KAR1 following wet–dry cycling and dry after-ripening. Light influenced the germination response of seeds to KAR1, with the majority of species germinating better in darkness. Germination with and without KAR1 fluctuated seasonally throughout the seed burial trial. Conclusions KAR1 responses are more complex than simply stating whether a species is responsive or non-responsive; light and temperature conditions, dormancy state and seed lot all influence the sensitivity of seeds to KAR1, and a response to KAR1 can be induced. Three response types for generalizing KAR1 responses are proposed, namely inherent, inducible and undetected. Given that responses to KAR1 were either inherent or inducible in all 15 seed lots included in this study, the Brassicaceae may be an ideal target for future application of KAR1 in weed management. [ABSTRACT FROM AUTHOR]