Characterisation and molecular basis of ALS inhibitor resistance in the grass weed Alopecurus myosuroides

Several UK populations of the grass weed Alopecurus myosuroides were identified where high proportions of individuals showed resistance to the acetolactate synthase (ALS)‐inhibiting herbicides, mesosulfuron‐methyl + iodosulfuron‐methyl sodium mixture and sulfometuron‐methyl. Screening with sulfometuron, followed by DNA sequencing of the ALS gene from resistant and susceptible individuals, led to the identification of eight populations where a single point mutation segregated with resistance to sulfometuron. All highly resistant individuals from seven of eight populations showed a single‐nucleotide polymorphism (SNP) in the first position of the Pro197 codon of an A. myosuroides ALS gene, conferring a predicted proline to threonine target‐site change. One population showed resistant individuals with single‐nucleotide polymorphism in the second position of the Trp574 codon, conferring a predicted tryptophan to leucine substitution. No other mutations segregating with resistance were found. Enzyme assays confirmed that resistance was due to an altered form of ALS enzyme, which was less susceptible to inhibition by sulfonylureas, making this one of the first fully characterised cases of ALS target‐site resistance in a European grass weed. Increased information regarding the nature and distribution of ALS target‐site mutation may help support sustainable management strategies, allowing continued use of mesosulfuron + iodosulfuron against this weed in the UK.     

Resistance to ACCase-inhibiting herbicides and isoproturon in UK populations of Lolium multiflorum: mechanisms of resistance and implications for control

Herbicide-resistant Lolium multiflorum (Italian rye-grass) was first reported in the UK in 1993 and had been confirmed on 25 farms by 1999. In this study, resistance to five herbicides belonging to the aryloxyphenoxypropionate, cyclohexanedione and phenyl-urea classes was determined in six populations of L multiflorum from the UK under glasshouse and simulated field conditions. Glasshouse conditions tended to exaggerate the degree of resistance, but experiments performed in both environments detected resistance in four populations of L multiflorum. Four populations (Essex A1, Lincs A1, Wilts B1, Yorks A2) were resistant to diclofop-methyl, fluazifop-P-butyl, tralkoxydim and partially resistant to isoproturon, but only the population from Yorkshire (Yorks A2) showed resistance to cycloxydim. Biochemical analyses of acetyl coenzyme A carboxylase (ACCase) activity, oxygen consumption by thylakoids, diclofop metabolism and glutathione S-transferase activity showed that, in three of the resistant populations, an enhanced rate of herbicide metabolism conferred resistance. This is the first report world-wide of an enhanced metabolism mechanism of diclofop resistance in L multiflorum. In the Yorks A2 population, an insensitive ACCase was detected (target-site resistance) which also conferred cross-resistance to all of the other ACCase inhibitors investigated.     

Ecophysiological factors contributing to the invasion of Panicum maximum into native Miscanthus sinensis grassland in Taiwan

Panicum maximum (guinea grass), a native grass of Africa, has invaded and displaced the native Miscanthus sinensis grassland at Dadu terrace in central Taiwan. The Dadu terrace has distinct wet and dry seasons and is prone to fire during the dry seasons. We compared photosynthesis and growth, as well as the growth response to two water treatments, between P. maximum and M. sinensis plants to understand ecophysiological factors contributing to the successful invasion of the grass. In comparison with M. sinensis, P. maximum had a significantly higher photosynthetic rate, larger specific leaf area and a higher leaf area/total biomass ratio, and thus grew faster and higher, and accumulated more biomass under well‐watered conditions. The growth of P. maximum was more susceptible than M. sinensis to drought stress. Faster growth of seedlings and more biomass accumulation of mature plants would give P. maximum a highly competitive advantage over M. sinensis for resource acquisition in the Dadu terrace during the wet season, which might contribute to the successful invasion of P. maximum into the grassland of M. sinensis.     

New, quick tests for herbicide resistance in black-grass (Alopecurus myosuroides Huds) based on increased glutathione S-transferase activity and abundance

Black-grass (Alopecurus myosuroides Huds) is a major grass weed in winter cereals in Europe. It reduces yields and can act as a secondary host for a range of diseases. Herbicide resistance in this species was first detected in the UK in the early 1980s, and has now been reported in thirty counties. To successfully manage herbicide resistance it is vital that suspect populations are tested so that appropriate action can be taken. Ideally, a test will be quick, cheap and easy to use. Furthermore, it should provide an unequivocal result before post-emergence herbicides are to be applied, allowing alternative strategies to be adopted where necessary. This paper reports the development of new tests for herbicide resistance based on our observation that the resistant black-grass biotype Peldon contains approximately double the activity of the enzyme glutathione S-transferase (GST) compared with susceptible biotypes. Data are presented on the production of a monoclonal antiserum to a novel 30 kDa GST polypeptide purified from the biotype Peldon. An ELISA using this antiserum is described and the utility of this assay to detect resistant black-grass biotypes in plants grown under glass and in the field is presented. In addition, a microtitre assay for GST activity is described, which allows the rapid assessment of GST activities of plants. Both abundance and activity of GSTs are discussed as markers for herbicide resistance in black-grass.     

Growth performance of Vallisneria spiralis under oligotrophic conditions supports its potential invasiveness in mid‐elevation freshwaters

Under the influence of human‐driven impacts, inland aquatic habitats of intermediate to mid‐high elevations may provide new areas for the establishment of lowland macrophytes that usually act as opportunistic species. Vallisneria spiralis, an obligate lowland macrophyte with an altitudinal range not exceeding 300 m, is a typical component of the plant diversity in meso‐ to eutrophic water habitats. This species is an engineering species in ecosystems, being able to modify the colonised sediments promoting its self‐maintenance under eutrophic conditions. Vallisneria spiralis delivers large amounts of oxygen to roots promoting the loss of nitrogen via denitrification and reducing the internal nutrient load of colonised environments. However, the establishment of the species results in rapid sediment accumulation and the progressive loss of more sensitive macrophytes. Furthermore, V. spiralis is one of the most widespread alien aquatic weeds in Europe. In this study, we evaluated the growth performance of V. spiralis under oligotrophic conditions at the upper limit of its altitudinal distribution using two different techniques: harvesting and leaf‐marking. Vallisneria spiralis proved to be able to grow under oligotrophic conditions, even if at rates lower than those recorded under eutrophic conditions. Accordingly, V. spiralis represents a potential threat for oligotrophic aquatic habitats at moderate elevations. However, the high variability observed in the growth responses highlights the need of further investigation to evaluate the key factors involved in the establishment and survival of the species.     

psbA Mutation (valine219 to isoleucine) in Poa annua resistant to metribuzin and diuron

The herbicide‐binding region of the chloroplast psbA gene from a total of 20 biotypes of Poa annua L resistant and susceptible to metribuzin and diuron was selectively amplified using PCR. Sequence analysis of the fragment from six herbicide‐resistant biotypes of P annua exhibited a substitution from valine to isoleucine at position 219 of the D1 protein encoded by the psbA gene. This is the same mutation as reported for Chlamydomonas and Synechococcus through site‐directed mutagenesis and in cell cultures of Chenopodium rubrum L. To our knowledge this is the first report of a higher plant exhibiting resistance in the field to photosystem II inhibitors due to a psbA mutation other than at position 264. The existence of additional biotypes of P annua resistant to diuron or metribuzin but lacking mutation in the herbicide‐binding region indicates that resistance to these herbicides can also be attained by other mechanisms. © 2000 Society of Chemical Industry     

Defining the habitat niche of Alopecurus myosuroides at the field scale

The distribution of Alopecurus myosuroides (black‐grass) in fields is patchy. The locations of these patches can be influenced by the environment. This presents an opportunity for precision management through patch spraying. We surveyed five fields on various types of soil using a nested sampling design and recorded both A. myosuroides seedlings in autumn and seed heads in summer. We also measured soil properties at those sampling locations. We found that the patches of seed heads within a field were smaller than the seedling patches, suggesting that techniques for patch spraying based on maps of heads in the previous season could be inherently risky. We also found that the location of A. myosuroides patches within fields can be predicted through their relationship with environmental properties and that these relations are consistent across fields on different soil types. This improved understanding of the relations between soil properties and A. myosuroides seedlings could allow farmers to use pre‐existing or suitably supplemented soil maps already in use for the precision application of fertilisers as a starting point in the creation of herbicide application maps.     

Interpopulation variability and adaptive potential for reduced glyphosate sensitivity in Alopecurus myosuroides

Glyphosate use in the United Kingdom has more than doubled in the last 20 years. Much of this increase is driven by efforts to control herbicide resistant weeds, particularly Alopecurus myosuroides, prior to crop drilling. There is precedent for evolution of glyphosate resistance in similar situations, raising concerns over the sustainability of glyphosate use in the UK. We used dose–response experiments to examine variation in glyphosate sensitivity amongst 40 field‐collected A. myosuroides populations. No populations were resistant to glyphosate, but ED₉₀ values ranged between 354 and 610 g a.i. ha^?1. Five populations had ED₉₀ values significantly higher than the unexposed control population collected from a site at Rothamsted Research with no previous glyphosate exposure. Recurrent selection experiments were performed to determine whether variation in glyphosate sensitivity had a heritable basis. Following two rounds of selection, five of six field populations evolved significantly reduced sensitivity to glyphosate, with R/S ratios, based on estimated ED₅₀ values, ranging from 1.2 to 1.5. These results confirm that there is a heritable basis to variation in glyphosate sensitivity. The response to selection was modest. Evolved populations were not highly resistant to glyphosate, although some twice‐selected individuals survived recommended field rates. These results do not represent definitive proof of the potential of A. myosuroides to evolve glyphosate resistance, although they do indicate caution is needed when considering the sustainability of increased glyphosate use to control this herbicide resistance‐prone species.     

The phytophagous insects on lantana in brazil and their potential for biological control in Australia

Abstract

A ten‐year survey of phytophagous insects associated with Lantana spp. (chiefly L. tiliaefolia Cham, and L. glutinosa Poepp.) in Brazil, yielded 345 species from eight orders. Half of the total species collected were Coleoptera, with Chrysomelidae being the most important family. Other important families were the Curculionidae and, in the Lepidoptera, Geometridae, Arctiidae and Noctuidae. More than 90% of insect species fed on the vegetative parts of the plant, with almost two‐thirds of these chewing holes in leaves, and fewer than 10% attacking the flowers and fruits. Woody stem borers were rare, constituting only 2% of the total insect species collected. The present status of biological control of lantana in Australia is reviewed together with the prospects of importing further biological control candidates from Brazil.     

Developmental changes in glutathione S‐transferase activity in herbicide‐resistant populations of Alopecurus myosuroides Huds (black‐grass) in the field

Herbicide‐resistant populations of Alopecurus myosuroides Huds (black‐grass) have become widespread throughout the UK since the early 1980s. Clear evidence suggests that more than one resistance mechanism exists, and glutathione S‐transferases (GSTs) have been implicated in resistance due to enhanced metabolism. This study reports the determination of GST activity in four UK black‐grass populations from field sites situated in the East Midlands. Data demonstrate that, as untreated plants in the field mature, there is an accompanying natural elevation of GST activity with natural environmental changes from winter to spring. We speculate that this endogenous change in enzyme activity with plant development in the field contributes to reduced efficacy of some graminicides applied in the spring. These observations are discussed in relation to predicting herbicide efficacy to achieve maximum control of this important grass weed. © 2001 Society of Chemical Industry     

Evaluating the potential of Unmanned Aerial Systems for mapping weeds at field scales: a case study with Alopecurus myosuroides

Mapping weed densities within crops has conventionally been achieved either by detailed ecological monitoring or by field walking, both of which are time‐consuming and expensive. Recent advances have resulted in increased interest in using Unmanned Aerial Systems (UAS ) to map fields, aiming to reduce labour costs and increase the spatial extent of coverage. However, adoption of this technology ideally requires that mapping can be undertaken automatically and without the need for extensive ground‐truthing. This approach has not been validated at large scale using UAS ‐derived imagery in combination with extensive ground‐truth data. We tested the capability of UAS for mapping a grass weed, Alopecurus myosuroides , in wheat crops. We addressed two questions: (i) can imagery accurately measure densities of weeds within fields and (ii) can aerial imagery of a field be used to estimate the densities of weeds based on statistical models developed in other locations? We recorded aerial imagery from 26 fields using a UAS . Images were generated using both RGB and R_mod (R_mod 670–750 nm) spectral bands. Ground‐truth data on weed densities were collected simultaneously with the aerial imagery. We combined these data to produce statistical models that (i) correlated ground‐truth weed densities with image intensity and (ii) forecast weed densities in other fields. We show that weed densities correlated with image intensity, particularly R_mod image data. However, results were mixed in terms of out of sample prediction from field‐to‐field. We highlight the difficulties with transferring models and we discuss the challenges for automated weed mapping using UAS technology.     

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.     

云南省六个国家级自然保护区外来入侵草本植物与本地植物的关系

为明确不同自然保护区外来入侵草本植物及其与本地植物多样性之间的关系,以我国生物多样性丰富的云南省为研究对象,于2014年对云南省6个国家级自然保护区的82个1 m2小样方(78个100 m2大样方)中外来入侵草本植物和本地植物物种丰富度和密度进行调查,并分析外来入侵草本植物物种丰富度与本地草本植物物种丰富度、外来入侵草本植物密度与本地草本植物密度的关系及对不同自然保护区外来入侵草本植物物种丰富度的差异进行分析。结果显示,共调查发现22种外来入侵草本植物,其中纳板河流域国家级自然保护区种类最多,达14种,金平分水岭国家级自然保护区仅有2种;6个国家级自然保护区本地草本植物的物种丰富度与外来入侵草本植物的物种丰富度之间、本地草本植物密度与外来入侵草本植物密度之间均呈现正、负2种相关关系,其中哀牢山国家级自然保护区本地草本植物的物种丰富度与外来入侵草本植物物种丰富度呈显著负相关,其它5个国家级自然保护区的相关性均不显著。简单回归分析表明自然保护区外来入侵草本植物物种丰富度与本地草本植物物种丰富度、本地草本植物盖度、本地乔灌木物种丰富度呈显著负相关,多元回归分析结果表明乔灌木物种丰富度和经纬度是造成不同自然保护区外来入侵草本植物物种多样性差异的主要原因。     

豆禾混播建植人工草地对牧草产量和草质的影响

以内蒙古锡林浩特内蒙古大学草地生态学研究基地为试验样地,通过对单播和混播情况下豆科牧草和禾本科牧草产量、竞争能力、牧草营养价值等测定分析,比较不同播种方式对牧草生长的影响。结果表明:豆禾混播情况下,牧草产量有较为明显的提高,尤其是第2年混播草地中禾本科牧草的产量较第1年有了提高。在豆禾播种比例1∶2的情况下,混播牧草在一定程度上避免了种间竞争,2 a内混播草地的相对产量总值均大于1,表明其生长较为协调,豆科和禾本科牧草双方均受益。豆禾混播时牧草的营养价值有所改变,豆禾混播草地粗蛋白含量增加,粗纤维含量下降,营养价值有所提高。因此,豆禾混播草地稳定性高,牧草产量和营养价值较高,有利于大面积推广,是建植人工草地的较好选择。     

Dormancy,germination and emergence of Urochloa panicoides regulated by temperature

Urochloa panicoides is an annual weed of summer crops. In Argentina, in subhumid areas with monsoon rainfall, it germinates and establishes in a single flush. To (i) identify the environmental factors that modify its seed dormancy level and germination and (ii) quantify the parameters describing the thermal behaviour of the germination and emergence dynamics of this weed under non‐limiting water conditions, we established a set of germination experiments performed (i) under controlled conditions using seeds after ripened for 3 or 6 months in different thermal and hydric conditions and (ii) under field conditions, where the soil temperature was modified by applying different shading levels. Seed dormancy level remained high with 3 months after ripening in all treatments. After 6 months, seeds stored at 4°C in dry conditions did not germinate at any temperature, while seeds stored at 25°C in dry conditions and in situ germinated c. 20% and 60% respectively. Germination percentage was higher in seeds harvested before their natural dispersal. The base, optimum and maximum temperatures for seed germination were 6, 35 and 45°C respectively. Shading reduced the number of emerged seedlings, possibly by reducing the soil thermal amplitude. The results explained the dormancy‐breaking mechanism of U. panicoides that allows a high germination rate in the field when rainfall occurs.     

Genetic analysis and molecular mapping of resistance to Puccinia striiformis f. sp. pseudo‐hordei in common wheat

This is the first genetic study reporting on the interaction and molecular mapping of resistance to the barley grass stripe rust pathogen (Puccinia striiformis f. sp. pseudo‐hordei, Psph) in common wheat. Seedlings of 638 wheat accessions were tested and it was determined that wheat is a near‐nonhost to Psph based on rare susceptibility observed in <2% of commercial cultivars and <5% of wheat landraces. As previously observed for P. striiformis f. sp. tritici (Pst), the Australian cultivar Teal was highly susceptible to Psph. In contrast, a selection of cv. Avocet carrying complementary resistance genes Yr73 and Yr74 (Avocet R; AvR) was resistant. The Teal × AvR (T/A) doubled haploid (DH) population was used to map resistance in AvR to Psph. Infection types on the T/A DH lines inoculated with Psph and Pst indicated that all DH lines carrying both Yr73 and Yr74 were also resistant to Psph; however, fewer DH lines were susceptible to Psph than expected, suggesting the resistance was more complex. QTL analysis using 9053 DArT‐Seq markers determined that resistance to Psph was polygenically inherited and mapped to chromosomes 3A, 3D, 4A and 5B. The 3DL and 5BL markers co‐located with Yr73 and Yr74, suggesting an overlap between host and non‐host resistance mechanisms.     

Combining mechanical rhizome removal and cover crops for Elytrigia repens control in organic barley systems

Mechanical weed control of perennial weeds in organic crop production over long post‐harvest periods is incompatible with the establishment of cover crops for improving soil quality and preventing nutrient leaching. We suggest a new concept that comprises uprooting and immediate removal of vegetative propagules located within the plough layer to allow for quick re‐establishment of a plant cover. A field experiment comparing the effects of conventional practices (stubble cultivation) with different combinations of rotary cultivation (One, Two or four passes) and cover crops (none vs. rye‐vetch‐mustard mixture) on Elytrigia repens rhizome removal, shoot growth and suppression of a subsequent barley crop was examined in two growing seasons. Four passes with a modified rotary cultivator, where each pass was followed by rhizome removal, reduced E. repens shoot growth in barley by 84% and 97%. In general, the cover crop developed poorly and did not affect barley or E. repens. Barley yield was only affected by treatments in the first season, where yield was negatively correlated with E. repens shoot biomass. The concept has potential for the control of severe E. repens infestations, but future research aimed at identifying more effective smother crops and less intensive methods of rhizome removal is needed.     

Assessing the efficacy and impact of management of an invasive species in a protected area: Poa annua on sub‐Antarctic Macquarie Island

Plant eradication is difficult, particularly in remote, protected areas. The Southern Ocean Islands are very isolated and highly protected, but the flora contains many alien plants. Small restricted populations have been eradicated, but eradication of established species has proven difficult. A better understanding of the efficacy of control methods at sub‐Antarctic temperatures and their off‐target impacts may increase eradication success. With interest in controlling non‐native Poa annua in the region, we aimed to determine if physical and chemical methods can control P. annua (the sub‐Antarctic biotype) in sub‐Antarctic conditions and examined their impact on native plants. We quantified the effectiveness of physical control methods on P. annua in situ on sub‐Antarctic Macquarie Island through field‐based experiments and assessed their selectivity on P. annua compared with native grasses. We also quantified the effectiveness of several herbicides on P. annua at sub‐Antarctic temperatures and assessed their selectivity on native grasses. Of the four physical disturbance methods tested, none effectively reduced P. annua cover as one‐off treatments. Of the herbicide treatments, glyphosate and trifloxysulfuron sodium were effective and were less damaging to native grass species, indicating potential selectivity. Physical control was of limited effectiveness, but did not affect native species richness. An integrated weed management programme utilising the strategic use of selective herbicides with follow‐up chemical and physical intervention may balance control and biodiversity outcomes. This research highlights the importance of site‐specific testing of control methods and understanding off‐target impacts of control when managing alien plant species in protected areas.