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

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

Effects of mixtures of allelopathic plant water extracts and a herbicide on weed suppression.

The present study investigated integrated effects of two allelopathic plant water extracts (WE) (Ambrosia artemisiifolia [AMBEL] and Xanthium strumarium [XANST]) and a herbicide (mesotrione) on morphological (height and fresh weight of plants) and physiological (pigments content) parameters of Abutilon theophrasti and Chenopodium album. Also, the study aimed to identify the main components of AMBEL and XANST WE and to evaluate their potential allelopathic effects. Of the 18 investigated compounds, 13 were detected in both tested WE, and p-coumaric acid was the leading component in AMBEL, while quinic acid was the predominant component of XANST. The WE of both weed species and their mixtures with the herbicide exhibited more powerful allelopathic effects on fresh weight and content of pigments than on the height of A. theophrasti and C. album. The results showed that all measured parameters of both weeds were inhibited in treatments with mesotrione and its mix with AMBEL and XANST WE. The data revealed a highly significant difference in effects (P < 0.05) between control weeds and those treated with AMBEL WE and mesotrione, where the inhibition of fresh weight was over 90%, while the inhibition of pigments content exceeded 80%, and plant height was inhibited by over 70%.

The response of Chenopodium album L. and Abutilon theophrasti Medik. to reduced doses of mesotrione.

The application of minimal doses of herbicides is very popular due to concerns about the negative impacts of herbicides on the environment and public health. Studies were conducted to estimate the possibility of using quick and non- destructive methods to investigate Chenopodium album L. and Abutilon theophrasti Medik. response to mesotrione. The studies were conducted in a controlled environment to determine the response of C. album and A. theophrasti to mesotrione using dose-response curves created based on plant dry weight, chlorophyll fluorescence parameters and chlorophyll content. The obtained effective dose values showed that the studied weeds were susceptible to reduced doses of mesotrione. ED95 values estimated for both species for dry weight and chlorophyll fluorescence parameters were lower than the recommended dose rate (120 g a.i. ha-1), with less than 85 g a.i. ha-1 needed to achieve a reduction of 95%, compared with untreated plants, while ED95 value (A. theophrasti: 182 g a.i. ha-1 and C. album: 180 g a.i. ha-1) for chlorophyll content for both species was above the recommended dose rates. Consequently, dry weight and the chlorophyll fluorescence parameters are suitable for estimating the plant response to mesotrione, while chlorophyll content is not.

Increased temperatures and elevated CO2 levels reduce the sensitivity of Conyza canadensis and Chenopodium album to glyphosate.

Herbicides are the most commonly used means of controlling weeds. Recently, there has been growing concern over the potential impacts of global climate change, specifically, increasing temperatures and elevated carbon dioxide (CO2) concentrations, on the sensitivity of weeds to herbicides. Here, glyphosate response of both Conyza canadensis and Chenopodium album was evaluated under different environmental conditions. Reduced glyphosate sensitivity was observed in both species in response to increased temperature, elevated CO2 level, and the combination of both factors. Increased temperature had greater effect on plant survival than elevated CO2 level. In combination, high temperature and elevated CO2 level resulted in loss of apical dominance and rapid necrosis in glyphosate-treated plants. To investigate the mechanistic basis of reduced glyphosate sensitivity, translocation was examined using 14C-glyphosate. In plants that were subjected to high temperatures and elevated CO2 level, glyphosate was more rapidly translocated out of the treated leaf to shoot meristems and roots than in plants grown under control conditions. These results suggest that altered glyphosate translocation and tissue-specific sequestration may be the basis of reduced plant sensitivity. Therefore, overreliance on glyphosate for weed control under changing climatic conditions may result in more weed control failures.

A global perspective on the biology, impact and management of Chenopodium album and Chenopodium murale: two troublesome agricultural and environmental weeds.

Chenopodium album and C. murale are cosmopolitan, annual weed species of notable economic importance. Their unique biological features, including high reproductive capacity, seed dormancy, high persistence in the soil seed bank, the ability to germinate and grow under a wide range of environmental conditions and abiotic stress tolerance, help these species to infest diverse cropping systems. C. album and C. murale grow tall and absorb nutrients very efficiently. Both these species are allelopathic in nature and, thus, suppress the germination and growth of native vegetation and/or crop plants. These weed species infest many agronomic and horticultural crops and may cause > 90% loss in crop yields. C. album is more problematic than C. murale as the former is more widespread and infests more number of crops, and it also acts as an alternate host of several crop pests. Different cultural and mechanical methods have been used to control these weed species with varying degrees of success depending upon the cropping systems and weed infestation levels. Similarly, allelopathy and biological control have also shown some potential, especially in controlling C. album. Several herbicides have been successfully used to control these species, but the evolution of wide-scale herbicide resistance in C. album has limited the efficacy of chemical control. However, the use of alternative herbicides in rotation and the integration of chemicals and biologically based control methods may provide a sustainable control of C. album and C. murale.

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

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

Ammonium 2,2'-thiodiacetates - Selective and environmentally safe herbicides.

2,2'-Thiodiacetic acid derivatives have a wide application potential, mainly in coordination chemistry. This research indicates that quaternary ammonium 2,2'-thiodiacetate salts may also be potent herbicidal agents used in agriculture. To provide a rationale for this statement, the toxic effect by a alkyl and aryl quaternary ammonium salts (QASs) on plant growth was investigated. The phytotoxicity of these compounds was tested against cultivated monocotyledonous (spring barley) and dicotyledonous (common radish) plants, whereas herbicidal activity was investigated in relation to popular weeds species (white goosefoot, sorrel and gallant-soldier). The results showed that aliphatic QASs possessed a low phytotoxicity to food crops and that some of them (in particular triethylammonium salt) had potent and selective herbicidal properties against common weeds, such as sorrel and gallant-soldier. However, the investigated compounds appeared to be ineffective herbicides against white goosefoot.

Herbicide hormesis can act as a driver of resistance evolution in weeds - PSII-target site resistance in Chenopodium album L. as a case study.

BACKGROUND: Herbicide hormesis may play a role in the evolution of weed resistance by increasing resistance selection. A standard herbicide rate may be subtoxic to resistant plants and make them more fit than untreated plants. If this increase in fitness is ultimately expressed in reproductive traits, resistance genes can accumulate more rapidly and exacerbate resistance evolution by magnifying the selection differential between resistant and sensitive plants. The hypothesis of hormetically enhanced reproductive fitness was studied for a photosystem II (PSII) target-site resistant (TSR) biotype of Chenopodium album exposed to the triazinone metamitron in comparison with its wild-type. RESULTS: Both biotypes showed an initial hormetic growth increase at different doses leading to fitness enhancements of between 19% and 61% above untreated plants. However, hormetic effects only resulted in higher fitness at maturity in resistant plants with a maximum stimulation in seed yield of 45% above untreated plants. Applying realistic metamitron rates, reproductive fitness of resistant plants was increased by 15-32%. CONCLUSIONS: Agronomically relevant doses of metamitron induced considerable hormesis in a PSII-TSR C. album genotype leading to enhanced relative fitness through reproductive maturity. This increase in relative fitness suggests an impact on resistance selection and can compensate for the oft-reported fitness costs of the mutation studied. Field rates of herbicides can, thus, not only select for resistant plants, but also enhance their reproductive fitness. The finding that herbicide hormesis can be eco-evolutionary important may have important implications for understanding the evolution of herbicide resistance in weeds. © 2018 Society of Chemical Industry.

New angular oxazole-fused coumarin derivatives: synthesis and biological activities.

Twelve angular oxazole-fused coumarin derivatives were designed, synthesised and characterised by 1H NMR, 13C NMR and HRMS. The structure of compound 4a was further confirmed by X-ray single-crystal diffraction. The bioassay experiment results indicated that compounds 4f and 4l have high antifungal activity on the mycelium growth of 4 plant disease fungi. Especially, compound 4l has a stronger antifungal activity compare to the commercial fungicide, Carbendazim. The herbicidal activity experiment showed that 4a and 4b can significantly inhibit the taproot and caulis development of Chenopodium album seedling and have better activities than the commercial herbicide, Acetochlor.

Herbicidal activity of slow-release herbicide formulations in wheat stands infested by weeds.

The present study reports the herbicidal activity of metribuzin and tribenuron-methyl embedded in the degradable matrix of natural poly-3-hydroxybutyrate [P(3HB)/MET and P(3HB)/TBM]. The developed formulations were constructed as films and microgranules, which were tested against the weeds such as white sweet clover Melilotus albus and lamb's quarters Chenopodium album in the presence of soft spring wheat (Triticum aestivum, cv. Altaiskaya 70) as the subject crop for investigation. The activity was measured in laboratory scale experiments by determining the density and weight of the vegetative organs of weeds. The study was also aimed at testing the effect of the experimental formulation on the growth of wheat crop as dependent on the method of herbicide delivery. The experimental MET and TBM formulations showed pronounced herbicidal activity against the weed species used in the study. The effectiveness of the experimental formulations in inhibiting weed growth was comparable to and, sometimes, higher than that of the commercial formulations (positive control). The amount of the biomass of the wheat treated with the experimental herbicide formulations was significantly greater than that of the wheat treated with commercial formulations.

Natural herbicide activity of Satureja hortensis L. essential oil nanoemulsion on the seed germination and morphophysiological features of two important weed species.

The aim of the present study was to obtain an oil/water (O/W) nanoemulsion (NE) containing garden savory (Satureja hortensis) essential oil (EO) and evaluating its herbicidal activity against Amaranthus retroflexus and Chenopodium album. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were employed to determine the chemical composition of the EO. Carvacrol (55.6%) and γ-terpinene (31.9%) were the major EO components. Low energy method was applied, allowing achievement of EO nanodroplets. The NE also presented low polydispersity, and the mean droplet was below 130nm even after storage for 30d. Laboratory tests showed that the NE at different concentrations (100, 200, 400, 800, and 1000µL.L-1) significantly (P≤0.05) reduced the germination indices and the seedling's growth in dose-response. The inhibitory effect was the greatest at 800µL.L-1 NE. Overall, root length was more inhibited as compared to shoot length. Post-emergence application of NE at different concentrations (1000, 2000, 3000, 4000 and 5000µL.L-1 of EO) on 2-4 true leaves' stage of the weeds caused significant (P≤0.05) decrease in the growth factors in dose-dependent manner. Complete lethality was observed by 4000µL.L-1 NE sprayed on the weeds. Spraying of NE significantly (P≤0.05) reduced chlorophyll content in the tested weeds. Increasing in relative electrolyte leakage (REL) 1 and 5d after treatment represented significant cell membrane disruption and increased cell membrane permeability. Transmission electron microscope (TEM) pictures confirmed NE droplet size and demonstrated membrane destruction. The study approved that the NE of S. hortensis EO has herbicidal properties as it has high phytotoxic effect, and interferes with the germination, growth and physiological processes of the weeds. The production of NE from S. hortensis EO is a low energy method that offers a promising practical natural herbicide for weed control in organic agricultural systems.

Alkyl(C16, C18, C22)trimethylammonium-Based Herbicidal Ionic Liquids.

In the framework of this study a synthesis methodology and characterization of long alkyl herbicidal ionic liquids (HILs) based on four commonly used herbicides (2,4-D, MCPA, MCPP, and dicamba) are presented. New HILs were obtained with high efficiency (>95%) using an acid-base reaction between herbicidal acids and hexadecyltrimethylammonium, octadecyltrimethylammonium, and behenyltrimethylammonium hydroxides in alcoholic medium. Among all synthesized salts, only three compounds comprising the MCPP anion were liquids at room temperature. Subsequently, the influence of both the alkyl chain length and the anion structure on their physicochemical properties (thermal decomposition profiles, solubility in 10 representative solvents, surface activity, density, viscosity, and refractive index) was determined. All HILs exhibited high thermal stability as well as surface activity; however, their solubility notably depended on both the length of the carbon chain and the structure of the anion. The herbicidal efficacy of the obtained salts was tested in greenhouse and field experiments. Greenhouse testing performed on common lambsquarters (Chenopodium album L.) and flixweed (Descurainia sophia L.) as test plants indicated that HILs were characterized by similar or higher efficacy compared to commercial herbicides. The results of field trials confirmed the high activity of HILs, particularly those containing phenoxyacids as anions (MCPA, 2,4-D, and MCPP).

Antifungal and Herbicidal Effects of Fruit Essential Oils of Four Myrtus communis Genotypes.

The chemical composition of the essential oils isolated by hydrodistillation from the fruits of four selected Myrtus communis L. genotypes from Turkey was characterized by GC-FID and GC/MS analyses. 1,8-Cineole (29.20-31.40%), linalool (15.67-19.13%), α-terpineol (8.40-18.43%), α-pinene (6.04-20.71%), and geranyl acetate (3.98-7.54%) were found to be the major constituents of the fruit essential oils of all M. communis genotypes investigated. The oils were characterized by high amounts of oxygenated monoterpenes, representing 73.02-83.83% of the total oil compositions. The results of the fungal growth inhibition assays showed that the oils inhibited the growth of 19 phytopathogenic fungi. However, their antifungal activity was generally lower than that of the commercial pesticide benomyl. The herbicidal effects of the oils on the seed germination and seedling growth of Amaranthus retroflexus L., Chenopodium album L., Cirsium arvense (L.) Scop., Lactuca serriola L., and Rumex crispus L. were also determined. The oils completely or partly inhibited the seed germinations and seedling growths of the plants. The findings of the present study suggest that the M. communis essential oils might have potential to be used as natural herbicides as well as fungicides.

Fungal Phytotoxins with Potential Herbicidal Activity to Control Chenopodium album.

This review deals with the isolation and chemical and biological characterization of phytotoxins produced by Ascochyta caulina and Phoma chenopodiicola proposed as mycoherbicides for the biological control of Chenopodium album, a worldwide spread weed which causes serious problems to some agrarian crops, including sugar beet and maize. Studies on the structure activity relationships and on the modes of actions of toxins isolated are also described, as well as the optimization of analytical methods focused on selection of the best fungal toxin producers. The attempts to scale up production of these phytotoxins aimed to obtain sufficient amounts for their application in greenhouse and field trials are also reported.

Identification of a new PSII target site psbA mutation leading to D1 amino acid Leu218 Val exchange in the Chenopodium album D1 protein and comparison to cross-resistance profiles of known modifications at positions 251 and 264.

BACKGROUND: Resistance of Chenopodium album to triazinones and triazines can be caused by two amino acid exchanges, serine-264-glycine (Ser(264) Gly) and alanine-251-valine (Ala(251) Val), in the chloroplast D1 protein. This paper describes the identification of a biotype with a leucine-218-valine (Leu(218) Val) switch found in German sugar beet fields with unsatisfactory weed control. A greenhouse experiment has been performed to compare the resistance profile of the newly identified biotype with biotypes that carry the Ser(264) Gly and Ala(251) Val mutations. RESULTS: Application rate-response curves obtained from the greenhouse experiment showed that the Leu(218) Val exchange induced significant resistance against the triazinones but not against terbuthylazine. The level of resistance against the triazinones was higher in the Ser(264) Gly and Ala(251) Val biotypes compared with the Leu(218) Val biotype. All biotypes tested were more resistant to metribuzin than to metamitron. Following terbuthylazine treatment, Ser264 Gly displayed a high level of resistance, Ala(251) Val showed moderate resistance. A PCR-RFLP assay for Ser(264) Gly has been extended to include detection of Ala251 Val and Leu(218) Val mutations. CONCLUSION: The D1 Leu(218) Val substitution in C. album confers significant resistance to triazinones. This suggests that Leu(218) Val is involved in the binding of triazinones. First establishment of the resistance profiles of the three psbA mutations suggests that these mutations have been independently selected.

Chenopodolin: a phytotoxic unrearranged ent-pimaradiene diterpene produced by Phoma chenopodicola, a fungal pathogen for Chenopodium album biocontrol.

A new phytotoxic unrearranged ent-pimaradiene diterpene, named chenopodolin, was isolated from the liquid culture of Phoma chenopodicola, a fungal pathogen proposed for the biological control of Chenopodium album, a common worldwide weed of arable crops such as sugar beet and maize. The structure of chenopodolin was established by spectroscopic, X-ray, and chemical methods as (1S,2S,3S,4S,5S,9R,10S,12S,13S)-1,12-acetoxy-2,3-hydroxy-6-oxopimara-7(8),15-dien-18-oic acid 2,18-lactone. At a concentration of 2 mg/mL, the toxin caused necrotic lesions on Mercurialis annua, Cirsium arvense, and Setaria viride. Five derivatives were prepared by chemical modification of chenopodolin functionalities, and some structure-activity relationships are discussed.

Physiological responses and tolerance mechanisms to Pb in two xerophils: Salsola passerina Bunge and Chenopodium album L.

Lead (Pb) has great toxicity to human beings and other livings. Although there are varied ways to rehabilitate the Pb contaminated area, phytoremediation of Pb pollution in arid lands is still a difficult task, it is therefore urgent to find and identify Pb tolerant plants in arid areas. The physiological responses and tolerance mechanisms to Pb stress (expressed as the Pb concentration, e.g., 0, 50, 150, 300, 600, 800, 1000 mg/L) were investigated for the xerophils Salsola passerina Bunge and Chenopodium album L. Results indicated that S. passerina exhibited higher Pb tolerance than Ch. album in terms of the seed germination rate, bio-activities of SOD and POD, and lower MDA production. There were two ways for S. passerina to reduce Pb toxicity in organism level, e.g., cell wall precipitation and state transfer of free Pb into anchorage. These findings demonstrate that S. passerina is a Pb tolerant species and may have potential application in phytoremediation of Pb contaminated arid lands.

The origin of metamitron resistant Chenopodium album populations in sugar beet.

Chenopodium album L. is a major weed in spring-planted crops in the temperate regions of the world. Since 2000, farmers have reported an unsatisfactory control of this weed in sugar beet fields in Belgium, France and The Netherlands. Frequently, the surviving C. album plants are resistant to metamitron, a key herbicide in this crop. Metamitron resistance in C. album is caused by a Ser264 to Gly mutation in the psbA gene on the chloroplast genome, which prevents binding of metamitron to its target site. This mutation causes also resistance to other herbicides with a similar mode of action, like metribuzin -applied in potato- and atrazine in particular. Atrazine has been applied very frequently in maize in the 1970s and the 1980s, but is now banned in Europe due to environmental reasons. The persistent use of atrazine in maize confronted Belgian and other European farmers in the early 1980s with atrazine resistant C. album with the same Ser264 to Gly mutation. The problems with atrazine resistant C. album disappeared when other herbicides were applied in maize. Unfortunately, this is not the case for metamitron resistant C. album in sugar beet, because no replacement herbicide is readily available. The history of atrazine use in maize brought up a question concerning the origin of the current metamitron resistant C. album populations. Have these populations been selected locally by regular use of metamitron in sugar beet or did the selection occur earlier by atrazine use when maize was grown in the same fields? This would have serious implications regarding the reversibility of herbicide resistance. Therefore, soil samples were collected on 16 fields with different histories: five fields with an organic management over 25 years, two fields with a history of atrazine resistant C. album, five fields with metamitron resistant C. album in sugar beet and four fields which were under permanent grassland for 10 years, preceded by a regular rotation in which sugar beet was a key crop. The seeds of C. album were extracted from the soil and germinated on a germination table. Germinated seeds were allowed to grow in a growth chamber. Metamitron resistance was determined by a chlorophyll fluorescence test and leaf material was sampled for AFLP-analysis. For all fields, estimations were made of the size of the seed bank (i.e. an indirect estimate of population size), the frequency of resistant plants and the genetic diversity of resistant and susceptible populations. The results indicate that herbicide-resistant C. album populations are persistent and maintain their adaptive capacity, challenging future management of metamitron resistant C. album.

Analysis of gene expression by ESTs from suppression subtractive hybridization library in Chenopodium album L. under salt stress.

To identify genes expression in Chenopodium album exposed to NaCl stress and screen ESTs related to salt stress, a subtractive suppression hybridization (SSH) library of C. album under salt stress was constructed in the present study. Random EST sequencing produced 825 high-quality ESTs with GenBank ID GE746311-GE747007, which had 301 bp of average size and were clustered into 88 contigs and 550 singletons. They were classified into 12 categories according to their function annotations. 635 ESTs (76.97%) showed similarities to gene sequences in the non-redundancy database, while 190 ESTs (23.03%) showed low or no similarities. The transcriptional profiles of 56 ESTs randomly selected from 347 unknown or novel ESTs of SSH library under varying NaCl concentration and at different time points were analyzed. The results indicated that a high proportion of tested ESTs were activated by salt stress. Four in 56 ESTs responded to NaCl were also enhanced in expression level when exposed to ABA and PEG stresses. The above four ESTs were validated by northern blotting which was consistent with the results of RT-PCR. The results suggested that genes corresponded to these ESTs might be involved in stress response or regulation. The complete sequences and detailed function of these ESTs need to be further studied.

Local spread of metamitron resistant Chenopodium album L. patches.

Molecular markers can provide valuable information on the spread of resistant weed biotypes. In particular, tracing local spread of resistant weed patches will give details on the importance of seed migration with machinery, manure, wind or birds. This study investigated the local spread of metamitron resistant Chenopodium album L. patches in the southwest region of the province West-Flanders (Belgium). During the summer of 2009, leaf and seed samples were harvested in 27 patches, distributed over 10 sugar beet fields and 1 maize field. The fields were grouped in four local clusters. Each cluster corresponded with the farmer who cultivated these fields. A cleaved amplified polymorphic sequence (CAPS) procedure identified the Ser264 to Gly mutation in the D1 protein, endowing resistance to metamitron, a key herbicide applied in sugar beet. The majority of the sampled plants within a patch (97% on average) carried this mutation. Amplified fragment length polymorphism (AFLP) analysis was performed with 4 primer pairs and yielded 270 molecular markers, polymorphic for the whole dataset (303 samples). Analysis of molecular variance revealed that a significant part of the genetic variability was attributed to variation among the four farmer locations (12 %) and variation among Chenopodium album patches within the farmer locations (14%). In addition, Mantel tests revealed a positive correlation between genetic distances (linearised phipt between pairs of patches) and geographic distances (Mantel-coefficient significant at p = 0.002), suggesting isolation-by-distance. In one field, a decreased genetic diversity and strong genetic relationships between all the patches in this field supported the hypothesis of a recent introduction of resistant biotypes. Furthermore, genetic similarity between patches from different fields from the same farmer and from different farmers indicated that seed transport between neighbouring fields is likely to have an important impact on the spread of metamitron resistant biotypes.