Herbicides Linuron Research Articles

OS03-07 Epigenetic transgenerational effects of environmental pollution: assessing the impact of the herbicide linuron on Xenopus tropicalis Frogs

OS03-07 Epigenetic transgenerational effects of environmental pollution: assessing the impact of the herbicide linuron on Xenopus tropicalis Frogs

Pesticide-induced transgenerational alterations of genome-wide DNA methylation patterns in the pancreas of Xenopus tropicalis correlate with metabolic phenotypes

The unsustainable use of manmade chemicals poses significant threats to biodiversity and human health. Emerging evidence highlights the potential of certain chemicals to cause transgenerational impacts on metabolic health. Here, we investigate male transmitted epigenetic transgenerational effects of the anti-androgenic herbicide linuron in the pancreas of Xenopus tropicalis frogs, and their association with metabolic phenotypes. Reduced representation bisulfite sequencing (RRBS) was used to assess genome-wide DNA methylation patterns in the pancreas of adult male F2 generation ancestrally exposed to environmentally relevant linuron levels (44 ± 4.7 μg/L). We identified 1117 differentially methylated regions (DMRs) distributed across the X. tropicalis genome, revealing potential regulatory mechanisms underlying metabolic disturbances. DMRs were identified in genes crucial for pancreatic function, including calcium signalling (clstn2, cacna1d and cadps2), genes associated with type 2 diabetes (tcf7l2 and adcy5) and a biomarker for pancreatic ductal adenocarcinoma (plec). Correlation analysis revealed associations between DNA methylation levels in these genes and metabolic phenotypes, indicating epigenetic regulation of glucose metabolism. Moreover, differential methylation in genes related to histone modifications suggests alterations in the epigenetic machinery. These findings underscore the long-term consequences of environmental contamination on pancreatic function and raise concerns about the health risks associated with transgenerational effects of pesticides.

In vitro antiandrogenic effects of the herbicide linuron and its metabolites

Although the herbicide linuron is banned for use in the EU due to its reproductive and developmental toxicity, it can still be found in randomly sampled foods grown in and outside the EU. It is not clear if metabolites of linuron can contribute to the endocrine disrupting effects following exposure to the parent compound. To address this gap, we analysed linuron and the metabolites 1-(3,4-dichlorophenyl) urea (DCU), 3,4-dichloroaniline (DCA) and 1-(3,4-dichlorophenyl)-3-methoxyurea (DCXU) for androgen receptor (AR) activities and effects on steroidogenesis. Generally, linuron and the metabolites showed qualitatively similar antiandrogenic profiles, but potencies varied. All compounds were AR antagonists, with linuron showing highest potency (IC50 of 2.8 μM). The overall picture of effects on steroidogenesis showed that linuron and metabolites increased the levels of estrogens and corticosteroids, whereas the synthesis of androgens was inhibited. The metabolite DCU was by far the most potent inhibitor of testosterone synthesis (IC50 of 6.7 μM compared to IC50 of 51.1 μM for linuron). We suggest that it is likely that the metabolites contribute to the antiandrogenic effects of linuron in vivo, especially by inhibiting testosterone synthesis.

Effects of Various Environmental Conditions on the Growth of Amaranthus patulus Bertol. and Changes of Herbicide Efficacy Caused by Increasing Temperatures

Understanding the effects of climate change on weed growth and herbicide activity is important for optimizing herbicide applications for effective weed control in the future. Therefore, this study examined how climate change affects the growth of Amaranthus patulus and the efficacy of soil and foliar herbicides at different temperatures. Although the control values for A. patulus differed between herbicides and temperature, the control values increased with increasing time after the herbicide treatments. Under growth conditions in which the temperature remained constant, the efficacy of soil-applied herbicides, ethalfluralin, metolachlor, linuron, and alachlor, on A. patulus was highest when the weeds were grown at high temperature. In particular, 100% control values of A. patulus were achieved in response to metolachlor treatments at the total recommended dosage in growth chambers at 35 °C. The efficacy of foliar herbicides, glufosinate-ammonium, bentazone, and mecoprop, on A. patulus was also highest when the plant was grown at high temperature, except for glyphosate isopropylamine, which had similar efficacy rates regardless of the temperature. A. patulus was 100% controlled in response to glufosinate-ammonium, bentazone, and mecoprop at the recommended dosages in growth chambers at 30 and 35 °C. Under growth conditions in which the temperature changed from day to night, the efficacy of soil-applied herbicides, alachlor and linuron, on A. patulus was highest when the weeds were grown at high temperature. On the other hand, the efficacy of the soil-applied herbicides metolachlor and linuron on A. patulus was similar regardless of the temperature. The efficacy of foliar herbicides, glyphosate isopropylamine, glufosinate-ammonium, bentazone, and mecoprop, on A. patulus was highest when the weeds were grown at high temperature. Although herbicide efficacy varied depending on whether the weeds were grown at constant or alternating temperatures, herbicide efficacy was generally highest when the temperature was high.

Changes in dry weight and starch content in potato under the effect of herbicides and biostimulants

The aim of the study was to determine the interaction of herbicides and herbicides with biostimulants on the accumulation of dry matter and starch in potato tubers. In a three-year field experiment based on the method of randomised sub-blocks, two factors were taken into account. The first factor were potato cultivars: Bartek, Gawin, Honorata. The second factor were five methods of herbicides and biostimulants application: (1) the control object without chemical protection (CO); (2) herbicide linuron + clomazone (Harrier 295 ZC) (H); (3) herbicide linuron + clomazone (Harrier 295 ZC) and biostimulant Ecklonia maxima (Kelpak SL) – (H + K); (4) herbicide metribuzin (Sencor 70 WG) – (S); (5) herbicide metribuzin (Sencor 70 WG) and biostimulant sodium para-nitrophenol, sodium ortho-nitrophenol, sodium 5-nitroguaiacol (Asahi SL) – (S + A). The cultivars and weather conditions significantly affected the content and yields of dry matter and starch. The herbicides and biostimulants used determined the starch accumulation as well as dry matter and starch yields. Most starch in tubers (more by 3.7 g/kg) and the highest dry matter and starch yields (more by 2.87 and 1.79 t/ha, respectively), compared to the control object, were obtained after the application of the herbicide Sencor 70 WG and biostimulant Asahi SL.

Culture-Independent Analysis of Linuron-Mineralizing Microbiota and Functions in on-Farm Biopurification Systems via DNA-Stable Isotope Probing: Comparison with Enrichment Culture.

Our understanding of the microorganisms involved in in situ biodegradation of xenobiotics, like pesticides, in natural and engineered environments is poor. On-farm biopurification systems (BPSs) treat farm-produced pesticide-contaminated wastewater to reduce surface water pollution. BPSs are a labor and cost-efficient technology but are still mainly operated as black box systems. We used DNA-stable isotope probing (DNA-SIP) and classical enrichment to be informed about the organisms responsible for in situ degradation of the phenylurea herbicide linuron in a BPS matrix. DNA-SIP identified Ramlibacter, Variovorax, and an unknown Comamonadaceae genus as the dominant linuron assimilators. While linuron-degrading Variovorax strains have been isolated repeatedly, Ramlibacter has never been associated before with linuron degradation. Genes and mobile genetic elements (MGEs) previously linked to linuron catabolism were enriched in the heavy DNA-SIP fractions, suggesting their involvement in in situ linuron assimilation. BPS material free cultivation of linuron degraders from the same BPS matrix resulted in a community dominated by Variovorax, while Ramlibacter was not observed. Our study provides evidence for the role of Variovorax in in situ linuron biodegradation in a BPS, alongside other organisms like Ramlibacter, and further shows that cultivation results in a biased representation of the in situ linuron-assimilating bacterial populations.

Community composition modifies direct and indirect effects of pesticides in freshwater food webs

For environmental risk assessment, the effects of pesticides on aquatic ecosystems are often assessed based on single species tests, disregarding the potential influence of community composition. We, therefore, studied the influence of changing the horizontal (the number of species within trophic levels) and vertical composition (number of trophic levels) on the ecological effects of the herbicide linuron and the insecticide chlorpyrifos, targeting producers and herbivores, respectively. We tested how adding, to a single primary producer, 4 selected competing producer species, 0–1-4 selected herbivore species, and one selected predator species resulting in 1, 2 and 3 trophic levels, changes the effects of the two pesticides.Linuron decreased producer biovolume less (17%) when the 4 producers were added, because insensitive producers compensated for the loss of sensitive producers. However, linuron decreased producer biovolume 42% and 32% more as we increased the number of herbivore species from 0 to 4 and as we increased trophic levels from 1 to 3, respectively. The indirect negative effect of linuron on herbivore biovolume was 11% and 15% lower when more producer and herbivores were added, respectively. Adding a predator increased this indirect negative effect by 22%.Chlorpyrifos decreased herbivore biovolume about 10% less when adding multiple herbivore or producer species. However, adding a predator magnified the direct negative impact on herbivores (13%). Increasing the number of producer, herbivore species and adding trophic levels increased the indirect positive impact on producer biovolume (between 10% and 35%).Our study shows that changing horizontal composition can both increase and decrease the effects of the selected pesticides, while changing vertical composition by adding number of trophic levels always increased these effects. Therefore, single species sensitivity will not always represent a worst case estimate of ecological effects. Protecting the most sensitive species may not ensure protection of ecosystems.

Characterization of a Linuron-Specific Amidohydrolase from the Newly Isolated Bacterium Sphingobium sp. Strain SMB.

The phenylurea herbicide linuron is globally used and has caused considerable concern because it leads to environmental pollution. In this study, a highly efficient linuron-transforming strain Sphingobium sp. SMB was isolated, and a gene (lahB) responsible for the hydrolysis of linuron to 3,4-dichloroaniline and N,O-dimethylhydroxylamine was cloned from the genome of strain SMB. The lahB gene encodes an amidohydrolase, which shares 20-53% identity with other biochemically characterized amidohydrolases, except for the newly reported linuron hydrolase Phh (75%). The optimal conditions for the hydrolysis of linuron by LahB were determined to be pH 7.0 and 30 °C, and the Km value of LahB for linuron was 37.3 ± 1.2 μM. Although LahB and Phh shared relatively high identity, LahB exhibited a narrow substrate spectrum (specific for linuron) compared to Phh (active for linuron, diuron, chlortoluron, etc.). Sequence analysis and site-directed mutagenesis revealed that Ala261 of Phh was the key amino acid residue affecting the substrate specificity. Our study provides a new amidohydrolase for the specific hydrolysis of linuron.

Marketable Yield of Potato and Its Quantitative Parameters after Application of Herbicides and Biostimulants

Potato (Solanum tuberosum L.) is grown in over 160 countries. Weed competition and environmental stressors during the vegetative growth stage significantly impact crop yields. An experiment was conducted from 2012 to 2014 in Poland to assess the effect of herbicides linuron + clomazone (L+CH) and metribuzin (M) as well as herbicides mixed with biostimulants (linuron + clomazone and algae extract of Ecklonia maxima—auxins and gibberellins (L+CH+E) and metribuzin + sodium p-nitrophenolate, sodium o-nitrophenolate and sodium 5-nitroguaiacolate (M+S)) on weed infestation, marketable yield and yield components of the following three table potato cultivars: Bartek, Gawin and Honorata. In plots where potato had been treated with herbicides and herbicides mixed with biostimulants, a decline in the fresh matter of weeds was observed, ranging from 72.4% to 96.1%, which was followed by an increase in potato marketable yield (from 27.5% to 61%) and improved parameters of S. tuberosum yield components, compared with the control. Linear correlation coefficients indicated that the following characteristics: marketable yield, weight of tubers per plant and average weight per tuber were associated with weed infestation determined prior to potato harvest.

PromA Plasmids Are Instrumental in the Dissemination of Linuron Catabolic Genes Between Different Genera.

PromA plasmids are broad host range (BHR) plasmids, which are often cryptic and hence have an uncertain ecological role. We present three novel PromA γ plasmids which carry genes associated with degradation of the phenylurea herbicide linuron, two of which originated from unrelated Hydrogenophaga hosts isolated from different environments (pPBL-H3-2 and pBPS33-2), and one (pEN1) which was exogenously captured from an on-farm biopurification system (BPS). Hydrogenophaga sp. plasmid pBPS33-2 carries all three necessary gene clusters (hylA, dca, ccd) determining the three main steps for conversion of linuron to Krebs cycle intermediates, while pEN1 only determines the initial linuron hydrolysis step. Hydrogenophaga sp. plasmid pPBL-H3-2 exists as two variants, both containing ccd but with the hylA and dca gene modules interchanged between each other at exactly the same location. Linuron catabolic gene clusters that determine the same step were identical on all plasmids, encompassed in differently arranged constellations and characterized by the presence of multiple IS1071 elements. In all plasmids except pEN1, the insertion spot of the catabolic genes in the PromA γ plasmids was the same. Highly similar PromA plasmids carrying the linuron degrading gene cargo at the same insertion spot were previously identified in linuron degrading Variovorax sp. Interestingly, in both Hydrogenophaga populations not every PromA plasmid copy carries catabolic genes. The results indicate that PromA plasmids are important vehicles of linuron catabolic gene dissemination, rather than being cryptic and only important for the mobilization of other plasmids.

After-effect of Some Herbicides and Their Mixtures with Growth Regulator and Foliar Fertilizer on the Primary Germ Weight of Cotton Seeds (Gossypium Hirsutum L.)

The trial was carried out during 2013-2015, with two cotton cultivars – Helius and Darmi (Gossypium hirsutum L.). Herbicides Goal 2 Е (oxifluorfen), Linuron45 SC (linuron), Wing P (pendimethalin + dimethenamid), Merlin 750 WG (izoxaflutole), Bazagran 480 SL (bentazone) were studied. These herbicides were used alone or in combinations with the growth regulator Amalgerol premium or with the foliar fertilizer Lactofol O during the budding stage of cotton. From the viewpoint of cotton growing technology, technologically the most valuable are combinations of all herbicides with Lactofol O, which are followed by Wing-P + Amalgerol, Bazagran 480 SL + Amalgerol and sole use of herbicide Wing-P on cultivar Helius. Technologically the most valuable are herbicides Goal 2 E and Wing-P and tank mixtures Goal 2 E + Amalgerol, Wing-P + Amalgerol, Merlin 750 WG + Amalgerol, Bazagran 480 SL + Amalgerol, Wing-P + Lactofol O, Merlin 750 WG + Lactofol O and Bazagran 480 SL + Lactofol O on cultivar Darmi. These variants combine biggest primary germ weight and high stability of this index during the different years. The alone use of the herbicides Linuron 45 SC and Merlin 750 WG has low assessment and should be avoided.

Effect of Some Herbicides and Their Mixtures with Growth Regulator and Foliar Fertilizer on Protein Content in Cotton (Gossypium hirsutum L.) Seeds

For the first time in the world we established that the highest protein content in seeds of cultivar Helius was obtained with vegetative treatment with herbicides Goal 2 E, Linuron 45 SC, Wing P and Merlin 750 WG. The highest protein content in seeds of cultivar Darmy was obtained with vegetative treatment with herbicides Goal 2 E, Wing P and Basagran 480 SL. The highest phytotoxicity on protein content in seeds of cultivar Helius was shown by the herbicide Basagran 480 SL and all herbicide combinations with the growth regulator Amalgerol and the foliar fertilizer Lactofol O. The strongest phytotoxic effects on seeds of cultivar Darmy was produced by the herbicides Linuron 45 SC and Merlin 750 WG and herbicide mixture Wing P + Lactofol O. For the first time, we established that, from the point of view of the technology for cotton growing of cultivar Helius, the most valuable are tank mixtures Goal 2 E + Amalgerol, Linuron 45 SC + Amalgerol, Goal 2 E + Lactofol O, as well as the single use of herbicides Goal 2 E, Linuron 45 SC, Wing P and Merlin 750 WG. Technologically the most valuable in cotton cultivar Darmy were herbicide combinations Goal 2 E + Amalgerol, Wing P + Amalgerol, Merlin 750 WG + Amalgerol, Basagran 480 SL + Amalgerol, Goal 2 E + Lactofol O, Linuron 45 SC + Lactofol O, Merlin 750 WG + Lactofol O and Basagrain 480 SL + Lactofol O, and the herbicides Goal 2 E, Wing P and Basagran 480 SL.

Environmental Control of Astrocyte Pathogenic Activities in CNS Inflammation

Genome-wide studies have identified genetic variants linked to neurologic diseases. Environmental factors also play important roles, but no methods are available for their comprehensive investigation. We developed an approach that combines genomic data, screens in a novel zebrafish model, computational modeling, perturbation studies, and multiple sclerosis (MS) patient samples to evaluate the effects of environmental exposure on CNS inflammation. We found that the herbicide linuron amplifies astrocyte pro-inflammatory activities by activating signaling via sigma receptor 1, inositol-requiring enzyme-1α (IRE1α), and X-box binding protein 1 (XBP1). Indeed, astrocyte-specific shRNA- and CRISPR/Cas9-driven gene inactivation combined with RNA-seq, ATAC-seq, ChIP-seq, and study of patient samples suggest that IRE1α-XBP1 signaling promotes CNS inflammation in experimental autoimmune encephalomyelitis (EAE) and, potentially, MS. In summary, these studies define environmental mechanisms that control astrocyte pathogenic activities and establish a multidisciplinary approach for the systematic investigation of the effects of environmental exposure in neurologic disorders.

Stars Are Not in Outer Space: Astrocytes Respond to Environmental Cues

By invivo screening in zebrafish larvae, Wheeler etal. identify environmental agents that directly modulate transcriptional programs of astrocytes. The herbicide linuron exploits the unfolded protein response pathway to induce a pro-inflammatory program in reactive astrocytes that potentiates inflammatory tissue damage in the CNS.

Nationwide screening of surface water toxicity to algae

According to the European Water Framework Directive (WFD), chemical water quality is assessed by monitoring 45 priority substances. However, observed toxic effects can often not be attributed to these priority substances, and therefore there is an urgent need for an effect-based monitoring strategy that employs bioassays to identify environmental risk. Algal photosynthesis is a sensitive process that can be applied to identify the presence of hazardous herbicides in surface water. Therefore, the aim of this study was to employ an algal photosynthesis bioassay to assess surface water toxicity to algae and to identify the compounds causing the observed effects. To this purpose, Raphidocelis subcapitata was exposed to surface water samples and after 4.5 h photosynthetic efficiency was determined using PAM fluorometry. In this rapid high throughput bioassay, algal photosynthesis was affected by surface water from only one of 39 locations. Single compounds toxicity confirmation elucidated that the observed effect could be solely attributed to the herbicide linuron, which occurred at 110 times the EQS concentration and which is not included in the WFD priority substances list. In conclusion, applying the algal photosynthesis bioassay enables more efficient and effective assessment of toxicity to primary producers because it: (i) identifies the presence of herbicides that would be overlooked by routine chemical WFD monitoring, and (ii) avoids redundant chemical analyses by focusing only on (non-)target screening in samples with demonstrated effects.

Adsorption of linuron by an Algerian palygorskite modified with magnetic iron

Abstract Three types of palygorskite nanoparticles for the retention of the herbicide linuron in water were tested: purified palygorskite (Pur Pal), palygorskite modified by magnetic iron oxides (FeO Pal 1 ) and this one synthesized by hydrothermal treatment (FeO Pal 2 ). The adsorbents were characterized by X-ray diffraction (XRD), infrared spectroscopy (FTIR), electron microscopy (TEM) and chemical analysis (XRF). XRD patterns indicate the presence of a magnetic Fe 2 O 3 compound, and TEM images evidence how these crystals cover the palygorskite fibers' surface with different crystallite sizes (7–15 nm in FeO Pal 1 and 30-50 nm in FeO Pal 2 ). The adsorbent mass, the contact time, the initial adsorbate concentration and the effect of temperature were evaluated for linuron retention. Adsorption kinetics revealed that adsorption followed a pseudo-second order equation. Adsorption isotherms were better fitted to the Freundlich than to the Langmuir equation. Removed linuron amounts were ranged as FeO Pal 2 (83%) > FeO Pal 1 (55%) > Pur Pal (27%). The adsorbents could be used as a cheap alternative for pesticide removal in environmentally-contaminated waters.

Expression of the human gene CYP1A2 enhances tolerance and detoxification of the phenylurea herbicide linuron in Arabidopsis thaliana plants and Escherichia coli

The phenylurea herbicide, linuron (LIN), is used to control various types of weeds. Despite its efficient role in controlling weeds, it presents a persistent problem to the environment. In the current study, phytoremediation properties of transgenic CYP1A2 Arabidopsis thaliana plants to LIN were assessed. CYP1A2 gene was firstly cloned and expressed in bacteria before proceeding to plants. In presence of LIN, The growth of CYP1A2 expressing bacteria was superior compared to control bacteria transformed with the empty bacterial expression vector pET22b(+). No clear morphological changes were detected on CYP1A2 transgenic plants. However, significant resistance to LIN herbicide application either via spraying the foliar parts of the plant or via supplementation of the herbicide in the growth medium was observed for CYP1A2 transformants. Plant growth assays under LIN stress provide strong evidence for the enhanced capacity of transgenic lines to grow and to tolerate high concentrations of LIN compared to control plants. HPLC analyses showed that detoxification of LIN by bacterial extracts and/or transgenic plant leaves is improved as compared to the corresponding controls. Our data indicate that over expression of the human CYP1A2 gene increases the phytoremediation capacity and tolerance of Arabidopsis thaliana plants to the phenylurea herbicide linuron.

Molecular processes underlying synergistic linuron mineralization in a triple-species bacterial consortium biofilm revealed by differential transcriptomics.

The proteobacteria Variovorax sp. WDL1, Comamonas testosteroni WDL7, and Hyphomicrobium sulfonivorans WDL6 compose a triple‐species consortium that synergistically degrades and grows on the phenylurea herbicide linuron. To acquire a better insight into the interactions between the consortium members and the underlying molecular mechanisms, we compared the transcriptomes of the key biodegrading strains WDL7 and WDL1 grown as biofilms in either isolation or consortium conditions by differential RNAseq analysis. Differentially expressed pathways and cellular systems were inferred using the network‐based algorithm PheNetic. Coculturing affected mainly metabolism in WDL1. Significantly enhanced expression of hylA encoding linuron hydrolase was observed. Moreover, differential expression of several pathways involved in carbohydrate, amino acid, nitrogen, and sulfur metabolism was observed indicating that WDL1 gains carbon and energy from linuron indirectly by consuming excretion products from WDL7 and/or WDL6. Moreover, in consortium conditions, WDL1 showed a pronounced stress response and overexpression of cell to cell interaction systems such as quorum sensing, contact‐dependent inhibition, and Type VI secretion. Since the latter two systems can mediate interference competition, it prompts the question if synergistic linuron degradation is the result of true adaptive cooperation or rather a facultative interaction between bacteria that coincidentally occupy complementary metabolic niches.

Efficacy and Selectivity of Pre- and Post-emergence Herbicides in Chia (Salvia hispanica L.) under Mediterranean Semi-arid Conditions

In the present study, the selectivity and efficacy of several post and pre-emergence herbicides in chia (Salvia hispanica L.) were examined. Field experiments were conducted at two locations with different soil types and environmental conditions. Our results showed that the pre-emergence application of pendimethalin, oxyfluorfen and linuron reduced total weed density by 71-74%, 74-82% and 53-55%, respectively. Despite their high effectiveness, the above-mentioned herbicides had a negative effect on plant density and biomass yield. In addition, herbicides bentazon and fluazifop-p-butyl which applied post-emergence did not affect significantly plant height and biomass yield. In conclusion, the herbicides linuron, pendimethalin and oyxfluorfen do not seem to be a viable option for weed control in chia crop, whereas the post-emergence applied herbicides did not affect significantly plants’ growth. Further evaluation of chia tolerance to herbicides is needed under different application rates in order to make safe suggestions for chemical control of weeds.

Catabolic task division between two near-isogenic subpopulations co-existing in a herbicide-degrading bacterial consortium: consequences for the interspecies consortium metabolic model.

Variovorax sp. WDL1 mediates hydrolysis of the herbicide linuron into 3,4-dichloroaniline (DCA) and N,O-dimethylhydroxylamine in a tripartite bacterial consortium with Comamonas testosteroni WDL7 and Hyphomicrobium sulfonivorans WDL6. Although strain WDL1 contains the dcaQTA1A2B operon for DCA oxidation, this conversion is mainly performed by WDL7. Phenotypic diversification observed in WDL1 cultures and scrutiny of the WDL1 genome suggest that WDL1 cultures consist of two dedicated subpopulations, i.e., a linuron-hydrolysing subpopulation (Lin + DCA-) and a DCA-oxidizing subpopulation (Lin-DCA+). Whole genome analysis of strains representing the respective subpopulations revealed that they are identical, aside from the presence of hylA (in Lin + DCA- cells) and the dcaQTA1A2B gene cluster (in Lin-DCA+ cells), and that these catabolic gene modules replace each other at exactly the same locus on a 1380 kb extra-chromosomal element that shows plasmid gene functions including genes for transferability by conjugation. Both subpopulations proliferate in consortium biofilms fed with linuron, but Lin + DCA- cells compose the main WDL1 subpopulation. Our observations instigated revisiting the interactions within the consortium and suggest that the physical separation of two essential linuron catabolic gene clusters in WDL1 by mutually exclusive integration in the same mobile genetic element is key to the existence of WDL1 in a consortium mode.