Colonization Of Surfaces Research Articles

Differentiated responses of the phyllosphere bacterial community of the yellowhorn tree to precipitation and temperature regimes across Northern China.

As an ephemeral and oligotrophic environment, the phyllosphere harbors many highly diverse microorganisms. Importantly, it is known that their colonization of plant leaf surfaces is considerably influenced by a few abiotic factors related to climatic conditions. Yet how the dynamics of phyllosphere bacterial community assembly are shaped by detailed climatological elements, such as various bioclimatic variables, remains poorly understood. Using high-throughput 16S rRNA gene amplicon sequencing technology, we analyzed the bacterial communities inhabiting the leaf surfaces of an oilseed tree, yellowhorn (Xanthoceras sorbifolium), grown at four sites (Yinchuan, Otogqianqi, Tongliao, and Zhangwu) whose climatic status differs in northern China. We found that the yellowhorn phyllosphere's bacterial community was generally dominated by four phyla: Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes. Nevertheless, bacterial community composition differed significantly among the four sampled site regions, indicating the possible impact of climatological factors upon the phyllosphere microbiome. Interestingly, we also noted that the α-diversities of phyllosphere microbiota showed strong positive or negative correlation with 13 bioclimatic factors (including 7 precipitation factors and 6 temperature factors). Furthermore, the relative abundances of 55 amplicon sequence variants (ASVs), including three ASVs representing two keystone taxa (the genera Curtobacterium and Streptomyces), exhibited significant yet contrary responses to the precipitation and temperature climatic variables. That pattern was consistent with all ASVs' trends of possessing opposite correlations to those two parameter classes. In addition, the total number of links and nodes, which conveys community network complexity, increased with rising values of most temperature variables. Besides that, remarkably positive relevance was found between average clustering coefficient and most precipitation variables. Altogether, these results suggest the yellowhorn phyllosphere bacterial community is capable of responding to variation in rainfall and temperature regimes in distinctive ways.

Calcium promotes persistent soil organic matter by altering microbial transformation of plant litter

Calcium (Ca) can contribute to soil organic carbon (SOC) persistence by mediating physico-chemical interactions between organic compounds and minerals. Yet, Ca is also crucial for microbial adhesion, potentially affecting colonization of plant and mineral surfaces. The importance of Ca as a mediator of microbe-mineral-organic matter interactions and resulting SOC transformation has been largely overlooked. We incubated 44Ca labeled soils with 13C15N labeled leaf litter to study how Ca affects microbial transformation of litter and formation of mineral associated organic matter. Here we show that Ca additions promote hyphae-forming bacteria, which often specialize in colonizing surfaces, and increase incorporation of litter into microbial biomass and carbon use efficiency by approximately 45% each. Ca additions reduce cumulative CO2 production by 4%, while promoting associations between minerals and microbial byproducts of plant litter. These findings expand the role of Ca in SOC persistence from solely a driver of physico-chemical reactions to a mediator of coupled abiotic-biotic cycling of SOC.

Lignan-containing maple products inhibit Listeria monocytogenes biofilms on fresh produce.

Major listeriosis outbreaks have been associated with fresh produce contaminated with Listeria monocytogenes. Strains that synthesize the Pss exopolysaccharide (EPS) have an estimated 102 to 104-fold advantage over nonsynthesizing strains in causing listeriosis. They more readily attach to the surfaces of fruit and vegetables forming EPS-biofilms that better withstand stresses associated with produce storage and consumption. Here, we show that the threat to fresh produce safety posed by the listerial EPS-biofilms may be countered by broadly available maple products. We serendipitously discovered that aqueous extracts of wood from several Acer (maple) and Carya (pecan, hickory) species inhibit the formation of listerial EPS-biofilms without affecting bacterial viability. One active ingredient in maple wood was identified as nortrachelogenin-8'-O-β-D-glucopyranoside (NTG). At 120 μM, this lignan decreased colonization of the EPS-synthesizing L. monocytogenes on cantaloupe pieces by approximately 150-fold, and on cut celery and lettuce by 10 to 11-fold. Another lignan, lariciresinol, which is abundant in a common food sweetener, maple syrup, had antibiofilm activity comparable to that of NTG. Diluted in the range of 1:200 to 1:800 maple syrup from two random manufacturers prevented formation of listeiral EPS-biofilms. Importantly, not only did maple products drastically decrease colonization of fresh produce by the EPS-synthesizing strains, they also decreased, by 6 to 30-fold, colonization by the L. monocytogenes strains that do not synthesize measurable EPS, including strains from the infamous 2011 cantaloupe listeriosis outbreak. Inhibition of surface colonization by various listerial strains, broad availability of maple sap and syrup as well as maple lumber processing waste position maple products as potential antibiofilm agents for protecting fresh produce from L. monocytogenes.

Metabolism-linked methylotaxis sensors responsible for plant colonization in Methylobacterium aquaticum strain 22A.

Motile bacteria take a competitive advantage in colonization of plant surfaces to establish beneficial associations that eventually support plant health. Plant exudates serve not only as primary growth substrates for bacteria but also as bacterial chemotaxis attractants. A number of plant-derived compounds and corresponding chemotaxis sensors have been documented, however, the sensors for methanol, one of the major volatile compounds released by plants, have not been identified. Methylobacterium species are ubiquitous plant surface-symbiotic, methylotrophic bacteria. A plant-growth promoting bacterium, M. aquaticum strain 22A exhibits chemotaxis toward methanol (methylotaxis). Its genome encodes 52 methyl-accepting chemotaxis proteins (MCPs), among which we identified three MCPs (methylotaxis proteins, MtpA, MtpB, and MtpC) responsible for methylotaxis. The triple gene mutant of the MCPs exhibited no methylotaxis, slower gathering to plant tissues, and less efficient colonization on plants than the wild type, suggesting that the methylotaxis mediates initiation of plant-Methylobacterium symbiosis and engages in proliferation on plants. To examine how these MCPs are operating methylotaxis, we generated multiple gene knockouts of the MCPs, and Ca2+-dependent MxaFI and lanthanide (Ln3+)-dependent XoxF methanol dehydrogenases (MDHs), whose expression is regulated by the presence of Ln3+. MtpA was found to be a cytosolic sensor that conducts formaldehyde taxis (formtaxis), as well as methylotaxis when MDHs generate formaldehyde. MtpB contained a dCache domain and exhibited differential cellular localization in response to La3+. MtpB expression was induced by La3+, and its activity required XoxF1. MtpC exhibited typical cell pole localization, required MxaFI activity, and was regulated under MxbDM that is also required for MxaF expression. Strain 22A methylotaxis is realized by three independent MCPs, two of which monitor methanol oxidation by Ln3+-regulated MDHs, and one of which monitors the common methanol oxidation product, formaldehyde. We propose that methanol metabolism-linked chemotaxis is the key factor for the efficient colonization of Methylobacterium on plants.

PlzD modifies Vibrio vulnificus foraging behavior and virulence in response to elevated c-di-GMP.

Many free-swimming bacteria propel themselves through liquid using rotary flagella, and mounting evidence suggests that the inhibition of flagellar rotation initiates biofilm formation, a sessile lifestyle that is a nearly universal surface colonization paradigm in bacteria. In general, motility and biofilm formation are inversely regulated by the intracellular second messenger bis-(3´-5´)-cyclic dimeric guanosine monophosphate (c-di-GMP). Here, we identify a protein, PlzD, bearing a conserved c-di-GMP binding PilZ domain that localizes to the flagellar pole in a c-di-GMP-dependent manner and alters the foraging behavior, biofilm, and virulence characteristics of the opportunistic human pathogen, Vibrio vulnificus. Our data suggest that PlzD interacts with components of the flagellar stator to decrease bacterial swimming speed and changes in swimming direction, and these activities are enhanced when cellular c-di-GMP levels are elevated. These results reveal a physical link between a second messenger (c-di-GMP) and an effector (PlzD) that promotes transition from a motile to a sessile state in V. vulnificus.

Identification of rare microbial colonizers of plastic materials incubated in a coral reef environment.

Plastic waste accumulation in marine environments has complex, unintended impacts on ecology that cross levels of community organization. To measure succession in polyolefin-colonizing marine bacterial communities, an in situ time-series experiment was conducted in the oligotrophic coastal waters of the Bermuda Platform. Our goals were to identify polyolefin colonizing taxa and isolate bacterial cultures for future studies of the biochemistry of microbe-plastic interactions. HDPE, LDPE, PP, and glass coupons were incubated in surface seawater for 11 weeks and sampled at two-week intervals. 16S rDNA sequencing and ATR-FTIR/HIM were used to assess biofilm community structure and chemical changes in polymer surfaces. The dominant colonizing taxa were previously reported cosmopolitan colonizers of surfaces in marine environments, which were highly similar among the different plastic types. However, significant differences in rare community composition were observed between plastic types, potentially indicating specific interactions based on surface chemistry. Unexpectedly, a major transition in community composition occurred in all material treatments between days 42 and 56 (p < 0.01). Before the transition, Alteromonadaceae, Marinomonadaceae, Saccharospirillaceae, Vibrionaceae, Thalassospiraceae, and Flavobacteriaceae were the dominant colonizers. Following the transition, the relative abundance of these taxa declined, while Hyphomonadaceae, Rhodobacteraceae and Saprospiraceae increased. Over the course of the incubation, 8,641 colonizing taxa were observed, of which 25 were significantly enriched on specific polyolefins. Seven enriched taxa from families known to include hydrocarbon degraders (Hyphomonadaceae, Parvularculaceae and Rhodobacteraceae) and one n-alkane degrader (Ketobacter sp.). The ASVs that exhibited associations with specific polyolefins are targets of ongoing investigations aimed at retrieving plastic-degrading microbes in culture.

A Candida auris-specific adhesin, Scf1, governs surface association, colonization, and virulence.

Candida auris is an emerging fungal pathogen responsible for health care-associated outbreaks that arise from persistent surface and skin colonization. We characterized the arsenal of adhesins used by C. auris and discovered an uncharacterized adhesin, Surface Colonization Factor (Scf1), and a conserved adhesin, Iff4109, that are essential for the colonization of inert surfaces and mammalian hosts. SCF1 is apparently specific to C. auris, and its expression mediates adhesion to inert and biological surfaces across isolates from all five clades. Unlike canonical fungal adhesins, which function through hydrophobic interactions, Scf1 relies on exposed cationic residues for surface association. SCF1 is required for C. auris biofilm formation, skin colonization, virulence in systemic infection, and colonization of inserted medical devices.

Bioadhesive Microcarriers Encapsulated with IL-27 High Expressive MSC Extracellular Vesicles for Inflammatory Bowel Disease Treatment.

Mesenchymal stem cell (MSC) therapy is a promising candidate for inflammatory bowel disease (IBD) treatment, while overcoming the limitations of naive seeding cells function and realizing efficient intestinal targeting remains a challenge. Here, a bioadhesive microparticle carrying interleukin-27 (IL-27) MSC-derived extracellular vesicles (MSCIL-27 EVs) is developed to treat IBD. The MSCIL-27 EVs prepared through lentivirus-mediated gene transfection technology show ideal anti-inflammatory and damage repair function. By encapsulating MSCIL-27 EVs into dopamine methacrylamide-modified hydrogel, a bioadhesive EVs microcarrier via microfluidic technology is fabricated. The resultant microcarriers exhibit ideal MSCIL-27 EVs sustained release effect and effective wet adhesion property. Furthermore, the therapeutic potential of MSCIL-27 EVs-loaded microcarriers in treating IBD is demonstrated. Through giving IBD rats a rectal administration, it is found that the microcarriers can firmly anchor to the surface of colon, reduce the inflammatory response, and repair the damaged barrier. Therefore, the bioadhesive MSCIL-27 EVs-loaded microcarriers provide a promising strategy for the biomedical application of MSC-derived EVs, and broaden the clinical potential of MSC therapy.

Antibacterial Activity of a Lysin LysP53 against Streptococcus mutans.

Dental caries is a common disease affecting quality of life globally. In the present study, we found that a bacteriophage lysin LysP53 against Acinetobacter baumannii possesses selective activity on Streptococcus mutans, the main etiological agent of dental caries, even in low pH caries microenvironments, whereas only minor LysP53 activity was detected against Streptococcus sanguinis, Streptococcus oralis, and Streptococcus mitis. Testing activity against S. mutans planktonic cells showed that 4 μM LysP53 could kill more than 84% of S. mutans within 1 min in buffer with optimal pHs ranging from 4.0 to 6.5. Daily application of LysP53 on biofilms formed in BHI medium supplemented or not with sucrose could reduce exopolysaccharides, expression of genes related to acid resistance and adhesion, and the number of live bacteria in the biofilms. LysP53 treatment also showed similar effects as 0.12% chlorhexidine in preventing enamel demineralization due to S. mutans biofilms, as well as effective removal of S. mutans colonization of tooth surfaces in mice without observed toxic effects. Because of its selective activity against main cariogenic bacteria and good activity in low pH caries microenvironments, it is advantageous to use LysP53 as an active agent for preventing caries.

Quantitative whole-tissue 3D imaging reveals bacteria in close association with mouse jejunum mucosa

Because the small intestine (SI) epithelium lacks a thick protective mucus layer, microbes that colonize the thin SI mucosa may exert a substantial effect on the host. For example, bacterial colonization of the human SI may contribute to environmental enteropathy dysfunction (EED) in malnourished children. Thus far, potential bacterial colonization of the mucosal surface of the SI has only been documented in disease states, suggesting mucosal colonization is rare, likely requiring multiple perturbations. Furthermore, conclusive proof of bacterial colonization of the SI mucosal surface is challenging, and the three-dimensional (3D) spatial structure of mucosal colonies remains unknown. Here, we tested whether we could induce dense bacterial association with jejunum mucosa by subjecting mice to a combination of malnutrition and oral co-gavage with a bacterial cocktail (E. coli and Bacteroides spp.) known to induce EED. To visualize these events, we optimized our previously developed whole-tissue 3D imaging tools with third-generation hybridization chain reaction (HCR v3.0) probes. Only in mice that were malnourished and gavaged with the bacterial cocktail did we detect dense bacterial clusters surrounding intestinal villi suggestive of colonization. Furthermore, in these mice we detected villus loss, which may represent one possible consequence that bacterial colonization of the SI mucosa has on the host. Our results suggest that dense bacterial colonization of jejunum mucosa is possible in the presence of multiple perturbations and that whole-tissue 3D imaging tools can enable the study of these rare events.

Viscoelasticity enhances collective motion of bacteria.

Bacteria form human and animal microbiota. They are the leading causes of many infections and constitute an important class of active matter. Concentrated bacterial suspensions exhibit large-scale turbulent-like locomotion and swarming. While the collective behavior of bacteria in Newtonian fluids is relatively well understood, many fundamental questions remain open for complex fluids. Here, we report on the collective bacterial motion in a representative biological non-Newtonian viscoelastic environment exemplified by mucus. Experiments are performed with synthetic porcine gastric mucus, natural cow cervical mucus, and a Newtonian-like polymer solution. We have found that an increase in mucin concentration and, correspondingly, an increase in the suspension's elasticity monotonously increases the length scale of collective bacterial locomotion. On the contrary, this length remains practically unchanged in Newtonian polymer solution in a wide range of concentrations. The experimental observations are supported by computational modeling. Our results provide insight into how viscoelasticity affects the spatiotemporal organization of bacterial active matter. They also expand our understanding of bacterial colonization of mucosal surfaces and the onset of antibiotic resistance due to swarming.

Cytocompatibility of titanium and poly(etheretherketone) surfaces after O2 non-thermal plasma sterilization.

The sterilization of medical devices is paramount to achieve an acceptable level of sterility assurance and to prevent hospital-acquired infections. However, some medical devices cannot be sterilized by usual processes such as autoclave (AC) and gamma-ray irradiation (GI). A new non-thermal plasma (NTP) process using sealed bag that preserves the sterile state of the devices could be used as an alternative sterilization method. The aim of the study was to assess the cytocompatibility of titanium and poly(etheretherketone) (PEEK) surfaces after O2-NTP sterilization compared to GI and AC. MG-63 osteoblast-like cells were seeded on titanium (TA6V) and PEEK disks sterilized by AC, GI and O2-NTP. The cells' viability and proliferation, determined by WST-1 and DNA quantification respectively, were enhanced whatever the material types from 3 to 10 days. When seeded on titanium, MG-63 cells showed a higher viability and proliferation after GI and O2-NTP treatment compared to AC treatment. When cultured on PEEK, MG-63 cells showed a higher viability after O2-NTP treatment. No difference of proliferation was observed whatever the sterilization processes. The cell colonization of the materials' surface was confirmed by scanning electron microscopy. Lactate dehydrogenase (LDH) assay revealed no cytotoxicity. Thus, O2-NTP led to similar cell responses to AC and GI and could be a cost-effective alternative process to the usual sterilization methods for fragile medical devices.

Early-life surface colonization with multi-drug resistant organisms in the neonatal intensive care unit

ObjectivesIn this study, we aim to describe the patterns of early-life surface colonization with multi-drug resistant (MDR) organisms (MDROs) among newborns admitted to the neonatal intensive care unit (NICU). MethodsWe conducted a retrospective descriptive study of infants with culture-positive external ear surface swabs performed immediately after admission to our NICU from January 1, 2017 – December 31, 2021. Clinical characteristics, culture and antibiotic susceptibility data were extracted from the department data collection and hospital electronic databases. ResultsA total of 314 infants were included - median 34 weeks gestation (interquartile range [IQR] 30, 38) and median birthweight 2147 g (IQR 1470, 2875). Of the 379 bacterial isolates obtained, 259 (68.3%) were gram-negative with Escherichia coli (149/379, 39.3%) and Klebsiella spp (57/379, 15.0%) the most common gram-negatives isolated. MDROs accounted for 17.4% (45/259) of gram-negative isolates. There was no methicillin-resistant Staphylococcus aureus (0/22 isolates) or vancomycin-resistant Enterococcus (0/68) detected among isolates tested. A total of 27 (8.6%) infants developed bacteremia, of which 21/27 (77.8%) had concordant bacteria isolated from surface cultures, with identical resistance patterns, and 4/21 (19.0%) isolates were MDROs. ConclusionIn our setting where gram-negative bacteria accounted for a high proportion of initial colonization, MDR gram-negatives accounted for up to 17% of colonizing gram-negative bacteria detected.

Magneto-mechanically induced antimicrobial properties of cone-like shaped surfaces

Hygienic surfaces that prevent the proliferation of harmful microorganisms are required in a large variety of environments, including medical areas. Novel strategies are being developed to impede microorganisms colonization of surfaces. In this work, Terfenol-D cone-like shaped nanopatterned surfaces are fabricated by sputtering. The bactericidal effect of such surfaces owed to their morphology is increased in combination with an alternating magnetic field, which boosts the mechanical injury caused to the planktonic cells. Bactericidal assays with Gram-negative Escherichia coli are carried out under static (i.e. without any external stimuli) and dynamic (under the application of an alternating magnetic field) conditions for control silicon substrates, Terfenol-D films and nanostructured surfaces. The nanostructured surfaces at the dynamic condition exhibit the larger bactericidal effect. Bacterial adhesion on the materials was analyzed, and results show a reduction of the attachment surface of bacterial cells on Terfenol-D surfaces in comparison with the control silicon that are attributed both to material properties and nanostructuration. Thus, this work exhibits a method to induce and/or improve the mechanical antimicrobial behavior of surfaces via application of a magnetic field, as an alternative or in combination with chemical methods, which are losing effectiveness due to the increase of antibiotic resistance.

Interaction of surfactants with Prunus laurocerasus leaf surfaces: time-dependent recovery of wetting contact angles depends on physico-chemical properties of surfactants

BackgroundSurfactants are added to spray solutions because they significantly improve foliar uptake of active ingredients (AIs) into the leaves. It was intended to investigate whether surfactant solutions forming a dried deposit on Prunus leaf surfaces after they were sprayed, lead to structural and functional changes of the cuticle/atmosphere interface. This could potentially result in irreversibly enhanced leaf surface wetting, which should be of major disadvantage. Enhanced wetting could promote leaching of ions and promote leaf surface colonization with microorganisms.ResultsPrunus laurocerasus leaf surfaces were sprayed with aqueous solutions of non-ionic alcohol ethoxylates, a cationic, an anionic and one large polar surfactant. Directly after spraying and drying of the different surfactant solutions, wetting contact angles of deionized water (without surfactant) were significantly lower (between 6 and 54°) compared to wetting contact angles on untreated leaves (77°). Leaf surface wettability with deionized water was more pronounced with non-ionic alcohol ethoxylates (wetting contact angles ranging between 6 and 22°) compared to the other 3 surfactants (wetting contact angles ranging between 42 and 54°). Wetting contact angles of deionized water on leaf surfaces treated with non-ionic alcohol ethoxylates continuously increased again over time resulting in final wetting contact angles not different from untreated leaf surfaces. The time-dependent recovery of wetting contact angles was dependent on the degree of ethoxylation of the non-ionic alcohol ethoxylates. The wetting contact angle recovery rate was lower the higher the degree of ethoxylation of the alcohol ethoxylates was. With the cationic, anionic and large polar surfactant a recovery of wetting contact angles was not observed. In addition, on fully dehydrated and dead leaves wetting contact angle recovery was not observed for any of the tested surfactants after spraying and drying. Analytical determinations of the amounts of alcohol ethoxylates on the leaf surfaces after spraying and drying showed that amounts of alcohol ethoxylates decreased over time on the surface (24–72 h).ConclusionOur results indicate that non-ionic alcohol ethoxylates diffused within hours from the leaf surface into the leaf over time and thus fully disappeared from the leaf surface. This was not the case with the cationic, anionic and the large polar surfactants remaining on the leaf surface.Graphical

Geopolymer Antimicrobial and Hydrophobic Modifications: A Review

The article summarizes the state of the art in increasing antimicrobial activity and hydrophobic properties of geopolymer materials. Geopolymers are inorganic polymers formed by polycondensation of aluminosilicate precursors in an alkaline environment and are considered a viable alternative to ordinary Portland cement-based materials, due to their improved mechanical properties, resistance to chemicals, resistance to high temperature, and lower carbon footprint. Like concrete, they are susceptible to microbially induced deterioration (corrosion), especially in a humid environment, primarily due to surface colonization by sulphur-oxidizing bacteria. This paper reviews various methods for hydrophobic or antimicrobial protection by the method of critical analysis of the literature and the results are discussed, along with potential applications of geopolymers with improved antimicrobial properties. Metal nanoparticles, despite their risks, along with PDMS and epoxy coatings, are the most investigated and effective materials for geopolymer protection. Additionally, future prospects, risks, and challenges for geopolymer research and protection against degradation are presented and discussed.

Diatom adhesive trail proteins acquired by horizontal gene transfer from bacteria serve as primers for marine biofilm formation.

Biofilm-forming benthic diatoms are key primary producers in coastal habitats, where they frequently dominate sunlit intertidal substrata. The development of gliding motility in raphid diatoms was a key molecular adaptation that contributed to their evolutionary success. However, the structure-function correlation between diatom adhesives utilized for gliding and their relationship to the extracellular matrix that constitutes the diatom biofilm is unknown. Here, we have used proteomics, immunolocalization, comparative genomics, phylogenetics and structural homology analysis to investigate the evolutionary history and function of diatom adhesive proteins. Our study identified eight proteins from the adhesive trails of Craspedostauros australis, of which four form a new protein family called Trailins that contain an enigmatic Choice-of-Anchor A (CAA) domain, which was acquired through horizontal gene transfer from bacteria. Notably, the CAA-domain shares a striking structural similarity with one of the most widespread domains found in ice-binding proteins (IPR021884). Our work offers new insights into the molecular basis for diatom biofilm formation, shedding light on the function and evolution of diatom adhesive proteins. This discovery suggests that there is a transition in the composition of biomolecules required for initial surface colonization and those utilized for 3D biofilm matrix formation.

Cell-autonomous diversification in bacteria arises from calcium dynamics self-organizing at a critical point.

How dynamic bacterial calcium is regulated, with kinetics faster than typical mechanisms of cellular adaptation, is unknown. We discover bacterial calcium fluctuations are temporal-fractals resulting from a property known as self-organized criticality (SOC). SOC processes are poised at a phase transition separating ordered and chaotic dynamical regimes and are observed in many natural and anthropogenic systems. SOC in bacterial calcium emerges due to calcium channel coupling mediated via membrane voltage. Environmental or genetic perturbations modify calcium dynamics and the critical exponent suggesting a continuum of critical attractors. Moving along this continuum alters the collective information capacity of bacterial populations. We find that the stochastic transition from motile to sessile lifestyle is partially mediated by SOC-governed calcium fluctuations through the regulation of c-di-GMP. In summary, bacteria co-opt the physics of phase transitions to maintain dynamic calcium equilibrium, and this enables cell-autonomous population diversification during surface colonization by leveraging the stochasticity inherent at a boundary between phases.

Contrasting interactions of the aggressive Verticillium longisporum lineage A1/D1 and the non‐aggressive A1/D2 lineage with roots of oilseed rape

AbstractVerticillium longisporum is a soilborne fungal vascular pathogen of Brassica crops. Three hybrid lineages originating from three independent hybridization events of four haploid Verticillium parents are known. V. longisporum lineage A1/D2 has only been found in horseradish and previous pathogenicity tests have shown that it is non‐aggressive on oilseed rape (Brassica napus). The aim of this study was to investigate the interaction of the non‐aggressive lineage A1/D2 with oilseed rape in comparison with the aggressive A1/D1 lineage. Although A1/D2 did not cause severe disease symptoms, the study revealed restricted root infection and systemic colonization by this lineage, as assessed by quantitative PCR of fungal DNA. Assessment of root surface colonization by confocal laser microscopy indicated early sporulation, disorganized loosely attached hyphal growth and hyphal coiling of A1/D2. By contrast, A1/D1 showed organized hyphal growth tight to the root surface, as well as hyphal swellings at points of penetration. Vascular colonization was studied with transmission electron microscopy, which revealed less vessel colonization, higher intensity of vascular responses and lower plant cell wall degradation in A1/D2‐treated plants in comparison with A1/D1. Vascular responses consisted of strong coating of vessel walls and fungal hyphae with an electron‐dense material, indicating higher intensity of plant defence in comparison with A1/D1. The results, for the first time, demonstrate stem colonization and thus a basic level of pathogenic interaction of non‐aggressive strains of A1/D2 with oilseed rape associated with a lack of severe disease symptoms, which indicate tolerance in oilseed rape to lineage A1/D2.

Environmental aging and biodegradation of tire wear microplastics in the aquatic environment

Tire wear microplastics (TWM) are formed by friction between tires and road surfaces during driving and they belong among the most abundant microplastics in the environment. However, the information about their fate in the environment is still unknown. The aim of this study was to investigate the aging of TWM in freshwater under controlled laboratory conditions over 12 weeks. The development of biofilm, changes in physical properties and chemical composition, leaching and biodegradation of TWM were followed. The results showed colonization of the TWM surface by microorganisms (up to 45 mg/g), which, however, began to detach from the particles after eight weeks, reducing the amount of biofilm. TWM initially leached zinc and organic compounds (expressed as dissolved organic carbon - DOC), but their concentrations were low and decreased with time. The increase in DOC was observed after 10 weeks, possibly due to the decomposition of the biofilm and the release of organic matter. Aging resulted in changes of density of TWM, but the morphology and chemical composition of the TWM surface did not change. This confirms the results of the biodegradability tests, which showed no biodegradation within 12 weeks. Overall, the results indicated that TWM are not readily biodegradable and therefore may accumulate and persist in the aquatic environment.