Search Results (19)

Orchid plants, due to their high aesthetic qualities of large inflorescences, long flowering period, and ease of care, have high commercial potential; however, when grown industrially in factories, they are susceptible to infectious diseases. In this study, we isolated from Phalaenopsis spp. plants epiphytic, rhizospheric, and endophytic bacteria associated with soft rot symptoms. Twenty-nine isolates exhibiting pectolytic activity were identified as strains of the genera Bacillus, Klebsiella, Microbacterium, Paenibacillus, Paracidovorax, Pseudomonas, and Psychrobacillus based on 16S rRNA analysis. These isolates were tested for their ability to produce cellulase, amylase, sucrase, proteinase, and lipase; to form biofilms; and to exhibit motility (swimming and swarming). Potato microplants under in vitro conditions were used as a model object for initial screening of the strains’ potential phytotoxicity. Most strains were shown to inhibit plant growth, particularly root development. Injection of suspensions of these strains into orchid leaves caused symptoms of soft rot. Thus, we isolated Gram-positive bacteria for the first time from orchid tissues with soft rot symptoms and demonstrated an association of these strains with plant tissue maceration in potato and orchids. Gram-positive bacteria with pectolytic activity are not typical pathogens of orchid soft rot and may require changes in approaches to the monitoring of phytopathogens for this group of plants. Full article

Epiphytic and leafless orchids possess specialized root structures and host diverse associated microorganisms, which may contribute to their adaptation to limited access to water, nutrients, and photosynthetic tissues. Microscopic, anatomical, and molecular analyses of aerial and substrate roots of Chiloschista lunifera (Rchb.f.) J.J.Sm. revealed consistent phototrophic microbial biofilms on the velamen surface. These biofilms comprised filamentous and unicellular cyanobacteria, singular bacterial cells, and green algae. Morphological characterisation identified cyanobacteria belonging to five taxonomic orders, which are Chroococcales, Chroococcidiopsidales, Nostocales, Leptolyngbyales, and Synechococcales. 16S rRNA amplicon sequencing confirmed cyanobacterial dominance, with Chroococcidiopsis thermalis PCC 7203 strongly prevalent in root wash samples (up to 99.99% relative abundance), while root homogenate samples harboured a more diverse assemblage including Phormidiaceae, Leptolyngbya, Scytonema, and Calothrix. In addition, a green alga from Watanabeales (Jaagichlorella sp.) was identified based on morphological characteristics. TEM showed diverse cyanobacterial forms and unicellular green algae with well-developed photosynthetic structures. Root anatomy differed between root types. Substrate roots exhibited an inverted kidney-shaped transverse profile, whereas aerial roots were circular, with differences in velamen distribution reflecting adaptation for water retention and substrate attachment. Branched root hairs occurred on substrate roots, while aerial roots possessed unbranched root hairs, indicating functional specialization. Autofluorescence analysis revealed lignified tissues and abundant cortical chloroplasts, suggesting a photosynthetic role of the roots that may compensate for leaflessness. These findings enhance our understanding of the anatomical and ultrastructural features of epiphytic leafless orchid roots and may support future conservation and propagation efforts. Full article

Cyanobacteria of the genus Tychonema are common inhabitants of freshwater bodies in a temperate zones. In Lake Baikal, Tychonema sp. was first reported in 2014. It grows excessively on the bottom on almost all substrates types, forming epiphytic and epizoic biofilms, and its role in the mass mortality events affecting of endemic Baikal sponges is discussed. The cyanobacterial strain BBK16 (=IPPAS B-2063^T), isolated from a biofilm on a log pier in the Bolshiye Koty settlement in 2016, was used for further taxonomic characterization. Key morphological features of strain BBK16 include its growth as a creeping mat, highly motile trichomes that are sometimes narrowed and hooked at the ends, and the presence of rounded-conical apical cells with a calyptra. Strain ultrastructure (fascicular parietal thylakoids and type D cell division) differs from Tychonema species with radial thylakoids but aligns with other genera in the Microcoleaceae family. A comprehensive analysis, including 16S rRNA gene phylogeny, conserved protein phylogeny, and whole-genome comparisons, confirmed its placement within the genus Tychonema. The average nucleotide identity, average amino acid identity and digital DNA–DNA hybridization values between strain BBK16 and T. bourrellyi FEM GT703 were 90.7%, 91.1% and 43.3%, respectively, indicating values below the standard thresholds for species delineation. Based on combined morphological, genomic, and ecological evidence, we propose the name Tychonema litorale sp. nov. for strain BBK16, a novel species described in accordance with the International Code of Nomenclature for Algae, Fungi, and Plants. Full article

Submerged plants are common bioindicators of heavy metal contamination in aquatic ecosystems. Traditional research often focused on the overall metal content within plants or their tissues, and overlooked the role of epiphytic biofilms, which may affect the accuracy of bioindicators’ function. This study conducted cadmium (Cd) exposure experiments on Vallisneria natans L. (V. natans) to investigate the Cd accumulation in the plant tissues and their associated epiphytic biofilms. It was observed that phyllosphere and rhizosphere biofilms accounted for 27.3–60.6% and 31.3–41.7% of the total Cd in the leaf and root areas, respectively. The phyllosphere biofilm rapidly accumulated Cd directly from the water column, showing a strong correlation with aqueous Cd levels, whereas Cd in leaf tissue was primarily root-derived. Microbial analysis revealed that Proteobacteria dominated the biofilm communities, and the relative abundance of specific genera (e.g., Nitrospira) correlated with Cd exposure. Our findings underscore a critical refinement for bioindication: distinguishing metal accumulation in epiphytic biofilms from that in plant tissues significantly enhances source attribution: phyllosphere biofilms reflect water pollution, while rhizosphere biofilms indicate sediment–root interface contamination. This refined approach provides a pathway toward more precise environmental monitoring, contributing to sustainable water quality management by enabling earlier detection of pollution and informing targeted remediation efforts to protect aquatic ecosystem health. Full article

Salinity is a key determinant governing microalgal growth, biochemical composition, and the structure of associated epiphytic bacterial communities. To investigate the effects of salinity on the structure and function of the epiphytic bacterial community in Desmodesmus intermedius, this study utilized 16S rRNA gene high-throughput sequencing to analyze the communities across the control (S0) and experimental groups (S5, S10, S15). The results demonstrated that salinity is a key environmental driver governing the structural and functional succession of the bacterial community. Alpha diversity analysis revealed that the control group exhibited the highest bacterial diversity and greater evenness. In contrast, the experimental groups showed a significant increase in the relative abundance of Thauera and a concurrent decrease in Roseococcus with increasing salinity. Beta diversity analysis revealed clear segregation of the epiphytic bacterial communities across the salinity groups. FAPROTAX functional prediction revealed that increasing salinity led to a reduction in chemoheterotrophy, photoheterotrophy, and aerobic chemoheterotrophy, while enhancing nitrogen respiration, nitrate reduction, and other denitrification processes. This shift indicates a substantial reconfiguration of carbon and nitrogen metabolic pathways. BugBase phenotype analysis further revealed that the experimental groups exhibited a higher proportion of Gram-positive bacteria and enhanced biofilm-forming capacity. Canonical correspondence analysis identified salinity as the predominant factor shaping bacterial community structure. This study comprehensively investigates the response mechanisms of the D. intermedius epiphytic bacterial community to salt stress, laying a foundation for understanding microbial functions within the phycosphere. Full article

Constructed wetlands play a critical role in mitigating aquaculture wastewater pollution. However, the comprehensive treatment performance of aquatic plants and microorganisms under various water treatment processes remains insufficiently understood. Here, a multi-stage surface flow constructed wetland (SFCW) comprising four different aquatic plant species, along with aeration and biofiltration membrane technologies, was investigated to explore the combined effects of aquatic plants and epiphytic biofilms on wastewater removal efficiency across different vegetation periods and treatment processes. The results demonstrated that the total removal efficiency consistently exceeded 60% in both vegetation periods, effectively intercepting a range of pollutants present in aquaculture wastewater. Changes in the vegetation period influenced the performance of the SFCW, with the system’s ability to treat total nitrogen becoming more stable over time. The removal efficiency of the treatment pond planted with submerged plants was highest in July, while the pond planted with emergent plants showed an increased removal rate in November. The aeration pond played a significant role in enhancing dissolved oxygen levels, thereby improving phosphorus removal in July and nitrogen removal in November. Additionally, the α-diversity of epiphytic bacteria in the aeration and biofiltration ponds was significantly higher compared to other ponds. In terms of bacterial composition, the abundance of Firmicutes was notably higher in July, whereas Nitrospirota and Acidobacteriota exhibited a significant increase in November. Furthermore, the functional genes associated with sulfur metabolism, nitrogen fixation, and oxidative phosphorylation displayed significant temporal variations in the aeration pond, highlighting that both growth period changes and treatment processes influence the expression of functional genes within biofilms. Our findings suggest that the integration of water treatment processes in SFCWs enhances the synergistic effects between aquatic plants and microorganisms, helping to mitigate the adverse impacts of vegetation period changes and ensuring stable and efficient wastewater treatment performance. Full article

Bacillus sp. G2112, an isolate from cucumber plants that inhibited plant pathogens, produces not only surfactins, iturins, and fengycins common to many Bacillus spp., but also a large variety of N-acyl-(depsi)peptides related to A-3302-B and nobilamides. Four known and fourteen previously unreported nobilamide peptides were characterized using high-resolution mass spectrometry, tandem mass spectrometry, and NMR. The stereochemistry of the amino acids of nobilamide peptides was determined using Marfey’s method. The diversity of nobilamide peptides from Bacillus sp. G2112 resulted from the incorporation of different acyl groups and amino acids in the sequence. The peptides occur in linear or cyclic form. In addition, a truncated N-acetylpentapeptide was produced. Agar diffusion assays with selected nobilamide peptides against plant pathogens and human pathogens revealed that A-3302-B and its N-acyl homologs, A-3302-A and nobilamide J, exhibited powerful antibiotic activity (at 5 µg/hole) against Lysinibacillus sphaericus that can cause severe sepsis and bacteremia in patients. Moreover, nobilamide peptides from Bacillus sp. G2112 strongly promoted biofilm formation in the Gram-positive Mycobacterium aurum and Gram-negative pseudomonads. Structurally diverse nobilamides from Bacillus sp. G2112, whether linear or cyclic, penta and heptapeptides, induced biofilm formation, suggesting that the common N-acetyl-D-Phe-D-Leu-L-Phe-D-allo-Thr-L-Val amino acid sequence motif is important for the biofilm-inducing activity. Full article

This study evaluated the biocontrol effect of isolated epiphytic yeasts (Papiliotrema terrestris, Hanseniaspora uvarum, and Rhodosporidium glutinis) against Botrytis cinerea and Alternaria alternata in blueberry fruits and its possible mechanisms. Our findings indicated that the three tested yeasts exerted a good biocontrol effect on postharvest diseases in blueberry, and that H. uvarum was the most effective. In addition, the three tested yeasts could improve the postharvest storage quality of blueberry fruits to some extent. H. uvarum demonstrated the strongest direct inhibitory effect on pathogens by suppressing spore germination, mycelial growth, and antifungal volatile organic compound (VOC) production. P. terrestris showed the highest extracellular lytic enzymes activities. It also had better adaptation to low temperature in fruit wounds at 4 °C. The biofilm formation capacity was suggested to be the main action mechanism of R. glutinis, which rapidly colonized fruit wounds at 20 °C. Several action mechanisms are employed by the superb biocontrol yeasts, while yeast strains possess distinctive characteristics and have substantially different action mechanisms. Full article

In order to investigate the effects of various nitrogen (ammonium) concentrations on the epibiotic bacterial community associated with Oocystis borgei, a metabarcoding sequencing method was employed. The 16S rDNA sequencing and bioinformatic analysis were conducted on cultures of O. borgei that were grown on four different nitrogen element concentrations (2.5, 10, 50, and 100 mg/L), and the differences in the epibiotic bacterial community and functions of O. borgei among different nitrogen concentrations were compared. The results showed that the chlorophyll a content of O. borgei increased with increasing nitrogen concentrations. A total of 43 operational taxonomic units (OTUs) were obtained from the four groups, which were categorized into 7 phyla, 9 classes, 18 orders, 21 families, and 26 genera. Winogradskyella was the dominant genus in the groups with 2.5 and 100 mg/L nitrogen, while Marinobacter and Winogradskyella were relatively abundant in the groups with 50 and 100 mg/L nitrogen. Functional analysis using PICRUST2 showed that the three most abundant gene functions were carbohydrate metabolism, amino acid transport and metabolism, ribosomal structure and biogenesis, and energy production and conversion. BugBase phenotypic analysis revealed that there were no significant differences in phenotypes between the groups with 2.5 and 10 mg/L nitrogen, while the group with 50 mg/L nitrogen exhibited higher abundance in aerobicity, biofilm formation, mobile genetic elements, and stress tolerance phenotypes. Most of the bacteria in this work belonged to the aerobic types. A redundancy analysis (RDA) of environmental factors demonstrated that nitrogen concentration showed a positive correlation with species changes in the groups with 10 and 50 mg/L nitrogen; chlorophyll a exhibited a positive correlation with species changes in the groups with 50 and 100 mg/L nitrogen. Nitrogen concentration significantly influenced the epibiotic bacterial community associated with O. borgei, leading to changes in dominant species and community structure. This study provides important references for understanding the functional characteristics of the epiphytic microbial community of O. borgei and the exploration of specific microorganisms. Full article

N-acyl homoserine lactones (AHLs) are small, diffusible chemical signal molecules that serve as social interaction tools for bacteria, enabling them to synchronize their collective actions in a density-dependent manner through quorum sensing (QS). The QS activity from epiphytic bacteria of the red macroalgae Porphyra haitanensis, along with its involvement in biofilm formation and regulation, remains unexplored in prior scientific inquiries. Therefore, this study explores the AHL signal molecules produced by epiphytic bacteria. The bacterium isolated from the surface of P. haitanensis was identified as Pseudoalteromonas galatheae by 16s rRNA gene sequencing and screened for AHLs using two AHL reporter strains, Agrobacterium tumefaciens A136 and Chromobacterium violaceum CV026. The crystal violet assay was used for the biofilm-forming phenotype. The inferences revealed that P. galatheae produces four different types of AHL molecules, i.e., C4-HSL, C8-HSL, C18-HSL, and 3-oxo-C16-HSL, and it was observed that its biofilm formation phenotype is regulated by QS molecules. This is the first study providing insights into the QS activity, diverse AHL profile, and regulatory mechanisms that govern the biofilm formation phenotype of P. galatheae. These findings offer valuable insights for future investigations exploring the role of AHL producing epiphytes and biofilms in the life cycle of P. haitanensis. Full article

Brownification denotes increasing water color, partly caused by increasing dissolved organic matter of terrestrial origin in freshwater. Brownification has become a wide-spread environmental problem because water color alters the physicochemical environment and biological communities in aquatic ecosystems. However, our understanding of its ecological effects on aquatic macrophytes is limited. Here, an indoor mesocosm experiment with a common submerged macrophyte, Potamogeton crispus, along an increasing gradient of brownification was conducted over a period of 42 days. Results showed that P. crispus was able to overcome low degrees of brownification owing to the plasticity in morphological and physiological traits and P. crispus growth even benefitted from the concomitant nutrients along with brownification. However, the macrophyte growth was negatively affected by a 10-fold increase in water color beyond its current level. Additionally, collapse in antioxidant systems and potent photosynthesis inhibition implied that P. crispus could not adapt to the low-light stress generated under the high degree of brownification. Epiphytic bacteria are more sensitive to brownification than their hosts. Any degree of brownification initially caused a decrease in microbial diversity of epiphytic biofilm, whereafter the concomitant nutrients under brownification favored the growth of epiphytic microorganisms. The shading effect of a large number of epiphytic biofilms under brownification may further aggravate the low-light stress on macrophytes. Overall, the study provides new insights into the comprehensive effects and underlying mechanisms of brownification on macrophytes. Full article

This paper investigates the effects of water flow on the growth and physiological indicators of the submerged macrophyte, Vallisneria natans, and the bacteria and algae community composition on its epiphytic biofilm-covered leaves. The authors set up a simulated flowing water laboratory experiment testing high nitrogen (N) and phosphorus (P) concentrations. Total chlorophyll and dissolved oxygen (DO) was significantly enhanced, and turbidity was reduced, thereby accelerating the growth of V. natans. These experiments were compared to another set of observations on a static group. The accumulation of malonaldehyde (MDA) in the dynamic groups was significantly higher than that in the static group. As an antioxidant stress response, the total superoxide dismutase (T-SOD) was also induced in plants exposed to nutrient-rich flowing water. The results of 16S rRNA high-throughput sequencing analyses showed that the water flow increased the bacteria community diversity of biofilm-producing bacteria with N and P removing bacteria, carbon cycle bacteria, and plant growth-promoting rhizobacteria on the epiphytic biofilm. This research determined that water flow alleviates the adverse effects of eutrophication when V. natans grows in water containing high N and P concentrations. Water flow also inhibits the growth of cyanobacteria (also referred to as blue-green algae) in epiphytic biofilm. The ecological factor of water flow, such as water disturbance and aeration measures, could alleviate the adverse effect of eutrophic water by providing a new way to restore submerged macrophytes, such as V. natans, in eutrophic water. Full article

The leafless orchids are rare epiphytic plants with extremely reduced leaves, and their aerial roots adopted for photosynthesis. The beneficial plant–microbial interactions contribute significantly to host nutrition, fitness, and growth. However, there are no data available on the bacterial associations, inhabiting leafless orchids. Here, we describe the diversity of cyanobacteria, which colonize the roots of greenhouse Microcoelia moreauae and Chiloschista parishii. The biodiversity and structure of the cyanobacterial community were analyzed using a complex approach, comprising traditional cultivable techniques, denaturing gradient gel electrophoresis (DGGE), and phylogenetic analysis, as well as the light and scanning electron microscopy (SEM). A wide diversity of associated bacteria colonize the root surface, forming massive biofilms on the aerial roots. The dominant populations of filamentous nitrogen-fixing cyanobacteria belonged to the orders Oscillatoriales, Synechococcales, and Nostocales. The composition of the cyanobacterial community varied, depending on the nitrogen supply. Two major groups prevailed under nitrogen-limiting conditions, belonging to Leptolyngbya sp. and Komarekiella sp. The latter was characterized by DGGE profiling and sequencing, as well as by its distinctive features of morphological plasticity. The leading role of these phototrophophic and diazotrophic cyanobacteria is discussed in terms of the epiphytic lifestyle of the leafless orchids. Full article

Planctomycetes are bacteria that were long thought to be unculturable, of low abundance, and therefore neglectable in the environment. This view changed in recent years, after it was shown that members of the phylum Planctomycetes can be abundant in many aquatic environments, e.g., in the epiphytic communities on macroalgae surfaces. Here, we analyzed three different macroalgae from the North Sea and show that Planctomycetes is the most abundant bacterial phylum on the alga Fucus sp., while it represents a minor fraction of the surface-associated bacterial community of Ulva sp. and Laminaria sp. Especially dominant within the phylum Planctomycetes were Blastopirellula sp., followed by Rhodopirellula sp., Rubripirellula sp., as well as other Pirellulaceae and Lacipirellulaceae, but also members of the OM190 lineage. Motivated by the observed abundance, we isolated four novel planctomycetal strains to expand the collection of species available as axenic cultures since access to different strains is a prerequisite to investigate the success of planctomycetes in marine environments. The isolated strains constitute four novel species belonging to one novel and three previously described genera in the order Pirellulales, class Planctomycetia, phylum Planctomycetes. Full article

Xanthomonas citri subsp. citri (X. citri) is a plant pathogenic bacterium causing citrus canker disease. The xanA gene encodes a phosphoglucomutase/phosphomannomutase protein that is a key enzyme required for the synthesis of lipopolysaccharides and exopolysaccharides in Xanthomonads. In this work, firstly we isolated a xanA transposon mutant (xanA::Tn5) and analyzed its phenotypes as biofilm formation, xanthan gum production, and pathogenesis on the sweet orange host. Moreover, to confirm the xanA role in the impaired phenotypes we further produced a non-polar deletion mutant (ΔxanA) and performed the complementation of both xanA mutants. In addition, we analyzed the percentages of the xanthan gum monosaccharides produced by X. citri wild-type and xanA mutant. The mutant strain had higher ratios of mannose, galactose, and xylose and lower ratios of rhamnose, glucuronic acid, and glucose than the wild-type strain. Such changes in the saccharide composition led to the reduction of xanthan yield in the xanA deficient strain, affecting also other important features in X. citri, such as biofilm formation and sliding motility. Moreover, we showed that xanA::Tn5 caused no symptoms on host leaves after spraying, a method that mimetics the natural infection condition. These results suggest that xanA plays an important role in the epiphytical stage on the leaves that is essential for the successful interaction with the host, including adaptive advantage for bacterial X. citri survival and host invasion, which culminates in pathogenicity. Full article