Epiphytic Microbial Communities Research Articles

Impacts of fertilization methods on Salvia miltiorrhiza quality and characteristics of the epiphytic microbial community

Plant epiphytic microorganisms have established a unique symbiotic relationship with plants, which has a significant impact on their growth, immune defense, and environmental adaptation. However, the impact of fertilization methods on the epiphytic microbial community and their correlation with the yield and quality of medicinal plant was still unclear. In current study, we conducted a field fertilization experiment and analyzed the composition of epiphytic bacterial and fungal communities employing high throughput sequencing data in different organs (roots, stems, and leaves) of Salvia miltiorrhiza, as well as their correlation with plant growth. The results showed that fertilization significantly affected the active ingredients and hormone content, soil physicochemical properties, and the composition of epiphytic microbial communities. After fertilization, the plant surface was enriched with a core microbial community mainly composed of bacteria from Firmicutes, Proteobacteria, and Actinobacteria, as well as fungi from Zygomycota and Ascomycota. Additionally, plant growth hormones were the principal factors leading to alterations in the epiphytic microbial community of S. miltiorrhiza. Thus, the most effective method of fertilization involved the application of base fertilizer in combination with foliar fertilizer. This study provides a new perspective for studying the correlation between microbial community function and the quality of S. miltiorrhiza, and also provides a theoretical basis for the cultivation and sustainable development of high-quality medicinal plants.

Epiphytic and endophytic bacteria on Camellia oleifera phyllosphere: exploring region and cultivar effect

The epiphytic and endophytic bacteria play an important role in the healthy growth of plants. Both plant species and growth environmental influence the bacterial population diversity, yet it is inconclusive whether it is the former or the latter that has a greater impact. To explore the communities of the epiphytic and endophytic microbes in Camellia oleifera, this study assessed three representative C. oleifera cultivars from three areas in Hunan, China by Illumina high-throughput sequencing. The results showed that the diversity and species richness of endophytic microbial community in leaves were significantly higher than those of microbial community in the epiphytic. The diversity and species richness of epiphytic and endophytic microbes are complex when the same cultivar was grown in different areas. The C. oleifera cultivars grown in Youxian had the highest diversity of epiphytic microbial community, but the lowest abundance, while the cultivars grown in Changsha had the highest diversity and species richness of endophytic microbes in leaves. It was concluded that the dominant phylum mainly included Proteobacteria, Actinobacteriota and Firmicutes through the analysis of the epiphytic and endophytic microbial communities of C. oleifera. The species and relative abundances of epiphytic and endophytic microbial community were extremely different at the genus level. The analysis of NMDS map and PERMANOVA shows that the species richness and diversity of microbial communities in epiphytes are greatly influenced by region. However, the community structure of endophytic microorganisms in leaves is influenced by region and cultivated varieties, but the influence of cultivars is more significant. Molecular ecological network analysis showed that the symbiotic interaction of epiphytic microbial community was more complex.

Accelerated nitrogen cycling on Mediterranean seagrass leaves at volcanic CO2 vents

Seagrass meadows form highly productive and diverse ecosystems in coastal areas worldwide, where they are increasingly exposed to ocean acidification (OA). Efficient nitrogen (N) cycling and uptake are essential to maintain plant productivity, but the effects of OA on N transformations in these systems are poorly understood. Here we show that complete N cycling occurs on leaves of the Mediterranean seagrass Posidonia oceanica at a volcanic CO2 vent near Ischia Island (Italy), with OA affecting both N gain and loss while the epiphytic microbial community structure remains largely unaffected. Daily leaf-associated N2 fixation contributes to 35% of the plant’s N demand under ambient pH, while it contributes to 45% under OA. Nitrification potential is only detected under OA, and N-loss via N2 production increases, although the balance remains decisively in favor of enhanced N gain. Our work highlights the role of the N-cycling microbiome in seagrass adaptation to OA, with key N transformations accelerating towards increased N gain.

Spatiotemporal Dynamics of Assyrtiko Grape Microbiota.

Vitis vinifera, an economically significant grapevine species, is known for wine, juice, and table grape production. The berries of wine grapes host a diverse range of microorganisms influencing both grapevine health and the winemaking process. Indigenous to Greece, the emblematic variety Assyrtiko, renowned for high-quality white wines, originated from Santorini and spread to various Greek regions. Despite existing studies on the microbiota of several varieties, the carposphere microbiota of Assyrtiko grapes remains unexplored. Thus, we conducted a spatiotemporal metagenomic study to identify the epiphytic microbial community composition of Assyrtiko grapes. The study was conducted in two consecutive vintage years (2019 and 2020) across three different and distinct viticulture regions in Greece (Attica, Thessaloniki, Evros). We performed amplicon sequencing, targeting the 16S rRNA gene for bacteria and the ITS region for fungi, with subsequent comprehensive bioinformatic analysis. Our data indicate that the distribution and relative abundance of the epiphytic carposphere microbial communities of the Assyrtiko variety are shaped both by vintage and biogeography.

A matter of choice: Understanding the interactions between epiphytic foraminifera and their seagrass host Halophila stipulacea

In sub/tropical waters, benthic foraminifera are among the most abundant epiphytic organisms inhabiting seagrass meadows. This study explored the nature of the association between foraminifera and the tropical seagrass species H. stipulacea, aiming to determine whether these interactions are facilitative or random. For this, we performed a "choice" experiment, where foraminifera could colonize H. stipulacea plants or plastic "seagrasses" plants. At the end of the experiment, a microbiome analysis was performed to identify possible variances in the microbial community and diversity of the substrates. Results show that foraminifera prefer to colonize H. stipulacea, which had a higher abundance and diversity of foraminifera than plastic seagrass plants, which increased over time and with shoot age. Moreover, H. stipulacea leaves have higher epiphytic microbial community abundance and diversity. These results demonstrate that seagrass meadows are important hosts of the foraminifera community and suggest the potential facilitative effect of H. stipulacea on epiphytic foraminifera, which might be attributed to a greater diversity of the microbial community inhabiting H. stipulacea.

Effects of Nitrogen on the Bacterial Microbiome Community of Oocystis borgei, an Alga Widely Used in Marine Aquaculture

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.

Seasonal dynamics of phyllosphere epiphytic microbial communities of medicinal plants in farmland environment.

The phyllosphere of plants is inhabited by various microorganisms, which play a crucial role in plant physiological metabolism. Currently, there is limited research on the dynamic effects of species and seasons on plant phyllosphere microbial community diversity and microbial interactions. In this study, high-throughput sequencing technology was used to sequence the leaf surface parasitic microorganisms of five medicinal plants (Bupleurum chinense, Atractylodes lancea, Salvia miltiorrhiza, Astragalus membranaceus, and Lonicera japonica). The results showed that bacteria and fungi clustered into 3,898 and 1,572 operational taxonomic units (OTUs), respectively. Compared to host species, seasons had a more significant impact on the a diversity of bacteria and fungi. The heterogeneity of phyllosphere microbial communities was greater in winter compared to summer. Key species analysis at the OTU level and Spearman correlation analysis demonstrated significant preferences in microbial interactions under plant and seasonal backgrounds. The network connections between bacterial and fungal communities significantly increased during seasonal transitions compared to connections with plants. This study enhances our understanding of the composition and ecological roles of plant-associated microbial communities in small-scale agricultural environments. Additionally, it provides valuable insights for assessing the biodiversity of medicinal plants.

Epiphytic microorganisms of submerged macrophytes effectively contribute to nitrogen removal

Submerged macrophytes play important roles in nutrient cycling and are widely used in ecological restoration to alleviate eutrophication and improve water quality in lakes. Epiphytic microbial communities on leaves of submerged macrophytes might promote nitrogen cycling, but the mechanisms and quantification of their contributions remain unclear. Here, four types of field zones with different nutrient levels and submerged macrophytes, eutrophic + Vallisneria natans (EV), eutrophic + V. natans + Hydrilla verticillata, mesotrophic + V. natans + H. verticillata, and eutrophic without macrophytes were selected to investigate the microbial communities that involved in nitrification and denitrification. The alpha diversity of bacterial community was higher in the phyllosphere than in the water, and that of H. verticillata was higher compared to V. natans. Bacterial community structures differed significantly between the four zones. The highest relative abundance of dominant bacterioplankton genera involved in nitrification and denitrification was observed in the EV zone. Similarly, the alpha diversity of the epiphytic ammonia-oxidizing archaea and nosZI-type denitrifiers were highest in the EV zone. Consist with the diversity patterns, the potential denitrification rates were higher in the phyllosphere than those in the water. Higher potential denitrification rates in the phyllosphere were also found in H. verticillata than those in V. natans. Anammox was not detected in all samples. Nutrient loads, especially nitrogen concentrations were important factors influencing potential nitrification, denitrification rates, and bacterial communities, especially for the epiphytic nosZI-type taxa. Overall, we observed that the phyllosphere harbors more microbes and promotes higher denitrification rates compared to water, and epiphytic bacterial communities are shaped by nitrogen nutrients and macrophyte species, indicating that epiphytic microorganisms of submerged macrophytes can effectively contribute to the N removal in shallow lakes.

Metabolomics and Microbiomics Insights into Differential Surface Fouling of Three Macroalgal Species of Fucus (Fucales, Phaeophyceae) That Co-Exist in the German Baltic Sea.

The brown algal genus Fucus provides essential ecosystem services crucial for marine environments. Macroalgae (seaweeds) release dissolved organic matter, hence, are under strong settlement pressure from micro- and macrofoulers. Seaweeds are able to control surface epibionts directly by releasing antimicrobial compounds onto their surfaces, and indirectly by recruiting beneficial microorganisms that produce antimicrobial/antifouling metabolites. In the Kiel Fjord, in the German Baltic Sea, three distinct Fucus species coexist: F. vesiculosus, F. serratus, and F. distichus subsp. evanescens. Despite sharing the same habitat, they show varying fouling levels; F. distichus subsp. evanescens is the least fouled, while F. vesiculosus is the most fouled. The present study explored the surface metabolomes and epiphytic microbiota of these three Fucus spp., aiming to uncover the factors that contribute to the differences in the fouling intensity on their surfaces. Towards this aim, algal surface metabolomes were analyzed using comparative untargeted LC-MS/MS-based metabolomics, to identify the marker metabolites influencing surface fouling. Their epiphytic microbial communities were also comparatively characterized using high-throughput amplicon sequencing, to pinpoint the differences in the surface microbiomes of the algae. Our results show that the surface of the least fouling species, F. distichus subsp. evanescens, is enriched with bioactive compounds, such as betaine lipids MGTA, 4-pyridoxic acid, and ulvaline, which are absent from the other species. Additionally, it exhibits a high abundance of the fungal genera Mucor and Alternaria, along with the bacterial genus Yoonia-Loktanella. These taxa are known for producing antimicrobial/antifouling compounds, suggesting their potential role in the observed fouling resistance on the surface of the F. distichus subsp. evanescens compared to F. serratus and F. vesiculosus. These findings provide valuable clues on the differential surface fouling intensity of Fucus spp., and their importance in marine chemical defense and fouling dynamics.

Impact of nitrogen application and crop stage on epiphytic microbial communities on silage maize leaf surfaces.

This study aimed to examine the impact of nitrogen (N) fertilization on phyllosphere microorganisms in silage maize (Zea mays) to enhance the production of high-quality silage. The effects of different N application rates (160, 240, and 320 kg ha-1) and maturity stages (flowering and dough stages) on microbial diversity, abundance and physiochemical properties of the leaf surfaces were evaluated in a field experiment. The results showed that N application rates did not significantly impact the abundance of lactic acid bacteria (LAB), aerobic bacteria (AB), yeasts, or molds on the leaf surfaces. However, these microbes were more abundant during the flowering stage compared to the dough stage. Furthermore, the N application rate had no significant impact on inorganic phosphorus, soluble sugar, free amino acids, total phenolic content, and soluble protein concentrations, or pH levels on the leaf surfaces. Notably, these chemical indices were lower during the dough stage. The abundance of Pantoea decreased with higher N application rates, while that of other microorganisms did not changes significantly. The abundance of AB, LAB, yeasts, and molds were positively correlated with soluble sugar, soluble protein, inorganic phosphorus, free amino acids, and total phenolic concentrations on leaf surfaces. Moreover, water loss was negatively correlated with the abundance of AB, LAB, yeasts, and molds, whereas water retention capacity and stomatal density were positively correlated with microbial abundance. We recommend applying an optimal N rate of 160 kg ha-1 to silage maize and harvesting at the flowering stage is recommended.

Coupling of submerged macrophytes and epiphytic biofilms reduced methane emissions from wetlands: Evidenced by an antibiotic inhibition experiment

Wetlands are the largest natural methane source, but how submerged macrophytes affect methane emission remains controversial. In this study, the impacts of submerged macrophytes on methane fluxes, water purification, and epiphytic microbial community dynamics were investigated in simulated wetlands (with and without Hydrilla verticillata) treated with norfloxacin (NOR) for 24 days. Mean methane fluxes were significantly lower in treatments with Hydrilla verticillata (56.84–90.94 mg/m2/h) than bulks (65.96–113.21 mg/m2/h) (p < 0.05) during the experiment regardless of NOR. The relative conductivity (REC) values, H2O2, and malondialdehyde (MDA) contents increased in plant leaves, while water nutrients removal rates decreased with increasing NOR concentration at the same sampling time. The partial least squares path model analysis revealed that plant physiological indices and water nutrients positively affected methane fluxes (0.72 and 0.49, p < 0.001). According to illumina sequencing results of 16S rRNA and pmoA genes, α-proteobacteria (type II) and γ-proteobacteria (type I) were the dominant methanotroph classes in all epiphytic biofilms. The ratio of type I/type II methanotrophs and pmoA gene abundance in epiphytic biofilm was considerably lower in treatment with 16 mg/L NOR than without it (p < 0.05). pmoA gene abundance was negatively correlated with methane fluxes (p < 0.05). Additionally, the assembly of epiphytic bacterial community was mainly governed by deterministic processes, while stochastic dispersal limitation was the primary assembly process in the epiphytic methanotrophic community under NOR stress. The deterministic process gained more importance with time both in bacterial and methanotrophic community assembly. Network analysis revealed that relationships among bacteria in epiphytic biofilms weakened with time but associations among methanotrophic members were enhanced under NOR stress over time. It could be concluded that submerged macrophytes-epiphytic biofilms symbiotic system exhibited potential prospects to reduce methane emissions from wetlands under reasonable management.

Growth promotion of Sargassum fusiforme by epiphytic microbes is dependent on the extent of interspecific interactions of the microbial community

Profound growth differences such as seedling length and biomass are often observed during the cultivation of Sargassum fusiforme despite the absence of detectable variance in abiotic factors that could have affected this process. This highlights the importance of biotic factors such as epiphytic microbiota in controlling seedling growth. Yet, how, and to what extent microbial activities can affect host growth in the presence of seawater flow and continuous erosion remains debatable. Particularly, the contribution of microbial network interactions to the growth of macroalgae remains poorly understood. This study aimed to compare the physicochemical properties of S. fusiforme seedlings via 16S rRNA gene Illumina sequencing-based profiling of the epiphytic microbial communities of seedlings with different lengths. Significantly different epiphytic bacterial communities were observed among S. fusiforme seedlings of different lengths. The result showed that community from longer seedlings maintained higher bacterial diversity with the taxa Gammaproteobacteria, Burkholderiales, Alteromonadales, Vibrionaceae, Ralstonia, Colwelliaceae, and Thalassotalea being selectively enriched. More importantly, microbial interspecific interactions, which were predominantly positive, were enhanced consistently in communities of the longer seedlings, indicative of reinforced prevalent and mutually cooperative relationships among the microorganisms associated with S. fusiforme seedlings of greater length. Furthermore, longer seedlings also displayed up-regulation of microbial functional potentials involved in N fixation and mineralization, P mineralization and transportation, and ion transportation compared with shorter ones. Lastly, stochastic processes dominated the community assembly of the epiphytic microorganisms. These findings could provide new insights into the relationship between microbial communities and growth in S. fusiforme seedlings and enable us to predict the community diversity and assembly of macroalgae-associated microbial communities. This could have important implications for linking microbial community diversity and network interactions to their host productivity.

Epiphytic common core bacteria in the microbiomes of co-located green (Ulva), brown (Saccharina) and red (Grateloupia, Gelidium) macroalgae

BackgroundMacroalgal epiphytic microbial communities constitute a rich resource for novel enzymes and compounds, but studies so far largely focused on tag-based microbial diversity analyses or limited metagenome sequencing of single macroalgal species.ResultsWe sampled epiphytic bacteria from specimens of Ulva sp. (green algae), Saccharina sp. (brown algae), Grateloupia sp. and Gelidium sp. (both red algae) together with seawater and sediment controls from a coastal reef in Weihai, China, during all seasons. Using 16S rRNA amplicon sequencing, we identified 14 core genera (consistently present on all macroalgae), and 14 dominant genera (consistently present on three of the macroalgae). Core genera represented ~ 0.7% of all genera, yet accounted for on average 51.1% of the bacterial abundances. Plate cultivation from all samples yielded 5,527 strains (macroalgae: 4,426) representing 1,235 species (685 potentially novel). Sequencing of selected strains yielded 820 non-redundant draft genomes (506 potentially novel), and sequencing of 23 sampled metagenomes yielded 1,619 metagenome-assembled genomes (MAGs), representing further 1,183 non-redundant genomes. 230 isolates and 153 genomes were obtained from the 28 core/dominant genera. We analyzed the genomic potential of phycosphere bacteria to degrade algal polysaccharides and to produce bioactive secondary metabolites. We predicted 4,451 polysaccharide utilization loci (PULs) and 8,810 biosynthetic gene clusters (BGCs). These were particularly prevalent in core/dominant genera.ConclusionsOur metabolic annotations and analyses of MAGs and genomes provide new insights into novel species of phycosphere bacteria and their ecological niches for an improved understanding of the macroalgal phycosphere microbiome.DnRc39z963CYrL7RdN5UNdVideo

The composition, function and assembly mechanism of epiphytic microbial communities on Gracilariopsis lemaneiformis

Gracilariopsis lemaneiformis is effective for bioremediation and important ecologically and economically in coastal ecosystems. However, little is known about the epiphytic microbes on it. We used the 16S rRNA gene and ITS amplicon sequencing, and a comprehensive functional gene array (GeoChip 5.0) to analyze both the epiphytic microbial communities on G. lemaneiformis and the free-living microbial communities in the surrounding seawater. The dominant epiphytic taxa, Aquimarina, Leucothrix, Formosa and Croceitalea, are associated with the seaweed and the degradation of algal polysaccharides. Genes enriched in the epiphytic community contribute to the seaweed surface life stress responses, quorum sensing, formation and stabilization of biofilms and antifouling. Moreover, homogeneous selection was the predominant deterministic process of epiphytic microbial community assembly. Our study provides new insights into understanding the G. lemaneiformis epiphytic communities and proposes guidance for future studies of other microbe-seaweed interactions in order to uncover the ecological mechanisms of epiphytes.

Potamogeton crispus restoration increased the epiphytic microbial diversity and improved water quality in a micro-polluted urban river

Special characterization and assembly of epiphytic microbial communities remain unclear in micro-polluted water column during submersed macrophytes restoration. In this study, an in-situ enclosure area sowing with turions of Potamogeton crispus (P. crispus) was conducted in a micro-polluted urban river to investigate the characterization of P. crispus and epiphytic microbial communities and their response to water environment under different water depths. Turions completely germinated in water column with <90 cm water depth and the germination speed decreased with increasing water depth within 18 days. There were obvious differences in morphological characteristics of P. crispus between deep and shallow water layers. P. crispus restoration decreased by 12–32%, 13–36%, 9–43% and 5–36% of COD, NH4+-N, TN and TP concentration, respectively, in enclosed overlying water compared to the river (P < 0.05) during 5 months of experiment. Illumina sequencing was employed to explore the epiphytic bacterial and microeukayotic communities at water depth 25–35 cm (shallow area) and 80–90 cm (deep area). A total of 9 bacterial and 12 microeukayotic dominant phyla were obtained in eight samples. It should be noted that the algae abundances were higher in shallow area than deep area but a reverse trend was observed for methanotrophs. Null model analysis revealed that dispersal limitation and undominated process was the most important assembly process, whereas stochastic processes gained more importance in shallow area than deep one. According to cooccurrence analysis (|r| > 0.6, P < 0.05), there were more strongly correlated edges in shallow area (456 edges) than deep area (340 edges). These results highlight that submerged macrophytes restoration can increase microbial diversity and improve water quality, and provide a “summer disease cured in winter” way by using could-resistant P. crispus for water purification in micro-polluted rivers in low-temperature season.

Environmental factors and host genotype control foliar epiphytic microbial community of wild soybeans across China.

The microbiome inhabiting plant leaves is critical for plant health and productivity. Wild soybean (Glycine soja), which originated in China, is the progenitor of cultivated soybean (Glycine max). So far, the community structure and assembly mechanism of phyllosphere microbial community on G. soja were poorly understood. Here, we combined a national-scale survey with high-throughput sequencing and microsatellite data to evaluate the contribution of host genotype vs. climate in explaining the foliar microbiome of G. soja, and the core foliar microbiota of G. soja were identified. Our findings revealed that both the host genotype and environmental factors (i.e., geographic location and climatic conditions) were important factors regulating foliar community assembly of G. soja. Host genotypes explained 0.4% and 3.6% variations of the foliar bacterial and fungal community composition, respectively, while environmental factors explained 25.8% and 19.9% variations, respectively. We further identified a core microbiome thriving on the foliage of all G. soja populations, including bacterial (dominated by Methylobacterium-Methylorubrum, Pantoea, Quadrisphaera, Pseudomonas, and Sphingomonas) and fungal (dominated by Cladosporium, Alternaria, and Penicillium) taxa. Our study revealed the significant role of host genetic distance as a driver of the foliar microbiome of the wild progenitor of soya, as well as the effects of climatic changes on foliar microbiomes. These findings would increase our knowledge of assembly mechanisms in the phyllosphere of wild soybeans and suggest the potential to manage the phyllosphere of soya plantations by plant breeding and selecting specific genotypes under climate change.

Mangroves leaves phyllosphere bacteria community and its ability to survive under pyrene stress during the acclimation process

Plants in general and mangroves in particular can harbor hyper-diverse microorganisms in their different compartments including the phyllosphere area. This study used the leaves of three mangrove species; black mangrove (Avicenia germinans), red mangrove (Rhizophora mangle) and mangrove apple (Sonneratia alba) in order to evaluate the phyllosphere epiphytic bacterial community on their leaves surface and assess the ability of some epiphytic bacteria to tolerate and survive under pyrene stress. Through the 16S rRNA genes sequencing, 380203, 405203 and 344863 OTUs were identified respectively in the leaves of mangroves apple, black and red mangroves. The identified OTUs was positively correlated with leaves-wax (p < 0.05, r2 = 0.904), nitrogen (r2 = 0.72), phosphorus content (r2 = 0.62) and the water factor (r2 = 0.93). It was however highly and negatively correlated with the canopy cover (r2 = 0.93). The pyrene degradation rate in the mineral salt medium (MSM) containing pyrene as external stress was different in each mangrove species and varied depending on various factors. Therefore, through the succession culture in MSM, several bacteria strain belonging to Rhizobiales and Enterobacteres were found to be abundant in red mangroves. Bacteria belonging to Bacilliales and Sphingobacteriales were more abundant in mangroves apples and bacteria from Xanthomonadales and Sphingomonadales were more presents in back mangroves. The important finding was to reveal that the black mangrove at the non-submerged substrate, recorded the highest number of OTU, coinciding with its highest leaf's nitrogen and phosphorus content and most importantly, its highest rate of pyrene degradation. The general result of this study join previous research results and get place in the mangrove agenda, as part of a better understanding insight into the role of plant identity in driving the phyllosphere epiphytic microbial community structures in mangrove ecosystems.

Varieties and ensiling: Impact on chemical composition, fermentation quality and bacterial community of alfalfa.

Six species of alfalfa commonly found in northern China were collected in the present study. The chemical composition and epiphytic microbial communities during the ensiling were analyzed; and their effects on fermentation quality and silage bacterial communities were assessed. The effects of physicochemical characteristics of alfalfa on the bacterial community were also investigated in terms of nutritional sources of microbial growth and reproduction. The results showed that the chemical composition was significantly different in various alfalfa varieties, yet, the dominant genera attached to each variety of alfalfa was similar, except for pantoea (p<0.05). After ensiling, both the fermentation quality and microbial community changed obviously (p<0.05). Specifically, ZM2 had lower pH and ammonia nitrogen (NH3-N) content but higher LA content than other varieties of alfalfa silage. Beneficial bacteria such as Lentilactobacillus and Lactiplantibacillus were predominant in ZM2, which accounted for the higher fermentation quality. Significant correlations between the chemical composition of silage, fermentation quality and bacterial communities composition were observed. Moreover, variations in bacteria community structure during the fermentation of alfalfa were mainly influenced by water-soluble carbohydrates (36.79%) and dry matter (21.77%). In conclusion, this study revealed the influence of chemical composition on microbial community and fermentation quality, laying the groundwork for future studies on high-quality silage.

Effect of Formic Acid and Inoculants on Microbial Community and Fermentation Profile of Wilted or Un-Wilted Italian Ryegrass Silages during Ensiling and Aerobic Exposure

Wilted (around 35% DM) or un-wilted (around 35% DM) Italian ryegrass treated with three additives (formic acid, FA; Lactobacillus plantarum, LP; Lactobacillus buchneri, LB) was utilized to evaluate the effects of the dry matter (DM) contents on the microbial community and fermentation characteristics, which was ensiled for 60 days in a laboratory-scale silo, followed by 3 days of aerobic exposure. Significantly lower pH and higher lactic acid (LA) contents were observed in the LP-treated group ensiled at both DM contents (differences were significant when p &lt; 0.05). The contents of LA, acetic acid (AA), numbers of lactic acid bacteria (LAB) and ammonia nitrogen (NH3-N) in the FA-treated group were significantly lower than those in other treatments (p &lt; 0.05). L. buchneri was the dominant bacteria after 60 days fermentation, while Enterobacteria became prevalent after 3 days of aerobic exposure. L. buchneri was found in the LB-treated group with higher acetic acid. Although the best fermentation quality was observed in the LP-treated silages, the aerobic stability was lowest compared to other groups (p &lt; 0.05). In conclusion, our findings suggest that the DM content of Italian ryegrass affected its epiphytic microbial community and the effectiveness of the different type of additives. Formic acid was more suitable for un-wilted Italian ryegrass silage, L. plantarum had a better effect in wilted Italian ryegrass silage, and L. buchneri prolonged the aerobic stability of Italian ryegrass. DM content and purpose of ensiling should be the key factors for choosing different types of additives for Italian ryegrass silage.

Discovering the Characteristics of Community Structures and Functional Properties of Epiphytic Bacteria on Spartina alterniflora in the Coastal Salt Marsh Area

The invasive submerged Spartina alterniflora is dominant in the coastal Yellow River Delta wetland. Although sediment microorganisms have been found to mediate the nutrient cycle in wetlands, the role of epiphytic bacteria on submerged S. alterniflora has rarely drawn attention. In the present study, we analyzed the characteristics of epiphytic microbial community diversity and functional properties related to S. alterniflora in summer and winter by Illumina MiSeq sequencing and functional prediction. Marked high abundances of Proteobacteria, Actinobacteriota, Planctomycetota, Cyanobacteria and Desulfobacterota were found in S. alterniflora epiphytic microbiome. Beta diversity based on NMDS and LDA analysis revealed that the distribution of these epiphytic microbial communities clustered according to the leaf locations and variation in seasons. Environmental factors, including temperature, salinity, DO and total organic matter, exert important roles in impacting the microbial community. Significantly higher abundances of chemoheterotrophy, aerobic_chemoheterotrophy, hydrocarbon degradation, fermentation, nitrate reduction and nitrate respiration were correlated with the submerged S. alterniflora epiphytic microbiome. Collectively, the results indicated that S. alterniflora epiphytic bacterial community diversity and functional guilds varied greatly with variations in leaf locations and seasons. These results will also provide guidance for the isolation of functional bacteria in controlling plant spread.