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The phyllosphere, or plant leaf surface, represents a microbial ecosystem of considerable size, holding extraordinary biodiversity and enormous potential for the discovery of new products, tools, and applications in biotechnology, agriculture, medicine, and elsewhere. This mini-review highlights the applied microbiology of the phyllosphere as an original field of study concerning itself with the genes, gene products, natural compounds, and traits that underlie phyllosphere-specific adaptations and services that have commercial and economic value for current or future innovation. Examples include plant-growth-promoting and disease-suppressive phyllobacteria, probiotics and fermented foods that support human health, as well as microbials that remedy foliar contamination with airborne pollutants, residual pesticides, or plastics. Phyllosphere microbes promote plant biomass conversion into compost, renewable energy, animal feed, or fiber. They produce foodstuffs such as thickening agents and sugar substitutes, industrial-grade biosurfactants, novel antibiotics and cancer drugs, as well as enzymes used as food additives or freezing agents. Furthermore, new developments in DNA sequence-based profiling of leaf-associated microbial communities allow for surveillance approaches in the context of food safety and security, for example, to detect enteric human pathogens on leafy greens, predict plant disease outbreaks, and intercept plant pathogens and pests on internationally traded goods. KEY POINTS: • Applied phyllosphere microbiology concerns leaf-specific adaptations for economic value • Phyllobioprospecting searches the phyllosphere microbiome for product development • Phyllobiomonitoring tracks phyllosphere microbial profiles for early risk detection.

Bacteria on and inside leaves can influence forest tree health and resilience. The distribution and limits of a tree species' range can be influenced by various factors, with biological interactions among the most significant. We investigated the processes shaping the bacterial needle community across the species distribution of limber pine, a widespread Western conifer inhabiting a range of extreme habitats. We tested four hypotheses: (i) Needle community structure varies across sites, with site-specific factors more important to microbial assembly than host species selection; (ii) dispersal limitation structures foliar communities across the range of limber pine; (iii) the relative significance of dispersal and selection differs across sites in the tree species range; and (iv) needle age structures bacterial communities. We characterized needle communities from the needle surface and tissue of limber pine and co-occurring conifers across 16 sites in the limber pine distribution. Our findings confirmed that site characteristics shape the assembly of bacterial communities across the host species range and showed that these patterns are not driven by dispersal limitation. Furthermore, the strength of selection by the host varied by site, possibly due to differences in available microbes. Our study, by focusing on trees in their natural setting, reveals real needle bacterial dynamics in forests, which is key to understanding the balance between stochastic and deterministic processes in shaping forest tree-microbe interactions. Such understanding will be necessary to predict or manipulate these interactions to support forest ecosystem productivity or assist plant migration and adaptation in the face of global change.

Currently, phylloremediation has emerged as a highly effective method for eliminating air pollutants, particularly polyaromatic hydrocarbons (PAHs). When PAHs accumulate on the phyllosphere, they significantly impact the fungal communities residing on leaf surfaces. This study aimed to investigate how pollution distribution patterns affect the diversity and PAH-degrading abilities of phyllosphere fungi, alongside identifying suitable mathematical models for PAH degradation. Leaf samples from two locations, Maradana and Sapugaskanda, were identified as having the highest PAH concentrations through principal component analysis. The fungal diversity in these highly contaminated regions was varied, with dominant species exhibiting greater PAH-degrading capabilities than those in less polluted areas. Thirty-five morphologically different epiphytic fungal strains were isolated on Potato Dextrose Agar (PDA) medium using the sample leaf wash. Two different fungal strains were selected as the best PAH degraders among those 35 different strains. These fungal strains were identified as Trichoderma harzianum P4M-16, and Fusarium solani P11M-46 based on ITS sequence data. Notably, these fungal species were more prevalent in highly polluted urban areas compared to less contaminated sites. High-Performance Liquid Chromatography (HPLC) analysis revealed that these two fungal species degrade PAHs more efficiently than others. Their kinetics assays demonstrated alignment with four degradation models when breaking down phenanthrene, naphthalene, pyrene, and anthracene. Scanning electron microscopy images showed that these fungi function as endophytes, extending their mycelium into the core leaf tissue layers beyond the epidermis. Gas Chromatography-Mass Spectrometry (GCMS) analysis indicated that F. solani P11M-46 produced non-toxic byproducts during the breakdown of phenanthrene and anthracene, confirming their eco-friendly nature. Overall, our findings suggest the promising potential of using phyllosphere fungi in bioremediation processes for environmental pollutants such as PAHs. [ABSTRACT FROM AUTHOR]

Despite that climate change is currently one of the most pervasive challenges, its effects on the plant-associated microbiome is still poorly studied. The aim of this study was to evaluate the impact of the independent and combinatory effect of climate warming and drought on the microbiome assembly of oak from seed to seedling. In a multifactorial experimental set up, acorns were subjected to different temperatures (15 °C, 20 °C, and 25 °C) and soil moisture levels (drought (15%) and control (60%)) from germination until the seedling stage, after which the bacterial and fungal communities associated to the rhizosphere and phyllosphere were characterized by amplicon sequencing and qPCR. The results showed a stronger effect of temperature on fungal than on bacterial diversity and the effect was more pronounced in the phyllosphere. Under drought condition, temperature had a significantly negative effect on phyllosphere fungal diversity. In the rhizosphere, temperature had a significant effect on the fungal community composition which was primarily caused by species turnover. Regardless of temperature, Actinobacteriota was significantly enriched in drought, a group of bacteria known to increase plant drought tolerance. This study provides new insights into the effect of climate change on the plant microbiome in natural ecosystems. [ABSTRACT FROM AUTHOR]

The structure of the leaf microbiome can alter host fitness and change in response to abiotic and biotic factors, like seasonality, climate, and leaf age. However, relatively few studies consider the influence of host age on microbial communities at a time scale of a few days, a short time scale relevant to microbes. To understand how host age modulates changes in the fungal and bacterial leaf microbiome on a short time scale, we ran independent field and greenhouse-based studies and characterized phyllosphere communities using next-generation sequencing approaches. Our field study characterized changes in the fungal and bacterial phyllosphere by examining leaves of different relative ages across individuals, whereas the greenhouse study examined changes in the fungal microbiome by absolute leaf age across individuals. Together, these results indicate that fungal communities are susceptible to change as a leaf ages as evidenced by shifts in the diversity of fungal taxa both in the field and the greenhouse. Similarly, there were increases in the diversity of fungal taxa by leaf age in the greenhouse. In bacterial communities in the field, we observed changes in the diversity, composition, and relative abundance of common taxa. These findings build upon previous literature characterizing host-associated communities at longer time scales and provide a foundation for targeted work examining how specific microbial taxa might interact with each other, such as fine-scale interactions between pathogenic and non-pathogenic species. [ABSTRACT FROM AUTHOR]

Epiphytic microbes are those that live for some or all of their life cycle on the surface of plant leaves. Leaf surfaces are a topologically complex, physicochemically heterogeneous habitat that is home to extensive, mixed communities of resident and transient inhabitants from all three domains of life. In this review, we discuss the origins of leaf surface microbes and how different biotic and abiotic factors shape their communities. We discuss the leaf surface as a habitat and microbial adaptations which allow some species to thrive there, with particular emphasis on microbes that occupy the continuum between epiphytic specialists and phytopathogens, groups which have considerable overlap in terms of adapting to the leaf surface and between which a single virulence determinant can move a microbial strain. Finally, we discuss the recent findings that the wheat pathogenic fungus Zymoseptoria tritici spends a considerable amount of time on the leaf surface, and ask what insights other epiphytic organisms might provide into this pathogen, as well as how Z. tritici might serve as a model system for investigating plant–microbe-microbe interactions on the leaf surface. [ABSTRACT FROM AUTHOR]

Developing an effective method for isolating bacterial genetic material from plants is a relatively challenging task and often does not yield adequately prepared material for further analyses. Previous studies often overlook connections, primarily focusing on laboratory investigations. With advancements in high-throughput sequencing techniques, we can now revisit and delve deeper into these interactions. Our study focuses on the initial phase of these investigations: genetic material isolation. Extracting bacterial DNA from aboveground plant parts, known as the phyllosphere, poses a significant challenge due to plant-derived contaminants. Existing isolation protocols frequently yield inconsistent results, necessitating continuous refinement and optimization. In our study, we developed an effective isolation protocol employing mechanical–chemical lysis, sonication, and membrane filtration. This approach yielded high-quality DNA at a concentration of 38.08 ng/µL, suitable for advanced sequencing applications. Our results underscore the effectiveness and necessity of these methods for conducting comprehensive microbiological analyses. Furthermore, our research not only lays the groundwork for further studies on lettuce microbiota, but also highlights the potential for utilizing our developed protocol in investigating other plants and their microbiomes. [ABSTRACT FROM AUTHOR]

Plant leaves harbor a rich diversity of bacteria and fungi that, through their interactions with host plants, assume an influential role in plant physiological and metabolic processes. The unique phyllosphere environment of different plant species may shape and select distinct phyllosphere microbial communities. While most academic research has focused on the phyllosphere microorganisms within the same plant variety, there is relatively limited research on the phyllosphere microbial communities between different varieties. Populus L. is a typical tree species in temperate monsoon climates, widely distributed in northern China, and it constitutes a crucial component of China's forestry resources. For the purpose of this study, we investigated the community structure and diversity of phyllosphere fungi and bacteria in different poplar varieties under identical growth conditions to elucidate the main factors contributing to differences in phyllosphere microbial communities among these varieties. Our findings revealed variations in nitrogen, phosphorus, starch, and soluble sugar contents among the three poplar species studied. Additionally, there were considerable disparities in both abundance and α diversity index of phyllosphere fungal and bacterial communities among these species. At the phylum level, Ascomycota and Basidiomycota have been identified as the dominant fungal communities; while Proteobacteria and Actinomycetes were dominant bacterial communities. The correlation analysis pointed out that chemical traits in the leaves, in particular the total phosphorus and the quantity of soluble sugar, had a significant correlation with the structure and diversity of the microbial community residing in the phyllosphere. Overall, our results demonstrate that even under identical site conditions, each poplar species harbors its own unique phyllosphere microbial community composition as well as distinct leaf characteristics—highlighting host plant diversity as a crucial factor driving differences in phyllosphere microbial composition. [ABSTRACT FROM AUTHOR]

The foliage of the small-leaved lime (Tilia cordata) is characterised by the nerve axils being grown by non-glandular trichomes, which trait contributes to the enhanced retention of the particulate matter (PM). This fact may disturb the ecological service of T. cordata related to the structure of its leaves, which is to provide acarodomatia (micro-shelters) for the predatory mites of the Phytoseiidae family. Phytoseiids are natural enemies of a variety of plant pests, widely applied in integrated pest management (IPM). Their occurrence is largely related to acarodomatia in which these mites hide, feed, reproduce, and develop. For the first time, the influence of PM deposition within spaces typically occupied by phytoseiids is investigated. Experimental populations of Euseius finlandicus were reared on T. cordata leaves in the progressive PM-pollution. The results showed that the values of life table parameters of the predator depended significantly on the level of PM deposition on leaves. Contrary to clean leaves from the control, the medium and high contamination intensities significantly reduced the daily (by 47% and 70%, respectively) and the total fecundity (by 62% and 77%, respectively) of females which, in turn, resulted in a decreased net reproductive rate (by 67% and 81%, respectively), intrinsic rate of increase (by 40% and 55%, respectively) and finite rate of increase (by 8% and 10%, respectively) of E. finlandicus. The pre-ovipositional period was prolonged, while the oviposition duration was shortened and the mites matured longer. In high pollution level the mortality of phytoseiids was boosted by 19% and some females were observed with pollutant lumps adhered to the idiosoma. Also, significant shares of juvenile forms (13%) and adult females (25%) made attempts to escape from highly contaminated experimental arenas. The implications of PM retention on the shelter vegetation are discussed in the context of IPM and ecological services. [ABSTRACT FROM AUTHOR]

Exploring the intricate relationships between plants and their resident microorganisms is crucial not only for developing new methods to improve disease resistance and crop yields but also for understanding their co-evolutionary dynamics. Our research delves into the role of the phyllosphere-associated microbiome, especially Actinomycetota species, in enhancing pathogen resistance in Theobroma grandiflorum, or cupuassu, an agriculturally valuable Amazonian fruit tree vulnerable to witches' broom disease caused by Moniliophthora perniciosa. While breeding resistant cupuassu genotypes is a possible solution, the capacity of the Actinomycetota phylum to produce beneficial metabolites offers an alternative approach yet to be explored in this context. Utilizing advanced long-read sequencing and metagenomic analysis, we examined Actinomycetota from the phyllosphere of a disease-resistant cupuassu genotype, identifying 11 Metagenome-Assembled Genomes across eight genera. Our comparative genomic analysis uncovered 54 Biosynthetic Gene Clusters related to antitumor, antimicrobial, and plant growth-promoting activities, alongside cutinases and type VII secretion system-associated genes. These results indicate the potential of phyllosphere-associated Actinomycetota in cupuassu for inducing resistance or antagonism against pathogens. By integrating our genomic discoveries with the existing knowledge of cupuassu's defense mechanisms, we developed a model hypothesizing the synergistic or antagonistic interactions between plant and identified Actinomycetota during plant-pathogen interactions. This model offers a framework for understanding the intricate dynamics of microbial influence on plant health. In conclusion, this study underscores the significance of the phyllosphere microbiome, particularly Actinomycetota, in the broader context of harnessing microbial interactions for plant health. These findings offer valuable insights for enhancing agricultural productivity and sustainability. [ABSTRACT FROM AUTHOR]

The abiotic environment exerts strong effects on plant‐associated microbes, shaping their interactions with plants and resulting ecosystem processes. However, these abiotic effects on plant–microbe interactions are often highly specific and contingent on the abiotic driver or microbial group, requiring synthesis work describing general patterns and from this generate hypotheses and guide mechanistic work. To address this, we conducted a meta‐analysis of the effects of climate change‐related abiotic factors, namely warming, drought, and eCO2, on plant‐associated microbes distinguishing by microbial taxonomic or biological group (bacteria, fungi or virus) and the plant part where microbes are found or associated with (phyllosphere or rhizosphere). We found abiotic driver‐specific patterns, whereby drought significantly reduced microbial abundance, whereas warming and eCO2 had no significant effects. In addition, these abiotic effects were contingent on the microbial taxonomic group, with fungi being negatively affected by drought but positively affected by warming (eCO2 enrichment had no effect), whereas bacteria and viruses were not significantly affected by any factor. Likewise, rhizospheric microbes were negatively affected by drought but positively affected by warming (eCO2 enrichment had no effect), whereas phyllospheric microbes were not significantly affected by any factor. Collectively, these findings point to important implications for global change research by highlighting contrasting effects of climate change‐related abiotic drivers on plant‐associated microbes and the contingency of such effects on microbe life histories and the nature of their interactions with plants. [ABSTRACT FROM AUTHOR]

Background: Nanotechnology holds revolutionary potential in the field of agriculture, with zinc oxide nanoparticles (ZnO NPs) demonstrating advantages in promoting crop growth. Enhanced photosynthetic efficiency is closely linked to improved vigor and superior quality in tea plants, complemented by the beneficial role of phyllosphere microorganisms in maintaining plant health. However, the effects of ZnO NPs on the photosynthesis of tea plants, the sprouting of new shoots, and the community of phyllosphere microorganisms have not been fully investigated. Results: This study investigated the photosynthetic physiological parameters of tea plants under the influence of ZnO NPs, the content of key photosynthetic enzymes such as RubisCO, chlorophyll content, chlorophyll fluorescence parameters, transcriptomic and extensive targeted metabolomic profiles of leaves and new shoots, mineral element composition in these tissues, and the epiphytic and endophytic microbial communities within the phyllosphere. The results indicated that ZnO NPs could enhance the photosynthesis of tea plants, upregulate the expression of some genes related to photosynthesis, increase the accumulation of photosynthetic products, promote the development of new shoots, and alter the content of various mineral elements in the leaves and new shoots of tea plants. Furthermore, the application of ZnO NPs was observed to favorably influence the microbial community structure within the phyllosphere of tea plants. This shift in microbial community dynamics suggests a potential for ZnO NPs to contribute to plant health and productivity by modulating the phyllosphere microbiome. Conclusion: This study demonstrates that ZnO NPs have a positive impact on the photosynthesis of tea plants, the sprouting of new shoots, and the community of phyllosphere microorganisms, which can improve the growth condition of tea plants. These findings provide new scientific evidence for the application of ZnO NPs in sustainable agricultural development and contribute to advancing research in nanobiotechnology aimed at enhancing crop yield and quality. [ABSTRACT FROM AUTHOR]

Abstract Despite that climate change is currently one of the most pervasive challenges, its effects on the plant-associated microbiome is still poorly studied. The aim of this study was to evaluate the impact of the independent and combinatory effect of climate warming and drought on the microbiome assembly of oak from seed to seedling. In a multifactorial experimental set up, acorns were subjected to different temperatures (15 °C, 20 °C, and 25 °C) and soil moisture levels (drought (15%) and control (60%)) from germination until the seedling stage, after which the bacterial and fungal communities associated to the rhizosphere and phyllosphere were characterized by amplicon sequencing and qPCR. The results showed a stronger effect of temperature on fungal than on bacterial diversity and the effect was more pronounced in the phyllosphere. Under drought condition, temperature had a significantly negative effect on phyllosphere fungal diversity. In the rhizosphere, temperature had a significant effect on the fungal community composition which was primarily caused by species turnover. Regardless of temperature, Actinobacteriota was significantly enriched in drought, a group of bacteria known to increase plant drought tolerance. This study provides new insights into the effect of climate change on the plant microbiome in natural ecosystems.

Abstract Epiphytic microbes are those that live for some or all of their life cycle on the surface of plant leaves. Leaf surfaces are a topologically complex, physicochemically heterogeneous habitat that is home to extensive, mixed communities of resident and transient inhabitants from all three domains of life. In this review, we discuss the origins of leaf surface microbes and how different biotic and abiotic factors shape their communities. We discuss the leaf surface as a habitat and microbial adaptations which allow some species to thrive there, with particular emphasis on microbes that occupy the continuum between epiphytic specialists and phytopathogens, groups which have considerable overlap in terms of adapting to the leaf surface and between which a single virulence determinant can move a microbial strain. Finally, we discuss the recent findings that the wheat pathogenic fungus Zymoseptoria tritici spends a considerable amount of time on the leaf surface, and ask what insights other epiphytic organisms might provide into this pathogen, as well as how Z. tritici might serve as a model system for investigating plant–microbe-microbe interactions on the leaf surface.

Abstract The foliage of the small-leaved lime (Tilia cordata) is characterised by the nerve axils being grown by non-glandular trichomes, which trait contributes to the enhanced retention of the particulate matter (PM). This fact may disturb the ecological service of T. cordata related to the structure of its leaves, which is to provide acarodomatia (micro-shelters) for the predatory mites of the Phytoseiidae family. Phytoseiids are natural enemies of a variety of plant pests, widely applied in integrated pest management (IPM). Their occurrence is largely related to acarodomatia in which these mites hide, feed, reproduce, and develop. For the first time, the influence of PM deposition within spaces typically occupied by phytoseiids is investigated. Experimental populations of Euseius finlandicus were reared on T. cordata leaves in the progressive PM-pollution. The results showed that the values of life table parameters of the predator depended significantly on the level of PM deposition on leaves. Contrary to clean leaves from the control, the medium and high contamination intensities significantly reduced the daily (by 47% and 70%, respectively) and the total fecundity (by 62% and 77%, respectively) of females which, in turn, resulted in a decreased net reproductive rate (by 67% and 81%, respectively), intrinsic rate of increase (by 40% and 55%, respectively) and finite rate of increase (by 8% and 10%, respectively) of E. finlandicus. The pre-ovipositional period was prolonged, while the oviposition duration was shortened and the mites matured longer. In high pollution level the mortality of phytoseiids was boosted by 19% and some females were observed with pollutant lumps adhered to the idiosoma. Also, significant shares of juvenile forms (13%) and adult females (25%) made attempts to escape from highly contaminated experimental arenas. The implications of PM retention on the shelter vegetation are discussed in the context of IPM and ecological services.

Leaves harbor distinct microbial communities that can have an important impact on plant health and microbial ecosystems worldwide. Nevertheless, the ecological processes that shape the composition of leaf microbial communities remain unclear, with previous studies reporting contradictory results regarding the importance of bacterial dispersal versus host selection. This discrepancy could be driven in part because leaf microbiome studies typically consider the upper and lower leaf surfaces as a single entity despite these habitats possessing considerable anatomical differences. We characterized the composition of bacterial phyllosphere communities from the upper and lower leaf surfaces across 24 plant species. Leaf surface pH and stomatal density were found to shape phyllosphere community composition, and the underside of leaves had lower richness and higher abundances of core community members than upper leaf surfaces. We found fewer endemic bacteria on the upper leaf surfaces, suggesting that dispersal is more important in shaping these communities, with host selection being a more important force in microbiome assembly on lower leaf surfaces. Our study illustrates how changing the scale in which we observe microbial communities can impact our ability to resolve and predict microbial community assembly patterns on leaf surfaces. IMPORTANCE Leaves can harbor hundreds of different bacterial species that form unique communities for every plant species. Bacterial communities on leaves are really important because they can, for example, protect their host against plant diseases. Usually, bacteria from the whole leaf are considered when trying to understand these communities; however, this study shows that the upper and lower sides of a leaf have a very different impact on how these communities are shaped. It seems that the bacteria on the lower leaf side are more closely associated with the plant host, and communities on the upper leaf side are more impacted by immigrating bacteria. This can be really important when we want to treat, for example, crops in the field with beneficial bacteria or when trying to understand host-microbe interactions on the leaves.

Abstract 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.

Elucidating the plant−microbiome network is of importance in understanding species coexistence in natural ecosystems. The phyllosphere, which is the aerial parts of terrestrial plants, is inhabited by diverse microbes. However, few studies have focused on the phyllosphere microbiome and plant-microbiome network in temperate grasslands. In this study, we explored the diversity, community structure, and network architecture of phyllosphere bacteria and fungi in 19 plant species native to the temperate grassland in Inner Mongolia, China. We obtained 3,313 and 758 phyllosphere bacterial and fungal operational taxonomic units, respectively, and found that the bacterial community was dominated by Proteobacteria, Actinobacteriota, and Firmicutes, whereas the fungal community was dominated by Ascomycota and Basidiomycota. Plant identity exerted significant impacts on α-diversities of both bacterial and fungal communities. The composition of bacterial and fungal communities differed among plant species. Plant identity had a greater effect on fungal than bacterial communities. Both bacterial and fungal network structures were characterized by specialized, modular, lowly connected, and no nested properties. The plant−fungal network had a high level of specification, modularity, antinestedness, and connectance compared with the plant−bacterial network. Our results suggest more intimate relationships between plants and phyllosphere fungi than between plants and phyllosphere bacteria and also that the phyllosphere fungal community in the temperate grassland ecosystem is more resistant to environmental disturbance than the phyllosphere bacterial community. These findings may contribute to our understanding of the mechanisms by which species coexist and communities stabilize in grassland ecosystems.

Abstract Background The phyllosphere microbiome is crucial for plant health and ecosystem functioning. While host species play a determining role in shaping the phyllosphere microbiome, host trees of the same species that are subjected to different environmental conditions can still exhibit large degrees of variation in their microbiome diversity and composition. Whether these intra-specific variations in phyllosphere microbiome diversity and composition can be observed over the broader expanse of forest landscapes remains unclear. In this study, we aim to assess the variation in the top canopy phyllosphere bacterial communities between and within host tree species in the temperate European forests, focusing on Fagus sylvatica (European beech) and Picea abies (Norway spruce). Results We profiled the bacterial diversity, composition, driving factors, and discriminant taxa in the top canopy phyllosphere of 211 trees in two temperate forests, Veluwe National Parks, the Netherlands and Bavarian Forest National Park, Germany. We found the bacterial communities were primarily shaped by host species, and large variation existed within beech and spruce. While we showed that there was a core microbiome in all tree species examined, community composition varied with elevation, tree diameter at breast height, and leaf-specific traits (e.g., chlorophyll and P content). These driving factors of bacterial community composition also correlated with the relative abundance of specific bacterial families. Conclusions While our results underscored the importance of host species, we demonstrated a substantial range of variation in phyllosphere bacterial diversity and composition within a host species. Drivers of these variations have implications at both the individual host tree level, where the bacterial communities differed based on tree traits, and at the broader forest landscape level, where drivers like certain highly plastic leaf traits can potentially link forest canopy bacterial community variations to forest ecosystem processes. We eventually showed close associations between forest canopy phyllosphere bacterial communities and host trees exist, and the consistent patterns emerging from these associations are critical for host plant functioning.

Coffee leaf rust, caused by the fungus Hemileia vastatrix , has become a major concern for coffee-producing countries. Additionally, there has been an increase in the resistance of certain races of the fungus to fungicides and breeding cultivars, making producers use alternative control methods. In this work, we transplanted the leaf surface microbiota of rust-resistant coffee species (Coffea racemosa and Coffea stenophylla) to Coffea arabica and tested whether the new microbiota would be able to minimize the damage caused by H. vastatrix. It was seen that the transplant was successful in controlling rust, especially from C. stenophylla , but the protection depended on the concentration of the microbiota. Certain fungi, such as Acrocalymma, Bipolaris, Didymella, Nigrospora, Setophaeosphaeria, Simplicillium, Stagonospora and Torula , and bacteria, such as Chryseobacterium, Sphingobium and especially Enterobacter , had their populations increased and this may be related to the antagonism seen against H. vastatrix. Interestingly, the relative population of bacteria from genera Pantoea, Methylobacterium and Sphingomonas decreased after transplantation, suggesting a positive interaction between them and H. vastatrix development. Our findings may help to better understand the role of the microbiota in coffee leaf rust, as well as help to optimize the development of biocontrol agents. [ABSTRACT FROM AUTHOR]

The phyllosphere, a reservoir of diverse microbial life associated with plant health, harbors microbial communities that are subject to various complex ecological processes acting at multiple scales. In this study, we investigated the determinants of the spatiotemporal variation in bacterial and fungal communities within the apple tree phyllosphere, employing 16S and ITS amplicon sequencing. Our research assessed the impact of key factors—plant compartment, site, time, and cultivar—on the composition and diversity of leaf and flower microbial communities. Our analyses, based on samples collected from three cultivars in three orchards in 2022, revealed that site and time are the strongest drivers of apple tree phyllosphere microbial communities. Conversely, plant compartment and cultivar exhibited minor roles in explaining community composition and diversity. Predominantly, bacterial communities comprised Hymenobacter (25%) and Sphingomonas (10%), while the most relatively abundant fungal genera included Aureobasidium (27%) and Sporobolomyces (10%). Additionally, our results show a gradual decrease in alpha-diversity throughout the growth season. These findings emphasize the necessity to consider local microbial ecology dynamics in orchards, especially as many groups worldwide aim for the development of biocontrol strategies (e.g., by manipulating plant–microbe interactions). More research is needed to improve our understanding of the determinants of time and site-specific disparities within apple tree phyllosphere microbial communities across multiple years, locations, and cultivars. [ABSTRACT FROM AUTHOR]

Heat stress has a substantial impact on black gram (Vigna mungo L.) by reducing growth, grain yield, and quality. Black gram is highly sensitive to temperature; it requires a range of 27–35 °C for flowering and pod formation. Therefore, to mitigate heat stress during the summer season in black gram, phyllosphere-associated methylotrophic bacteria (Methylobacterium brachiatum (KD1 and SD3), M. thiocyanatum (SD2), and Methylorubrum populi (KD5) were chosen from the previously identified strains. Screening for plant growth-promoting properties revealed that all strains (KD1, KD5, SD2, and SD3) were able to grow on nitrogen-free (N2) medium, produced siderophore (12.57 to 69.10%), indole-acetic acid (IAA) like substances (91.93 to 94.14 µg/mL), and 1-aminocyclopropane-1-carboxylate deaminase (ACC deaminase) (0. 34 to 1.33 nmol α-ketobutyrate/mg protein/h). A pot study experiment during the summer season (average temperature 42 °C) revealed that black gram seeds treated with methylotrophic bacteria had a higher biomass. Moreover, the chlorophyll content was significantly enhanced, ranging from least 188.00% to maximum 264.94%, and carotenoid pigments ranging from least 104.82% to maximum 155.10% compared to the control plants group. Similarly, the inoculation with methylotrophs also enhanced antioxidant enzymes such as POD in leaves (with least 7.14 and maximum 66.66%) and stems (with least 2.94 and maximum 191.17%), SOD (with least 92.30 and maximum 607.69%) and (with least 30.76 and maximum 150.00%), and PAL (with least 1.33 and maximum 89.33%) and (with least 25.58 and maximum 60.46%) compared to the control plants. In addition, Methylobacterium strains reduced oxidative stress in plants by managing the production of hydrogen peroxide (H2O2) and superoxide radicals (O2.−). These results showed that methylotrophs have a strong ability to protect black gram plants, and enhance their biomass under heat stress. [ABSTRACT FROM AUTHOR]

Given the importance of biological nitrogen (N) fixation in agroecosystems, using inoculants with phyllosphere N fixers effective across various crops would revolutionize agriculture. In this study, the application of an inoculant prepared from Methylobacterium symbioticum was tested on young olive trees. The pot experiment was arranged in a factorial design, with inoculant (Yes and No) and mineral N applied to the soil [0 (N0), 25 (N25), 50 (N50), and 100 (N100) kg ha−1], and four replicates. The inoculant application did not increase plant dry matter yield (DMY), whereas the application of mineral N had a significant and pronounced effect. The inoculant also did not significantly increase N concentration in tissues, unlike the strong increase observed with N applied to the soil. The inoculant significantly increased plant N recovery, a cumulative effect resulting from small increases in DMY and N concentration in tissues. This increase represented 5.2% more N in plants receiving the inoculant compared to untreated ones. However, only treatments receiving mineral N recorded positive values of fixed N, with the highest value observed in the N50 treatment (12.4%), whereas a negative value (−7.7%) was observed in the N0 treatment. Overall, these low values of fixed N question the economic rationale of using this inoculant by farmers and especially render it unsuitable for organic farming systems, where plants tend to have lower N levels in tissues. [ABSTRACT FROM AUTHOR]

The American cranberry is a perennial North American fruit plant that is grown successfully on commercial plantations in Poland. The purpose of this study was to recognize filamentous fungi that colonize roots, leaves, and fruits without visible disease symptoms. Pure fungal cultures were isolated from disinfected plant fragments in agar media and identified by sequencing common taxonomic DNA markers such as the ITS region, the TEF-1α, or RPB2 genes. Of the 141 isolates studied, 59% were identified as closely related to soil saprotrophs. They were classified primarily as showing the greatest similarity to type strains of Trichoderma amoenum, Trichoderma dorothopsis, Paraphaeosphaeria sporulosa, and Penicillium murcianum. Additionally, isolates that are most similar to strains of Penicillium crustosum, Aspergillus flavus, and Aspergillus versicolor that produced mycotoxins were detected. The fungi identified as closest to Alternaria geophila, Alternaria senecionicola, Paraphoma radicina, Pestalotiopsis unicolor, Pestalotiopsis scoparia, and Neopestalotiopsis spp., whose hosts are plants other than American cranberry, represented 33.81% of the isolates tested. Only 7.2% of the isolates corresponded to the species of Physalospora vaccinia, Diaporthe vaccinii, and Diaporthe eres, known cranberry pathogens. The results of this study can be used to identify latent plant infection and potential disease risks. [ABSTRACT FROM AUTHOR]

Plant microbiomes play important roles in plant health and fitness. Bacterial horizontal gene transfer (HGT) can influence plant health outcomes, driving the spread of both plant growth-promoting and phytopathogenic traits. However, community dynamics, including the range of genetic elements and bacteria involved in this process are still poorly understood. Integrons are genetic elements recently shown to be abundant in plant microbiomes, and are associated with HGT across broad phylogenetic boundaries. They facilitate the spread of gene cassettes, small mobile elements that collectively confer a diverse suite of adaptive functions. Here, we analysed 5,565 plant-associated bacterial genomes to investigate the prevalence and functional diversity of integrons in this niche. We found that integrons are particularly abundant in the genomes of Pseudomonadales, Burkholderiales, and Xanthomonadales. In total, we detected nearly 9,000 gene cassettes, and found that many could be involved in plant growth promotion or phytopathogenicity, suggesting that integrons might play a role in bacterial mutualistic or pathogenic lifestyles. The rhizosphere was enriched in cassettes involved in the transport and metabolism of diverse substrates, suggesting that they may aid in adaptation to this environment, which is rich in root exudates. We also found that integrons facilitate cross-species HGT, which is particularly enhanced in the phyllosphere. This finding may provide an ideal opportunity to promote plant growth by fostering the spread of genes cassettes relevant to leaf health. Together, our findings suggest that integrons are important elements in plant microbiomes that drive HGT, and have the potential to facilitate plant host adaptation. [ABSTRACT FROM AUTHOR]

Many microorganisms inhabit the aboveground parts of plants (i.e. the phyllosphere), which mainly comprise leaves. Understanding the structure of phyllosphere microbial communities and their drivers is important because they influence host plant fitness and ecosystem functions. Despite the high prevalence of ant–plant associations, few studies have used quantitative community data to investigate the effects of ants on phyllosphere microbial communities. In the present study, we investigated the effects of ants on the phyllosphere fungal communities of Mallotus japonicus using high‐throughput sequencing. Mallotus japonicus is a myrmecophilous plants that bears extrafloral nectaries, attracting several ant species, but does not provide specific ant species with nest sites like myrmecophytes do. We experimentally excluded ants with sticky resins from the target plants and collected leaf discs to extract fungal DNA. The ribosomal DNA internal transcribed spacer 1 (ITS1) regions of the phyllosphere fungi were amplified and sequenced to obtain fungal community data. Our results showed that the exclusion of ants changed the phyllosphere fungal community composition; however, the effect of ants on OTU richness was not clear. These results indicate that ants can change the community of phyllosphere fungi, even if the plant is not a myrmecophyte. [ABSTRACT FROM AUTHOR]

Objetive/Background. The aim of this was to assess the diversity and antibiotic resistance of bacteria isolated from 19 crops with bacterial infection symptoms. Material and Methods: This collection was identified using 16S rRNA gene sequencing and the Biolog system. Susceptibility and minimum inhibitory concentration (MIC) for streptomycin, tetracycline, and gentamicin were determined using disk diffusion and E-test methods, respectively. Results: A total of 55 species belonging to 20 bacterial genera were identified, with Pseudomonas, Serratia, Pantoea, and Stenotrophomonas being the most abundant. Approximately 27% of the isolates were categorized as pathogenic through the hypersensitivity reaction test, including phytopathogenic species like Pseudomonas syringae, P. cichorii, Pantoea anthophila, P. stewartii, Stenotrophomonas maltophilia, Dickeya oryzae, Erwinia billingiae, Pectobacterium aroidearum, and Enterobacter cloacae subsp. dissolvens. Resistance to at least one antibiotic was detected in 60% of isolates from 17 crops, with tomatoes, heart of palm, and lettuce exhibited the highest proportion of resistant bacteria (>80%). Streptomycin resistance was most common (35%), followed by tetracycline (28%) and gentamicin (9%). Conclusions: The findings indicate the presence of antibiotic resistance in saprophytic and pathogenic bacteria associated with 17 out of 19 assessed crops, posing risks to the environment, phytosanitary conditions, and public health. [ABSTRACT FROM AUTHOR]

Background: The phyllosphere microbiome is crucial for plant health and ecosystem functioning. While host species play a determining role in shaping the phyllosphere microbiome, host trees of the same species that are subjected to different environmental conditions can still exhibit large degrees of variation in their microbiome diversity and composition. Whether these intra-specific variations in phyllosphere microbiome diversity and composition can be observed over the broader expanse of forest landscapes remains unclear. In this study, we aim to assess the variation in the top canopy phyllosphere bacterial communities between and within host tree species in the temperate European forests, focusing on Fagus sylvatica (European beech) and Picea abies (Norway spruce). Results: We profiled the bacterial diversity, composition, driving factors, and discriminant taxa in the top canopy phyllosphere of 211 trees in two temperate forests, Veluwe National Parks, the Netherlands and Bavarian Forest National Park, Germany. We found the bacterial communities were primarily shaped by host species, and large variation existed within beech and spruce. While we showed that there was a core microbiome in all tree species examined, community composition varied with elevation, tree diameter at breast height, and leaf-specific traits (e.g., chlorophyll and P content). These driving factors of bacterial community composition also correlated with the relative abundance of specific bacterial families. Conclusions: While our results underscored the importance of host species, we demonstrated a substantial range of variation in phyllosphere bacterial diversity and composition within a host species. Drivers of these variations have implications at both the individual host tree level, where the bacterial communities differed based on tree traits, and at the broader forest landscape level, where drivers like certain highly plastic leaf traits can potentially link forest canopy bacterial community variations to forest ecosystem processes. We eventually showed close associations between forest canopy phyllosphere bacterial communities and host trees exist, and the consistent patterns emerging from these associations are critical for host plant functioning. [ABSTRACT FROM AUTHOR]

Summary: DNA contamination can critically confound microbiome studies. Here, we take a systematic approach to review the current literature and investigate the prevalence of contamination controls in phyllosphere microbiome research over the past decade. By utilising systematic review principles for this review, we were able to conduct a thorough investigation, screening 450 articles from three databases for eligibility and extracting data in a controlled and methodical manner. Worryingly, we observed a surprisingly low usage of both positive and negative contamination controls in phyllosphere research. As a result, we propose a set of minimum standards to combat the effects of contamination in future phyllosphere research. [ABSTRACT FROM AUTHOR]

Despite rapid progress in plant–microbe interaction research within terrestrial environments, our knowledge of aquatic plant and macroalgae microbiome structure, function, and ecology remains scarce, even though these hosts are key players in structuring the lacustrine environments. Here, we used the co-occurring, fast-growing hosts Hydrilla verticillata (Hydrocharitaceae) and Cladophora spp. (Chlorophyta), which dominate the littoral zones of a nitrogen-limited, hard-water lake, Lake Atitlán (Guatemala). The aim of this study was: (1) to assess the structure of Hydrilla phyllosphere-associated and Cladophora filament-associated bacterial and fungal assemblages in the context of host specificity; (2) to predict microbial potential to contribute to biogeochemical cycling in the lake littoral; and (3) to compare the aquatic microbiome structure to available datasets from terrestrial ecosystems, using next-generation amplicon sequencing, co-occurrence network analysis, and N2-fixation activity measurements. We show that the microbiomes associated with the phyllosphere of aquatic macrophytes and macroalgal filaments are surprisingly similar, with taxonomic and functional complexity analogous to that of rhizospheric assemblages in terrestrial plants, and have a potential to efficiently recycle nutrients from organic matter. We suggest that especially the fungal associations with these hosts represent an untapped research area of microbial ecology that warrants further attention. [ABSTRACT FROM AUTHOR]

Leaves are colonized by a complex mix of microbes, termed the leaf microbiota. Even though the leaf microbiota is increasingly recognized as an integral part of plant life and health, our understanding of its interactions with the plant host is still limited. Here, mature, axenically grown Arabidopsis thaliana plants were spray inoculated with six diverse leaf‐colonizing bacteria. The transcriptomic changes in leaves were tracked over time and significant changes in ethylene marker (ARL2) expression were observed only 2–4 days after spray inoculation. Whole‐transcriptome sequencing revealed that 4 days after inoculation, leaf transcriptional changes to colonization by nonpathogenic and pathogenic bacteria differed in strength but not in the type of response. Inoculation of plants with different densities of the nonpathogenic bacterium Williamsia sp. Leaf354 showed that high bacterial titers resulted in disease phenotypes and led to severe transcriptional reprogramming with a strong focus on plant defense. An in silico epigenetic analysis of the data was congruent with the transcriptomic analysis. These findings suggest (1) that plant responses are not rapid after spray inoculation, (2) that plant responses only differ in strength, and (3) that plants respond to high titers of nonpathogenic bacteria with pathogen‐like responses. [ABSTRACT FROM AUTHOR]

The structure of the leaf microbiome can alter host fitness and change in response to abiotic and biotic factors, like seasonality, climate, and leaf age. However, relatively few studies consider the influence of host age on microbial communities at a time scale of a few days, a short time scale relevant to microbes. To understand how host age modulates changes in the fungal and bacterial leaf microbiome on a short time scale, we ran independent field and greenhouse-based studies and characterized phyllosphere communities using next-generation sequencing approaches. Our field study characterized changes in the fungal and bacterial phyllosphere by examining leaves of different relative ages across individuals, whereas the greenhouse study examined changes in the fungal microbiome by absolute leaf age across individuals. Together, these results indicate that fungal communities are susceptible to change as a leaf ages as evidenced by shifts in the diversity of fungal taxa both in the field and the greenhouse. Similarly, there were increases in the diversity of fungal taxa by leaf age in the greenhouse. In bacterial communities in the field, we observed changes in the diversity, composition, and relative abundance of common taxa. These findings build upon previous literature characterizing host-associated communities at longer time scales and provide a foundation for targeted work examining how specific microbial taxa might interact with each other, such as fine-scale interactions between pathogenic and non-pathogenic species.

ABSTRACT Host-associated microbiomes are shaped by both their environment and host genetics, and often impact host performance. The scale of host genetic variation important to microbes is largely unknown yet fundamental to the community assembly of host-associated microbiomes, with implications for the eco-evolutionary dynamics of microbes and hosts. Using Ipomoea hederacea, ivyleaf morning glory, we generated matrilines differing in quantitative genetic variation and leaf shape, which is controlled by a single Mendelian locus. We then investigated the relative roles of Mendelian and quantitative genetic variation in structuring the leaf microbiome and how these two sources of genetic variation contributed to microbe heritability. We found that despite large effects of the environment, both Mendelian and quantitative genetic host variation contribute to microbe heritability and that the cumulative small effect genomic differences due to matriline explained as much or more microbial variation than a single large effect Mendelian locus. Furthermore, our results are the first to suggest that leaf shape itself contributes to variation in the abundances of some phyllosphere microbes.IMPORTANCEWe investigated how host genetic variation affects the assembly of Ipomoea hederacea’s natural microbiome. We found that the genetic architecture of leaf-associated microbiomes involves both quantitative genetic variation and Mendelian traits, with similar contributions to microbe heritability. The existence of Mendelian and quantitative genetic variation for host-associated microbes means that plant evolution at the leaf shape locus or other quantitative genetic loci has the potential to shape microbial abundance and community composition.

Abstract Phyllosphere endophytes play a critical role in a myriad of biological functions, such as maintaining plant health and overall fitness. They play a determinative role in crop yield and quality by regulating vital processes, such as leaf functionality and longevity, seed mass, apical growth, flowering, and fruit development. This study conducted a comprehensive bibliometric analysis aiming to review the prevailing research trajectories in phyllosphere endophytes and harness both primary areas of interest and emerging challenges. A total of 156 research articles on phyllosphere endophytes, published between 2002 and 2022, were retrieved from the Web of Science Core Collection (WoSCC). A systematic analysis was conducted using CiteSpace to visualize the evolution of publication frequency, the collaboration network, the co-citation network, and keywords co-occurrence. The findings indicated that initially, there were few publications on the topic of phyllosphere endophytes. However, from 2011 onwards, there was a notable increase in the number of publications on phyllosphere endophytes, gaining worldwide attention. Among authors, Arnold, A Elizabeth is widely recognized as a leading author in this research area. In terms of countries, the USA and China hold the highest rankings. As for institutional ranking, the University of Arizona is the most prevalent and leading institute in this particular subject. Collaborative efforts among the authors and institutions tend to be confined to small groups, and a large-scale collaborative network needs to be established. This study identified the influential journals, literature, and hot research topics. These findings also highlight the interconnected nature of key themes, e.g., phyllosphere endophyte research revolves around the four pillars: diversity, fungal endophytes, growth, and endophytic fungi. This study provides an in-depth perspective on phyllosphere endophytes studies, revealing the identification of biodiversity and microbial interaction of phyllosphere endophytes as the principal research frontiers. These analytical findings not only elucidate the recent trajectory of phyllosphere endophyte research but also provide invaluable insights for similar studies and their potential applications on a global scale.

Abstract Symbiotic Methylobacterium comprise a significant portion of the phyllospheric microbiome, and are known to benefit host plant growth, development, and confer tolerance to stress factors. The near ubiquitous use of the broad-spectrum herbicide, glyphosate, in farming operations globally has necessitated a more expansive evaluation of the impacts of the agent itself and formulations containing glyphosate on important components of the plant phyllosphere, including Methylobacterium. This study provides an investigation of the sensitivity of 18 strains of Methylobacterium to glyphosate and two commercially available glyphosate-based herbicides (GBH). Nearly all strains of Methylobacterium showed signs of sensitivity to the popular GBH formulations WeatherMax® and Transorb® in a modified Kirby Bauer experiment. However, exposure to pure forms of glyphosate did not show a significant effect on growth for any strain in both the Kirby Bauer test and in liquid broth, until polysorbate-20 (Tween20) was added as a surfactant. Artificially increasing membrane permeability through the introduction of polysorbate-20 caused a 78–84% reduction in bacterial cell biomass relative to controls containing glyphosate or high levels of surfactant only (0–9% and 6–37% reduction respectively). Concentrations of glyphosate as low as 0.05% w/v (500 µg/L) from both commercial formulations tested, inhibited the culturability of Methylobacterium on fresh nutrient-rich medium. To better understand the compatibility of important phyllospheric bacteria with commercial glyphosate-based herbicides, this study endeavours to characterize sensitivity in multiple strains of Methylobacterium, and explore possible mechanisms by which toxicity may be induced.

Plants interact with complex microbial communities in which microorganisms play different roles in plant development and health. While certain microorganisms may cause disease, others promote nutrient uptake and resistance to stresses through a variety of mechanisms. Developing plant protection measures requires a deeper comprehension of the factors that influence multitrophic interactions and the organization of phyllospheric communities. High-throughput sequencing was used in this work to investigate the effects of climate variables and bacterial wildfire disease on the bacterial community's composition and assembly in the phyllosphere of tobacco (Nicotiana tabacum L.). The samples from June (M1), July (M2), August (M3), and September (M4) formed statistically separate clusters. The assembly of the whole bacterial population was mostly influenced by stochastic processes. PICRUSt2 predictions revealed genes enriched in the M3, a period when the plant wildfire disease index reached climax, were associated with the development of the wildfire disease (secretion of virulence factor), the enhanced metabolic capacity and environmental adaption. The M3 and M4 microbial communities have more intricate molecular ecological networks (MENs), bursting with interconnections within a densely networked bacterial population. The relative abundances of plant-beneficial and antagonistic microbes Clostridiales, Bacillales, Lactobacillales, and Sphingobacteriales, showed significant decrease in severally diseased sample (M3) compared to the pre-diseased samples (M1/ M2). Following the results of MENs, we further test if the correlating bacterial pairs within the MEN have the possibility to share functional genes and we have unraveled 139 entries of such horizontal gene transfer (HGT) events, highlighting the significance of HGT in shaping the adaptive traits of plant-associated bacteria across the MENs, particularly in relation to host colonization and pathogenicity. [ABSTRACT FROM AUTHOR]

Plants are associated with a large diversity of microbes, and these complex plantassociated microbial communities are critical for plant health. Welsh onion (Allium fistulosum L.) is one of the key and oldest vegetable crops cultivated in Taiwan. The leaf of the Welsh onion is one of the famous spices in Taiwanese cuisine, thus, it is crucial to control foliar diseases. In recent years, Welsh onion cultivation in Taiwan has been severely threatened by the occurrence of leaf blight disease, greatly affecting their yield and quality. However, the overall picture of microbiota associated with the Welsh onion plant is still not clear as most of the recent etiological investigations were heavily based on the isolation of microorganisms from diseased plants. Therefore, studying the diversity of fungal communities associated with the leaf blight symptoms of Welsh onion may provide information regarding key taxa possibly involved in the disease. Therefore, this investigation was mainly designed to understand the major fungal communities associated with leaf blight to identify key taxa potentially involved in the disease and further evaluate any shifts in both phyllosphere and rhizosphere mycobiome assembly due to foliar pathogen infection by amplicon sequencing targeting the Internal Transcribed Spacer (ITS) 1 region of the rRNA. The alpha and beta-diversity analyses were used to compare the fungal communities and significant fungal groups were recognized based on linear discriminant analyses. Based on the results of relative abundance data and co-occurrence networks in symptomatic plants we revealed that the leaf blight of Welsh onion in Sanxing, is a disease complex mainly involving Stemphylium and Colletotrichum taxa. In addition, genera such as Aspergillus, Athelia and Colletotrichum were abundantly found associated with the symptomatic rhizosphere. Alphadiversity in some fields indicated a significant increase in species richness in the symptomatic phyllosphere compared to the asymptomatic phyllosphere. These results will broaden our knowledge of pathogens of Welsh onion associated with leaf blight symptoms and will assist in developing effective disease management strategies to control the progress of the disease. [ABSTRACT FROM AUTHOR]

In previous studies, two strains isolated from the maize phyllosphere were identified as Bacillus subtilis (EM-A7) and Bacillus velezensis (EM-A8) and selected as potential biocontrol agents against Exserohilum turcicum. This study aimed to assess the ability of EM-A7 and EM-A8 to form biofilm and have antagonistic activity under varying light conditions. LED sources were custom-designed so that each corresponded to a given spectrum at a specific photosynthetically active photon flux density. Significant differences were observed in growth parameters (generation time and constant growth rate) under different LED sources. Blue light inhibited the growth of both strains. Red increased k rate in EM-A8, while the g values increased in EM-A7. Red and white light generally increased biofilm formation, and blue light inhibited it. EM-A7 and EM-A8 significantly reduced their ability to swim under blue LED, but it was not affected by red, green, or white light. The ability to swarm was negatively affected. Fungal growth decreased significantly compared to the control when the bacterium growing on the same plate had been previously incubated under red and white light or in the dark. These results indicate that different light wavelengths clearly influenced the aspects assessed in B. subtilis and B. velezensis, with the effects of blue light being overall negative and those of red and white overall positive. Given that, all these factors can be important for the establishment and survival of Bacillus strains on leaves, as well as for their effectiveness against pathogens, light could be a significant factor to consider in the design of biocontrol strategies. [ABSTRACT FROM AUTHOR]

Aims: The phyllosphere and soil are crucial and distinct microbial spheres in forests, connected through trees that interact with both. As part of the tree's holobiont, these communities are vital to the fitness and evolution of the host. Differences between the spheres may be particularly evident at the two extreme ends of tall and long-lived trees of natural temperate forest; the top-canopy and the soil. Here, we evaluated the connectivity between the top-canopy and soil microbial communities of European beech and Norway spruce trees to determine the significance of tree-sphere and host-species identity, and to assess the contribution of taxa inhabiting both spheres. Methods: Bacterial and fungal community composition was determined through metabarcoding analysis of linked top-canopy leaf and bulk soil samples collected from tall (old) trees in the natural forest of Bavarian Forest National Park. Results: This study shows sphere-specific communities in European temperate forests, characterized by low connectivity. Results highlight that spheres exert stronger influence than host identity. Only a few taxa inhabited both spheres, yet they accounted for the bulk of the (relative) abundance in each sphere. Conclusion: Analysing the divergence and shared characteristics of these interlinked communities redefines the tree holobiont concept and enhances our understanding of the evolution of plant-associated microbial communities in a sphere-specific manner. This study emphasizes the importance of examining multiple microbiome components for a thorough understanding of temperate forest ecology, while also highlighting the existence of a small group of overlapping taxa that may play a bigger role than previously anticipated. [ABSTRACT FROM AUTHOR]

Phyllosphere microbial communities are increasingly experiencing intense pulse disturbance events such as drought. It is currently unknown how phyllosphere communities respond to such disturbances and if they are able to recover. We explored the stability of phyllosphere communities over time, in response to drought stress, and under recovery from drought on temperate forage grasses. Compositional or functional changes were observed during the disturbance period and whether communities returned to non-stressed levels following recovery. Here, we found that phyllosphere community composition shifts as a result of simulated drought but does not fully recover after irrigation is resumed and that the degree of community response to drought is host species dependent. However, while community composition had changed, we found a high level of functional stability (resistance) over time and in the water deficit treatment. Ecological modeling enabled us to understand community assembly processes over a growing season and to determine if they were disrupted during a disturbance event. Phyllosphere community succession was characterized by a strong level of ecological drift, but drought disturbance resulted in variable selection, or, in other words, communities were diverging due to differences in selective pressures. This successional divergence of communities with drought was unique for each host species. Understanding phyllosphere responses to environmental stresses is important as climate change-induced stresses are expected to reduce crop productivity and phyllosphere functioning. IMPORTANCE Leaf surface microbiomes have the potential to influence agricultural and ecosystem productivity. We assessed their stability by determining composition, functional resistance, and resilience. Resistance is the degree to which communities remain unchanged as a result of disturbance, and resilience is the ability of a community to recover to pre-disturbance conditions. By understanding the mechanisms of community assembly and how they relate to the resistance and resilience of microbial communities under common environmental stresses such as drought, we can better understand how communities will adapt to a changing environment and how we can promote healthy agricultural microbiomes. In this study, phyllosphere compositional stability was highly related to plant host species phylogeny and, to a lesser extent, known stress tolerances. Phyllosphere community assembly and stability are a result of complex interactions of ecological processes that are differentially imposed by host species. [ABSTRACT FROM AUTHOR]

Mitogen-activated protein kinase (MAPK) cascades are highly conserved signaling modules that coordinate diverse biological processes such as plant innate immunity and development. Recently, MAPK cascades have emerged as pivotal regulators of the plant holobiont, influencing the assembly of normal plant microbiota, essential for maintaining optimal plant growth and health. In this review, we provide an overview of current knowledge on MAPK cascades, from upstream perception of microbial stimuli to downstream host responses. Synthesizing recent findings, we explore the intricate connections between MAPK signaling and the assembly and functioning of plant microbiota. Additionally, the role of MAPK activation in orchestrating dynamic changes in root exudation to shape microbiota composition is discussed. Finally, our review concludes by emphasizing the necessity for more sophisticated techniques to accurately decipher the role of MAPK signaling in establishing the plant holobiont relationship. [ABSTRACT FROM AUTHOR]

Peru is one of the leading countries that produce and export specialty coffees, favorably positioned in the international markets for its physical and organoleptic cup qualities. In recent years, yellow coffee rust caused by the phytopathogenic fungus Hemileia vastatrix stands out as one of the main phytosanitary diseases that affect coffee culture yields. Many studies have demonstrated bacteria antagonistic activity against a number of phytopathogen fungi. In this context, the aim of this work was to select and characterize phyllospheric bacteria isolated from Coffea arabica with antagonistic features against coffee rust to obtain biocontrollers. For that purpose, a total of 82 phyllospheric bacteria were isolated from two coffee leaf rust-susceptible varieties, typica and caturra roja, and one tolerant variety, catimor. Of all the isolates, 15% were endophytic and 85% were epiphytes. Among all the isolates, 14 were capable of inhibiting the mycelial radial growth of Mycena citricolor, and Colletotrichum sp. 16S rRNA gene sequence-based analysis showed that 9 isolates were related to Achromobacter insuavis, 2 were related to Luteibacter anthropi and 1 was related to Rodococcus ceridiohylli, Achromobacter marplatensis and Pseudomonas parafulva. A total of 7 representative bacteria of each group were selected based on their antagonistic activity and tested in germination inhibition assays of coffee rust uredinospores. The CRRFLT7 and TRFLT8 isolates showed a high inhibition percentage of urediniospores germination (81% and 82%, respectively), similar to that obtained with the chemical control (91%). An experimental field assay showed a good performance of both strains against rust damage too, making them a promising alternative for coffee leaf rust biocontrol. [ABSTRACT FROM AUTHOR]

Symbiotic Methylobacterium comprise a significant portion of the phyllospheric microbiome, and are known to benefit host plant growth, development, and confer tolerance to stress factors. The near ubiquitous use of the broad-spectrum herbicide, glyphosate, in farming operations globally has necessitated a more expansive evaluation of the impacts of the agent itself and formulations containing glyphosate on important components of the plant phyllosphere, including Methylobacterium. This study provides an investigation of the sensitivity of 18 strains of Methylobacterium to glyphosate and two commercially available glyphosate-based herbicides (GBH). Nearly all strains of Methylobacterium showed signs of sensitivity to the popular GBH formulations WeatherMax® and Transorb® in a modified Kirby Bauer experiment. However, exposure to pure forms of glyphosate did not show a significant effect on growth for any strain in both the Kirby Bauer test and in liquid broth, until polysorbate-20 (Tween20) was added as a surfactant. Artificially increasing membrane permeability through the introduction of polysorbate-20 caused a 78–84% reduction in bacterial cell biomass relative to controls containing glyphosate or high levels of surfactant only (0–9% and 6–37% reduction respectively). Concentrations of glyphosate as low as 0.05% w/v (500 µg/L) from both commercial formulations tested, inhibited the culturability of Methylobacterium on fresh nutrient-rich medium. To better understand the compatibility of important phyllospheric bacteria with commercial glyphosate-based herbicides, this study endeavours to characterize sensitivity in multiple strains of Methylobacterium, and explore possible mechanisms by which toxicity may be induced. [ABSTRACT FROM AUTHOR]

Phyllosphere endophytes play a critical role in a myriad of biological functions, such as maintaining plant health and overall fitness. They play a determinative role in crop yield and quality by regulating vital processes, such as leaf functionality and longevity, seed mass, apical growth, flowering, and fruit development. This study conducted a comprehensive bibliometric analysis aiming to review the prevailing research trajectories in phyllosphere endophytes and harness both primary areas of interest and emerging challenges. A total of 156 research articles on phyllosphere endophytes, published between 2002 and 2022, were retrieved from the Web of Science Core Collection (WoSCC). A systematic analysis was conducted using CiteSpace to visualize the evolution of publication frequency, the collaboration network, the co-citation network, and keywords co-occurrence. The findings indicated that initially, there were few publications on the topic of phyllosphere endophytes. However, from 2011 onwards, there was a notable increase in the number of publications on phyllosphere endophytes, gaining worldwide attention. Among authors, Arnold, A Elizabeth is widely recognized as a leading author in this research area. In terms of countries, the USA and China hold the highest rankings. As for institutional ranking, the University of Arizona is the most prevalent and leading institute in this particular subject. Collaborative efforts among the authors and institutions tend to be confined to small groups, and a large-scale collaborative network needs to be established. This study identified the influential journals, literature, and hot research topics. These findings also highlight the interconnected nature of key themes, e.g., phyllosphere endophyte research revolves around the four pillars: diversity, fungal endophytes, growth, and endophytic fungi. This study provides an in-depth perspective on phyllosphere endophytes studies, revealing the identification of biodiversity and microbial interaction of phyllosphere endophytes as the principal research frontiers. These analytical findings not only elucidate the recent trajectory of phyllosphere endophyte research but also provide invaluable insights for similar studies and their potential applications on a global scale. [ABSTRACT FROM AUTHOR]

Aims This study aimed to evaluate and describe the functional differences of cultivable bacteria and fungi inhabiting the leaves of Gevuina avellana Mol. (Proteaceae) in an urban area with high levels of air pollution and in a native forest in the southern Andes. Methods and results Phyllosphere microorganisms were isolated from the leaves of G. avellana , their plant growth-promoting capabilities were estimated along with their biocontrol potential and tolerance to metal(loid)s. Notably, plants from the urban area showed contrasting culturable leaf-associated microorganisms compared to those from the native area. The tolerance to metal(loid)s in bacteria range from 15 to 450 mg l−1 of metal(loid)s, while fungal strains showed tolerance from 15 to 625 mg l−1, being especially higher in the isolates from the urban area. Notably, the bacterial strain Curtobacterium flaccumfaciens and the fungal strain Cladosporium sp. exhibited several plant-growth-promoting properties along with the ability to inhibit the growth of phytopathogenic fungi. Conclusions Overall, our study provides evidence that culturable taxa in G. avellana leaves is directly influenced by the sampling area. This change is likely due to the presence of atmospheric pollutants and diverse microbial symbionts that can be horizontally acquired from the environment. [ABSTRACT FROM AUTHOR]

Introduction: 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. Methods: 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). Results: 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. Discussion: 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. [ABSTRACT FROM AUTHOR]

During our ongoing survey of basidiomycetous yeasts associated with plant leaves in virgin forest, five Colacogloea strains were isolated in the Baotianman Nature Reserve, Henan Province, central China. Phenotypes from cultures and a phylogeny based on the internal transcribed spacer (ITS) regions and the D1/D2 domains of the large subunit (LSU) rRNA gene were employed to characterize and identify these isolates. As a result, two new species, namely Colacogloea celtidis sp. nov. and C. pararetinophila sp. nov., are introduced herein. In the phylogeny of combined ITS and LSU dataset, the new species C. celtidis sp. nov. formed a clade with the unpublished Colacogloea strain (KBP: Y-6832), and together these formed the sister group to C. armeniacae, while C. pararetinophila sp. nov. was retrieved as a sister to C. retinophila. A detailed description and illustration of both new species, as well as the differences between them and their closest relatives in the genus are provided. Results from the present study will add to our knowledge of the biodiversity of Colacogloea in China. [ABSTRACT FROM AUTHOR]