Extramatrical mycelial biomass is mediated by fine root mass and ectomycorrhizal fungal community composition across tree species.

In many ecosystems, a large fraction of gross primary production is invested in mycorrhiza. Ectomycorrhizal (ECM) mycelium is involved in regulating soil carbon and nutrient cycling. However, little is known about how mycelial biomass, production and turnover differ depending on ECM fungal community composition and associated tree species. We quantified fine root biomass and length using soil cores, and mycelial traits (biomass, production, and turnover) using mesh-bags and ergosterol analysis, and identified ECM exploration types by Illumina MiSeq sequencing of four ECM-dominated tree species (Picea asperata, Larix gmelinii, Quercus aquifolioides and Betula albosinensis) in subalpine forest. The ECM fungal community composition separated between needle-leaved and broadleaved species, and between evergreen and deciduous species. The ratio of mycelial to fine root biomass was similar across the species regardless of genus-scale community composition and the relative abundance of exploration types. Compared to the other species, Q. aquifolioides displayed higher fine root biomass and mycelial biomass and production, dominated by contact-short exploration type. Mycelial turnover rate tended to be lowest in P. asperata, dominated by medium-long exploration type. Much higher production of mycelium and only slightly higher turnover rate in Q. aquifolioides suggests that its steady-state mycelial biomass would be higher than of the other species. Moreover, compared to the two deciduous species, with similar production but somewhat lower turnover rate, the standing crop of mycelium in P. asperata may stabilize at a higher value. Our findings, that exploration type may affect production and turnover, highlight the importance of characterizing ECM fungal communities by exploration types when estimating the contribution of mycelium biomass to forest carbon sink and storage.

Overexpression of Ginkbilobin-2 homologous domain gene improves tolerance to Phytophthora cinnamomi in somatic embryos of Quercus suber.

In recent decades an extensive mortality and decline of Quercus suber populations mainly caused by Phytophthora cinnamomi has been observed. In the current study, a chestnut gene homologous to ginkbilobin-2 (Cast_Gnk2-like), which in Ginkgo biloba codifies an antifungal protein, was transferred into cork oak somatic embryos of three different embryogenic lines by Agrobacterium mediated transformation. The transformation efficiency varied on the genotype from 2.5 to 9.2%, and a total of 22 independent transformed lines were obtained. The presence of Cast_Gnk2-like gene in transgenic embryos was verified in all lines by PCR. The number of transgene copies was estimated by qPCR in embryogenic lines with high proliferation ability and it varied between 1 and 5. In addition, the expression levels of Cast_Gnk2-like gene were determined in the embryogenic lines, with higher levels in lines derived from the genotype ALM6-WT. Transgenic plants were obtained from all transgenic lines and evaluated after cold storage of the somatic embryos for 2 months and subsequent transfer to germination medium. In vitro tolerance tests made under controlled conditions and following zoospore treatment showed that plants overexpressing Cast_Gnk2-like gene improved tolerance against Pc when compared to wild type ones.

Seasonal variations in soil microbial community co-occurrence network complexity respond differently to field-simulated warming experiments in a northern subtropical forest.

Global warming may reshape seasonal changes in microbial community diversity and co-occurrence network patterns, with significant implications for terrestrial ecosystem function. We conducted a 2-year in situ field simulation of the effects of warming on the seasonal dynamics of soil microbial communities in a northern subtropical Quercus acutissima forest. Our study revealed that warming had no significant effect on the richness or diversity of soil bacteria or fungi in the growing season, whereas different warming gradients had different effects on their diversity in the nongrowing season. Warming also changed the microbial community structure, increasing the abundance of some thermophilic microbial species and decreasing the abundance of some symbiotrophic microorganisms. The co-occurrence network analysis of the microbial community showed that warming decreased the complexity of the intradomain network in the soil bacterial community in the growing and nongrowing seasons but increased it in the fungal community. Moreover, increasing warming temperatures increased the complexity of the interdomain network between bacteria and fungi in the growing season but decreased it in the nongrowing season, and the keystone species in the interdomain network changed with warming. Warming also reduced the proportion of positive microbial community interactions, indicating that warming reduced the mutualism, commensalism, and neutralism of microorganisms as they adapted to soil environmental stress. The factors affecting the fungal community varied considerably across warming gradients, with the bacterial community being significantly affected by soil temperature, MBC, NO3--N and NH4+-N, moreover, SOC and TN significantly affected fungal communities in the 4 °C warming treatment. These results suggest that warming increases seasonal differences in the diversity and complexity of soil microbial communities in the northern subtropical region, significantly influencing soil dynamic processes regulating forest ecosystems under global warming.

Superhydrophobic coatings reduce human bacterial foodborne pathogen attachment to woods used in fresh produce harvest and postharvest packing.

Wood is reportedly more difficult to maintain in hygienic condition versus other food contact materials, yet its use in produce packing and retail warrants efforts to reduce the risk of microbial pathogen contamination and attachment. This study characterized antifouling capabilities of fluorinated silanes applied to wood used in fresh edible produce handling to render the wood superhydrophobic and less supportive of bacterial pathogen attachment. Pine and oak cubic coupon surfaces were treated with 1% (w/w) silane or left untreated. Treated and untreated coupons were inoculated with Salmonella enterica or Listeria monocytogenes and held to facilitate pathogen attachment for 1, 4, or 8 h. Silane treatment of wood produced significant reductions in the proportions of strongly attaching cells for both pathogens versus loosely attaching cells (P < 0.01). Salmonella attachment demonstrated a dependency on wood treatment; silane-treated wood supported a lower fraction of strongly adhering cells (1.87 ± 1.24 log CFU/cm2) versus untreated wood (3.72 ± 0.67 log CFU/cm2). L. monocytogenes demonstrated significant declines in strongly attaching cells during extended exposure to silane-treated wood, from 7.59 ± 0.14 to 5.27 ± 0.68 log CFU/cm2 over 8 h post-inoculation. Microscopic analysis demonstrated silane treatment increased the surface roughness of both woods, leading to superhydrophobic conditions on wood surfaces, consequently decreasing strong attachment of pathogenic bacteria.

Genotypic Identification of Trees Using DNA Barcodes and Microbiome Analysis of Rhizosphere Microbial Communities.

DNA barcodes can provide accurate identification of plants. We used previously reported DNA primers targeting the internal transcribed spacer (ITS1) region of the nuclear ribosomal cistron, internal transcribed spacer (ITS2), and chloroplast trnL (UAA) intron to identify four trees at Bergen Community College. Two of the four trees were identified as Acer rubrum and Fagus sylvatica. However, Quercus was only identified at the genus level, and the fourth tree did not show similar identification between barcodes. Next-generation sequencing of 16S rRNA genes showed that the predominant bacterial communities in the rhizosphere mainly consisted of the Pseudomonadota, Actinomycetota, Bacteroidota, and Acidobacteriota. A. rubrum showed the most diverse bacterial community while F. sylvatica was less diverse. The genus Rhodoplanes showed the highest relative bacterial abundance in all trees. Fungal ITS sequence analysis demonstrated that the communities predominantly consisted of the Ascomycota and Basidiomycota. Quercus showed the highest fungi diversity while F. sylvatica showed the lowest. Russula showed the highest abundance of fungi genera. Average similarity values in the rhizosphere for fungi communities at the phylum level were higher than for bacteria. However, at the genus level, bacterial communities showed higher similarities than fungi. Similarity values decreased at lower taxonomical levels for both bacteria and fungi, indicating each tree has selected for specific bacterial and fungal communities. This study confirmed the distinctiveness of the microbial communities in the rhizosphere of each tree and their importance in sustaining and supporting viability and growth but also demonstrating the limitations of DNA barcoding with the primers used in this study to identify genus and species for some of the trees. The optimization of DNA barcoding will require additional DNA sequences to enhance the resolution and identification of trees at the study site.

Recover of Soil Microbial Community Functions in Beech and Turkey Oak Forests After Coppicing Interventions.

Forest management influences the occurrence of tree species, the organic matter input to the soil decomposer system, and hence, it can alter soil microbial community and key ecosystem functions it performs. In this study, we compared the potential effect of different forest management, coppice and high forest, on soil microbial functional diversity, enzyme activities and chemical-physical soil properties in two forests, turkey oak and beech, during summer and autumn. We hypothesized that coppicing influences soil microbial functional diversity with an overall decrease. Contrary to our hypothesis, in summer, the functional diversity of soil microbial community was higher in both coppice forests, suggesting a resilience response of the microbial communities in the soil after tree cutting, which occurred 15-20 years ago. In beech forest under coppice management, a higher content of soil organic matter (but also of soil recalcitrant and stable organic carbon) compared to high forest can explain the higher soil microbial functional diversity and metabolic activity. In turkey oak forest, although differences in functional diversity of soil microbial community between management were observed, for the other investigated parameters, the differences were mainly linked to seasonality. The findings highlight that the soil organic matter preservation depends on the type of forest, but the soil microbial community was able to recover after about 15 years from coppice intervention in both forest ecosystems. Thus, the type of management implemented in these forest ecosystems, not negatively affecting soil organic matter pool, preserving microbial community and potentially soil ecological functions, is sustainable in a scenario of climate change.

Modelling suggests Wolbachia-induced cytoplasmic incompatibility in oak gall wasps with cyclical parthenogenesis.

Oak gall wasps typically exhibit a life cycle with one sexual and one asexual generation each year. These wasps can carry various endosymbionts, one of which is the maternally inherited bacterium Wolbachia that can induce several reproductive manipulations on its host. Cytoplasmic incompatibility (CI) has been described as the most prominent of these manipulations. CI leads to embryonic mortality in the hosts' offspring when infected males mate with either uninfected females or with females that harbour different Wolbachia strains. It has been hypothesized that Wolbachia can induce CI in oak gall wasps. To address this hypothesis, we derived a mathematical model to investigate the spread of a bacterial infection in naive populations and to determine the plausibility of CI occurrence. To validate our model, we used published data from Wolbachia-infected Belonocnema kinseyi populations in two approaches. Our first approach uses measurements of infection frequencies and maternal transmission in the sexual generation. For the second approach, we extended the model to compare predictions to estimates of mtDNA-haplotypes, which, like Wolbachia, are maternally inherited, and can therefore be associated with the infection. Both approaches indicate that CI is present in these populations. Our model can be generalized to investigate the occurrence of CI not only for oak gall wasps but also for other species.

Effect of Oak Powdery Mildew on Ascorbate-Glutathione Cycle and Other Antioxidants in Plant-Erysiphe alphitoides Interaction.

Erysiphe alphitoides is a species of powdery mildew responsible for the major foliar disease of oak trees, including Quercus robur. Infection with E. alphitoides leads to a reduction in the growth of the trees and in their ability to survive. This paper reports on the biochemical changes characteristic of defence responses in oak leaves with different infection area sizes, collected in July, August, and September during three growing seasons. The study highlights the effect of E. alphitoides infection on changes in the ascorbate-glutathione cycle, phenolic compound profile, and metal content (mineral distribution). Visible symptoms of pathogen infection appeared gradually in July, but the most intense biochemical plant responses in oak leaves were detected mainly in August and September. These responses included increased ascorbate-glutathione enzyme activities, phenolic compounds, and metal contents. In addition, microscopic analyses revealed a strong fluorescence signal of lignin in the epidermis of pathogen-infected leaves. The involvement of the studied compounds in the basic defence mechanisms of oak against E. alphitoides infection is discussed in the paper.

Pathogens of the oak processionary moth Thaumetopoea processionea: Developing a user-friendly bioassay system and metagenome analyses for microorganisms.

The oak processionary moth (OPM) Thaumetopoea processionea is a pest of oak trees and poses health risks to humans due to the urticating setae of later instar larvae. For this reason, it is difficult to rear OPM under laboratory conditions, carry out bioassays or examine larvae for pathogens. Biological control targets the early larval instars and is based primarily on commercial preparations of Bacillus thuringiensis ssp. kurstaki (Btk). To test the entomopathogenic potential of other spore-forming bacteria, a user-friendly bioassay system was developed that (i) applies bacterial spore suspensions by oak bud dipping, (ii) targets first instar larvae through feeding exposure and (iii) takes into account their group-feeding behavior. A negligible mortality in the untreated control proved the functionality of the newly established bioassay system. Whereas the commercial Btk HD-1 strain was used as a bioassay standard and confirmed as being highly efficient, a Bacillus wiedmannii strain was ineffective in killing OPM larvae. Larvae, which died during the infection experiment, were further subjected to Nanopore sequencing for a metagenomic approach for entomopathogen detection. It further corroborated that B.wiedmannii was not able to infect and establish in OPM, but identified potential insect pathogenic species from the genera Serratia and Pseudomonas.

Potential distribution of Biscogniauxia mediterranea and Obolarina persica causal agents of oak charcoal disease in Iran's Zagros forests.

In Iran, native oak species are under threat from episodes of Charcoal Disease, a decline syndrome driven by abiotic stressors (e.g. drought, elevated temperature) and biotic components, Biscogniauxia mediterranea (De Not.) Kuntze and Obolarina persica (M. Mirabolfathy). The outbreak is still ongoing and the country's largest ever recorded. Still, the factors driving its' epidemiology in time and space are poorly known and such knowledge is urgently needed to develop strategies to counteract the adverse effects. In this study, we developed a generic framework based on experimental, machine-learning algorithms and spatial analyses for landscape-level prediction of oak charcoal disease outbreaks. Extensive field surveys were conducted during 2013-2015 in eight provinces (more than 50 unique counties) in the Zagros ecoregion. Pathogenic fungi were isolated and characterized through morphological and molecular approaches, and their pathogenicity was assessed under controlled water stress regimes in the greenhouse. Further, we evaluated a set of 29 bioclimatic, environmental, and host layers in modeling for disease incidence data using four well-known machine learning algorithms including the Generalized Linear Model, Gradient Boosting Model, Random Forest model (RF), and Multivariate Adaptive Regression Splines implemented in MaxEnt software. Model validation statistics [Area Under the Curve (AUC), True Skill Statistics (TSS)], and Kappa index were used to evaluate the accuracy of each model. Models with a TSS above 0.65 were used to prepare an ensemble model. The results showed that among the different climate variables, precipitation and temperature (Bio18, Bio7, Bio8, and bio9) in the case of O. persica and similarly, gsl (growing season length TREELIM, highlighting the warming climate and the endophytic/pathogenic nature of the fungus) and precipitation in case of B. mediterranea are the most important influencing variables in disease modeling, while near-surface wind speed (sfcwind) is the least important variant. The RF algorithm generates the most robust predictions (ROC of 0.95; TSS of 0.77 and 0.79 for MP and OP, respectively). Theoretical analysis shows that the ensemble model (ROC of 0.95 and 0.96; TSS = 0.79 and 0.81 for MP and OP, respectively), can efficiently be used in the prediction of the charcoal disease spatiotemporal distribution. The oak mortality varied ranging from 2 to 14%. Wood-boring beetles association with diseased trees was determined at 20%. Results showed that water deficiency is a crucial component of the oak decline phenomenon in Iran. The Northern Zagros forests (Ilam, Lorestan, and Kermanshah provinces) along with the southern Zagros forests (Fars and Kohgilouyeh va-Boyer Ahmad provinces) among others are the most endangered areas of potential future pandemics of charcoal disease. Our findings will significantly improve our understanding of the current situation of the disease to pave the way against pathogenic agents in Iran.

Effects of four bolete species on ectomycorrhizae formation and development in Pinus thunbergii and Quercus acutissima.

BACKGROUND: Bolete cultivation is economically and ecologically valuable. Ectomycorrhizae are advantageous for plant development and productivity. This study investigated how boletes affect the formation of Pinus thunbergii and Quercus acutissima ectomycorrhizae using greenhouse-based mycorrhizal experiments, inoculating P. thunbergii and Q. acutissima with four species of boletes (Suillus bovinus, Suillus luteus, Suillus grevillei, and Retiboletus sinensis). RESULTS: Three months after inoculation, morphological and molecular analyses identified S. bovinus, S. luteus, S. grevillei and R. sinensis ectomycorrhizae formation on the roots of both tree species. The mycorrhizal infection rate ranged from 40 to 55%. The host plant species determined the mycorrhiza morphology, which was independent of the bolete species. Differences in plant growth, photosynthesis, and endogenous hormone secretion primarily correlated with the host plant species. Infection with all four bolete species significantly promoted the host plants' growth and photosynthesis rates; indole-3-acetic acid, zeatin, and gibberellic acid secretion increased, and the abscisic acid level significantly decreased. Indole-3-acetic acid was also detected in the fermentation broths of all bolete species. CONCLUSIONS: Inoculation with bolete and subsequent mycorrhizae formation significantly altered the morphology and hormone content in the host seedlings, indicating growth promotion. These findings have practical implications for culturing pine and oak tree species.

Detecting Pathogenic Phytophthora Species Using Volatile Organic Compounds.

There are several highly damaging Phytophthora species pathogenic to forest trees, many of which have been spread beyond their native range by the international trade of live plants and infested materials. Such introductions can be reduced through the development of better tools capable of the early, rapid, and high-throughput detection of contaminated plants. This study utilized a volatilomics approach (solid-phase microextraction coupled to gas chromatography-mass spectrometry) to differentiate between several Phytophthora species in culture and discriminate between healthy and Phytophthora-inoculated European beech and pedunculate oak trees. We tentatively identified 14 compounds that could differentiate eight Phytophthora species from each other in vitro. All of the Phytophthora species examined, except Phytophthora cambivora, uniquely produced at least one compound not observed in the other species; however, most detected compounds were shared between multiple species. Phytophthora polonica had the most unique compounds and was the least similar of all the species examined. The inoculated seedlings had qualitatively different volatile profiles and could be distinguished from the healthy controls by the presence of isokaurene, anisole, and a mix of three unknown compounds. This study supports the notion that volatiles are suitable for screening plant material, detecting tree pathogens, and differentiating between healthy and diseased material.

Ectomycorrhizal fungal communities in natural and urban ecosystems: Quercus humboldtii as a study case in the tropical Andes.

Worldwide urban landscapes are expanding because of the growing human population. Urban ecosystems serve as habitats to highly diverse communities. However, studies focusing on the diversity and structure of ectomycorrhizal communities are uncommon in this habitat. In Colombia, Quercus humboldtii Bonpl. is an ectomycorrhizal tree thriving in tropical montane forests hosting a high diversity of ectomycorrhizal fungi. Q. humboldtii is planted as an urban tree in Bogotá (Colombia). We studied how root-associated fungal communities of this tree change between natural and urban areas. Using Illumina sequencing, we amplified the ITS1 region and analyzed the resulting data using both OTUs and Amplicon Sequence Variants (ASVs) bioinformatics pipelines. The results obtained using both pipelines showed no substantial differences between OTUs and ASVs for the community patterns of root-associated fungi, and only differences in species richness were observed. We found no significant differences in the species richness between urban and rural sites based on Fisher's alpha or species-accumulation curves. However, we found significant differences in the community composition of fungi present in the roots of rural and urban trees with rural communities being dominated by Russula and Lactarius and urban communities by Scleroderma, Hydnangium, and Trechispora, suggesting a high impact of urban disturbances on ectomycorrhizal fungal communities. Our results highlight the importance of urban trees as reservoirs of fungal diversity and the potential impact of urban conditions on favoring fungal species adapted to more disturbed ecosystems.

Vibrational spectroscopic profiling of biomolecular interactions between oak powdery mildew and oak leaves.

Oak powdery mildew, caused by the biotrophic fungus Erysiphe alphitoides, is a prevalent disease affecting oak trees, such as English oak (Quercus robur). While mature oak populations are generally less susceptible to this disease, it can endanger young oak seedlings and new leaves on mature trees. Although disruptions of photosynthate and carbohydrate translocation have been observed, accurately detecting and understanding the specific biomolecular interactions between the fungus and the leaves of oak trees is currently lacking. Herein, via hybrid Raman spectroscopy combined with an advanced artificial neural network algorithm, the underpinning biomolecular interactions between biological soft matter, i.e., Quercus robur leaves and Erysiphe alphitoides, are investigated and profiled, generating a spectral library and shedding light on the changes induced by fungal infection and the tree's defence response. The adaxial surfaces of oak leaves are categorised based on either the presence or absence of Erysiphe alphitoides mildew and further distinguishing between covered or not covered infected leaf tissues, yielding three disease classes including healthy controls, non-mildew covered and mildew-covered. By analysing spectral changes between each disease category per tissue type, we identified important biomolecular interactions including disruption of chlorophyll in the non-vein and venule tissues, pathogen-induced degradation of cellulose and pectin and tree-initiated lignification of cell walls in response, amongst others, in lateral vein and mid-vein tissues. Via our developed computational algorithm, the underlying biomolecular differences between classes were identified and allowed accurate and rapid classification of disease with high accuracy of 69.6% for non-vein, 73.5% for venule, 82.1% for lateral vein and 85.6% for mid-vein tissues. Interfacial wetting differences between non-mildew covered and mildew-covered tissue were further analysed on the surfaces of non-vein and venule tissue. The overall results demonstrated the ability of Raman spectroscopy, combined with advanced AI, to act as a powerful and specific tool to probe foliar interactions between forest pathogens and host trees with the simultaneous potential to probe and catalogue molecular interactions between biological soft matter, paving the way for exploring similar relations in broader forest tree-pathogen systems.

Comparative Spatial Modeling of Bretziella fagacearum Distribution in Northern Wisconsin, United States.

Bretziella fagacearum, the ascomycete fungus oak wilt, is considered a virulent threat to North American oaks, but the influence of the physical environment on this pathosystem remains unclear, particularly at the forest scale. This study explored the influence of terrain and soil factors on B. fagacearum infections, applying discrete and continuous spatial models to investigate the question, besides proximity to other infections, which environmental factors influenced B. fagacearum incidence? Locations of infections were recorded from 586 confirmed B. fagacearum sites, identified from 2004 through 2021 in a 76 km2 area of deep, sandy glacial outwash in Chequamegon-Nicolet National Forest, northern Wisconsin. Public datasets derived from remote sensing were incorporated as covariates, describing terrain elevation (USGS 10-m DEM), soil physical and chemical properties (POLARIS), and forest composition (WiscLand2). Spatial models included generalized additive models (GAMs) and Neyman-Scott cluster process models. The results indicated that spatial dependence and the distribution of oak forests were the most important drivers of B. fagacearum distribution in this area, with more minor influence from elevation, hill shade, and drainage patterns. Comparison between modeling approaches indicated that-at this scale and in this area-the most accurate models were those that included host distribution, spatial dependence, and quantitative terrain and soil descriptions. However, a close approximation could be attained using nonlinear models (GAMs) that incorporated only host distribution and spatial dependence.

Sequential application of inoculation methods improves mycorrhization of Quercus ilex seedlings by Tuber melanosporum.

The use of mycorrhized seedlings has been critical to the success of modern truffle cultivation, which nowadays supplies most European black truffles (Tuber melanosporum) to markets. Ascospore inoculation has been traditionally used to produce these seedlings, but little scientific information is publicly available on the inoculation methods applied or on the possibility of combining them. We evaluated the potential of sequential inoculation for the controlled colonization of holm oak fine roots by T. melanosporum, with two different nursery assays and a full factorial design. Three inoculation methods were sequentially applied: radicle inoculation, inoculation of the substrate in seedling trays and inoculation of the substrate in the final pot. Despite the differences in the results of the two assays, which suggest that cultivation conditions and/or the timing of nursery operations may influence the relative effectiveness of inoculation methods, the sequential application appeared as an effective and realistic alternative for commercial inoculation of holm oak seedlings with T. melanosporum. The increase in the amount of inoculum applied with each inoculation method improved the mycorrhizal colonization of seedlings, whereas separately none of the inoculation methods appeared clearly superior to the other ones. The depth distribution of truffle mycorrhizae pointed that the inoculation in the final pot was more effective than other methods in lower parts of the root system, whereas the early inoculation appeared more effective to reduce the occurrence of the opportunist ectomycorrhizal fungus Sphaerosporella brunnea.

Into the void: ECM fungal communities involved in the succession from rockroses to oak stands.

Oak forests accompanied by Cistus species are a common landscape in the Mediterranean basin. It is argued that Cistus dominated fields serve as recruitment areas for Quercus seedlings, as they help in the transmission of the fungal community through vegetative succession in these ecosystems. To test these assumptions, we analyzed the fungal community in terms of its richness and composition, taking into account the effects of host (Oaks vs. Cistus) and forest structure, mainly based on age. Edaphic variables related to the different structures were also analyzed to examine how they evolve through succession and relate to shifts in the fungal community. No differences in fungal richness were observed between old Cistus stands and younger Quercus, while a brief increase in ECM richness was observed. Community composition also showed a greater overlap between old Cistus and young Quercus stands. We suggest that the most important step in fungal transfer from one host to another is the shift from the oldest Cistus fields to the youngest Quercus stands, with the genera Amanita, Cortinarius, Lactarius, Inocybe, Russula, and Tomentella probably playing a major role. In summary, our work has also revealed the network of fungal community structure in the succession of Cistus to Oak stands, it would suggest that the fungi share niches and significantly enhance the ecological setting of the transition from Cistus to Oak stands.

Description of Dryocola gen. nov. and two novel species, Dryocola boscaweniae sp. nov. and Dryocola clanedunensis sp. nov. isolated from the rhizosphere of native British oaks.

While investigating the role of the rhizosphere in the development of Acute Oak Decline, bacterial strains belonging to the family Enterobacteriaceae were isolated from rhizosphere soil following enrichment for the Enterobacterales. Partial sequencing of several housekeeping genes showed that these strains could not be assigned to an existing genus. Overall, 16 strains were investigated using a polyphasic approach to determine their taxonomic status. This involved phenotypic testing and fatty acid analysis paired with phylogenetic analyses of 16S rRNA and housekeeping gene sequences, as well as phylogenomic analysis of whole genome sequences. Phylogenomic and phylogenetic analyses consistently demonstrated that the 16 isolates could be separated into two distinct clusters in a monophyletic clade situated between the genera Cedecea and Buttiauxella. The two clusters could be genotypically and phenotypically differentiated from each other and from their closest neighbours. As such we propose the description of Dryocola boscaweniae gen. nov. sp. nov. (type strain H6W4T = CCUG 76177T = LMG 32610T) and Dryocola clanedunesis sp. nov. (type strain H11S18T = CCUG 76181T = LMG 32611T).

Host Specificity and Seasonal Variation in the Colonization of Tubakia sensu lato Associated with Evergreen Oak Species in Eastern Japan.

Foliar fungal endophytes are ubiquitous and hyperdiverse, and tend to be host-specific among dominant forest tree species. The fungal genus Tubakia sensu lato is comprised of foliar pathogens and endophytes that exhibit host preference for Quercus and other Fagaceae species. To clarify interspecific differences in ecological characteristics among Tubakia species, we examined the endophyte communities of seven evergreen Quercus spp. at three sites in eastern Japan during summer and winter. Host tree species was the most significant factor affecting endophyte community composition. Tubakia species found at the study sites were divided into five specialists and three generalists according to their relative abundance in each host species and their host ranges. Specialists were dominant on their own host in summer, and their abundance declined in winter. To test the hypothesis that generalists are more widely adapted to their environment than specialists, we compared their spore germination rates at different temperatures. Spores of generalist Tubakia species were more tolerant of colder temperatures than were spores of specialist Tubakia species, supporting our hypothesis. Seasonal and site variations among Tubakia species were also consistent with our hypothesis. Host identity and ecology were significantly associated with endophyte community structure.

Intraspecific Competition Results in Reduced Evenness of Tuber melanosporum Mating-Type Abundance from the Nursery Stage.

The highly prized black truffle is a fungus mostly harvested in orchards planted with mycorrhizal seedlings. It is an obligatory outcrossing fungus with a single MAT locus containing two alternative mating-type idiomorphs. In the orchards, at the mycorrhizal level, these mating types are frequently spatially segregated. Some studies found that this segregation was pronounced from the nursery stage, whereas others did not find such a marked segregation. Besides, information on the host tree species and nursery conditions used in Spain, one of the main truffle-producing countries, are very scarce. In this study, we investigated the temporal dynamics of mating types in nursery seedlings of Quercus ilex and Quercus faginea, as well as the influence of cultural conditions in the nursery. Our results indicated that at the plant level, there was a trend for one of the mating types to dominate over the other from the first to the second year in the nursery, in both host species and both nursery conditions tested. However, this segregation process was not so sharp as previously reported. Our results support the hypothesis that intraspecific competition results in reduced evenness of mating-type abundance from the nursery stage, although almost all seedlings maintained both mating types and, at the seedling batch scale, the occurrence of both mating types was roughly balanced.