Fungal Partners Research Articles

Plant pests influence the movement of plant‐fixed carbon and fungal‐acquired nutrients through arbuscular mycorrhizal networks

Abstract Plants typically interact with multiple, co‐occurring symbionts, including arbuscular mycorrhizal (AM) fungi which can form networks, connecting neighbouring plants. A characteristic aspect of the mycorrhizal symbiosis is the bidirectional exchange of nutrients between host plants and fungal partners. Concurrent interactions with competing organisms such as aphids or potato cyst nematodes (PCN) can disrupt the carbon‐for‐nutrient exchange between plants and AM fungi. However, the role of mycorrhizal networks (MNs) in mediating these interactions remains unclear. Using isotope tracing in multi‐plant experimental systems, we investigated the movement of plant photosynthates and fungal‐acquired soil phosphorus through MNs and the interactive effects of PCN infection on this. We found evidence of preferential allocation of fungal‐acquired phosphorus to plants that were not infected by PCN compared with infected neighbours. Contrary to previous findings using single plants, we did not detect a PCN‐induced reduction in the amounts of plant carbon delivered to AM fungi in multi‐plant systems. However, the MN(s) moved plant‐fixed carbon away from PCN‐infected host plants, regardless of the PCN infection status of the neighbouring plant host. Our work highlights the responsiveness of MNs to interactions with below‐ground organisms. It also strengthens the argument for a more mycocentric view of AM–plant symbioses. Experimental designs of increasing ecological complexity are needed for a more comprehensive understanding of the carbon‐for‐nutrient dynamics in AM fungi–plant networks. This will, in turn, elucidate the role of AM fungi in terrestrial carbon cycling and their function in agricultural systems. Read the free Plain Language Summary for this article on the Journal blog.

Bacterial phylotypes associated with rock-dwelling Umbilicaria Lichens from Arctic/Subarctic areas in North America and Northern Europe

AbstractThe diversity of bacteria associated with lichens has received increasing attention. However, studies based on next-generation sequencing of microbiomes have not yet been conducted in the Arctic and Subarctic regions. In this study, rock-dwelling lichens belonging to the Umbilicariaceae family were sampled from the Arctic and Subarctic biological zones. The primary research purpose was to undertake a comparative investigation of the bacterial composition and diversity, identify potential indicators, and explore their potential metabolic pathways. 18S rRNA gene sequences of the fungal partner belonging to the genus Umbilicaria (Ascomycota) and the algal partner affiliated with the lineage Trebouxia (Chlorophyta). Comparing Umbilicaria spp. with a previous study in the Antarctic zone, the fungal partners were more inclined to cluster by sampling region. Operational taxonomic units (OTUs) were established based on a predetermined similarity threshold for V3-V4 sequences, which were ascribed to 19 bacterial phyla, and ten of them were consistently present in all samples. The most distinct zonal indicator genera based on OTU frequencies from Arctic and Subarctic lichens were Capsulimonas (Armatimonadota) and Jatrophihabitans (Actinomycota), respectively. Although the Subarctic zone had higher biodiversity and species richness based on alpha-diversity, the beta-diversity showed that the main species of bacterial communities were not significantly different, and the predictions of metabolic pathways based on the bacterial microbiome in lichen samples from the two zones were similar. These findings provide evidence that the geographical and/or bioclimatic environment and the different lichen-forming fungal species mainly and partially influence bacterial microbiomes and metabolic pathways.

The decision for or against mycoparasitic attack by Trichoderma spp. is taken already at a distance in a prey-specific manner and benefits plant-beneficial interactions

BackgroundThe application of plant-beneficial microorganisms as bio-fertilizer and biocontrol agents has gained traction in recent years, as both agriculture and forestry are facing the challenges of poor soils and climate change. Trichoderma spp. are gaining popularity in agriculture and forestry due to their multifaceted roles in promoting plant growth through e.g. nutrient translocation, hormone production, induction of plant systemic resistance, but also direct antagonism of other fungi. However, the mycotrophic nature of the genus bears the risk of possible interference with other native plant-beneficial fungi, such as ectomycorrhiza, in the rhizosphere. Such interference could yield unpredictable consequences for the host plants of these ecosystems. So far, it remains unclear, whether Trichoderma is able to differentiate between plant-beneficial and plant-pathogenic fungi during the process of plant colonization.ResultsWe investigated whether Trichoderma spp. can differentiate between beneficial ectomycorrhizal fungi (represented by Laccaria bicolor and Hebeloma cylindrosporum) and pathogenic fungi (represented by Fusarium graminearum and Alternaria alternata) in different confrontation scenarios, including a newly developed olfactometer “race tube”-like system. Using two independent species, T. harzianum and T. atrobrunneum, with plant-growth-promoting and immune-stimulating properties towards Populus x canescens, our study revealed robustly accelerated growth towards phytopathogens, while showing a contrary response to ectomycorrhizal fungi. Transcriptomic analyses identified distinct genetic programs during interaction corresponding to the lifestyles, emphasizing the expression of mycoparasitism-related genes only in the presence of phytopathogens.ConclusionThe findings reveal a critical mode of fungal community interactions belowground and suggest that Trichoderma spp. can distinguish between fungal partners of different lifestyles already at a distance. This sheds light on the entangled interactions of fungi in the rhizosphere and emphasizes the potential benefits of using Trichoderma spp. as a biocontrol agent and bio-fertilizer in tree plantations.

The interactive effect of tree mycorrhizal type, mycorrhizal type mixture and tree diversity shapes rooting zone soil fungal communities in temperate forest ecosystems

Abstract The underlying processes of plant‐microbe associations particularly their interactions with their mycorrhizal fungal partners have been extensively studied. However, considerably less is known about the consequences of tree‐tree interactions on rooting zone soil microbiota when tree species of different mycorrhizal type (myco‐type) grow together as mono and mixed myco‐type mixtures along a tree diversity gradient. Using the MyDiv tree diversity experiment, where arbuscular mycorrhizal (AM) and ectomycorrhizal (EcM) trees and their mixtures were planted in monocultures, two‐species and four‐species mixture plots, we investigated the interplay of target tree myco‐type, myco‐type mixture, tree diversity and rooting zone compartment (target tree dominated and its interaction zones with neighbour trees) on the rooting zone soil mycobiota employing meta‐barcoding of the ITS2 rDNA fragment of the fungal internal transcribed spacer (ITS). Our results revealed significant individual and interaction effects of tree myco‐type, myco‐type mixture and tree diversity but not rooting zone compartment on the fungal taxonomic and functional alpha and beta diversity. This implies intermingling of roots of target and neighbouring tree species there by reducing the target tree species effect in its rooting zone. As tree diversity increases, we found convergence of the fungal community in general, where the fungal community dissimilarity varies depending on the co‐existing tree species myco‐type and tree species diversity. Furthermore, the fungal community composition in the two and four species mixtures were consistently influenced by soil pH, whereas in the mixed multi‐species stands basal respiration, N, PO4−, NO3− were found to be equally important unlike in AM and EcM multi‐species stands. Comparative analysis of the fungal taxa specialisation between mono and mixed myco‐type multi‐species stands revealed that the mixed myco‐type plots shared 23.5% (AM) and 19.7% (EcM) of the generalist fungal communities However, the percentage of specialised fungal community in mixed myco‐type plots (13.2%) was significantly higher as compared to EcM (9.5%), and significantly lower (9%) as compared to AM (11.7%) plots, resulting in myco‐type and myco‐type mixture specific fungal communities and functional guild patterns Our results provide novel insights on the significance of tree species and its co‐existing trees preferred mycorrhizal association in shaping the target tree rooting zone soil mycobiome along a tree diversity gradient. Furthermore, it highlights the significance of generalist and specialist fungal communities in mono and mixed myco‐type stands in modulating tree‐tree interaction, tree species co‐existence and regulating soil properties and ecosystem functions. Read the free Plain Language Summary for this article on the Journal blog.

De Novo Genome Assembly of Toniniopsis dissimilis (Ramalinaceae, Lecanoromycetes) from Long Reads Shows a Comparatively High Composition of Biosynthetic Genes Putatively Involved in Melanin Synthesis.

Lichens have developed numerous adaptations to optimize their survival in various environmental conditions, largely by producing secondary compounds by the fungal partner. They often have antibiotic properties and are involved in protection against intensive UV radiation, pathogens, and herbivores. To contribute to the knowledge of the arsenal of secondary compounds in a crustose lichen species, we sequenced and assembled the genome of Toniniopsis dissimilis, an indicator of old-growth forests, using Oxford Nanopore Technologies (ONT, Oxford, UK) long reads. Our analyses focused on biosynthetic gene clusters (BGCs) and specifically on Type I Polyketide (T1PKS) genes involved in the biosynthesis of polyketides. We used the comparative genomic approach to compare the genome of T. dissimilis with six other members of the family Ramalinaceae and twenty additional lichen genomes from the database. With only six T1PKS genes, a comparatively low number of biosynthetic genes are present in the T. dissimilis genome; from those, two-thirds are putatively involved in melanin biosynthesis. The comparative analyses showed at least three potential pathways of melanin biosynthesis in T. dissimilis, namely via the formation of 1,3,6,8-tetrahydroxynaphthalene, naphthopyrone, or YWA1 putative precursors, which highlights its importance in T. dissimilis. In addition, we report the occurrence of genes encoding ribosomally synthesized and posttranslationally modified peptides (RiPPs) in lichens, with their highest number in T. dissimilis compared to other Ramalinaceae genomes. So far, no function has been assigned to RiPP-like proteins in lichens, which leaves potential for future research on this topic.

Colonization by orchid mycorrhizal fungi primes induced systemic resistance against necrotrophic pathogen.

Orchids and arbuscular mycorrhiza (AM) plants evolved independently and have different structures and fungal partners, but they both facilitate nutrient uptake. Orchid mycorrhiza (OM) supports orchid seed germination, but unlike AM, its role in disease resistance of mature plants is largely unknown. Here, we examined whether OM induces systemic disease resistance against a necrotrophic pathogen in a similar fashion to AM. We investigated the priming effect of mycorrhizal fungi inoculation on resistance of a terrestrial orchid, Bletilla striata, to soft rot caused by Dickeya fangzhongdai. We found that root colonization by a compatible OM fungus primed B. striata seedlings and induced systemic resistance against the infection. Transcriptome analysis showed that priming was mediated by the downregulation of jasmonate and ethylene pathways and that these pathways are upregulated once infection occurs. Comparison with the reported transcriptome of AM fungus-colonized rice leaves revealed similar mechanisms in B. striata and in rice. These findings highlight a novel aspect of commonality between OM and AM plants in terms of induced systemic resistance.

Halophilomyces hongkongensis, a Novel Species and Genus in the Lulworthiaceae with Antibacterial Potential, Colonizing the Roots and Rhizomes of the Seagrass Halophila ovalis.

Seagrass serves as a quintessential reservoir for obligate marine Lulworthiaceae fungi. Our current knowledge of the mycological diversity associated with seagrass in Hong Kong remains poor. We analyzed the diversity of fungi associated with the most widely distributed seagrass species in Hong Kong Halophila ovalis (Hydrocharitaceae), using a combination of culture-based methods and high-throughput amplicon sequencing. Halophilomyces hongkongensis, a novel fungal species in a newly proposed genus within the Lulworthiaceae family, was isolated from H. ovalis roots and rhizomes. The novel fungus showed distinct morphological characteristics, while both combined 18S-28S and internal transcribed spacer (ITS) phylogenetic trees based on maximum likelihood and Bayesian methods supported its discrimination from other existing Lulworthiaceae members. The ITS2 region in the Illumina sequencing results of multiple H. ovalis compartments, water, and adjacent non-seagrass sediments revealed continuous recruitment of H. hongkongensis by H. ovalis throughout the year despite dramatically fluctuating environmental conditions, with remarkably high proportions of this taxon found in root and rhizome internal tissues, possibly indicating a strong and specialized relationship established between the Lulworthiaceae fungal partner and its seagrass host. The inhibitory abilities exhibited by H. hongkongensis against Staphylococcus aureus SA29213 and ATCC 43300 (methicillin-resistant) may imply its capacity in producing (novel) antibacterial compounds. The discovery of H. hongkongensis as the first novel Lulworthiaceae taxon in Hong Kong, along with its distributional pattern in the seagrass meadow, provides valuable insights into the systematics and ecology of this strictly marine fungal family.

Exploring mycorrhizal diversity in sympatric mycoheterotrophic plants: a comparative study of Monotropastrum humile var. humile and M. humile var. glaberrimum.

Mycoheterotrophic plants (MHPs) rely on their mycorrhizal fungus for carbon and nutrient supply, thus a shift in mycobionts may play a crucial role in speciation. This study aims to explore the mycorrhizal diversity of two closely related and sympatric fully MHPs, Monotropastrum humile var. humile (Mhh) and M. humile var. glaberrimum (Mhg), and determine their mycorrhizal associations. A total of 1,108,710 and 1,119,071 ectomycorrhizal fungal reads were obtained from 31 Mhh and 31 Mhg, and these were finally assigned to 227 and 202 operational taxonomic units, respectively. Results show that sympatric Mhh and Mhg are predominantly associated with different fungal genera in Russulaceae. Mhh is consistently associated with members of Russula, whereas Mhg is associated with members of Lactarius. Associating with different mycobionts and limited sharing of fungal partners might reduce the competition and contribute to their coexistence. The ectomycorrhizal fungal communities are significantly different among the five forests in both Mhh and Mhg. The distinct mycorrhizal specificity between Mhh and Mhg suggests the possibility of different mycobionts triggered ecological speciation between sympatric species.

Mode of carbon gain and fungal associations of Neuwiedia malipoensis within the evolutionarily early-diverging orchid subfamily Apostasioideae.

The earliest-diverging orchid lineage, Apostasioideae, consists only of two genera: Apostasia and Neuwiedia. Previous reports of Apostasia nipponica indicated a symbiotic association with an ectomycorrhiza-forming Ceratobasidiaceae clade and partial utilization of fungal carbon during the adult stage. However, the trophic strategy of Neuwiedia throughout its development remains unidentified. To further improve our understanding of mycoheterotrophy in the Apostasioideae, this study focused on Neuwiedia malipoensis examining both the mycorrhizal association and the physiological ecology of this orchid species across various development stages. We identified the major mycorrhizal fungi of N. malipoensis protocorm, leafy seedling and adult stages using molecular barcoding. To reveal nutritional resources utilized by N. malipoensis, we compared stable isotope natural abundances (δ13C, δ15N, δ2H, δ18O) of different developmental stages with those of autotrophic reference plants. Protocorms exhibited an association with saprotrophic Ceratobasidiaceae rather than ectomycorrhiza-forming Ceratobasidiaceae and the 13C signature was characteristic of their fully mycoheterotrophic nutrition. Seedlings and adults were predominantly associated with saprotrophic fungi belonging to the Tulasnellaceae. While 13C and 2H stable isotope data revealed partial mycoheterotrophy of seedlings, it is unclear to what extent the fungal carbon supply is reduced in adult N. malipoensis. However, the 15N enrichment of mature N. malipoensis suggests partially mycoheterotrophic nutrition. Our data indicated a transition in mycorrhizal partners during ontogenetic development with decreasing dependency of N. malipoensis on fungal nitrogen and carbon. The divergence in mycorrhizal partners between N. malipoensis and A. nipponica indicates different resource acquisition strategies and allows various habitat options in the earliest-diverging orchid lineage, Apostasioideae. While A. nipponica relies on the heterotrophic carbon gain from its ectomycorrhizal fungal partner and thus on forest habitats, N. malipoensis rather relies on own photosynthetic carbon gain as an adult, allowing it to establish in habitats as widely distributed as those where Rhizoctonia fungi occur.

Evaluation the Ingredients and Influence of Thermal Treatment on Physicochemical Properties and Bioactive Compounds of Evernia prunastri from Algeria

Background: Lichen is a symbiotic organism with unique structure composed of a fungal partner and an algal partner. It has been utilized in folk foods and medicines. The objective of the present study was to analyze the ingredients and the impact of heating on various physicochemical and phytochemical characteristics of a 500 g sample of lichen Evernia prunastri from Jijel, Algeria collected in May 2021. Methods: The nutritional value of the lichen was determined by studying various parameters. The metal contents were analyzed using an Atomic Absorption Spectrophotometer. The effect of heating (70 and 100 °C) on the physicochemical parameters, and bioactive compounds of aqueous extracts obtained by maceration in distilled water was studied. Statistical analysis was performed using ANOVA. Results: A high percentage of carbohydrates (64.00%) were observed in the lichen material. Pectin, fats, and crude proteins levels were low. High ash content (10.85%) was obtained with the presence of metal elements. Preliminary phytochemical analysis indicated the presence of polyphenols and flavonoids with 51.13 mg Gallic Acid Equivalent (GAE)/100 g and 17.87 mg Quercetin Equivalent (QE)/100 g of lichen, respectively. The lichen sample subjected to 70 °C demonstrated an increase in the parameters examined, with the exception of proteins and vitamin C. Conversely, the lichen sample treated at 100 °C exhibited a reduction in vitamin C, protein, and bioactive compound content, as well as an increase in sugar content. Titratable acidity and pH parameters were unaffected. Conclusion: The appreciable quantities of carbohydrates, crude fibers, and other compounds in this lichen suggest a potential use for these active ingredients in the preparation of functional food. Heating to 70 °C produces a preparation enriched in bioactive substances and sugars. However, heating to 100 °C, exposes the various tissues to degradation, potentially leading to a decrease in nutrients

Evolutionary history of arbuscular mycorrhizal fungi and genomic signatures of obligate symbiosis

BackgroundThe colonization of land and the diversification of terrestrial plants is intimately linked to the evolutionary history of their symbiotic fungal partners. Extant representatives of these fungal lineages include mutualistic plant symbionts, the arbuscular mycorrhizal (AM) fungi in Glomeromycota and fine root endophytes in Endogonales (Mucoromycota), as well as fungi with saprotrophic, pathogenic and endophytic lifestyles. These fungal groups separate into three monophyletic lineages but their evolutionary relationships remain enigmatic confounding ancestral reconstructions. Their taxonomic ranks are currently fluid.ResultsIn this study, we recognize these three monophyletic linages as phyla, and use a balanced taxon sampling and broad taxonomic representation for phylogenomic analysis that rejects a hard polytomy and resolves Glomeromycota as sister to a clade composed of Mucoromycota and Mortierellomycota. Low copy numbers of genes associated with plant cell wall degradation could not be assigned to the transition to a plant symbiotic lifestyle but appears to be an ancestral phylogenetic signal. Both plant symbiotic lineages, Glomeromycota and Endogonales, lack numerous thiamine metabolism genes but the lack of fatty acid synthesis genes is specific to AM fungi. Many genes previously thought to be missing specifically in Glomeromycota are either missing in all analyzed phyla, or in some cases, are actually present in some of the analyzed AM fungal lineages, e.g. the high affinity phosphorus transporter Pho89.ConclusionBased on a broad taxon sampling of fungal genomes we present a well-supported phylogeny for AM fungi and their sister lineages. We show that among these lineages, two independent evolutionary transitions to mutualistic plant symbiosis happened in a genomic background profoundly different from that known from the emergence of ectomycorrhizal fungi in Dikarya. These results call for further reevaluation of genomic signatures associated with plant symbiosis.

Effect of Rootstock Genotype and Arbuscular Mycorrhizal Fungal (AMF) Species on Early Colonization of Apple.

The effect of plant cultivar on the degree of mycorrhization and the benefits mediated by arbuscular mycorrhizal fungi (AMF) have been documented in many crops. In apple, a wide variety of rootstocks are commercially available; however, it is not clear whether some rootstock genotypes are more susceptible to mycorrhization than others and/or whether AMF species identity influences rootstock compatibility. This study addresses these questions by directly testing the ability/efficacy of four different AMF species (Rhizophagus irregularis, Septoglomus deserticola, Claroideoglomus claroideum or Claroideoglomus etunicatum) to colonize a variety of commercially available Geneva apple rootstock genotypes (G.11, G.41, G.210, G.969, and G.890). Briefly, micropropagated plantlets were inoculated with individual species of AMF or were not inoculated. The effects of the rootstock genotype/AMF interaction on mycorrhization, plant growth, and/or leaf nutrient concentrations were assessed. We found that both rootstock genotype and the identity of the AMF are significant sources of variation affecting the percentage of colonization. However, these factors largely operate independently in terms of the extent of root colonization. Among the AMF tested, C. etunicatum and R. irregularis represented the most compatible fungal partners, regardless of apple rootstock genotype. Among the rootstocks tested, semi-dwarfing rootstocks appeared to have an advantage over dwarfing rootstocks in regard to establishing and maintaining associations with AMF. Nutrient uptake and plant growth outcomes were also influenced in a rootstock genotype/AMF species-specific manner. Our findings suggest that matching host genetics with compatible AMF species has the potential to enhance agricultural practices in nursery and orchard systems.

Genotype-by-genotype interkingdom cross-talk between symbiotic nitrogen fixing Sinorhizobium meliloti strains and Trichoderma species

In the understanding of the molecular interaction between plants and their microbiome, a key point is to identify simplified models of the microbiome including relevant bacterial and fungal partners which could also be effective in plant growth promotion. Here, as proof-of-concept, we aim to identify the possible molecular interactions between symbiotic nitrogen-fixing rhizobia and soil fungi (Trichoderma spp.), hence shed light on synergistic roles rhizospheric fungi could have in the biology of symbiotic nitrogen fixation bacteria. We selected 4 strains of the model rhizobium Sinorhizobium meliloti and 4 Trichoderma species (T. velutinum, T. tomentosum, T. gamsii and T. harzianum). In an experimental scheme of 4 ×4 strains x species combinations, we investigated the rhizobia physiological and transcriptomic responses elicited by fungal spent media, as well as spent media effects on rhizobia-host legume plant (alfalfa, Medicago sativa L.) symbiosis. Fungal spent media had large effects on rhizobia, specific for each fungal species and rhizobial strains combination, indicating a generalized rhizobia genotype x fungal genotype interaction, including synergistic, neutral and antagonistic effects on alfalfa symbiotic phenotypes. Differential expression of a high number of genes was shown in rhizobia strains with up to 25% of total genes differentially expressed upon treatment of cultures with fungal spent media. Percentages over total genes and type of genes differentially expressed changed according to both fungal species and rhizobial strain. To support the hypothesis of a relevant rhizobia genotype x fungal genotype interaction, a nested Likelihood Ratio Test indicated that the model considering the fungus-rhizobium interaction explained 23.4% of differentially expressed genes. Our results provide insights into molecular interactions involving nitrogen-fixing rhizobia and rhizospheric fungi, highlighting the panoply of genes and genotypic interactions (fungus, rhizobium, host plant) which may concur to plant symbiosis.

Three helmet-orchid species share abundant fungi of Serendipitaceae regardless of altitude

The tiny terrestrial orchids (plant height less than 8 cm) in the genus Corybas dependent on mycorrhizal fungal (OMF) partners for seed germination and seedling development. The OMF community of the Corybas remains poorly understood, although the relevant knowledge is very important for in situ and ex situ conservation of these orchids. In this study, we characterized OMF richness and compositions of three helmet-orchid species, i.e., Corybas geminigibbus, C. himalaicus, and C. shanlinshiensis, from their natural habitats by using Illumina sequencing of the internal transcribed spacer 2 region. We found that fungal colonization was restricted in the rhizomes of the helmet-orchids instead of their tuberoids, and serendipitoid fungi were predominant, while tulasnelloid were absent in the three investigated Corybas species regardless of their altitude. The three Corybas species shared 27 serendipitoid operational taxonomic units that are different to those of their related orchids, the genera of Cyrtostylis and Stigmatodactylus. Corybas shanlinshiensis alone had a range of ectomycorrhizal fungi (mainly russuloid and thelephoroid) broader than C. himalaicus and C. geminigibbus. Our study provides new information about terrestrial orchid–fungi associations and may further contribute to orchid conservation.

Life history costs drive the evolution of mycoheterotrophs: Increased sprouting and flowering in a strongly mycoheterotrophic Pyrola species

Abstract Land plants are typically photosynthetic, but some species have lost the ability to photosynthesize, instead relying on mycorrhizal fungi to obtain carbon. Increasing levels of partial mycoheterotrophy, in which seemingly autotrophic plants receive fungal carbon from their fungal partners, and reduced sprouting in concert with greater reproduction when sprouting, may be intermediate steps in the evolution of this trait. We studied the microevolutionary demography of Pyrola japonica and the closely related species P. subaphylla, which are currently considered sister species. While these species are both partially mycoheterotrophic, P. subaphylla is more strongly so, indicating that it may be closer to evolving full mycoheterotrophy. We tracked individuals in two nearby populations in Fukushima, Japan from 2015 to 2020. We analysed vital rates, population trends and long‐run population structure with historical function‐based matrix projection models. We assessed how shifts in P. subaphylla demography relate to fitness using a stochastic life table response experiment (SLTRE). P. subaphylla exhibited strong costs of growth to survival and of reproduction to sprouting, size and fecundity. It sprouted more than P. japonica but to smaller stages. P. subaphylla's flowering frequency was approximately four times that of P. japonica, but after flowering it was smaller and more likely to die. These life history costs appeared to drive differences in fitness, measured as stochastic population growth rate. Relative to P. japonica, shifts in fitness in P. subaphylla were due more to shifts in means than standard deviations of matrix elements. Shifts in growth transitions had the strongest negative impacts on fitness but also had the strongest positive impacts. Increased shifts to small non‐flowering stages were associated with increased fitness, while transitions involving dormancy were associated more strongly with drops in fitness. Synthesis. As far as we are aware, we found the first evidence that costs of growth and reproduction drive the evolution of increased sprouting and smaller size as a species becomes more intensely mycoheterotrophic. Surprisingly, vegetative dormancy did not evolve with mycoheterotrophy. We suggest further studies assessing whether these seemingly maladaptive trends result from genetic drift, genetic linkage or another mechanism.

Microbiomic Analysis of Bacteria Associated with Rock Tripe Lichens from Alpine Areas in Eastern Alps and Equatorial Africa

The diversity of bacteria associated with alpine lichens was profiled. Lichen samples belonging to the Umbilicariaceae family, commonly known as rock tripe lichens, were gathered from two distinct alpine fellfields: one situated on Mt. Brennkogel located in the Eastern European Alps (Austria), and the other on Mt. Stanley located in the Rwenzori mountains of equatorial Africa (Uganda). The primary aim of this research was to undertake a comparative investigation into the bacterial compositions, and diversities, identifying potential indicators and exploring their potential metabolisms, of these lichen samples. Bulk genomic DNA was extracted from the lichen samples, which was used to amplify the 18S rRNA gene by Sanger sequencing and the V3-V4 region of the 16S rRNA gene by Illumina Miseq sequencing. Examination of the fungal partner was carried out through the analysis of 18S rRNA gene sequences, belonging to the genus Umbilicaria (Ascomycota), and the algal partner affiliated with the lineage Trebouxia (Chlorophyta), constituted the symbiotic components. Analyzing the MiSeq datasets by using bioinformatics methods, operational taxonomic units (OTUs) were established based on a predetermined similarity threshold for the V3-V4 sequences, which were assigned to a total of 26 bacterial phyla that were found in both areas. Eight of the 26 phyla, i.e. Acidobacteriota, Actinomycota, Armatimonadota, Bacteroidota, Chloroflexota, Deinococcota, Planctomycetota, and Pseudomonadota, were consistently present in all samples, each accounting for more than 1% of the total read count. Distinct differences in bacterial composition emerged between lichen samples from Austria and Uganda, with the OTU frequency-based regional indicator phyla, Pseudomonadota and Armatimonadota, respectively. Despite the considerable geographic separation of approximately 5430 km between the two regions, the prediction of potential metabolic pathways based on OTU analysis revealed similar relative abundances. This similarity is possibly influenced by comparable alpine climatic conditions prevailing in both areas.

Volatile organic compounds emitted by Megaplatypusmutatus associated fungi: chemical identification and temperature-modulated responses by the ambrosial beetle

BackgroundIn ambrosia and bark beetles–fungi interaction, volatile organic compounds (VOCs) play a central role in mediating various aspects of community dynamics of beetles and/or fungi. These functions include facilitating beetle habitat location, mate identification, and fungal partner differentiation. However, the understanding on this context remains limited, especially in the globally distributed subfamily Platypodinae, which comprises predominantly ambrosia beetles. There is a lack of chemical data on ambrosia fungi from native South American species. This study addresses this gap by characterizing VOCs from twelve fungal species associated with Megaplatypusmutatus and assessing species-specific behavioral responses during dispersal.MethodsFungal VOCs were collected by gas chromatography–mass spectrometry combined with solid-phase microextraction and Y-olfactometry assays of males and females were performed at dispersal stage. Statistical analyses involved: non-metric multidimensional scaling multivariate plot and PERMANOVA test, a cluster analysis through unweighted pair group method with Jaccard index, and finally, a chi-square goodness-of-fit test for beetle behavioral assays.ResultsWe identified 72 VOCs from the fungal species isolated from M.mutatus galleries, exocuticle, and gut. The olfactory behavior of M.mutatus demonstrated its capacity to discriminate between volatile profiles, showing a preference for either the fungus or the control source. Our results also enhance the understanding in a chemotaxonomic context and in the behavioral responses of M.mutatus revealing the beetle's remarkable low temperature tolerance and its capability to maintain mobility and orientation toward volatile sources even after zero-degree Celsius exposure.ConclusionThis study presents a comprehensive insight into fungal VOC profiles, emphasizing the sources of isolation within pest associated fungi, as well as its symbiotic species from the Raffaelea genus. In conclusion, our findings suggest that Megaplatypusmutatus exhibits a general aversion to its fungal VOCs symbiont. However, a notable exception arises when the beetles are pre-exposed for 48 h to freezing conditions, highlighting the beetles' ability to withstand freezing conditions as adults and to exhibit altered responses to their fungal associates under these circumstances.

Mycorrhizal diversity in Spiranthes spiralis (L.) Chevall

For effective conservation of threatened orchids, identifying root-associated fungi and assessing their activity in seed germination is important for establishing conservation protocols. Therefore, our study investigated the diversity of Spiranthes spiralis‘s root-associated fungi. According to the culture-dependent approach, 37 endophytic fungi were isolated from the roots and morphologically and molecularly identified. It was determined that the dominant fungal species in the roots was the genus Tulasnella. For the first time in Türkiye, the Thanatephorus fusisporus species was isolated from roots. The germination efficiency of the isolated fungi in the symbiotic culture of S. spiranthes seeds was evaluated. VY 25 (Tulasnella) isolate isolated in April showed the highest germination rate (73.77%). VY 4, VY 18, VY 25, (Tulasnella) isolates promoted germination and seedling development. Thanatephorus (46.79%) and Ceratobasidium (32.42%) were not effective in germinating seeds. The study revealed that the fungal partner varied according to developmental stages and months. This study contains the first molecular data for organisms isolated from roots in Türkiye. According to these results, fungi that promote seed germination and plant growth can be recommended for the conservation and reintroduction of endangered temperate orchids.

Fungal communities associated with early immature tubers of wild Gastrodia elata.

Full myco-heterotrophic orchid Gastrodia elata Bl. is widely distributed in Northeast Asia, and previous research has not fully investigated the symbiotic fungal community of its early immature tubers. This study utilized Illumina sequencing to compare symbiotic fungal communities in natural G. elata immature tubers and their habitats. LEfSe (Linear Discriminant Analysis Effect Size) was used to screen for Biomarkers that could explain variations among different fungal communities, and correlation analyses were performed among Biomarkers and other common orchid mycorrhizal fungi. Our results illustrate that the symbiotic fungal communities of immature G. elata tubers cannot be simply interpreted as subsets of the environmental fungal communities because some key members cannot be traced back to the environment. The early growth of G. elata was related to a small group of fungi, such as Sebacina, Thelephora, and Inocybe, which were also common mycorrhizal fungi from other orchids. In addition, Mycena, Auricularia, and Cryptococcus were unique fungal partners of G. elata, and many new species have yet to be discovered. Possible symbiotic Mycena should be M. plumipes and its sibling species in this case. Our results provide insight into the symbiotic partner switch and trophic pattern change during the development and maturation of G. elata.

Endolichenic Fungi: A Promising Medicinal Microbial Resource to Discover Bioactive Natural Molecules-An Update.

Lichens are some of the most unique fungi and are naturally encountered as symbiotic biological organisms that usually consist of fungal partners (mycobionts) and photosynthetic organisms (green algae and cyanobacteria). Due to their distinctive growth environments, including hot deserts, rocky coasts, Arctic tundra, toxic slag piles, etc., they produce a variety of biologically meaningful and structurally novel secondary metabolites to resist external environmental stresses. The endofungi that live in and coevolve with lichens can also generate abundant secondary metabolites with novel structures, diverse skeletons, and intriguing bioactivities due to their mutualistic symbiosis with hosts, and they have been considered as strategically significant medicinal microresources for the discovery of pharmaceutical lead compounds in the medicinal industry. They are also of great importance in the fundamental research field of natural product chemistry. In this work, we conducted a comprehensive review and systematic evaluation of the secondary metabolites of endolichenic fungi regarding their origin, distribution, structural characteristics, and biological activity, as well as recent advances in their medicinal applications, by summarizing research achievements since 2015. Moreover, the current research status and future research trends regarding their chemical components are discussed and predicted. A systematic review covering the fundamental chemical research advances and pharmaceutical potential of the secondary metabolites from endolichenic fungi is urgently required to facilitate our better understanding, and this review could also serve as a critical reference to provide valuable insights for the future research and promotion of natural products from endolichenic fungi.