Abstract

Several Ipomoea species and related plants in the morning glory family (Convolvulaceae) harbor vertically-transmitted, symbiotic fungi in the genus Periglandula that produce ergot alkaloids. Many more Ipomoea species have been surveyed, and approximately one-third of them contain ergot alkaloids indicating symbiosis with uncharacterized Periglandula species. The effects of the Periglandula species on plant host biology has not been well characterized. Moreover, the question of whether the ergot alkaloid-negative Ipomoea species lack Periglandula species symbionts or associate with cryptic Periglandula species that simply do not produce ergot alkaloids has not been investigated extensively. We investigated both of these issues through a metabolomics approach. We studied the metabolomes of Ipomoea tricolor seeds collected from Periglandula sp.-infected plants (P+) or plants from the same lineage that had been cured of the Periglandula sp. by treatment with fungicide (P-). Methanol extractions were performed on weight-normalized, pulverized seeds. Seed extracts were screened for ergot alkaloids by HPLC with fluorescence detection. For high resolution accurate mass analysis of metabolites, extracts were analyzed by UHPLC-ESI-QTOF mass spectrometry. Chromatographic peak alignment was performed using quality control criteria, and aligned peaks were filtered by fold-change and minimum abundance. Peak abundances were assessed by a T-test (p < 0.05). The ergot alkaloids lysergic acid α-hydroxyethylamide (LAH), ergine, ergonovine, and chanoclavine-I were present in high concentrations in P+ seeds but not detected in the P- seeds. Plant stress hormones jasmonic acid and abscisic acid did not differ significantly between treatments; salicylic acid was not detected. Similarly, amino acid concentrations did not differ significantly between treatments. Several unidentified analytes that were significantly more abundant in P+ seeds compared to P- seeds were compared to metabolomes of a collection of seed extracts from ergot alkaloid-positive and ergot alkaloid-negative seeds of Ipomoea parasitica and Ipomoea pes-caprae–plants previously shown to contain or lack Periglandula-species symbionts on a plant-by-plant basis. Only four metabolites tracked the presence of symbiont in this survey; the identity and symbiont of origin of these compounds remain to be determined. A collection of seeds from nine ergot alkaloid-deficient Ipomoea species were screened for the presence of these four non-ergot alkaloid biomarkers of Periglandula sp. presence. Extracts of all nine species were negative for the presence of the Periglandula-associated biomarkers. The data indicate that apart from the accumulation of abundant ergot alkaloids, Periglandula species have a minimal impact on the metabolome of seeds of their host plants. Furthermore, we found no evidence of cryptic, non-ergot alkaloid producing Periglandula species in the seeds of the nine additional Ipomoea species analyzed.

Abstract

The agricultural relevance of commensal yeasts inhabiting the phylloplane of leafy green vegetables is still poorly understood. Our current research describing the culturable and total fungal communities associated with Romaine lettuce reported that basidiomycetous yeasts in Sporidiobolales and Tremellales are the most common fungal groups found in this broadly consumed vegetable. We also discovered that a single undescribed species, the sister of Sporobolomyces roseus (S. cfr. roseus), was constantly present in the majority of lettuce plants examined. Here, we present preliminary results of greenhouse experiments designed to evaluate the capabilities of S. cfr. roseus as a bioprotective agent against the establishment of fungal pathogens on Romaine lettuce. All of the experiments were conducted in a BL-2 biosecurity green house, where lettuce seedlings were grown in 12 h photoperiod. First, we demonstrated that S. cfr. roseus is not an inhabitant of lettuce grown under greenhouse conditions. Second, we demonstrated successful colonization of the lettuce phylloplane by S. cfr. roseus and it did not induce any antagonistic immune response in the host. Third, in co-inoculation experiments, we found that the well-known plant pathogen Botrytis cinerea significantly did not induce a hypersensitive response (HR) on lettuce when seedlings were previously inoculated with S. cfr. roseus. In sum, our results suggested that S. cfr. roseus could be used as a protective agent against the establishment of fungal pathogens on lettuce. The inoculation of commensal red yeasts to leafy green vegetables may also increase the nutritional value of pro-vitamin A, a compound obtained from the digestion of the carotenoid pigments present in vacuoles in the Sporidiobolales yeasts.

Abstract

As in many grass-endophyte symbionts, Epichloë grass endophyte produce active alkaloids, found in at least ten of the four major classes. One is pyrrolopyrazine, represented by peramine species that are toxic to certain nematodes and herbivores. To explore the optimal conditions for the production peramine by Epichloë grass edophyte in vitro and to screen their pest resistant activities in grass endophyte strains, peramine contents of eight strains of Epichloë grass edophyte (ATZ, SN, S3-3, PJ, NFS6, ZHYM, YZH, AKH) from Achnatherum inebrians, Elymus tangutorum, Festuca sinensis and Hordeum brevisubulatum were studied under different pH (8, 9, 10, 11, 12) and their pest resistant activities tested through feeding trail of detached leaves. It was found that: (1) The peramine content of eight Epichloë grass endophyte all had peak values at pH 8-9 in vitro, and the ZHYM's strain was 2-6 times that of other strains.(2) Compared with the control, eight strains of Epichloë grass edophyte had different effects on pest resistant. After 48h incubation, the ZHYM's pest resistance activity was significantly higher than the other seven groups, and the mortality rate reached 46.67%. (3) Compared with the stock solution, the pest resistance activity of the fermented liquid diluted 5, 10 and 15 times were significantly lower than that of the original liquid, and the mortality of Rhopalosiphum padi was also increased with the prolonging of the treatment time.

Abstract

Rapid, reliable and cost effective assessments of microbiological and chemical soil properties are highly essential for monitoring soil quality. Spectroscopy is known for its inexpensiveness, rapidity and accuracy and may be a useful tool for the assessment of soil quality. While chemical properties in agricultural systems can often be reliably assessed via spectroscopy, it is unknown to what extent changes in soil microbial communities can be detected with this technique. Soil fungi play a major role in several soil processes, e.g. organic matter decomposition, aggregate formation, disease suppression and plant growth promotion. The fungal community composition is mainly driven by soil C:N ratio, soil carbon content, pH, soil texture, land use and management. Therefore, it seems plausible to detect shifts in soil fungal communities by measuring soil characteristics measured by spectroscopy. In this study 1) we explain properties of the fungal community composition (obtained with Illumina sequencing) through soil chemistry data (obtained with conventional wet-chemistry analyses) and 2) try to predict elements of the fungal community composition using soil spectroscopy data (obtained with near-infrared reflectance – NIR, mid-infrared reflectance – MIR, and X-ray fluorescence – XRF). Preliminary results show that the fungal diversity and community structure varies in different soils. Moreover, fungal communities are susceptible to chemical factors, as pH, macro- and micronutrients (as P, Ca, Mg and Mn). Lastly, different fungal functional groups, as mycorrhizae, saprotrophs and plant pathogens, are affected by the land use and management. The next step is hence to explain through the combined used of diverse sensor data the variations of composition and functions of the fungal communities. We will discuss our findings in light of advantages and limitations for detecting changes in fungal communities measured through spectroscopy. Implementations of our results, with the delivery of geo-referenced and management advice, are of great interest for farmers, policy makers and nature conservationists.

Abstract

Mango, Mangifera indica L., is the most economically important fruit crop in the Anacardiaceae family. It ranks 2nd as the most important fruit in the country in terms of value and 3rd based on export volume and value. The Philippines is exporting fresh mango to 48 countries all over the world and its major export markets are Japan and Hong Kong with 52.9 % and 35.7% respective shares in terms of value. Recent studies suggest that the total performance of mango industry is declining as shown by the country’s production volume and export performance. One attribution of the decline is due to fungal attack of major and rapidly emerging new fungal species that generally affect the fruit development and the over-all quality of the harvested crops. Early detection for the infection of these fungi is necessary to implement necessary control for the spread of these pathogens. This project intends to develop the loop-mediated isothermal amplification (LAMP) detection assay for the anthracnose, stem-end rot and scab disease on Philippine ‘Carabao’ mango. Collections of infected stems, leaves, flowers and fruits were conducted in Zambales and Guimaras, Philippines. Collected samples were brought, processed and preliminary identified at the Polytechnic University of the Philippines, Manila. Results from the field survey and preliminary identification revealed the presence of several species of Colletotrichum, Diaporthe, Elsinoë, Lasiodiplodia, Neofusicoccum and Pestalotiopsis on the infected leaves and fruits of carabao mango. Isolates related to those symptoms were brought to Mie University, Japan for rigorous identification and for the development of LAMP detection method. Initial development of LAMP was made for the detection anthracnose. For designing the LAMP primers for generic and specific level, genomic DNA extracted from the isolates of Colletotrichum spp. on Mango were analysed those sequences of ITS, act, gapdh, and chs-1. And also, for evaluating the primer sets, representative strains from species of the genus Colletotrichum, including the major species complex of “boninense”, “acutatum”, “caudatum”, “dematium”, “destructivum”, “gloeosporioides”, “graminicola”, “orbiculare”, “truncatum”, and “spaethianum”, were examined. The sets of LAMP primers designed in this study successfully amplified species of Collectotrichum gloeosporioides species-complex on the anthracnose symptoms. Future works will include morphological and molecular analysis, and the development of LAMP for Stem-end rot and scab diseases.

Abstract

Isaria javanica is a commonly isolated entomopathogenic fungi with activity against many insect species. Over the years, strains of this species have been commonly misidentified as Isaria fumosorosea. We have developed a simple high resolution DNA melting assay to discriminate Isaria javanica from other Isaria species. The assay utilizes a previously identified 103 base pair PCR amplicon, which was reported to be selective for Isaria fumosorosea. Our study finds the amplicon selective for Isaria javanica and Isaria poprawskii, when assayed against all members of the genus. The practical application of this technique was confirmed using a bioassay on whitefly nymphs (Bemisia tabaci biotype B) inoculated with I. javanica.

Abstract

Ergot alkaloids are specialized metabolites produced by certain fungi, including species of Metarhizium. Many important ergot alkaloids are lysergic acid amides, such as ergonovine and lysergic acid alpha-hydroxyethylamide (LAH). Lysergic acid amides are important in agriculture, where they can contaminate food and feed but also deter insect feeding and have insecticidal properties. They also are important in medicine, where they serve as the basis of drugs used to treat dementia and migraines. The pathway to create ergonovine is established, but the pathway to LAH is unknown. Based on genome sequence comparisons we hypothesized that a FAD monooxygenase gene (easO) was involved in the production of LAH. Using PCR methods, we prepared a gene knock out construct and introduced it into the LAH-producing fungus Metarhizium anisopliae by protoplast transformation. Four independent knock outs were identified by PCR strategies that showed the knock-out construct had integrated into the easO locus. High performance liquid chromatography (HPLC) and HPLC-mass spectrometry (LC-MS) analyses demonstrated that the knock-out fungal strains lacked LAH and retained ergonovine. The data supported our hypothesis that easO was involved in the production of LAH. The ergonovine-accumulating easO knockout strain was injected into larvae of Galleria mellonella to investigate its virulence relative to wild-type (LAH-accumulating) M. anisopliae. Ergonovine-accumulating knockout strains killed larvae faster than the LAH-accumulating wild type; however, the easO knockout fungus rarely emerged from dead larvae, whereas the wild-type fungus sporulated profusely on larval cadavers. These data indicate that ergot alkaloids play a role in the interaction of M. anisopliae with insects. An understanding of the production of lysergic acid amides is beneficial because of their positive (anti-insect) and negative (anti-mammalian) effect in agriculture and because of the medical uses of lysergic acid derivatives.

Abstract

Even though the pathogenic fungi associated with invertebrates are well-known for centuries and have been intensively studied for a long time, certain groups of fungi, and in particular the spider parasitic fungi have not yet received much attention. Gibellula and Hevansia are classified in the Cordycipitaceae which are known to be obligate parasites of spiders. Most of them have been investigated based solely on morphological descriptions. Over a few decades there has been an increasing interest in the study of phylogenetic relationship between them and their allies, it is nevertheless limited to specific research groups. Besides the limited molecular data, their production of secondary metabolites also remains largely unexplored. In the course of a study on invertebrate-pathogenic fungi in Thailand where is accepted to be a rich source for a diverse range of microorganisms particularly invertebrate-pathogenic fungi, more than a hundred of fungal specimens were collected from various areas and encountered in the study. To extend our understanding of the taxonomic relationship among Gibellula, Hevansia and allied genera as well as to investigate and explore their secondary metabolites, HPLC- and PCR-based techniques were employed to generate the fungal chemoprofiles and molecular data, respectively. The species identification was relied on morphological features and multigene phylogenetic analysis. Five nuclear gene regions including the internal transcribed spacer (ITS), nuclear ribosomal large subunit DNA (nrLSU), elongation factor 1α (EF-1α), the largest and second largest subunits of RNA polymerase II (RPB1 and RPB2), were sequenced and the phylogenetic tree was subsequently reconstructed. The fungal secondary metabolite profiles were examined using analytical HPLC coupled with diode array and mass spectrometric detection (HPLC-DAD/MS) and compared within species according to the multigene tree. Some species remarkably revealed a unique pattern of secondary metabolite production in particular H. novoguineensis in which all nine unprecedented secondary metabolites are specifically produced by the species. This is the first report on secondary metabolite profiling of Gibellula and Hevansia showing the species-specific distribution pattern of their secondary metabolites as well as unprecedented components that might eventually be used as powerful chemotaxonomic markers in species identification.

Abstract

According to a widely held view, fungi do not produce parenchymatous tissues. Following up on a recent TEM study that challenged this paradigm in three lichens representing different orders within two classes of ascomycetes, we apply SEM here to determine the orientation of cell divisions in the single-layered cortex of six species of Collemataceae. This family of gelatinous cyanolichens includes both corticated and uncorticated species showing diverse surface morphologies. Examination of thallus surfaces in four species of Leptogium (L. austromericanum, L. burnetiae, L. chloromelum, L. marginellum) and two species of Scytinium (S. gelatinosum, S. lichenoides) revealed the positions of recently formed cortical septa in the six morphologically distinct taxa. Septa adjoined to preceding septa, usually perpendicularly, indicating parenchymatous division. Such divisions were involved the growth and development of characteristic surface structures, such as thallus wrinkles, folds, isidia, and lobules. Tomentum, by contrast, arose as filamentous outgrowths of the cortical cells. For comparison, we also examined the surfaces of the uncorticated Collema furfuraceum, which closely resembles species of Leptogium morphologically. We conclude that the monostromatic cellular cortex in Collemataceae is actively involved in growth and morphogenesis by means of parenchymatous cell divisions, in a remarkable parallel to plant meristems. However, cortical cell divisions are not likely to be the underlying cause of morphogenesis, as very similar morphologies develop in the closely related genus Collema, which lacks a cortex altogether.

Abstract

The primal transition from vegetative hyphae to conidiophore development in Aspergillus nidulans involves a set of proteins which are expressed in vegetative hyphae (Upstream Developmental Activators). One of the earliest acting UDA factors is fluG (An4819), along with the G protein regulator flbA (Lee and Adams, 1996). Deletion of fluG results in colonies which fail to produce conidiophores and accumulate aerial vegetative hyphae, resulting in a colony morphology commonly known as fluffy (Lee et al, 1994). In addition, ΔfluG mutants show defects in autolysis (Pocsi et al, 2009) and secretion (Wang et al, 2015). Despite these findings, FluG has not been attributed a specific role in conidiophore development. In this investigation, we have conducted a sequence and structural analysis of the FluG protein, establishing an N-terminal amidohydrolase-like region with sequence and structural similarity to the amidohydrolase 2QPX of Lactobacillus casei, and a C-terminal region with sequence and structural similarity to a γ-glutamyl ligase (PauA7) from Pseudomonas aeruginosa (Ladner et al., 2012). Alanine substitutions of the predicted key catalytic residues in each region yielded distinct loss of function phenotypes, and the separate expression of both regions yielded a partially functional phenotype, indicating that coupling of both predicted enzyme activities is required for full biological function. Our findings situate FluG as a putative bifunctional enzyme that may exert changes in the levels of nitrogen-containing signal intermediates that are required for development.

Abstract

Pythiogeton is a cryptic genus in Pythiaceae. A total of 15 species have been reported in this genus and most of the species are saprophytic. Pythiogeton spp. are characterized by their unique way to form the sporangium and zoospores. The shape of sporangia are globose, ovoid, ellipsoidal, bursiform or multilobate, and the long axis of sporangia are mostly at the right angles to the supporting hypha. At the late stage of sporangial development, the incomplete differentiated protoplasm mass was emitted into the water through a discharge tube. Afterwhile, the protoplasm mass differentiated into zoospores. The ultrastructures of the sporangial development and zoospores releasing process of some Pythiogeton species will be reported in this study.

Abstract

Ascomycetes are mostly saprobic and/or plant pathogenic fungi, feeding on living or dead plant material. Hemicellulose is a major component of plant biomass. The pentoses D-xylose and L-arabinose, and the hexose D-galactose are frequent building monomers of all hemicelluloses as well as pectin. The Pentose Catabolic Pathway (PCP) is comprised of subsequent reduction and oxidation steps, ultimately followed by a phosphorylation to xylulose-5-phosphate that enters the pentose phosphate pathway (PPP). In Aspergillus nidulans, the oxido-reductive pathway (ORP) for D-galactose catabolism employs a similar biochemical strategy, with the resulting fructose-6-phosphate entering glycolysis. In contrast, the Leloir pathway of D-galactose breakdown is specifically epimerizing D-galactose into D-glucose. While there is a palpable overlap between PCP and ORP in A. nidulans, genes, enzymes and intermediates of the Leloir pathway are (mostly) exclusive for the downstream catabolism of D-galactose. However, A. nidulans conidiospores do not germinate on D-galactose alone, but need to be stimulated first with low concentration of other carbon sources such as D-xylose. Due to these overlapping functions pentoses and D-galactose play in the nutrition of fungi, we hypothetized that each catabolic gene of the ORP, PCP and Leloir pathway will be responding by increased levels of expression to the presence of either sugar, irrespective of whether the enzymes encoded are actually involved in the catabolism of the given sugar. As a proof of evidence, we will demonstrate that the first two genes of the Leloir pathway, i.e., D-galactose kinase (galE) and galactose-1-P uridyltransferase (galD), as well as a D-galactose mutarotase (galmB) recently shown to be an integral part of D-galactose breakdown are all induced by D-xylose and L-arabinose, but the respective deletion mutants display no (growth) phenotype on these pentoses.

Abstract

Species of Phyllosticta occur on numerous plant hosts, including mangoes, citrus, grapes and bananas, where they have been recorded as both endophytes and plant pathogens. On citrus, Phyllosticta spp. can cause disease symptoms ranging from small freckles on leaves (P. citrichinaensis), to necrotic lesions on fruits (P. citricarpa). The latter has been reported specifically in citrus species in Asia, Australia, Africa and North America, and is considered a serious pathogen of this host. On the contrary, P. capitalensis is found as a widespread endophyte in more than 70 different hosts, and is rarely recorded as plant pathogen. The ability to degrade cell walls through Plant Cell Wall Degrading Enzymes (PWCDEs) is considered to play a role in the host preference of fungal species. The difference in host and tissue specificity of Phyllosticta spp. makes them interesting subjects to study in relation to carbon utilization. The objective of this study is therefore to analyse the repertoire of PWCDEs present in selected Phyllosticta spp., and their ability to grow on different carbon sources. This gives insight as to whether the basis of the interaction and lifestyle mode between these fungi and their plant hosts lies in the ability of Phyllosticta spp. to take advantage of its host’s carbon sources.

Abstract

Herbicides are frequently used to control weeds in both agricultural settings and around residential dwellings. Little is known about the interaction that herbicides have with truffles (Tuber species), morels (Morchella species), and Mortierella species. Understanding how weed management tools such as glyphosate or glufosinate interact with these fungi may better inform the use of these herbicides especially in morel or truffle production locations. For this experiment, Morchella americana, Morchella importuna, Tuber borchii, Tuber gibbosum, and Mortierella elongata were each grown in stepwise serial dilutions of herbicide containing agar media ranging in concentrations above and below the working ratio recommended by the manufactures. Two brands of herbicide (Roundup® – glyphosate and Liberty® – glufosinate) were diluted and mixed into agar media to determine if there is an interaction between different concentrations of glyphosate or glufosinate-based herbicides and mycelial growth of the aforementioned fungi. The mycelial growth front the fungi was routinely measured and recorded in four quadrants of each replicate. None of the tested fungi grew at the upper range of herbicide working concentrations used in this experiment but grew unimpeded at the lower range. Differences in susceptibility between the tested genera were negligible as the concentration range where the treatment effect was seen in each isolate varied; however, in each instance susceptibility was near the working concentration recommended for application. Though the working herbicide concentration is near the point of fungal growth inhibition, the herbicide becomes further diluted within the environment during broadcast application. As such, the use of glyphosate or glufosinate at recommended rates would not be expected to interfere with the mycelial growth of the fungi used in this experiment; however, use of these herbicides in higher than recommended concentrations may inhibit mycelial growth.

Abstract

Soft-rot (type II) fungi belonging to the ascomycetous family Xylariaceae are known to substantially degrade hardwood by means of a poorly understood lignocellulolytic enzyme system, which comprises various hydrolases (e.g. glycosidases, esterases) and some oxidoreductases (e.g. laccases). Here, we report on the genomes of four Xylaria species, X. polymorpha, X. longipes, X. hypoxylon and X. grammica, with the aim to better understand this enzyme system. The complete genomes were sequenced using an Ion Torrent® PGM NGS platform. Assembling and gene prediction was performed based on MIRA, Geneious R11 and AUGUSTUS software; gene annotation was done by Blast2GO and dbCAN or via reference-based annotation.

The genome size of the four ascomycetes ranged between 42.8 (X. hypoxylon) and 47.0 Mb (X. grammica). The highest number of predicted genes was found for X. longipes (12,638), the smallest number for X. polymorpha (10,285). Among them, is a large number of putative carbohydrate-active enzymes (CAZymes) and several carbohydrate-binding modules (CBM) as evidenced in all four genomes (e.g. 691 and 58 genes, respectively, in the X. polymorpha genome). The CAZymes are represented by glycoside hydrolases (GH; e.g. 296 for X. polymorpha), carbohydrate esterases (CE; e.g. 117 for X. grammica), polysaccharide lyases (PL; e.g. 18 for X. grammica and X. longipes) and glycosyl transferases (GT; e.g. 95 for X. hypoxylon). CBM1 members, twelve of them were found in all four species, may foster the attack on crystalline cellulose. On the other hand, the four Xylaria genomes revealed the absence of ligninolytic class-II peroxidases (i.e. lignin, manganese or versatile peroxidases), which are characteristic for white-rot basidiomycetes. Instead, sequences of putative unspecific peroxygenases (UPOs; EC1.11.2.1) and dye-decolorizing peroxidases (DyPs; EC1.11.1.19) as well as of lytic polysaccharide monooxygenases (LPMOs; AA9 formerly GH61) were identified. Furthermore, numerous genes encoding for other accessory enzymes (AA) of lignocellulose degradation such as cellobiose dehydrogenase (CDH; AA3_1 with one to three genes) and H₂O₂-supplying enzymes (e.g. galactose, glyoxal and alcohol oxidases) were present in the four Xylaria genomes. Our genomic findings have been verified by a proteomic approach with X. polymorpha grown on beech wood for several weeks. In total, 125 secreted proteins (18.1%) were identified as CAZymes. Most of them belong to GH families (73; 58%) like GH4 and GH78. The latter contains unique enzymes, which combine catalytic features of α-L-rhamnosidase and feruloyl esterase. Secreted members of the AA group have either cellulolytic activities, generate H₂O₂ (e.g. GMC oxidoreductases, AA3_2 or glucooligosaccharide oxidases, AA7) or oxidize phenolics (laccases, AA1_1). No extracellular peroxidase was found in the fungal wood cultures.

Our data give new insights into the enzyme machinery of soft-rot fungi that seemingly ranges – from the biocatalytic point of view – between those of white-rot and brown-rot fungi.

Abstract

Unknown endophytic fungi from forests are one of substantial resources beneficial substances for our future. Fungal terpenes are bioactive functions and also recently found as potential drop-in biofuels compatible to internal combustion energy. An endophytic fungus identified as Annulohypoxylon sp.FPYF3050 from a tree, Neolitsea pulchella (Meissn.) Merr., in southern China was reported to produce 1,8-cineole at high quality and major quantity in its VOCs using methods of headspace solid phase microextraction combined with gas chromatography-mass spectrometry. It grew well on seven media with different carbon sources and five raw agro-forest residues. The content of 1,8-cineole in the mixed VOCs maintained up to 94.95% relative area only with four compounds growing in PDA, and 91.25% relative area only with five compounds growing in raw poplar sawdust. The fungus preferred starch and lignocelluloses to cellulose and sucrose to facilitate 1,8-cineole production. To our knowledge, the strain produced exclusively VOCs with 1.8-cineole as the major product even growing lignocelluloses biomass. 1,8-cineole is as a monoterpene is an ideal fuel additive for both diesel and gasoline engines. Comparing to environmental concern on extraction the oil from Eucalyptus plants, Annulohypoxylon sp. FPYF3050 would be a feasible organism to maximize 1,8-cineole production at industrial scale in a more economically and environmentally friendly way, and also to be an ideal organism-producing 1,8-cineole or monoterpene metabolic pathways for fungi. The genome of the strain was sequenced using PacBio RS II instrument and de novo genome assembly size was 42.3 Mbp. In this genome, three metabolic pathways related to terpene biosynthesis were identified according to KEGG pathway annotation. Among the pathways is one to synthesize the other jet biofuel sesquiterpene, bisabolene. In addition, twelve members were determined in terpene synthase gene family in the genome through InterPro database.

Abstract

Carbohydrate active enzymes (CAZymes) are responsible for synthesis, modification and degradation of glycosidic bonds and some are associated with pathogenicity and host range of fungal pathogens. The aim of this study was to infer the host range of Colletotrichum tanaceti, a major foliar pathogen of Tanacetum cineraiifolium, using a CAZyme pathogenicity profile consisting of plant cell wall (lignocellulose, pectin, and hemicellulose) degrading enzymes, cutinases and host immunity disrupting chitin-binding domain (CBM50). The recently sequenced genome of BRIP57314 isolate was used to predict the proteome of C. tanaceti. Publically available proteomes of 13 Colletotrichum species were used for comparison. Each proteome was searched for proteins encoding domains homologous to CAZyme-specific Hidden Markov Model profiles in the dbcan database V.6, using cut-off values for fungi (e value-1e-17, coverage-0.45) to identify CAZyme-like proteins across the representative taxa. The gene copy numbers of selected CAZyme families were recorded to generate CAZyme profiles for each taxon, which were then subjected to principal component analysis (PCA) and hierarchical clustering. The taxa were also grouped based on previous knowledge of their host-type (dicot/dicot and monocot/monocot) and host-range (broad/Intermediate/narrow). The “narrow” host range was defined for pathogens that are host-specific or infecting few host species within a plant family. Pathogens that infect many host species within a family or few hosts across families were classified under “intermediate”. The “broad” category includes pathogens infecting many host species across plant families. No obvious separation was observed among the host type classes “dicot” and “dicot and monocot” in the PCA scatter plot. However, the two species that exclusively infect monocot plants, C. graminicola and C. sublionela clustered independently in both PCA plot and the dendrogram, from the rest of the Colletotrichum spp which have been reported to infect dicots, exclusively or not. In this instance, C. tanaceti grouped with the latter. The two graminaceous pathogens exhibited major contractions in pectinase families, supporting the fact that the pectin content in cell walls of monocot is less than dicots. The three classes of host range were separated in both the scatter plot and the heatmap. Interestingly the clustering appeared to complement the host range more than the host type and the taxonomy. The Colletotrichum spp with a broad host range separated from the rest and included two sub-clusters of the acutatum complex and gloeosporioides complex with expansions of certain hemicellulose degrading enzymes and pectinases. Colletotrichum tanaceti clustered with C. chlorophyti which is known to infect herbaceous plants. Pathogens with an intermediate host range, with which C. tanaceti clustered, exhibited expansions in pectinase families PL1, PL4, PL9 and contractions in hemicellulose degrading GH29, GH36, GH43 families with respect to those with a broad host range. These findings suggested that comparison of CAZyme profiles within genera is a good approach to infer the host range of a pathogen and that C. tanaceti is likely to have an intermediate host range although to date C. tanaceti has only been detected in T. cineraiifolium.

Abstract

Fungal morphogenesis depends on accurate cell cycle progression. GTPase activating protein complex CoBub2-CoBfa1 interacts with downstream factor, GTPase CoTem1, and is required for G1/S progression and pathogenesis in Colletotrichum orbiculare (Fukada and Kubo. 2015). To elucidate the signal cascade of CoTem1, we performed the screening of physical interaction factor with CoTem1 by Yeast Two-Hybrid (Y2H) system and identified CoNPC2, that encodes a putative phosphatidylglycerol phosphatidylinositol transfer protein. The amino acid sequence of CoNPC2 showed high homology to sterol transporter Saccharomyces cerevisiae NPC2 reported in mammals and yeast. To analyze the function of CoNpc2 in infection related morphogenesis, we obtained Δconpc2 mutant. On the cellulose membrane, Δconpc2 formed appressoria and penetration hyphae indistinguishable from those of the wild type. Whereas, on cucumber cotyledons, Δconpc2 formed normal appressoria, however, formation of penetration hyphae was attenuated, thus the lesion formation of the host plant was markedly reduced. To test whether CoNpc2 is required for invasive growth in plant, we conducted wounded inoculation of Δconpc2 to the cucumber cotyledon. Δconpc2 formed lesions similar to that of the wild type, suggesting that CoNpc2 is not essential for invasive hyphae growth, but is essential for the host cuticle infection. In S. cerevisiae, Npc2 acts as a sterol transporter (Bergeret et al., 2005), thus to test whether CoNpc2 involved in sterol transport, we observed sterol accumulation in conidia by FilipinⅢ, a sterol staining during appressorium development. In Δconpc2, sterol accumulation in vacuoles was shown with high frequency compared with the wild type. Furthermore, to elucidate the functional similarity of CoNpc2 to Npc2, we obtained the CoNpc2-Npc2 partial replacement strain. In this strain, sterol accumulation was reduced compared with Δconpc2. Therefore, it was suggested that CoNpc2 is involved in sterol transport as a functional orthologue of S. cerevisiae Npc2 in C. orbiculare. In addition, the CoNpc2-Npc2 partial replacement strain restored disease symptom formation compared with Δconpc2. Thus, CoNpc2 could function as a sterol transporter that involves in appressorial infection.

Abstract

Raffaelea lauricola, an invasive fungal pathogen, has caused widespread mortality to redbay (Persea borbonia) and other species of the Lauraceae family in the southeastern United States. We produced a draft assembly of the R. lauricola genome and compared this genome to the genomes of other closely related species in the Ophiostomatales including R. quercus-mongolicae, Grosmannia clavigera, Ophiostoma piceae, O. novo-ulmi, and O. ulmi. Structural and functional annotations and analyses were performed for the assembled genome to determine genes that are potentially involved in disease development by this wilt pathogen. The R. lauricola genome assembly resulted in 1,535 scaffolds and a total length of 35 Mb, which is larger than the other genomes evaluated. It also has a larger number of secreted proteins and small secreted proteins, ABC transporters, cytochrome P450, and CAZYmes. Taken together, the number of proteins related to pathogenicity encoded in the R. lauricola genome suggests that this species is well adapted as a pathogen, and possibly better equipped for pathogenicity than other related species. These findings explain, at least in part, the success of R. lauricola as a lethal pathogen with a wide host range within the Lauraceae.

Abstract

Currently, the Aspergillus versicolor clade of the sect. Nidulantes harbours 17 species all of which are considered anamorphic, and no sexual stage is known. The common occurrence of these species in the indoor environment is associated with various respiratory problems, some species have been also reported as a cause of opportunistic infections, including invasive aspergillosis. In this project, we assembled approximately 300 strains from various substrates (indoor and cave environment, indoor air, house dust, various food, clinical material, soil) and continents (mostly North America, Europe and Asia) and revised the species boundaries using a multidisciplinary approach combining phylogenetic analysis with phenotypic data. We analyzed DNA sequences of five protein-coding genes with species delimitation methods based on the multispecies coalescent model. New microsatellite typing scheme was developed from available genomes to investigate the population structure. Both mating type gene idiomorphs (MAT1-1-1 and MAT1-1-2) were found in genomes of A. versicolor clade members suggesting the heterothallic reproductive mode. Preliminary results from different species delimitation methods showed that number of species within A. versicolor clade should be reduced significantly. There is large genetic and morphological intraspecific variability within these species, that we are able to detect due to our large sampling. It permits the detection of recombination and thus gives us chance to understand the species boundaries within this economically important group of fungi.

Abstract

Abstract

Discovering and describing the world’s biodiversity is of paramount importance to biologists. Conservative estimates of fungal diversity put the total number of species worldwide at about 1.5 to 5.1 million, yet only 1-10% of these have been described. Predicting where the estimated “missing fungi” are, is being debated among mycologists and one likely hypothesis is that the majority of unrecorded fungi are microfungi especially those inhabiting specialty niches. However, data from poorly studied geographic regions such as the tropics are limited. Not only may these regions harbor unexpected fungal diversity but studies of tropical fungi could even lead to a revised estimate of total fungal species, given that the current estimate is based on data from temperate regions alone. For this study, fresh collections of saprotrophic fungal species were made from previously established study plots within the Dja Biosphere Reserve (DBR) in Cameroon, West Africa. Descriptions of macromorphological features were made from fresh material in the field. Colors were compared with plates in the Online Auction Color Chart. Fresh specimens were also photographed in situ and in the field laboratory. We dried the sporocarps in the field using silica gel and duplicates were deposited in the National Herbarium of Yaoundé (YA) and the Kriebel Herbarium at Purdue University (PUL). DNA was extracted and analyzed using established protocols. After two years of intensive sampling from this single region in the Congo Basin of Cameroon, we have documented more than 200 species or morphospecies of saprotrophic fungi. Approximately one quarter of these species appear to be new to science. Phylogenetic analyses of these fungi will also aid in species delimitation, as many of these fungi are part of species complexes. These data indicate that tropical regions may not only be host to many of the world’s “missing fungi”, but that estimates of total number of fungal species may need to be revised.

Abstract

The macrofungi exploration in Turgo tropical forest ecosystem of Mount Merapi National Park aimed to document the diversity and distribution of edible and inedible macrofungi in that region. Macrofungi sampling was distinguished by different forest elevations ie Turgo forest, Tritis forest, and Bingungan forest using adaptive sampling method of plot making. Data collection was conducted in May-June where the month is categorized as the wettest month in Turgo area. The total number of macrofungi obtained 133 specimens, in the Turgo forest found 48 species, in the forest Bingungan got 36 species and in the forest Tritis got 49 species. Species with a high level of macrofungi diversity based on the Shannon-Weiner index was owned by Tritis forest with the lowest elevation among the three is 2.521 followed by the Bingungan forest with an index value of 2.51 and the last of the Turgo forest was worth the index of 2.01 which means the three tracks have a level of diversity of moderate-high. Comparing similarity between the three tracks based on the Sorenson index value found that the three tracks have a much different similarity. Measuring from the important values, the highest value morphogroups was owned by macrofungi agarics group with 82% -169% value, followed by other groups ie jelly, polypore, cup, club, coral, and tooth which are calculated based on relative frequency and relative density values. Found 8 species that are edible, 12 species are inedible 6 species are medicinal 106 still remaining unknown. Some key or important species in Bingungan forest were detected like Xylaria spp. and Auricularia delicata; Marasmiellus sp. in Turgo forest and no species can be concluded as the most importan in Tritis forest. The map of macrofungal distribution has been made for each forest tracking.

Abstract

Mycology has experienced a rapid development during the last two decades through the application of molecular techniques and phylogenetics to fungal taxonomy, ecology and evolution. Many fungal species display a limited number of morphological and anatomical characters, making species demarcation difficult. It has been demonstrated that misidentification, mostly of cryptic species, has led to the deaths of many people, whilst traditional taxonomical methods hamper our ability to assess global diversity of fungi. To get a clear picture of fungal diversity and community phylogenetics, systematic sampling of fruit bodies of ectomycorrhizal fungi was carried out in species-rich ecosystems of Benin. We recorded a total of 110 morphological species in 33 genera. DNA was extracted from representative specimens of each morphological species using either the QuiaGen DNeasy Plant Mini kit or a protocol of cryogenic disruption followed by extraction in CTAB buffer, cleaning with chloroform, and alcohol precipitation. The internal transcribed spacer (ITS) region of the rDNA was amplified by PCR using, variously, the primer pairs ITS1–F/ITS4, ITS1-F/ITS4-B, or ITS1/LB-w, and sequenced using the Sanger method. We generated a total of 117 sequences sorted into Russulaceae (33 sequences), Amanataceae (43 sequences), Boletaceae (37 sequences), Cortinariaceae (3 sequences) and Tricholomataceae (1 sequence). Similar sequences were downloaded from Genbank to generate a dataset of 3304 sequences. In the present talk, we will showcase the placement of our sequences within the global phylogenetic context, whilst the consistency of traditional delimitation of species and sections within core genera will be tested phylogenetically.

Relevance/Significance: We expect to depict strong phylogenetic argument to support the description of numerous putative new species and to support delimitations within cryptic taxa. Our investigations will increase our understanding of species limits within taxonomically complex genera. Results from the present study will nourish the Fungi DNA Centre under construction at the University of Parakou.

Abstract

The Gulf Coast region of the Southeastern USA is known as a hotspot of plant and animal diversity, but the fungi of the region are still highly understudied. The most prolific collector and taxonomic expert in the region was William Alphonso Murrill, who collected from the 1920’s until the 1950’s. Murrill described approximately 800 fleshy basidiomycetes, mostly within walking distance of the University of Florida in Gainesville. Many taxa that Murrill described have not been re-collected since their original discovery. Sadly, many of Murrill’s type specimens are degraded so they are challenging to use for phylogenetic analyses and many of Murrill’s original collecting sites near the campus have been lost due to development. As part of a long-term goal to document the mycoflora of Florida and to recollect Murrill’s Florida species, we systematically collected fleshy macrofungi over a one-year period at the Ordway-Swisher Biological Station (OSBS). Ordway-Swisher is located approximately 25 miles east of Gainesville and contains many habitats similar to those where Murrill collected. Here we present an overview of our DNA barcoding project and we discuss some of the findings and challenges from this work. We collected approximately 1000 samples and generated >600 ITS sequences, with additional 28S sequences for select genera. The most heavily collected groups were Boletales (111 collections), Amanita (94), Russula (89), Cortinarius (53), Lactarius (38), Pluteus (31), and Inocybe (19). Preliminary morphological analyses of our collections indicates that we successfully recollected several taxa that are thus far known only from the type specimens (e.g. Inocybe taedophila) as well as several putative new species (e.g. Tuber and Elaphomyces spp.). The high number of Amanita and Boletales specimens and molecular OTUs confirms that the local biodiversity is high for these groups. BLAST analysis of our ITS sequences from OSBS specimens revealed that more than a third of all sequences had no close match in GenBank (<97% similarity) and many taxa had a closest match that was from an unnamed environmental sample. When sequences did match closely with something in GenBank, they were mostly from eastern states (Georgia north to Massachusetts) or from nearby countries across the Caribbean region. Diversity was also highly structured by season; genera such as Cortinarius and Tricholoma were most abundant in the winter months (December-February) while Amanita, Boletales, Cantharellus, Inocybe, Lactarius, and Russula were more diverse and abundant in summer (May-September). This suggests that Florida’s winter mycoflora is dominated by temperate taxa that normally fruit during Fall in sites further north. In the summer, Florida’s mycoflora is dominated by tropical and subtropical taxa, many of which appear to be restricted to the Gulf Coast and Caribbean regions. The strong seasonal temperature fluctuations and potential for rain throughout the year in Florida may contribute to a high diversity of fungal species. This project serves as a model for how local herbaria can contribute to the MycoFlora 2.0 initiative.

Abstract

Ecuador is one of the few megadiverse countries in the world. There is a rich history in the exploration of plants and animals in Ecuador and its fairly well documented. Nevertheless, new species are being described every year even in the better studied groups, such as plants and tetrapods. Fungi, in contrast, are amongst the least studied groups and therefore harbor a large proportion of undescribed species. Access to Ecuadorian fungal species lists and collection data is very limited. Lack of basic information hinders efforts of mycologists in the private and public sectors to develop long-term research programs on Ecuadorian fungi. To alleviate this problem, we are launching FungiWeb, a free, open-access web portal dedicated specifically to publish information on fungal diversity with image guides and lists of species generated as PDFs. FungiWeb also provides access to the database of the biggest fungal collection in Ecuador, the QCAM Fungarium at Pontifical Catholic University of Ecuador (PUCE) in Quito. The collection contains over 7400 specimens with more than 60 orders, 155 families and 472 genera. Most samples are macrofungi from protected areas including understudied National Parks. FungiWeb is part of BIOWEB, a web portal aiming to discover, administer and publish general information about the biological diversity of Ecuador. BIOWEB has been developed by PUCE under the premise that free and open access to biological information is the best incentive for its study, conservation and sustainable use. BIOWEB gives access to information on over 452,000 specimens, which include plants, animals and fungi. The portal details taxonomic classifications, descriptions, distribution maps, images, and more.

Abstract

Found in virtually every ecological niche, fungi are key organisms to the environment and our lives. From nutrient recyclers, decomposers and beneficial symbiotic partners with plants and animals, to food and medicine providers, they can also represent a devastating threat to ecosystems, our food resources and even to our health. Nevertheless, we still suffer from inadequate sampling and study of the kingdom Fungi in many parts of the world. This is the case for Colombia, and specifically in the region of Boyacá, a virgin land for fungal exploration. For the first time, this project aims to close the gap and assess the unknown fungal diversity in the department of Boyacá, Colombia, by using a field-to-lab approach merging expeditions, standard barcoding techniques, and genomic approaches with traditional specimen-based identification methods. During the first phase of this project, we collected and documented fungal diversity in the six types of habitats present in Boyacá: páramo, sub-páramo, Weinmannia-pine forest, dry acacia forest, oak forest, and tropical rainforest. Using plots and transects, we systematically collected specimens grouped in four categories: macrofungi and non-lichenised ascomycetes, lichenised fungi, plant roots for mycorrhizal fungi, and leaf material for endophytic fungi. In total, we collected more than 1,500 samples (more than 400 macrofungi and non-lichenised ascomycetes, 760 lichenised fungi, 160 roots, and 200 leaves). Given that 79 species of macrofungi and 261 species of lichenised fungi have been cited so far for Boyacá region, our exhaustive sampling will surely provide many new records as well as potentially new discoveries to science. Furthermore, species accumulation curves show that more sampling is needed to cover the full diversity of the habitats surveyed in this study as well as other habitats present in the region.

Abstract

A review of literature indicated that little mycological work had been done in Mozambique, Africa. Therefore, we conducted a fungal survey of Gorongosa National Park (GNP) for one month in June-July of 2016. The Park had been a frequent battleground during the years of the Mozambican Civil War (1977-1992) and was left unprotected for another decade. In that time, the Park’s fauna was decimated. Today, restoration of the wildlife and tourist infrastructure is advancing apace. The establishment of the E. O. Wilson Biodiversity Laboratory in the Park, modeled along the lines of the Smithsonian Tropical Research Institute on Barro Colorado Island in Panama, provided an attractive research destination. We collected voucher specimens at random localities and habitats within the Park, focusing on discomycetes, polypores and anamorphic fungi. Despite a severe drought had affected the region that year, we were able to make over 500 collections. We are currently making determinations of these and will publish our results in the form of a checklist. Since most fleshy fruiting bodies occur during the rainy season, and as our survey was conducted in the beginning of the dry season, our collections represent species in fruit during a season that is not often sampled. Voucher specimens will be deposited at the Biodiversity Collection at the E. O. Wilson Biodiversity Laboratory at GNP, the Herbarium of Eduardo Mondlane University in Maputo (LMU), the Farlow Herbarium (FH) at Harvard University and the University of Illinois Herbarium (ILL). Our work will stimulate future studies of the Park’s mycobiota and become the cornerstone of a systematic fungarium within the Biodiversity Collection. This would be the first such collection housed at any African National Park. Furthermore, we hope that a baseline understanding of fungal species and their activities in GNP will help to inform current decisions around Park conservation and allotment of resources.

Abstract

Hurricanes generate disturbances in forests such as canopy opening, fallen trees and leaves which alter physicochemical characteristics of the habitat. Litter decomposition depends primarily on the interaction among climate, litter quality and biota, as a consequence any change in habitats will result in changes in these factors. Understanding the mechanistic processes of litter decay is essential for predicting nutrient cycling dynamics in tropical forests. While models of litter decay mostly rely on climate and litter chemistry, it is increasingly apparent that the decomposer communities (fungi, bacteria and arthropods) interact during leaf litter decomposition significantly influencing decay rates and mineralization of nutrients. White-rot basidiomycetes are the most efficient biodegraders of lignin, breaking down bonds to expose the assimilable cellulose and hemicelluloses surrounding lignin. This assimilable cellulose and hemicellulose is available for the bacterial community to degrade. Litter with more lignocellulose promotes the fungal decomposition pathway, which in turn favors soil and litter food webs dominated by arthropods. Our objective is to evaluate the effects of hurricane driven changes to forests on green litter decomposition, decomposer communities and nutrient mineralization. For this study, three blocks were selected, each with two plots of 20 m x 20 m, one plot was used as unmanipulated control and the other for canopy trimming. In each subplot, litterbags with different mesh sizes were placed. Each of these litterbags were used as the sampling unit. In each one, decomposer fauna and nutrients were measured, and the weight of green litter from the litterbags was used to measure mass loss through time. Microbial diversity was documented using TRFLP. Diversity and abundance of arthropods was determine using the Berlese funnel technique. Nutrient release was documented using Plant Root Simulators (PRS probes). Preliminary results suggest significant differences in abundance of decomposer fauna and in available nutrient concentration between trim plus debris and unmanipulated control plots, and among litterbags mesh sizes. The fungal community structure and nutrients differed significantly between unmanipulated control and trim plus debris. For example nitrogen and phosphorous were significantly higher in trim plus debris plots and in large mesh litterbags. Also, decomposer arthropod abundance was higher in large mesh litterbags and the TRFLP showed diversity of fungi in the unmanipulated control. In conclusion, there was a trend for higher arthropod abundance, higher nutrient availability and larger mass loss in large mesh litterbags, suggesting trophic dynamics mediated by all decomposer communities. Arthropod abundance increased with the increase in the diversity of fungi. Nutrient release was higher in the trim plus debris plots during the first 5 weeks after treatment and when all the trophic groups were present. In the future, samples will be sent for Illumina sequencing. These results will be further analyzed, and interpreted in the context of food web dynamics.

Abstract

Forested stream networks play a significant role in the storing and processing of terrestrial organic carbon (OC), such as leaves and wood. Fungi are at the forefront of stream OC processing by way of plant litter decomposition, mediating the flow of energy and nutrients to higher trophic levels. Increases in temperature under climate change predictions are expected to affect microbial activity, as well as carbon dynamics in aquatic ecosystems. The objectives of our study are (i) to determine the responses of litter associated decomposers to temperature and (ii) to test whether fungal responses can be explained by the Metabolic Theory of Ecology (MTE) within the temperature ranges found in temperate streams. We performed an experiment in laboratory microcosms simulating stream conditions and measured physiological responses of natural stream microbial assemblages, including fungi and bacteria, to increased temperature. Sterilized leaf disks (Liriodendron tulipifera) were inoculated in a stream at the Coweeta Hydrologic Laboratory, NC during peak litter fall. Leaf disks were then incubated at five temperatures (4-20°C) in laboratory microcosms. We determined leaf litter decomposition rates, fungal biomass accrual (from ergosterol), fungal and bacterial production (using radiolabeled tracers), sporulation rates and cumulative spore production by aquatic fungi, microbial respiration rates and activity of enzymes involved in carbon sequestration (β-1,4-glucosidase, β-1,4-xylosidase and phenol oxidase). We found that responses of aquatic litter-associated microorganisms to increases in temperature are more complex than simple predictions of the MTE, with more pronounced responses (higher estimates of apparent activation energy, E) at lower temperatures. For some parameters, our estimates of E at lower temperatures were higher than canonical values often reported for respiration (~0.65 eV), suggesting that fungi and microbial OC processing in streams during the cold months could be especially sensitive to temperature increases. These trends may have important implications for stream ecosystems under climate change scenarios, since bulk leaf litter inputs and peak fungal activity coincide with the coldest season (autumn-winter) in temperate streams.

Abstract

Fungi such as mushrooms occupy some of the most diverse ecosystems on the planet and have been found to grow and proliferate in extreme environments. Fungi thriving in harsh environments are of particular interest since they produce enzymes which possess higher thermostability resulting in activity at higher temperatures. In the current study we describe the sampling and isolation of a possible new fungus species from the South African deep subsurface (3.1 kmbls). The deep subsurface is characterized as an extreme environment due to elevated temperatures, humidity, radiation and limited nutrients. The fungus formed brackets and was isolated growing on wood packs supporting the rocks during the mining process in a stope gully. Stope gullies are found in areas were the reef is being actively mined resulting in high temperatures as well as blasting by explosives on a regular interval. With the aid of morphological and cultural observations, and DNA sequence comparisons the fungus was identified as a possible new species in Ganoderma. ITS sequencing (internal transcribed spacer region) resulted in a distinct phylogenetic signature clustering separately from all other Ganoderma species, including those from South Africa. This bracket fungus has displayed thermotolerant behaviour as maximum radial growth was achieved at the temperature of 35°C (45 mm), followed by 30°C (42.2 mm), 25°C (31.1 mm) and 40°C (23.8 mm) after 12 days of incubation. The ability to produce fruiting bodies was also observed at 35°C. This bracket fungus has also displayed above average lignolytic abilities degrading supporting wood stumps in days. The origin of this Ganoderma species is unknown but it is more likely to have originated as an inhabitant of the woody material than from the mine. Further studies to determine its correct origin, and whether its tolerance to high temperatures are natural or could maybe due to environmental pressures in the mine environment, have important implications for biosecurity within mines.

Abstract

In Pakistan climate change has emerged as a serious concern for agriculture especially for vegetable and ornamental plant. Investigations conducted against regular field crops like sunflower, sesame, maize and perishable crops Gladiolus, Tomato, Bitter Gourd and onion against most commonly occurring pathogens viz Macrophomina phaseolina, Alternara alternata and A porri spp, Fussarium spp and Myrothecium roridium from 2007-2016 on disease distribution pattern of Root rot of Peanut, Stalk rot of maize, Myrothecium leaf spot of Bitter Gourd, Charcoal rot of Sesame and Purple Blotch of Onion (ongoing).Comprehensive surveys were conducted toto identify changes in the distribution pattern of major disesase in relation to different geographical zones. Main objective of these studies was to acquire knowledge about the variation in terms of prevalence, diseases incidence and severity of the diseases with relation to the environmental conditions of the specific zone. The climate change also affected on Sporulation, morphology, new areas of introduction and lesser occur, adaptability physiology. The severity was measured on a 0-5 visual rating scale where 0 stands for no disease and 5 stands for 80% ⁺ disease severity. Some of the studies for post harvest quality deterioration in fruit and vegetable markets were also co related with changing climate. A structured questionnaire was also distributed among crop production and protection stake holders from public and privates sector. Data was collected from five Agro ecological zones in Punjab province in Pakistan that are designated as rice, cotton, mix cropping and rain fed or pothohar zone. In these zone farmers concern was noted as Change in inputs requirement: especially irrigation, pesticides and fertilizers, Marketing instability, Post harvest processing, Crop phenology. There is need for regular monitoring of the cropping systems and isolates of the most commonly occurring pathogens of the country. It would help in designing cropping patterns in accordance with the need.

Key words: Climate change, soil born fungi, air born fungi, perishable crops, field crops

Abstract

Climate change is already having significant impacts on forest ecosystems and these seem likely to be greater in the future. Understanding pest and pathogen dynamics and their impacts on tree health is essential to predict and potentially mitigate the effects of changing climate on forest stability. Most studies have focused on host tree and pest distribution under changing climate scenarios, but only a few have focused on the modulation of pathogenicity within host-pathogen systems. The European spruce bark beetle (Ips typographus) is one of the most aggressive insect pests of conifers in Europe. A warming climate has been shown to enhance I. typographus infestations by promoting swarming activity and reproduction, which are mainly temperature-dependent. This insect pest vectors the necrotrophic fungus, Endoconidiophora polonica, which is regarded as the most aggressive pathogen of Norway spruce (Picea abies). In this study, two powerful inoculation experiments were conducted to test in vivo the effects of a changed growing environment on Norway spruce seedlings infected with different strains of the E. polonica. The first experiment compared seedling performance under ambient summer temperatures and CO₂ levels with those predicted for the years 2030 and 2100 in Finland. The second compared seedling performance under high and low water availability treatments. For both experiments, seedling mortality was monitored throughout the annual growing season. Seedling growth and lesion length indices were measured at the end of the experiment. The results showed that increased temperatures coupled with elevated CO₂ concentration, and water availability can both enhance disease severity in P. abies. Higher temperature increases are likely to be the most detrimental to tree health, and interestingly, disease severity varied markedly among the fungal strains. Our results indicate that predicted climate change has the potential to alter the damage caused to Norway spruce by E. polonica. They also highlight the importance for a strain-specific level of understanding of the disease agents. Managing increasingly disturbed forests will be an important challenge in the future. There is consequently an urgent need for systems-based research to better understand the impacts of interactions between biotic agents and climate change in forest ecosystems.

Abstract

The Earth’s climate has undergone major changes in the past, with massive oscillations between cold glacial and warm interglacial periods during the last 2.6 Myr (Late Quaternary). Late Quaternary climate change can drive migration, extinction and speciation and therefore may influence the modern diversity and composition of macroorganism communities, but how it structures belowground microbial communities is less well known. As a major component of belowground microbial communities, soil fungi exhibit a wide range of strategies including mycorrhizal, pathogenic and saprotrophic species, and make a large contribution to biogeochemical processes and ecosystem functioning. The objective of this study is to reveal the relative effect of late Quaternary climate change and contemporary environment on fungal community in different forest ecosystems. We examined soil fungal communities in temperate, subtropical and tropical forests in China, using Illumina Miseq sequencing of ITS1 sequences. The fungi were divided into different trophic groups, such as saprotrophic, ectomycorrhizal and pathogenic fungi. The total, saprotrophic, ectomycorrhizal and pathogenic fungal richness were influenced primarily by the contemporary environment, but not by the late Quaternary climate change in the temperate, subtropical and tropical forests. By contrast, the late Quaternary climate change acted in concert with the contemporary environment to shape total, saprotrophic, ectomycorrhizal and pathogenic fungal community compositions and with a stronger effect in temperate forest than in tropic-subtropical forest ecosystems. Some contemporary environmental factors influencing total, saprotrophic, ectomycorrhizal and pathogenic fungal communities in the temperate and tropic-subtropical forests were different. In conclusion, we demonstrate that late Quaternary climate change can help to explain the mechanisms maintaining current soil fungal community composition and argue that climatic legacies can help to predict soil fungal responses to climate change in ecosystems.

Abstract

Fungal acquisition of metabolic carbon and its subsequent partitioning between biomass production and respiration is a central aspect of forest soils. A frequently used concept to express energy and carbon flow in the ecosystem is carbon use efficiency (CUE), defined as the fraction of acquired carbon that is incorporated in biomass. Fungal communities are central below-ground drivers of ecosystem process, especially in boreal forests, and estimating their CUE is important for understanding their role in the ecosystem. However, current available techniques for estimating CUE in natural systems, such as isotope labelling, biomass markers, and respiration measurements, are all associated with practical and theoretical shortcomings, making assessments unreliable. In this project, we propose that metatranscriptomics can be used to assess CUE of fungi in natural systems. In laboratory experiments, we aim to explore transcriptome profiles of fungal cultures, to select genetic markers of resource acquisition, mycelial growth and respiration. Transcriptomic indexes will be related to measurements of CUE. These indexes will then be used to analyze soil metatranscriptome profiles from boreal pine forest stand, in order to address questions about fungal adaptations and ecological strategies.

Abstract

Despite covering vast areas of boreal North America, the ecological factors structuring mycorrrhizal fungal communities in peatland forests are relatively poorly understood. To assess how these communities vary by age (younger v. mature), habitat (fen v. bog), and host (conifer trees v. ericaceous shrub), we sampled the roots of two canopy trees (Larix laricina and Picea mariana) and an ericaceous shrub (Ledum groenlandicum) at four sites in northern Minnesota, USA. To characterize the specific influence of host co-occurrence on mycorrhizal fungal community structure, we also conducted a greenhouse bioassay using the same three hosts. Root samples were assessed using Illumina-based high-throughput sequencing (HTS) of the ITS1 rRNA gene region. As expected, we found that the relative abundance of ectomycorrhizal fungi was high on both Larix and Picea, whereas ericoid mycorrhizal fungi had high relative abundance only on Ledum. Ericoid mycorrhizal fungal richness was significantly higher in mature forests, in bogs, and on Ledum hosts, while ectomycorrhizal fungal richness did not differ significantly across any of these three variables. In terms of community composition, ericoid mycorrhizal fungi were more strongly influenced by host while ectomycorrhizal fungi were more influenced by habitat. In the greenhouse bioassay, the presence of Ledum had consistently stronger effects on the composition of ectomycorrhizal, ericoid, and ericoid ectomycorrhizal fungal communities than either Larix or Picea. Collectively, these results suggest that partitioning HTS-based datasets by mycorrhizal type in boreal peatland forests is important, as their responses to rapidly changing environmental conditions are not likely to be uniform.

Abstract

Although the response of microbial diversity to environmental factors has been well-studied, it remains unclear how the traits of soil fungi and bacteria determine their biogeographical distributions. With the emergence of functional databases like FUNGuild for fungi and Silva for bacteria, we can extend beyond the fungal-bacterial dichotomy to investigate the relative effects of traits such as growth morphology on soil microbial diversity. Here we assess how both phylogenetic placement (fungi or bacteria) and growth morphology (unicellular or filamentous) independently affect changes in microbial diversity across regional environmental gradients. Fungal and bacterial soil communities from multiple ecosystems across an environmentally heterogeneous region of the Midwestern US were characterized by high-throughput sequencing. We hypothesized that microbial communities would be less similar to one another as geographic distance between them increased, but that microbes with unicellular growth morphologies would be less sensitive to changes in geographic distance or environmental factors. We found that fungal communities, on average, turned over much more quickly (i.e. have higher beta diversity) over a given geographic distance relative to bacterial communities. Additionally, we found that growth morphology was equally or more important than geographic distance in predicting microbial community diversity. Specifically, unicellular fungal communities (i.e., yeasts) turned over at rates more similar to bacteria than to other fungal communities, whereas filamentous bacterial communities had higher rates of turnover than unicellular bacterial communities. These results suggest that both fungal and bacterial filamentous microbes have higher beta diversity than unicellular microbes at a regional scale. Further, we found that fungal communities were more sensitive to plant community composition while bacterial communities had a higher sensitivity to abiotic environmental characteristics. Collectively, our results support the idea that fungi and bacteria are differentially affected by geographic distance and environment, and also that growth morphology is an important factor determining landscape level patterns of diversity.

Abstract

Microbial communities are the engines of decomposition, a fundamental process regulating the carbon cycle. While much is understood about how changes in abiotic conditions and substrate quality affect decomposition rates, the role of microbial community composition remains elusive. This knowledge gap may be key for predicting how ecosystems will respond to climate change. Here, we first surveyed the microbial community composition of decaying leaf litter along an elevation gradient in southern California, USA. The gradient included five sites with precipitation increasing, and temperature decreasing with elevation. To test the importance of climate for microbial composition, we conducted a transplant experiment. We used “microbial cages” to transplant leaf litter communities to different climates while preventing microbial exchange with the environment. We inoculated transplant communities onto a common, irradiated, grassland litter. In contrast with observational data, this manipulation isolates the effects of climatic conditions versus that of microbial composition on litter decomposition. To characterize fungal and bacterial community composition, we extracted DNA from the inoculum, intact litter at sites, and litter in the microbial cages, and amplified and Illumina sequenced part of the ITS and 16S rRNA regions. We also analyzed microbial biomass with fungal hyphal abundance counts and bacterial flow cytometry. We investigated the functional consequences of these transplants by measuring decomposition as mass loss and analyzing nutrient content. We hypothesized that communities from along the climatic gradient would differ in their abundance and composition, and that communities at the extremes of the gradient would be most affected by climate. We found that microbial communities did indeed differ greatly between the five sites (PERMANOVA; Fungi: R² = 0.58, P < 0.01; Bacteria: R² = 0.61, P < 0.01) with fungi primarily dominated by the Ascomycota. The main axis of community separation appeared to be between the colder and wetter sites versus the hotter and drier sites. After transplantation, fungal communities retained a strong signature of the inoculum whereas bacteria were quickly influenced by local climate. Inoculum source was the strongest factor influencing fungi (R² = 0.57, P < 0.01), but site was also significant (R² = 0.14, P < 0.01). In contrast, site was the strongest factor affecting bacterial composition (R² = 0.34, P < 0.01), although inoculum source was also significant (R² = 0.16, P < 0.01). However, a strong site by inoculum interaction effect for both fungi (R² = 0.15, P < 0.01) and bacteria (R² = 0.19, P < 0.01) indicates that the strength of the inoculum effect varied by site, meaning that not all communities responded similarly to the climate gradient. Unexpectedly, communities from the extremes of the gradient responded similarly. In contrast, communities from the intermediate elevation sites had significantly different impacts on decomposition and types of carbon compounds degraded when transplanted. Moreover, significant site by inoculum interactions impacting decomposition lasted a year after transplantation. These results demonstrate that microbial communities affect decomposition, but bacteria will shift more rapidly in response to climate than fungi, with fungi retaining a strong inoculum signal even after 1.5 years.

Abstract

It is clear that distribution of fungi is not governed by dispersal limitations only, but also by the availability of suitable ecological niches reflecting the abiotic and biotic conditions of the environment. The abiotic factors, especially temperature, precipitation, habitat type, pH, nitrogen and calcium contents were reported to be the most important drivers of fungal distribution. In this study we have focused on the identification of climatic drivers on the distribution of individual fungal taxa based on the records of their occurrences and abundances across the globe. For the study, we have selected top 200 globally distributed species from environmental samples taking the 98.5% species hypothesis (SH) as a proxy of fungal species. To explore the influence of climatic drivers on the distribution of fungi, CHELSA climatic dataset was combined with GPS coordinates of samples and Gradient Forest algorithm was used. This method extends Random Forest, which fits an ensemble of regression tree models between individual species abundance and environmental variables. Based on this analysis we proved that for more than 75% of dominant fungal species, their distribution is significantly affected by climate. Mean Diurnal Range (Mean of monthly (max temp - min temp)), Mean Temperature of the Wettest Quarter (a quarter is a period of three months), Precipitation of the Coldest Quarter and Precipitation Seasonality (Coefficient of Variation) were most often among the important bioclimatic variables that shape fungal distribution. In average, temperature-related variables explained 18% of the total variation, followed by precipitation variables (9%) and combined variables (7%), but the share of explained total variation ranged up to more than 70%. No difference in the share of the total variation in distribution explained by climatic variables was observed between fungi belonging to different ecophysiological groups, e.g. saprotrophs, ectomycorrhizal fungi, plant pathogens, etc. This study shows that climate belongs among the important factors that shape fungal distribution at a global scale.

Abstract

Interactions between fungi and other living organisms resulted in fungal adaptations to various trophic strategies. Occurrence of such fungi primarily depends on presence of suitable substrate or associated plant (host or symbiotic partner). Fungal distribution areas are known not to copy distribution of hosts (partners) and they are limited by ecological, climatic and geographic factors. However, the relation of trophic strategy and corresponding adaptations with flexibility of fungi to inhabit various climatic zones or geographical areas is unknown. We hypothesize that occurrence of fungi with different trophic strategies depends not only on local ecological conditions, but also on bioclimatic and geographic influences and that they response differently with various environmental factors. These aspects have been studied on three model fungal groups in Europe: parasitic powdery mildews (family Erysiphaceae) growing on selected forest trees, lichenized fungi of the genus Solenopsora and ectomycorrhizal fungi of the genus Russula. We have used known records on occurrence of precisely defined fungal taxa (genetically, morphologically) and selected environmental variables to model habitat suitability maps showing potential areas of distribution. Based on this, we identified geographical elements within each trophic group and assigned bioclimatic and geographical factors influencing their area of distribution. In addition, we identified barriers or geographic, ecological or climatic disjunctions limiting migration of fungi of studied trophic groups. The project is funded by the Slovak Research and Development Agency under the grant no. APVV-15-0210.

Abstract

Global patterns of fungal richness have been associated with edaphic variables and current climate, and it is specific for trophic group: ectomycorrhizae (EcM), saprobiont, and parasitic fungi. In specific, global richness in Agaricales sensu lato is high in tropics, and it tends to decrease towards the poles, correlated with precipitation and carbon availability. The southern of South America is dominated by the genus Nothofagus, and it is widely distributed in Chile (32° - 55 °S), with 10 species, where N. macrocarpa and N. obliqua are the northernmost species. Nothofagus has obligated ectomycorrhizal associations, being most of the species of the Agaricales s.l. group. Little is known about factors that determine in the distribution of this group along a latitudinal gradient. For N. obliqua it has been described that both richness and abundance of the associated EcM species decrease at high nitrogen levels, however the factors that influence N.macrocarpa distribution, remains unknown. In this study we describe both Agaricales s.l. richness and composition in N. obliqua – N. macrocarpa dominated forests, depending on plant composition, edaphic variables and climate. This will be evaluated along an increasing precipitation gradient towards the south and along increasing plant diversity, also towards the south. We sample in 6 localities, from Region Metropolitana (33 °S) to Region de la Araucanía (39 °S). We made on average 3 vegetation transects per locality where fungi fruiting bodies were collected. We also measured soil pH, organic matter content and soil nutrients. We found around 420 fungi species, where ~130 species correspond to EcM, ~290 species correspond to saprobiont, and 1 species to parasitic fungi. We also found an increasing diversity gradient toward the south and that variables that positively correlates with total species richness, EcM species richness, saprobiont species richness and wood decomposers richness are: 1) Mean annual precipitation; 2) Total plant species richness; 3) Woody plants species richness index and total soil carbon content. On the other hand, we found that variables that negatively correlate with richness factors mentioned before are: 1) soil pH; 2) percentage of uncovered sky in autumn and winter; 3) mean temperature of summer. Finally, our results also show that saprobiont species richness increase at higher mean annual temperatures, higher mean summer temperatures and minors soil pH. Acknowledgments: FONDECYT , PFB-23 (IEB).

Abstract

In the lowland tropics, monospecifically dominant forests are patchily distributed, and their persistently dominant trees are usually ectomycorrhizal (ECM). In such forests, supra-annual mast seeding results in dense cohorts of shade tolerant seedlings that ultimately replace conspecific adults, thereby maintaining monodominance. High seedling survivorship and mitigated density-dependent mortality may depend on rapid ectomycorrhization and resulting enhanced nutrient uptake, protection from root pathogens and rhizovores, and possible resource networking with adult trees. However, both the identities of early-colonizing ECM fungi and their influences on seedling establishment and survival are poorly known. To study these factors, we sampled hundreds of seedlings from an even-aged cohort of the monodominant ECM tree Dicymbe corymbosa (Fabaceae subfam. Detarioideae) in a Neotropical lowland forest. Seedlings were sampled at < 1 month, 6 months, and 12 months after a large mast seeding event in 2016. For each age class, the percentage of fine roots colonized by ECM fungi was estimated on a per-seedling basis. Root tips were sorted into distinct ECM fungal morphotypes and Sanger sequenced at the ITS region. To compare seedling and adult ECM fungal communities, ECM morphotypes of 30 nearby adult D. corymbosa trees were bulk DNA extracted and sequenced at the ITS locus on an Illumina platform. Ectomycorrhization occurred rapidly on seedlings, covering 26–50% of the roots at ~2 weeks of age, and increased with seedling age over 12 months. Early-colonizing ECM fungi on seedlings < 1 month old were primarily from the /tomentella-thelephora, /boletus, and /clavulina lineages, while the /russula-lactarius lineage appeared later in ECM development. While numerous ECM morphotypes were shared between seedlings and adults, two species of Tomentella (Thelephorales) dominated root tips of D. corymbosa at all age classes. The ecological impacts of rapid mycorrhization and ECM fungal community assembly on seedling recruitment will be discussed.

Abstract

Studies investigating patterns of fungal biogeography typically focus on one or two main groups, namely ectomycorrhizal (EcM) and saprotrophic fungi. However, few studies have explored biogeographic patterns of soil and root-associated fungi in arbuscular-mycorrhizal (AMF) compared to these other groups. Additionally, AMF research is generally biased towards herbaceous plant species making it difficult to compare these patterns to the forests in which EcM patterns are observed. AMF play important roles in both plant ecology and ecosystem function, however, the patterns of AMF biogeography and how these patterns compare to other fungal guilds remain largely understudied. Here we report the first comprehensive study of fungal communities associated with coast redwood (Sequoia sempervirens) where we tested biogeographic patterns of both soil and root fungal communities to compare patterns of distance-decay across different guilds. Using a nested study design, we sampled throughout the redwood range across 8 different sites spanning a 600km distance. Paired root and rhizosphere soils were collected and sequenced using Illumina amplicon sequencing. Soil chemistry, physical distance, and climatic variables were all collected in addition to these samples and used to create distance matrices to test the effects of each of these different physical factors using a Multiple-Matrix Regression (MMR). We also used FUNguild, a program designed to recognize different fungal guilds to analyze biogeographic patterns of these different fungal groups. We found that while soil fungal communities showed strong trends of distance-decay, there was no significant distance-decay demonstrated in fungi colonizing roots. After breaking these sample types into their different fungal guilds, we then compared different fungal guilds to see which taxa were driving these patterns. We found that saprobes, endophytes, and other mycorrhizal taxa showed strong patterns of distance-decay in both roots and soils. However, AMF showed no patterns of distance-decay in either sample type across the 600km geographic distance and large climatic gradient. It is clear, however, that AMF communities show different patterns of distance-decay than EcM at similar scales. Additionally, we found that certain AMF taxa were disproportionately selected by roots (33% of AMF taxa were significantly enriched in roots compared to soils). We also found numerous EcM taxa and ericoid taxa associated with coast redwood root samples (root DNA was confirmed via sequencing). Though we have not observed any EcM structures, to our knowledge, these findings mark the first discovery of historically EcM taxa in or on roots of an AMF-associated conifer. We assert that these patterns need to be tested at larger (continental, global) scales using high-resolution primers. Additionally, the occurrence of EcM taxa in or on roots of AMF-associated gymnosperms represents an area of future research in understand how common this might be and if there is any potential functional relationship.

Abstract

The ectomycorrhizal symbiosis has been largely understudied in the Neotropics, probably due to the rare observations of ectomycorrhizal root tips and fruiting bodies. Ectomycorrhizal hosts are probably rare, and localized, but curiously, recent studies from taxonomists have revealed a scattered distribution of ectomycorrhizal fungi in the Neotropics. In parallel, since the development of next generation sequencing, the number of studies sequencing soil fungi is growing in the Neotropics, increasing the chance of detecting ectomycorrhizal fungi in soil. However, the lack of reference sequences from fruiting bodies, and the short length of environmental sequences still limit bridges between taxonomists and ecologists. In French Guiana, we spent six years collecting fruiting bodies to increase the number of reference sequences, and environmental sequencing began in parallel in diverse forest habitats. We used new phylogenetic methods developed for short sequences to compare these two sources of sequences. We show that part of short sequences are still difficult to assign, and that environmental sequencing can detect putative new species. This study confirms that ectomycorrhizal fungi can be widespread in French Guiana, but are always rare compare to other fungi. We hope that such an approach could be followed by both ecologists and taxonomists, to deeper investigate the distribution and diversity of ectomycorrhizal symbiosis in the Neotropics.

Abstract

Weatherization typically focuses on air-sealing buildings to reduce energy use, which has the potential to alter indoor microbial communities through reduction of a primary source of indoor microbiota: the outdoors. Indoor sources of microbes may be enriched for human-associated organisms with increased probabilities of potential immune dysfunction and infection. Tightening of the building envelope by weatherization measures may also affect the indoor air quality, as reduced air exchange rates may concentrate harmful pollutants indoors. It is known that ventilation strategy alters indoor microbial signatures, but the role of building operation by occupants has not been well studied. We set out to study the ways that weatherization alters indoor air quality, including the composition and structure of indoor fungal and bacterial communities. Sixty-six households across the greater Portland, OR, metro area were recruited through our local partners to obtain paired weatherized and comparison homes. For both comparison and treatment homes, we collected data in three core research domains: bioaerosol microbial community composition, air quality and building science, and home occupant behaviour and perception. Briefly, each home was equipped for one week with dual sampling units (one indoors, and one out-of-doors), including standard air quality sensors and settling plates for bioaerosol microbial community sampling. Each home was sampled twice: before and after weatherization, or a similar interval for comparison homes; at each sampling, we administered a survey of home operational behaviours and health indicators to residents. Microbial communities were assessed using standard Illumina metabarcoding methods for bacteria (16S) and fungal (ITS) components; analysis used DADA2, phyloseq, vegan, and codyn in R. We observed no gross effect of weatherization treatment, indicating that microbiomes of weatherized homes were not more human-associated after treatment. We found that the community dissimilarity between paired indoor/outdoor bioaerosol samples is related to seasonal patterns of window operation, and that the synchrony between indoor/outdoor air quality measures helps explain the dissimilarity in microbial communities. These results, taken together, demonstrate the impact that seasonal choices in home operational mode has on the balance of microbial sourcing, which may play a role in the health of building occupants. While weatherization may impact the air quality and microbial communities of buildings when they are operated in an entirely ‘closed’ mode, the effect of choice of operational mode, which varies seasonally, has by far the greater impact on the indoor microbiota.

Abstract

Indoor environments are distinctive man-made habitats for fungi. Whenever water or moisture is available, building materials, such as wood, clothing, fabric, paper products, foodstuffs, etc. are suitable substrates for fungal infestation. Even house dust is an unusual habitat in which some xerophilic fungi can survive. Despite the fact that a number new taxa had been described from indoor environments in the past, fungal diversity in indoor environments has not been sufficiently studied. A unique fungus isolated from a swap sample collected from a residence in Massachusetts, USA was found to be new to science. The objective of the study is to describe it and determine its phylogenetic relationship with allied genera. Both morphological methods and phylogenetic analyses using ITS, LSU and SSU ribosomal sequences were used to study this fungus. The fungus was named Desertella microspora. The newly named species has thick-walled conidia globose to subglobose, 17.7 ± 2.6 × 17.3 ± 2.5 μm, and belongs to Chaetomiaceae, Sordariales.

Abstract

Following work was focused on physiology wood rotting fungi with special emphasis on potential inhibition of their growth with newly developed chemical agents based on metal nanoparticles. In our survey strains Serpula lacrymans, Coniophora puteana, Fibroporia vaillantii and Gloeophyllum sepiariumce have been tested for their ability to grow, colonize new substrate and produce extracelular enzymes in presence of mentioned chemical agents. The work was supported by the GAČR (Czech Science Foundation) grant No.17-05497S.

Abstract

Wood utility poles have a long record of providing excellent service for supporting electric transmission and distribution lines under a variety of environmental conditions. The major factors driving fungal decay of utility poles are not restricted to the in-service environment; they can be influenced before pole installation by factors during tree harvest, processing, seasoning, and preservative treatment. Air seasoning is often used for wood species with durable or moderately-durable heartwoods because it is simple and effective. However, long-term exposure increases the risk of fungal colonization. Improper sterilization during preservative treatment can result in utility poles with a high susceptibility to premature decay. An excellent potential example of this problem can be found in the electrical distribution system in Ireland. The Irish electrical system was largely rebuilt in the period between 2000 and 2007 using Scots pine (Pinus sylvestris) poles that were air-seasoned for 6 to 12 months in various Scandinavian countries, before being pressure treated with creosote. The specification called for 100% penetration of the sapwood, but there was no requirement for sterilization. This study investigated the presence of decay fungi in 963 creosote-treated Scots pine utility poles from Ireland. Nearly 4,000 increment cores were taken at five heights from groundline to pole-top. Cores were plated on malt benlate agar and unique decay fungi were identified using Sanger sequencing. Fungal data were combined with environmental and preservative treatment metadata, as well as visible pole decay, to identify wood protection problems. Sixty-seven percent of poles contained at least one decay fungus, with the most common isolates being Phlebiopsis gigantea (77%), Neolentinus lepideus (7%), Sistotrema brinkmannii (7%), and Stereum sanguniolentum (3%). The remaining 20 fungal species accounted for 6% of total isolations. Isolation frequencies suggested no differences with seasoning method, depth of sapwood treatment penetration, or pole age. It is worth noting that most isolations occurred in poles treated after 1996, which correlated with changes in pole procurement practices. Air seasoning for longer than 6-months also resulted in higher incidence of decay. Decay fungi were isolated from 96% of poles that contained visible decay and 49% of poles where decay was not visible, suggesting a larger percentage of poles are at risk of decay development. The results led to increased quality control and the inclusion of sterilization procedures prior to or during treatment. These data will be used to help guide process-management decisions in climates conducive to fungal decay.

Abstract

The order Phacidiales reflects the chaotic systematic situation within the Leotiomycetes, a class where generic and higher delimitations are subject to ongoing changes. Since the order was established by Bessey in 1907, its taxonomic concept has varied through the time, with authors such as von Höhnel 1917, Korf 1973, Dennis 1978, Crous et al. 2014, and Baral 2016, changing the number of species, genera and families that they accepted. Each author seems to have had a different morphological concept of the order. Nowadays, between 27 and 37 of the genera that have at some stage been placed in the Phacidiales are still accepted, but the majority of them are now belong in other families or orders (Baral 2016, Wijayawardene et al. 2017, Index Fungorum accessed I.2018). Only 24 of them have DNA sequences available in public repositories. We have carried out morphological and phylogenetic analyses to better characterize the order Phacidiales and the relationships among its members. Our molecular data set includes 31 species and three regions of rDNA (SSU, ITS, LSU), covering representative members of Phacidiales according to Baral 2016: Phacidiaceae (5 seq.), Helicogoniaceae (5 seq.) and Tympanidaceae (12 seq.). Also, five sequences of the Mniaecia lineage were included, and two sequences representing the genus Epithamnolia, which was recently placed in Phacidiales as Incertae sedis (Suija et al. 2017). As a result of our investigation, the new genus Aotearoamyces is proposed to accommodate specimens from New Zealand previously identified as Claussenomyces. Aotearoamyces belongs to Tympanidaceae, a recently erected family in Phacidiales. It is differentiated from other Tympanidaceae by its phragmospores with lipid bodies, the lack of conidia or ascoconidia, plectenchymatous excipulum with hyphae strongly spaced and gelatinized, and curved or helicoid paraphyses. Our bibliographic revision resulted in the correction of the authority of Phacidiales. The phylogenetic analyses of combined SSU, ITS and LSU strongly support the relationship between the sexual morph Aotearoamyces and the Collophorina clade containing species with endoconidia, which resulted in the recombination of C. paarla in Aotearoamyces due to the molecular similarities. Also, based on our molecular results, we placed the genus Epithamnolia in the Mniaecia lineage.

Abstract

Fusarium occurs ubiquitously in the world and probably in all known niches. Species have a diversity of functions, and include some of the most important beneficial and detrimental fungi influencing humans. Systematic studies revealed an increasing number of new species, species complexes and molecular locus sequence types (MLST) as more and more species from soils, plants, humans, animals and other niches are characterized. Many Fusarium species are known from South Africa, but these mostly include plant, human and animal pathogens, and those related to food storage problems. Those species known from natural environments are largely understudied, and their ecological roles and potential uses for humans are unknown. In a study comparing Fusarium spp. occurring inside plant tissues and the rhizospheres from six plant species occurring in one location in Bloemfontein (Free State province) a large number of isolates were obtained from a relatively small sample size. When focusing only on isolates grouping in the Fusarium fujikuroi species complex, DNA sequence comparisons of the Translation Elongation Factor 1α and β-Tubulin genes revealed several new species. These included species from the grasses Eragrostis capensis, Themeda triandra and Sporobolus fimbriatus, and the sedge Cyperus dives. This unprecedented diversity, including especially new species reports, emphasizes how poor our knowledge of our native fungi and their geographic and substrate associations are. Bioexploration of these Fusarium species will also significantly improve the international understanding of the classification of this complex group as the addition of more species will improve species resolutions.

Abstract

Ergot fungi (Claviceps spp.) have been noted for their toxicity to animals. In Japan, at least 13 species and seven varieties of Claviceps species have been recorded on gramineous plants. Most of these species are probably indigenous to Japan or East Asia. However, the classification of these Claviceps species needs to be reevaluated based on new data that have emerged from recent studies. Unfortunately, almost all type specimens or cultures of the Claviceps species in Japan have been lost. Therefore, we have collected hundreds of new Claviceps specimens from across Japan to characterize their relationships with their hosts and their phylogenetic positions. To reveal phylogenetic relationships, molecular phylogenetic studies were conducted using ITS-5.8S rDNA, D1/D2 region of 28S rDNA, and TEF 1α gene regions. So far, we have already found 16 distinct groups. One of them is a worldwide C. purpurea var. purpurea, found mainly on pasture grasses. Ergot fungus on Alopecurus, Poa, Dactylis and other some grasses in Japan were included in the C. humidiphila clade, which probably originated from Europe. They were previously recorded as C. purpurea var. alopecuri and C. purpurea var. dactylidis. In the present study, ergot fungus on reed canary grass (Phalaris arundinacea) or timothy (Phleum pratense) in Japan, which was recorded as C. purpurea var. phalaridis, was not included in the European C. humidiphila clade, although it is a type of C. humidiphila. Ergot fungus on Elymus tsukushiensis in Japan, which was previously recorded as C. purpurea var. agropyri, should be a distinct species, althoug it was synonymized with C. purpurea var. purpurea by previous study. Ergot fungus on Leymus mollis in Japan should be a distinct species, althoug it was synonymized with C. purpurea var. purpurea by previous study. Ergot fungus on Sasa spp., which was previously recorded as C. purpurea var. sasae, should be a distinct species. Ergot fungi on reed (Phragmites australis) in Japan are present as two distinct species that are clearly separate from C. arundinis on reeds in Europe. Ergot fungus on Oplismenus in Japan is closely related to Indian C. viridis on Oplismenus. Ergot fungus on Sorghum in Japan is a distinct indigenous species, C. sorghicola. Ergot fungus on Isachne globosa in Japan is a distincet indigenous species, C. panicoidearum. Ergot fungus mainly on Miscanthus spp. should be a distinct species, although it has been treated as C. panicoidearum. However, this fungus is closely related to C. sorghicola on Sorghum. Ergot fungi on Paspalum in Japan are present worldwide, known as C. paspali and C. paspali var. queenslandica. Ergot fungus on Arundinella hirta in Japan is an indigenous species, C. microspora. Ergot fungus on Spodiopogon, which was recorded as C. microspora var. kawatanii, should be a distinct species. We are yet to find speciemens of five further recorded species: C. amamiensis, C. bothriochloae, C. imperatae, and C. yanagawaensis. We will conduct a taxonomic study of these fungi in the future.

Abstract

Isaria Pers. is one of the oldest asexually typified genera of invertebrate fungal pathogens, with its species characterized by the formation of branched synnemata, that give rise to flask-shaped phialides produced in whorls. The phialides tend to possess swollen bases that narrow abruptly forming a distinct neck. Conidia are one-celled, hyaline, smooth, subglobose to subcylindrical and are produced in divergent chains. Aspects of this morphology are shared with other genera, resulting in a turbulent taxonomic history. With Isaria being polyphyletic throughout Cordycipitaceae and rejected in favor of Cordyceps, there is a need to identify the taxonomic placement of isarioid species collected in Thailand. We collected isarioid morphs of invertebrate-pathogenic fungi and other Cordyceps and Torrubiella species infecting coleopterans, lepidopterans and spiders. Sexually reproductive species that produce isarioid morphs in culture and a Torrubiella sp. on a spider, were also included in this study. The purposes of these investigations are to re-examine the taxonomic position of these specimens and to describe new taxa to accommodate the phylogenetic diversity of isarioid fungi. Phylogenetic analyses based on a combined dataset comprising ITS and 28S rDNA, partial sequences of translation elongation factor 1-α gene (TEF1) and the genes for RNA polymerase II largest (RPB1) and second largest (RPB2) subunits were used to clarify their relationships within Cordycipitaceae. A new genus and eight new species, all with isarioid phialides, are described in Cordycipitaceae from Thailand. The new genus, Samsoniella, is segregated from Akanthomyces based on morphological and molecular evidence. Samsoniella differs from Akanthomyces in producing orange cylindrical to clavate stromata with superficial perithecia and orange conidiophores with isarioid phialides and white to cream conidia. A new combination for CBS 240.32 and CBS 262.58, originally identified as Paecilomyces farinosus (Isaria farinosa) and Penicillium alboaurantium, respectively, is made in Samsoniella. Two new species, Samsoniella aurantia and S. inthanonensis are described from lepidopteran larvae. Two new species in Cordyceps, C. blackwelliae and C. lepidopterorum, are also found on coleopteran and lepidopteran larvae. Both species produce isarioid morphs with globose phialides and attenuated long necks and white mycelium in culture. We have established sexual-asexual link of Cordyceps javanica (=Isaria javanica) on lepidopteran larvae. Four new species described in Akanthomyces are pathogenic to spiders or unidentified insect larvae. Akanthomyces kanyawimiae, A. sulphureus, A. thailandicus and A. waltergamsii were found on spiders with some strains of A. kanyawimiae also found on unidentified insect larvae. These four species of Akanthomyces are found on the underside of leaves and produce white to cream white powdery conidia, while S. aurantia and S. inthanonensis were found in the leaf litter and produce bright orange stromata and synnemata with white conidia. Another new combination, Akanthomyces ryukyuensis is also proposed. Our results from molecular phylogenetic analyses strongly support these new species in Cordyceps, Akanthomyces, and in a new genus Samsoniella in Cordycipitaceae.

Abstract

The Neotropical region is considered one of the biological hot spots of the planet. This recognition has been made based on records of amphibians, birds, butterflies, orchids, palms and some groups of insects. However, as in other regions, a consideration of the diversity of fungi has been relegated to the background. Despite the efforts of Latin-American researchers in recent decades to assess the fungal diversity of the region, it is still a distant goal. Within the great diversity of fungal genera there is one which has aroused more interest recently due to its medicinal properties, its high diversity and its curious way of manipulating its hosts. This is the arthropodopathogenic fungus Cordyceps sensu lato (s. l.), an ascomycete with more than 500 species located in three families Cordycipitaceae, Clavicipitaceae and Ophiocordycipitaceae. This group comprises obligate endoparasites that show a high diversity related to the high diversity of its hosts. Because the Neotropical region is rich in diversity of arthropods, one would expect to find a high diversity of Cordyceps. However, to date there has been no survey of the diversity or abundance of species found in the region. The objective of this work was to pull together the large number of records of Cordyceps s.l. from herbariums, public repositories and publications. This review drew information from nine countries, namely, Argentina, Bolivia, Brazil, Colombia, Costa Rica, Ecuador, Guyana, Mexico, Peru, as well as from some collections from Caribbean islands and Central America. Brazil is the country with the greatest number of specimens and records of species, followed by Ecuador and Colombia. The region has yielded 2700 specimens, including 1444 specimens from Brazil of Metarhizium anisoplae and Beauveria bassiana found on crops. Mexico and Colombia have 342 and 238 specimens, respectively, coming from natural forest. In total, 133 different species were reported, of which Brazil has 50 % of the species followed by Ecuador, Bolivia and Colombia with 37%, 35% and 34%, respectively. Ophiocordyceps amazonica, which parasitizes young grasshoppers, is the unique species collected in all the area reviewed except by Mexico where it is not yet reported. In the countries with Amazon rain forest, the pathogens of ants, Ophiocordyceps australis and the Ophiocordyceps unilateralis complex are the most common species found, while Cordyceps militaris, which parasitizes pupae of Lepidoptera, and Tolypocladium capitatum, which parasitizes the false truffle Elaphomyces, have more records in Mexico and Costa Rica where oak forest is a dominant vegetation. 55% of the species reported in the region are representing by a single report for the region, and of them more than half are known only from their original description in the beginning of the 19^th century. Although, this revision does not reflect the natural diversity of the species of Cordyceps s. l. in the Neotropics if shows the dynamics of research in the region. Moreover, this exercise offers a prescription as to where we should prioritize Cordyceps s.l. research in the Neotropical zone and which particular areas require greater emphasis.

Abstract

Four new species of Prolixandromyces (Laboulbeniales, Ascomycota) found on Veliidae (Heteroptera) from Bolivia, Brazil, Peru, and Venezuela are described and illustrated. These four species, Prolixandromyces anseris, P. tritici, P. blackwelliae and P. bromelicola, represent the first records of this genus from South America and their discovery requires emendation of the original generic circumscription. The newly described fungi are compared to known species and a new key to identification is provided.

Abstract

The Zygomycota is a group of related basal clades comprising the subphyla Mucoromycotina, Entomophthoromycotina, Mortierellomycotina, Zoopagomycotina, and Kickxellomycotina. The Mucorales is the largest order of fungi, and classified into the subphylum Mucoromycotina. Many species within the order are used in various important biotechnological areas, such as the production of enzymes and antifungal proteins. Especially, some species are known as causal fungi of human mucormycosis. However, the knowledge about the taxonomy of mucoralean fungi in Korea is limited. In this study, 72 isolates representing 31 species belonging to 13 genera were isolated from different sources, including dung, insect, fruits, freshwater, and soil by using the dilution plating and baiting technique, and standard moist blotter technique. Among of these genera, Mucor presented with highest number of species, and followed by Absidia. The dominant species was Mucor circinelloides and M. hiemalis. Herein, the isolates were identified at the level of species based on the morphological characteristics and sequence analyses of internal transcribed spacer (ITS) region, small subunit (18S rDNA), D1-D2 region of large ribosomal subunit (28S rDNA), translation elongation factor-1α (EF-1α), and actin (act-1) genes. Sixteen species were represented as new records and nine species as new species. Especially, Backusella, Blakeslea, Gilbertella, and Piliobolus species known as rare species were found in Korea.

Abstract

Phaeochorella is a genus of biotrophic leaf-parasitic fungus of tropical distribution. Traditionally the genus was treated as a member of the family Phyllachoraceae/Phyllachoralles, considering exclusively morphological characteristics, without any molecular evidence. The fungus is morphologically well defined producing bright black epiphyllous pseudostromata of irregular shape, often coalescing to cover major portions of the host leaf; the ascospores are characteristically dark brown with an equatorial hyaline band. To elucidate the phylogenetic relationship of Phaeochorella species, with members of the Phyllachorales and other Sordariomycetes, in the present study multilocus analyses were performed. Nuclear rDNA sequences (small subunit nuc 18S, large subunit nuc 28S, and internal transcribed spacer nuc ITS) were used in combination with partial sequences of the nuclear second largest subunit of RNA polymerase II (RPB2), and the translation elongation factor 1 (TEF1). The results clearly showed that Phaeochorella species belong in Diaporthales, forming a clade with high support with the recently described genus Phaeoappendicospora, but without close phylogenetic connection with the other families in Diaporthales. Thus, this new position of the genus suggests the possibility of even a future establishment of a new family, depending on the expansion of the diversity base with the addition of sequences of other Phyllachora-like specimens that may fit in the Diaporthales, as it happened recently with the genus Apiosphaeria, long considered a member of the Phylachoraceae.

Abstract

Many species previously recognized as Phyllachora based exclusively on morphological traits were recently transferred to Telimena Racib. (Telimenaceae, Phyllachorales) according to molecular data, but no specimens from Brazil were included in previous studies. Two species, Phyllachora salaciae Henn. found on Salacia crassifolia (Celastraceae) and an unidentified species found on Serjania sp. (Sapidaceae) from the Brazilian Cerrado were morphologically examined. In addition, had the rDNA (nuclear small subunit nuc 18S, nuclear large subunit nuc 28S and nuclear internal transcribed spacer nuc ITS), the second largest subunit of RNA polymerase II (RPB2), and the translation elongation factor 1 (TEF1) nuclear loci partially sequenced. We investigate the phylogenetic relationship between these species and the currently accepted Phyllachorales with sequences publicly available on GenBank to perform multilocus analyses. The results indicated that both species belong in Telimena. Although the original description of P. salaciae did not identify the host at species level, the specimen presently studied matches perfectly its detailed description shown in the literature. Thus, P. salaciae will be recombined into Telimena salaciae, showing dark amphigenous lesions, up to 7 cm diam., occupying large parts of the leaf blade; pseudostromata dark-brown to black, occupying all the mesophyll extending between both epidermis; internal mycelium well developed, inter- and intra-cellularly, composed by septate hyphae with melanized, thick-walled toruloid to polyhedric cells (6–8 µm diam.); ascomata perithecial, subglobose, conoid to pyriform (114–195 µm diam.), with perifisate short-neck, ostiolate; asci unitunicate, thin-walled, cylindrical to clavate (69–89 × 10–15,5 µm), with rounded apex and inconspicuous apical structures, with 8 uni- or biseriate spores, paraphysate; paraphyses hyaline, filiform; ascospores hyaline, aseptate, fusiform (9–13 × 3–5 µm), guttulate. The specimen found on Serjania sp. proved to be morphologically and phylogenetically completely distinct from the Phyllachora species recorded on members of the family Celastraceae, and from all Telimena species currently recognized and, therefore it constitutes a new species of Telimena.

Abstract

Rice (Oryza sativa L.) is one of the most important crops in Uruguay covering around 170.000 ha with a total production of 1.350.000 tons, which represents approximately 7% of the national agricultural GDP over the past several years. Losses due to diseases are important constraints for rice production with some sclerotia-forming fungi considered at the most important pathogens in Uruguay. This fact is due to the intensification of the crop in the last years and the accumulation of inoculum in rice soils. The pathogens overwinter as sclerotia or mycelium in the soil or in rice crop residues and infect the next crop after flooding. One of these important diseases in Uruguay and other temperate rice producing areas is the sheath spot of rice. Yield losses for sheath spot can reach 9% in susceptible rice cultivar INIA Tacuarí, a japonica tropical type cultivar that covered around 20% of the rice area. This disease is caused by species of basidiomycete fungi in the genus Rhizoctonia or allied genera. Thus, a survey of Rhizoctonia spp. associated with sheath spot of rice was conducted in Uruguay to elucidate the species present, distribution and pathogenicity to Uruguayan rice cultivars. In total, 52 isolates of Rhizoctonia spp. were recovered from diseased rice plants exhibiting sheath spot symptoms. Isolates were obtained in the last 4 years from symptomatic rice plants from all the geographical rice areas of Uruguay. Also, a collection of isolates obtained in the last 15 years from rice soils was studied. These isolates were characterized molecularly, phenotypically and pathogenically. The species identity of the isolates was determined using phylogenetic analysis of the internal transcribed spacer (ITS) region and large sub-unit (LSU) of ribosomal DNA. Pathology studies were conducted under greenhouse conditions. Plants of 12 common cultivars, including testers, were sowed in plastic pots and plants were inoculated with a disc of potato-dextrose agar containing mycelia of each strain. Three pots with five plants by pot were inoculated with each strain. Plants were maintained under greenhouse conditions and evaluated for symptom development in a 0-9 severity degree scale. Two species were identified and confirmed for the rice area of Uruguay by means of ITS sequencing and phenotypical characterization and resulted Rhizoctonia oryzae-sativae and Waitea circinata (=R. oryzae). Other species suspected to occur in rice in Uruguay, including the commonest R. solani, were not found and its occurrence in the country is discarded. Waitea circinata was more common in most rice growing areas, although R. oryzae-sativae was common in certain areas of eastern Uruguay. Rhizoctonia oryzae-sativae was found to be more pathogenic for the cultivars studied in artificial inoculations. Both species were more pathogenic to japonica tropical type cultivars than to indica type cultivars. Waitea circinata isolates exhibited a wide phenotypical variation that deserves a more complete study to recognize types (pathology groups) within the varieties present in rice growing areas of Uruguay.

Abstract

Species within the Xylariaceae (Ascomycota, Sordariomycetes) are predominantly endophytes and/or saprophytes involved in wood decay, although a few species are recognized as plant pathogens. A recently characterized disease of soybean [Glycine max (L.) Merr], referred to as taproot decline, is caused by a Xylaria species nested within the Xylaria arbuscula species complex. However, little is known about the identity, origin, and life history of the species responsible for this emerging disease, and to date no fertile teleomorph has been observed. In order to understand the origin and distribution of this pathogen, isolates of Xylaria were recovered from soybean plant samples exhibiting symptoms of taproot decline collected across the currently known range of the disease in the southern United States (Alabama, Arkansas, Louisiana, Mississippi, and Tennessee). Twenty isolates were used to study the phylogenetic position and genotypic diversity of Xylaria species associated with taproot decline. Pure cultures were obtained from root tissue, and mycelia were harvested for DNA extraction, PCR amplification, and Sanger sequencing of four informative loci: nuclear rDNA internal transcribed spacer (ITS1, 5.8S, ITS2 = ITS), partial α-actin, partial β-tubulin, and partial RNA Polymerase II second largest subunit (RPB2). Sequences were aligned with MAFFT and alignments filtered with GBLOCKS prior to maximum likelihood and Bayesian phylogenetic inference. All of the Xylaria isolates associated with taproot decline are monophyletic and share a most recent common ancestor with Xylaria striata. Specimens of Xylaria from the southern United States deposited in several herbaria are being sequenced to see if a connection can be made between historical teleomorph specimens and isolates from soybean. This work should provide insight into the origin of the taproot decline lineage in the southern United States.

Abstract

The species of Diaporthe (Diaporthaceae, Diaporthales) are well-known as plant pathogens, endophytes or saprobes on the wide range of crops, ornamentals, forest trees and fruit trees. In 2017, Phomopsis stem blight on Tomato (Solanum lycopersicum L.) were occurred in Japan. The initial symptoms of the disease are characterized by necroses on cleavage regions of defoliation, and the disease plants are eventually wilted. Two different Diaporthe species from seven isolates (Diaporthe sp.1: KTH-c11, KTH-c12, KTH-c13; Diaporthe sp.2: KTH-c41, KTH-c42, KTH-c43, KTH-c44) were obtained from the symptoms. The results of our pathogenicity tests using these isolates showed that Diaporthe sp.2 developed more serious symptoms than Diaporthe sp.1. Diaporthe sp.1 are characterized by conidiomata with short necks and cream conidial droplets exuding from central ostioles. Its alpha conidia are cylindrical to ellipsoidal and 4.6-8.0 × 1.9-3.2 µm in size. Beta and gamma conidia are not observed. Diaporthe sp.2 are characterized by conidiomata with long necks and creamy-luteous conidial droplets exuding from central ostioles. Its alpha conidia are ellipsoidal to fusiform and 4.1-7.0 × 1.8-2.9 µm in size. Beta and gamma conidia are frequently observed. Morphological characters of Diaporthe sp.1 and 2 were clearly different from length of conidiomatal neck, shape and length of conidiogenous cells, shape of alpha conidia, and absence/presence of beta and gamma conidia. Our phylogenetic tree of five loci: ITS, tef1, tub2, his3 and cal, also showed that the two species were distantly related, and each species made a monophyletic clade in the genus of Diaporthe. Diaporthe sp.1 was phylogenetically close to D. compacta. However, they were distinguishable from absence/presence of beta conidia, width of conidiogenous cells and shape of alpha conidia. Diaporthe sp.2 was closely related to D. longicolla, but absence/presence of beta and gamma conidia, and the size of alpha conidia of the two species were useful characters to distinguish them. Previously, D. eres, D. vexans, D. phaseolorum, D. sojae and Phomopsis fusiformis have been reported from Tomato. Our two species are, however, clearly different from the five species based on morphological and phylogenetic evidence. We thus introduce two new species of Diaporthe on the stem blight of Tomato.

Abstract

Ixora is a genus of flowering plants in the family Rubiaceae native to the tropical and subtropical regions all over the world. However, its centre of diversity is in tropical Asia. Pestalotiopsis-like species occur commonly as plant pathogens and represent a fungal group known to produce a wide range of chemically novel, diverse metabolites. During a survey of fungal diseases associated with Ixora species in Taiwan, several Pestalotiopsis-like species causing leaf spot were isolated. Based on morphology coupled with single- and multi-gene sequence data of internal transcribed spacer (ITS), partial β-tubulin (TUB) and partial translation elongation factor 1-alpha (TEF) gene regions, revealed two novel taxa of Pseudopestalotiopsis namely Pseudopestalotiopsis ixorae and Ps. taiwanensis. Pseudopestalotiopsis ixorae and Ps. taiwanensis fit well with the generic concept of Pseudopestalotiopsis in having dark concolourous median cells with knobbed apical appendages but differ from the other species of Pseudopestalotiopsis based on the size of the conidiogenous cells, conidia and the number of apical appendages. Pathogenicity tests confirmed that the isolated species are the causal agent of the leaf spots on Ixora.

Keywords: Foliar pathogen, New species, Multi-gene, Phylogeny, Pseudopestalotiopsis, Taxonomy

Abstract

The objective of this work was to perform a scientometric analysis of the publications involving Inonotus rickii (anamorph: Ptychogaster cubensis Pat.), an important pathogen of woody species, causing white rot. The research consisted of a review of the works available in the virtual data bases Periódicos CAPES, Scielo and Science Direct. As search terms, it was used the name of the species, both in the anamorphic and teleomorphic stages. All papers published in English, Spanish or Portuguese were considered, totaling 42 publications, being two books and 40 articles. The articles were distributed in 33 journals, highlighting Forest Pathology (9% of total publications), Plant Disease (7%), Mycotaxon (5%), Mexican Journal of Biodiversity (5%), Mycological Progress (5%). The distribution of the works according to the time included the interval from 1942 to 2016, and shows a linear pattern, ranging from 0 to 5 publications per year. A total of 96 authors were found, most participating in a single study. Only six authors presented more than three papers, with emphasis on Annesi, T., with seven, Motta, E. and Lopes, S.E, with six papers each. The main countries to host the surveys were Argentina, Italy and China, representing 19, 19 and 14% of them, respectively. According to the main focus of the approach, the papers were distributed in five categories: morphological, molecular and biochemical characterization (36.4%); geographical distribution and / or new records of locality (33.3%); Phytopathology, new host records and / or economic losses (19.6%); biotechnology application (9.1%); others (1.5%). The data show that despite five more decades of the first studies, I. rickii is still little studied and devoid of specialists. Most of the work is focused on basic research, usually for taxonomic purposes and restricted to a few countries, thus revealing a gap to be explored, considering its worldwide importance as a pathogen and decomposer of wood, or its potential for application in biotechnological processes.

Abstract

Exobasidium camelliae Shirai and E. camelliae var. gracilis Shirai (current name: E. gracile) described from Japan are well known as pathogenic fungi causing unique symptoms on Camelliae spp. in the world. Exobasidium camelliae infects young flower buds of Camellia and transforms the buds into galls like ball or glove covered by white-powdery hymenium. Exobasidium gracile infects new leaves or shoots of Sasanqua and forms conspicuous watery hypertrophy with white-powdery hymenium inside abaxial epidermis. Although extensive explorations in several Japanese herbaria were conducted, neither any type nor authentic specimens of the two species collected by Shirai were found. Based on art 9.7 in ICN (International Code of Nomenclature for Algae, Fungi, and Plants http://www.iapt-taxon.org/nomen/main.php?page=art9), we therefore neotypified both species with the fresh specimens of E. camelliae and E. camelliae var. gracilis collected from type locality, Tokyo, Japan. Since the description of the two species, several mycologists have pointed out that E. camelliae may produce a few muriform basidiospores and relatively divergent sterigmata on basidia, which were not mentioned in its original descriptions. Currently, exobasidiaceous fungi having muriform basidiospores produced from two or more divergent sterigmata on basidia with probasidial swellings were transferred into the new genus, Muribasidiospora, in Exobasidiaceae. Members of Muribasidiospora are commonly isolated from red leaf spots on Rhus (Anacardiaceae) and are phylogenetically closely related to species of Exobasidium. Our neotype specimens of E. camelliae and E. gracile showed muriform basidiospores from two to four divergent sterigmata on basidia with probasidial swellings. In addition, their neotype cultures were included in the genus Muribasidiospora clade based on ITS and LSU phylogenetic analyses. Therefore, new combinations, Muribasidiospora camelliae and Muribasidiosora gracile, are proposed.

Abstract

To assess the risk of fumonisin contamination in cereals, we isolated Fusarium fujikuroi species complex (FFSC) strains from barley, maize, rice and soybean samples from 2011 to 2015. A total of 878 FFSC isolates were mostly from maize and rice, and species of the isolates were determined using the DNA sequence of the translation elongation factor 1-α (TEF-1α) and RNA polymerase II (RPB2) genes. The rice samples were dominated by F. fujikuroi (47.4%), F. proliferatum (27.3%), and F. concentricum (15.1%), whereas maize samples were dominated by F. verticillioides (33.9%), F. fujikuroi (25.3%), and F. proliferatum (21.1%). Seventy representative isolates were analyzed using a maximum likelihood method, showing each species was genealogically exclusive in the phylogenetic tree. Fumonisin production potential was tested using a PCR assay for FUM1, one of the fumonisin biosynthesis genes. Most of the isolates tested (94%) were positive for FUM1 and all the isolates of F. fujikuroi, F. proliferatum, F. verticillioides and F. thapsinum were positive regardless of their host origin. Seventy-seven representative isolates positive for FUM1 were examined for fumonisin production in rice medium. The majority of F. proliferatum (26/27, 96.3%), F. verticillioides (16/17, 94.1%) and F. fujikuroi (19/25, 76.0%) produced both FB1 and FB2. Out of 19 fumonisin-producing F. fujikuroi, 16 produced> 1000 μg/g of fumonisins (FB1+FB2) in rice medium. These results suggest that F. fujikuroi can produce high levels of fumonisins similar to F. verticillioides and F. proliferatum.

Abstract

Today’s agriculture requires fast, precise and non destructive methods to quantify and discriminate pathogens in plants before the symptoms of disease are visible. VIS/NIR spectroscopy is a non-invasive analytic method which is very appropriate for various agricultural applications due to its fast data acquisition and capacity to determine multiple parameters. The object of this study was to evaluate a quantification and discrimination method of two isolates of Fusarium oxysporum (F05 and F07) in tomato plants using reflectance spectroscopy in the VIS/NIR region. The methodology used shows evidence of an increase of Colony Forming Units (UFC) in the root and leaves of the plant as the incubation time increases, with findings of similar concentrations of the pathogens up to 12 days post infection (dpi). After the first two weeks, a larger growth rate in the roots was observed up to 1590 UFC/gr, but maintaining the high correlations between the UFC measured on both organs (R²=95.60, p=0.01). There were 9 regions in the spectral range of 380-1000nm that showed a high correlation with the increase of the pathogen in the plant, but of the many wavelengths that had high coefficients of determination (R²) with the UFC, only a few also had low p values (<0.05), for example, at 994 nm (R²=89.74 y p=0,01). Finally, the Discriminating Linear Analysis allowed the classification of two strains of F. oxysporum isolated from different plant species, increasing the percentage of precision by increasing the incubation time, with strain F05 the most clearly isolated one through the use of this method. The results suggest a high correlation between the concentrations of the pathogen and the changes in the reflectance spectra in the incubation period of the plant disease, when symptoms are still not visible, and that it is possible to quantify and discriminate the pathogen using few wavelengths determined through reflectance spectroscopy.

Abstract

Soil-borne diseases such as those caused by Fusarium species are of great concern in tomato production worldwide. Two major soilborne diseases, Fusarium wilt (caused by F. oxysporum f. sp. lycopersici (FOL) and Fusarium crown and root rot of tomato (caused by F. oxysporum f. sp. radicis-lycopersici (FORL), are widespread and destructive in major tomato-growing regions worldwide. Based on molecular markers and pathogenicity tests, we isolated and characterized a novel genotype of F. oxysporum f. sp. lycopersici race 3. Under certain conditions, this novel genotype can cause symptoms identical to Fusarium crown and root rot, making field diagnosis of Fusarium wilt and Fusarium crown and root rot difficult. This novel genotype, which most likely came from Florida via-seedborne transmission, belongs to VCG 0033. We demonstrated that FOL race 3 was readily recovery from seeds harvested from infected plants in commercial fields. The incidence of FOL race 3 from tomato seed batches was reduced by ~98% without affecting seed germination or vigor when FOL race 3-infected tomato seed was immersed in a hot water bath for 10 min at 55°C. FOL race 3 was eradicated from seed without reducing seed germination or vigor when seeds were immersed in a fungicide (Azoxystrobin or Fludioxonil) hot-water bath for 10 min at 55°C. Currently, two genotypes exist of FOL race 3, as well as the existence of FOL race 2, and FORL in California. We also demonstrated that Fusarium wilt is an inoculum-dependent disease. At a high inoculum level, a FOL race 3 resistant cultivar was asymptomatically colonized by FOL race 3. Host range experiments revealed that California and Florida genotypes of FOL race 3 are able to sustain themselves on the roots of many plant species, including weeds, but their capability of invading xylem tissue is limited to tomato.

Abstract

Understanding the interactions between plant pathogenic fungi and host plants is vital for the management of disease symptoms. Fusarium graminearum is a fungal plant pathogen and the primary causal agent of Fusarium Head Blight (FHB) on wheat and barley, which reduces quality and quantity of grain yields. FHB is partially controlled by fungicides, with no strong resistance available in host crops. Because of the inefficiency of fungicides in controlling some plant diseases, new methods for disease prevention and treatment need to be established. Plant penetration and colonization are two important stages of infection, which is where we focused our study. Barley has a moderate resistance response where F. graminearum cannot spread from individual infection sites. The plant produces a focal accumulation of plant defense compounds (cellulose and lignin) at the infection sites of trichomes on the surface of barley florets in response to F. graminearum inoculation. Previous work has shown greater numbers of focal accumulations (foci) on barley varieties with small, dome trichomes than varieties with long, prickle-like trichomes. The genetic basis of the differential response seen in trichome morphologies is being investigated through the use of near-isogenic barley varieties. In vitro and in planta assays characterize important plant and fungal responses during infections stages leading to disease development. A locus has been identified in barley as important for the defense response to F. graminearum. Light and confocal laser scanning microscopies are used to visualize these interactions. Genes of interest have been identified as important for F. graminearum – barley interactions. The molecular interactions studied in this work provide new areas to develop disease management tools.

Abstract

Fusarium head blight (FHB) is a worldwide relevant disease that causes important losses in grain yield, quality and safety. FHB in Argentina is mainly caused by Fusarium graminearum sensu stricto when high humidity conditions prevail during wheat anthesis stage. Management strategies to control the disease includes fungicide use, crop rotation, soil tilling and planting of less susceptible varieties, none of them being completely effective. Biological control offers an environmentally friendly tool that could be incorporated in the frame of an integrated pest management. Previous studies by our research group have showed the effectiveness of Bacillus velezensis RC 218 on FHB severity and deoxynivalenol (DON) accumulation. The aim of the present study was to analyze the impact of B. velezensis RC 218 over yield and quality parameters of wheat infected by F. graminearum. Two field trials with a complete randomized design were carried out in Marcos Juárez, Córdoba and Necochea, Buenos Aires, Argentina. During anthesis stage the following treatments were applied: i) Fusarium graminearum; ii) B. velezensis RC 218; iii) F. graminearum + B. velezensis RC 218 and iv) water + 0.05% tween (control). At 21 days post inoculation, FHB incidence and severity were evaluated and grains were harvested at maturity. Yield components, quality parameters, Fusarium-damage kernel (FDK) and DON content were measured. B. velezensis RC 218 showed effectiveness as biocontrol agent in both trials carried out in Marcos Juarez and Necochea. A disease incidence reduction of 29% and 20% was observed in both field trials. Severity was also significantly reduced by 52% and 58%. No significant differences were observed in grain yield nor in 1000-grain weight. A significant reduction of FDK was also observed. The results suggest that the application of B. velezensis RC 218 is a valuable tool to maintain good quality and safety of harvested wheat grains.

Abstract

The coffee berry borer (CBB) causes great damage to coffee crops. When the CBB attacks coffee fruits, it also facilitates the entrance of fungi that may cause coffee berry disease (CBD). It is not known if the fungi colonize through the opening made by the CBB or if the CBB is a vector that introduces them to the fruit. Also, it is not clear which fungi cause CBD; only Colletotrichum spp. have been identified. To determine if CBB is a vector, CBBs were collected from the field and plated on PDA. A total of 158 fungal colonies were isolated and identified using taxonomic keys and DNA sequences. ITS and EF-1 alpha genes were sequenced and compared using BLASTn tool in GenBank. The fungi identified included several species of Fusarium, often considered pathogenic but not known to be pathogens of coffee fruits. Several Fusarium spp. were also isolated from fruits with CBD in the field suggesting that the CBB may be a vector of these fungi. These results are important for the management of CBD in Puerto Rico and other coffee-producing countries.

Abstract

Verticillium dahliae is a soil-bourne ascomycete responsible for Verticillium wilt, a disease that impacts the billion-dollar cotton industry in Australia. Over the years V. dahliae has been classified in several ways including defoliating (D) and non-defoliating (ND) pathotypes, and vegetative compatibility groups (VCGs). In cotton it has appeared that the defoliating VCG1A causes the most severe disease symptoms, while the non-defoliating VCG2A causes less disease symptoms. However, in Australian cotton fields the D VCG1A is causing minimal disease, while the ND VCG2A has been responsible for more severe disease and crop loss. The reason for this difference in virulence expression in Australian isolates compared to global isolates is currently not understood. To get a better understanding of the variation within the NSW V. dahliae collection, PCR analysis of inter-simple sequence repeats (ISSR) was undertaken and the results used to map a dendrogram visualising the relationships between samples, while isolate virulence was analysed in a glasshouse over seven weeks. Australian V. dahliae VCG1A and VCG2A isolates were inoculated into four different cotton varieties at a concentration of 1x10⁶ conidia/ml by root dipping, and disease severity scored twice weekly. The assay revealed that while Australian VCG1As do cause severe damage and mortality, it is not exclusive to the 1A VCG type, with VCG2A also causing plant mortality. The resulting dendrogram showed that NSW V. dahliae isolates do not entirely cluster according to VCG, but seem to also group according to virulence. These results suggest that the disease potential of an isolate is not tied to VCG, and that perhaps a better classification is needed to fully describe V. dahliae isolates. The dendrogram also allows for the identification of virulent and non-virulent V. dahliae samples from the NSW historical collection. This capability will allow for a selection of diverse strains for genome sequencing to help us understand the differences between isolates. This will hopefully allow for the development of better diagnostics for a more rapid on farm response to Verticillium wilt.

Abstract

Eugenia uniflora (Myrtaceae), also known as ‘pitanga’, is a native fruit tree from Brazil cultivated throughout the country in domestic gardens and also commercially in the Brazilian Northeast. Recently, anthracnose has been frequently observed on leaves, flowers and fruits from E. uniflora in Brazil. In 2017 symptomatic plants were collected in the Brazilian region of Distrito Federal, and 9 isolates of Colletotrichum sp. were obtained from lesions affecting leaves, flowers and fruits. Initially, the isolates were identified as a species of the C. gloeosporioides complex, according to the characteristics of conidia (hyaline, aseptate, fusiform with obtuse ends, smooth-walled, guttulate, 13 to 18 µm long and 4.5 to 8 µm wide). After the genomic DNA of all 9 isolates were extracted, sequences were generated for the GAPDH and APMAT regions for an indication of identity of each isolate by means of Bayesian analyses. Subsequently, one isolate of each organ (leaf, flower and fruit) was selected, and the sequences from CAL, TUB2, CHS-1 and ACT were obtained for the phylogenetic analyses. For prove of pathogenicity, Koch’s postulates were performed. Fruits and leaves (adaxial and abaxial surface) of E. uniflora were inoculated with a 5 µl-drop of a suspension of 10⁶ conidia/ml. A drop (5 μl) of distilled water was deposited on control leaves. Fruit rot symptoms were observed on inoculated fruits three days after inoculation and on inoculated leaves five days after inoculation. The control fruits and leaves did not present any symptoms. The fungus was reisolated from the lesion and identified as C. siamense using the methods described above. Although C. siamense has a wide host range, this pathogen was reported only on Acca sellowiana, Coffea sp. and Fragaria × ananassa in Brazil. To our knowledge, this is the first report of C. siamense causing anthracnose on E. uniflora in Brazil. Financial support: FAP-DF, Capes, CNPq and UnB.

Abstract

Colletotrichum is one of the most important groups of pathogenic fungi causing anthracnose in crop plants all over the world. Correct identification and knowledge of the pathogenicity of Colletotrichum spp has great significance to the Australian fruit and vegetable industries, where there are serious biosecurity implications from incursion by new exotic pathogens entering from SE Asia. Recent redefined taxonomy using multigene phylogenetics of the Colletotrichum spp infecting chili fruit (Capsicum) in Australia identified a novel species, C. cairnsense, unique to Australia. The aim of this study was to identify Colletotrichum species isolated from diseased chili fruits collected from Thailand, Indonesia, Malaysia, Sri Lanka and Taiwan. The taxonomy of isolates was confirmed using morphological characters, multi gene phylogenetic analysis and pathogenicity based on disease development in inoculated chili fruit. Taxonomic analysis of isolates from SE Asian countries showed that Colletotrichum scovillei and C. truncatum were the predominant pathogens causing anthracnose in chili. Other Colletotrichum species causing anthracnose in chili included C. fructicola, C. karstii, C. siamense, and four new species from the C. gloeosporioides complex and one potential new species closely related to C. cliviae. The dominant species causing anthracnose of chili in SE Asia was C. scovillei which is yet to be identified in Australia. Although C. siamense had been reported from many plant species, this is the first report of identification of C. siamense causing anthracnose in Thailand, Indonesia and Sri Lanka.This is also the first report of C. fructicola infecting chili from Thailand and Taiwan. Identification of several new species from SE Asia increases the risk of incursion of exotic species to Australia which will have a significant impact on agriculture, biosecurity and quarantine. These results encourage more surveys to be made in chili anthracnose in the future and emphasizes the importance of correct disease diagnostic methods and management practices.

Abstract

There are almost 300 described species of the chytrid genus Synchytrium but currently there is only reference DNA sequence data for 18 of these species. The last systematic treatment of Synchytrium was published by Karling in 1964, pre-dating molecular technologies. This genus includes the species S. endobioticum, which is the causal agent of potato wart disease. This pathogen is a serious threat to trade and is included on the list of select agents of the United States of America. The identification and the publication of sequence data of the most closely related species to this high risk pathogen is essential in order to avoid false positive detection in metagenomics testing that could lead to trade disputes. To accomplish these tasks, tissue of herbarium specimens was sampled from both the Canadian National Mycological Herbarium (DAOM) and the United States National Fungus Collections (BPI). This sampling effort covered 75% of all known species of Synchytrium as well as some unidentified specimens. All available type specimens and a number of specimens observed and utilised by Karling in his morphological examinations of the genus were included. A DNA extraction protocol for herbarium specimens, previously developed for rusts and downy mildews, was optimised for Synchytrium. Portions of the ribosomal cistron (SSU and ITS regions) were amplified and sequenced using Chytridiomycota specific primers. To confirm the identity of the plant host the chloroplastic ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene was also amplified and sequenced. Finally, phylogenetic analyses were performed to assess the relationships of species in the genus and to provide a molecular taxonomic framework for further systematic work on Synchytrium.

Abstract

We examined genetic characterization of 19 isolates identified as Pseudomonas tolaasii by the sequence analysis of 16S rRNA and WLA assay. Also two antagonists against P. tolaasii, HC1 and HC5 were selected and their control efficacy of brown blotch disease was investigated in this study. From the mushrooms that shows symptoms of bacterial brown blotch disease, 180 bacterial strains were isolated, and 19 isolates of them were identified as P. tolaasii with 16S rRNA analysis. Two isolates (CHM01, CHM02) of the 19 isolates did not form white line in white line test. The 19 isolates formed in the same group with analysis of nucleotide sequence similarity of 16S rRNA gene. However, 2 strains (CHM01, CHM02) formed in different group to the rest 17 isolates with analysis of nucleotide sequence similarity of rpoB gene. The patterns of RAPD and REP-PCR of the 2 strains (CHM01, CHM02) were different to the 17 strains. Also, PCR with the primers designed from tolaasin biosynthesis gene did not show any DNA band from the two isolates while specific band was appeared on the 17 isolates by the PCR. By thin layer chromatography (TLC) analyzing of lipopeptide, the 2 isolates (CHM01, CHM02) showed spots of different Rf value to P. tolaasii's. The CHM01 and CHM02 isolates were weaker than the 17 isolates in inhibition of hyphal growth of Pleurotus ostreatus and Flammulina velutipes. Based on these results together, 17 isolates are typical P. tolaasii, whereas CHM01 and CHM02 are considered as low virulent variants of P. tolaasii. Further taxonomic research is needed for correct identification of these two isolates. Control efficacy of brown blotch disease by HC1 treatment was 69% on Agaricus bisporus, 68% on Flammulina velutipes and 55% on pleurotus ostreatus respectively. Control efficacy of brown blotch disease by HC1 treatment was 73% on Agaricus bisporus, 78% on Flammulina velutipes and 71% on Pleurotus ostreatus, respectively.

Abstract

The acquisition of mutualistic hosts may provide symbionts with ecological opportunities, which serve to drive evolutionary diversification. However, it remains unclear and controversial whether the origination of ectomycorrhizal (ECM) symbiosis promotes adaptive diversification of fungi. Importantly, a recent empirical study suggested that the evolution of ECM symbiosis did not necessarily result in adaptive diversification. On the basis of a high-resolution phylogenetic analysis, we herein show how timing of ECM evolution can determine the scale of adaptive divergence in a species-rich fungal lineage. For achieving this, we used nucleotide sequences of 87 single-copy genes and reconstructed the evolutionary history of the fungal order Boletales, which comprise over 1300 species. High-resolution phylogeny of Boletales indicated ECM symbiosis independently evolved at least four times in Boletales, specifically in the stem positions of Suillineae, Sclerodermatineae sensu strict, Paxillaceae, and the clade containing ECM lineages of Boletaceae. Among the four origins, a rapid increase in the net diversification rates (speciation rate minus extinction rate) and a rapid ecological divergence (transition from saprotrophic to mycorrhizal trophic state) appeared to occur only in the clade of Boletaceae, which acquired ECM symbiosis third in time in Boletales. These results contradict the hypothesis that the earliest ECM fungal lineages could have obtained broader niche spaces, as expected by the theory of evolutionary priority effect. Rather, owing to heterogeneity in host availability over space and time, high potentials for allopatric speciation may be available only for "timely innovators", which successfully expanded their geographical ranges via initiating ECM symbiosis with novel hosts (e.g., ECM rosids) that had not been associated with older fungal lineage. Although further studies are still required to corroborate our results and interpretations, the present study contributes to our understanding of how acquisition of ECM hosts could accelerate the evolutionary diversification of fungi.

Abstract

‘Sanghuang’ is a popular fungus used as a Chinese traditional medicine, but little is known about its origin and biogeography. The aim of this study was to characterize the molecular relationships, origin and biogeographical distribution of Sanghuangporus. We used multi-locus phylogenetic analyses to infer the phylogenetic relationships. In addition, based on Bayesian evolutionary analysis using sequences from the internal transcribed spacer (ITS), nuclear large subunit rDNA (nrLSU), translation elongation factor 1-α (EF1-α), and the largest and second largest subunits of RNA polymerase II (RPB1 and RPB2), we used a fungus fossil-based approach to gain insight into the divergence time of species in Sanghuangporus.

The molecular phylogeny strongly supports monophyly of Sanghuangporus (MP = 100%, BS = 100% and BPP = 1.00), and thirteen species are recognized in the genus. The Bayesian uncorrelated lognormal relaxed molecular clock using BEAST and reconstructed ancestral areas indicate that the maximum crown age of Sanghuangporus is approximately 30.85 million years. East Asia is the likely ancestral area (38%). Dispersal and differentiation to other continents then occurred during the late Middle Miocene and Pliocene.

The ancestor of Sanghuangporus probably originated in palaeotropical Northeast Asia and covered Northeast Asia and East Africa during the Oligocene-Miocene, hosted by plants that expanded via the ‘Gomphotherium Landbridge’. Six kinds of dispersal routes are proposed, including intercontinental dispersal events of three clades between Northeast Asia and East Africa, between East Asia and North America, and between Northeast Asia and Europe.

Abstract

Somatic mutation is the process by which novel genetic variation arises within an individual. For most multicellular organisms, somatic mutations are unlikely to be passed along through the germline because of irreversible differentiation. Filamentous fungi, whose cells are totipotent, hold the potential for an exaggerated rate of somatic mutations to enter the germline. This presents a scenario that could result in differential fitness across an individual, i.e. a mosaic of fitness. At small spatial scales, the potential for somatic mutation leading to differences in fitness across and individual is insignificant. However, at large spatial scales, this potential becomes increasingly probable, resulting in the possibility of a selective mosaic across an individual. The ‘Humongous Fungus’ in eastern Oregon is an individual genet of Armillaria ostoyae that covers an estimated 8.8 km², presenting a unique opportunity to test the hypothesis of mosaic fitness and selection in the world’s largest individual organism. Initial sample collection of the Humongous Fungus and a nearby genet was conducted in October 2017. Samples were taken from 15 sites within the area of genet D (Humongous Fungus) to canvas as much of the organism as possible. Along with samples from genet D, 10 samples from genet E, another large Armillaria ostoyae individual nearby, was also sampled for comparison. Samples of infected wood were collected from diseased host trees and fungi cultured from them on 2% malt agar containing chloramphenicol and Benomyl. Each of these cultures represent a section of The Humungous Fungus that occupies a distinct spatial area that is different from the other collected samples. Somatic incompatibility between isolates is being assessed using complementation tests to reproduce earlier results. Whole genome sequencing of these isolates is utilizing a combination of short- (Illumina) and long-read (Oxford Nanopore) technologies to generate a highly contiguous assembly that can be used as a reference to identify SNPs from low-coverage strain-level sequencing using Illumina. The occurrence of SNPs in coding regions will be determined from genome annotations, and synonymous and non-synonymous changes will be recorded for evidence of differences in phenotype.

Abstract

While ectomycorrhizal symbioses are among the most prominent mutualisms in the tree of life repeatedly and convergently evolved among fungi, the genetic mechanisms enabling the origin of an ectomycorrhizal symbiosis remain a mystery. In particular, little is known about the gain of any gene family actively enabling ectomycorrhizal symbiosis maintained across all species of an ectomycorrhizal lineage. Instead, the focus has been gene loss, especially the loss of decomposition genes. Here we identified 109 gene families unique to three diverse ectomycorrhizal Amanita species not found in decomposer Amanita species. When expression profiles are quantified for the ectomycorrhizal Amanita muscaria, genes in these families were more highly expressed during symbiosis compared to the genes in the families shared by both ectomycorrhizal and saprotrophic species suggesting these unique gene families play a functional role in the symbiosis. To understand the origins of these unique gene families, we built a pipeline to identify mechanisms of acquisition. We tested whether each gene family resulted from horizontal gene transfer (HGT), de novo gene creation, or simply represent quickly evolving gene families whose homologs remain unrecognized in decomposer lineages. Two, six and eight gene families were successfully categorized as resulting from de novo gene creation, HGT, or fast-evolving gene families. We also detected high proportions of genes that originated from HGT and fast-evolving gene families, highly expressed in ectomycorrhizal root tips. De novo genes were present in ectomycorrhizal root tips but not highly expressed. Specifically, one gene family derived from HGT encodes a putative 1-aminocyclopropane-1-carboxylate (ACC) deaminase, which can convert ethylene’s precursor, ACC, to alpha-ketobutyrate, and reduce the concentration of ethylene in a plant. In past research, ethylene can reduce ectomycorrhizal symbiosis. This evidence supports that acquiring a new gene family may help fungi transition into a new niche.

Abstract

A systematic investigation of North American pyrophilous species of Pholiota is presented based on morphological and molecular examination of primarily type and other historical collections. Much taxonomic confusion surrounds application of the names P. brunnescens, P. carbonaria, P. castanea, P. fulvozonata, P. highlandensis, P. molesta, and P. subsaponacea with multiple names applied to single species, and multiple species described more than once. Molecular annotations using ITS and rpb2 are used to clarify application of these names in a phylogenetic context. As a result the following heterotypic synonymies are proposed: P. highlandensis (syn. P. carbonaria and P. fulvozonata); P. molesta (syn. P. subsaponacea); and P. brunnescens (syn. P. luteobadia). In addition the species P. castanea, known previously only from the type collection, is found commonly on burned sites in the Gulf Coast and southeast regions of the U.S. Historical collections of this autonomous species were previously referred to as P. highlandensis. Overall, the burn habit appears to have evolved independently at least three times in Pholiota in P. brunnescens, P. castanea, and the complex containing P. highlandensis and P. molesta. Endophytic and endolichenic stages have been deduced for P. highlandensis, the most widely distributed of the four species. Lastly, the ‘body snatcher’ hypothesis is presented that explains the maintenance of some pyrophilous fungi in ecosystems.

Abstract

The enigmatic Ustilaginomycotina lineage Ceraceosorales consists of only one described genus Ceraceosorus. The genus currently has three known species of plant-associated fungi—two producing a plant pathogenic teleomorphic stage and one known only from its saprobic anamorphic stage—each of which is rarely found with limited known geographical distributions. The plant pathogenic species (C. africanus and C. bombacis) are associated with Bombax costatum and B. ceiba (Malvaceae) in Africa and India, respectively. A previous study has shown extreme intragenomic variation of the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA) repeat in C. bombacis and C. guamensis, seemingly presenting a case where concerted evolution of rDNA sequences has failed. The same study also showed complete mismatch between the primers commonly used for ITS amplification with both tested species of Ceraceosorus. Since these findings may have broad implications for metagenomic studies as well as fungal barcoding efforts, it is interesting to deeply investigate this phenomenon on a broader scale by expanding taxon sampling and examined rDNA regions. In this study, we will present a recently discovered anamorphic Ceraceosorus species collected from a healthy leaf phylloplane in Louisiana, USA. We incorporated all known Ceraceosorus species, as well as representatives of other Ustilaginomycotina lineages to investigate the intragenomic variation of the rDNA region through PCR-cloning and high-throughput sequencing. Our analyses revealed that the sequence heterogeneity of the ITS region exists in all examined Ceraceosorus species and thus most likely emerged in the common ancestor to these. The intragenomic variation in Ceraceosorus also expands into the small subunit (SSU) and large subunit (LSU) regions, but detected variant sites in the SSU and LSU regions were significantly less than those found in the ITS region. Similar intragenomic variation of the rDNA region was not found in any other investigated Ustilaginomycotina lineage. We will present findings from this study, as well as discuss a hypothesis for rDNA evolution in Ceraceosorales.

Abstract

Natural rocks and anthropogenic substrates are often colonized by black microcolonial rock-inhabiting fungi (RIF). Due to their polyextremotolerance, RIF can colonize the harshest environments on earth; it was even shown that they can tolerate long-term exposure in outer space. Lichenothelia and Saxomyces are two genera of RIF within the class Dothideomycetes and for which the evolutionary relationships have been recently clarified. Here two Lichenothelia and two Saxomyces species were sequenced on an Illumina platform to generate high-throughput data for draft genome assemblies. The four genomes were assembled and together with 250 Dothideomycetes assemblies (retrieved from NCBI and JGI portals) were mined for orthologous single copy genes using Benchmarking Universal Single Copy Orthologs (BUSCO) pipeline. A comprehensive, genome-based phylogeny of the class is reconstructed both with a maximum likelihood concatenation approach (IQ-TREE) and a multispecies coalescent approach (ASTRAL-II). The minimum amount of genetic information needed to produce a reliable phylogeny is also assessed by multiple runs of an increasing number of random selected orthologs. Tree topologies are evaluated and compared with the concatenation tree (3004 genes, ~2*10⁶ bp alignment) using Internode Certainty (IC), Tree Certainty (TC) and Robinson-Foulds distance metrics. Further, the overall evolutionary rates of fungal genomes are here compared among fungi with different lifestyles (lichens, plant and human pathogens, saprotrophs, RIF) to test whether ecological niches trigger faster or slower genome evolutionary rates. Comparative genomic techniques are also applied using the genomes of the most extremophile fungus Cryomyces antarticus and the lichenized fungus Trypethelium eluteriae to assess whether features related to extremotolerance and lichenization are present in Lichenothelia genome.

Abstract

Cladonia is one of the world’s most charismatic and cosmopolitan lineage of lichens. However, efforts towards explaining why the genus is so widespread and diverse have been stymied due to the lack of a phylogeny with global representation and extensive sampling. This presentation focuses on exploring preliminary research ideas regarding (1) centers of biodiversity for the genus, (2) the use of Next Generation Sequencing to reconstruct evolutionary relationships among species with metagenomic DNA, and (3) future pathways to examine ecological, evolutionary, or historical factors contributing to species radiations.

Abstract

The principal goals of this study are to describe the population structure of lichen Physconia muscigena (Lecanorales) and to investigate possible phylogenetic relationships among the closely related species P. muscigena and P. isidiomuscigena. The project is focused on clarification of the mating system of this lichen (homothalic or heterothalic), on characterization and population analysis of the mating-type genes within the populations. Are there reasons for loss of sexual reproduction in many populations? Additionally we investigate phylogenetic relationships within the genus Physconia (multigene analysis).

Abstract

A major challenge to evolutionary biologists is to understand how biodiversity is distributed through space and time and across the tree of life. Diversification of organisms is influenced by many factors that act at different times and geographic locations but it is still not clear which have a significant impact and how drivers interact. To study diversification, we chose the lichen genus Sticta, by sampling through most of the global range and producing a time tree. We estimate that Sticta originated about 30 million years ago, but biogoegraphic analysis was unclear in estimating the origin of the genus. Furthermore, we investigated the effect of dispersal ability finding that Sticta has a high dispersal rate, as collections from Hawaii showed that divergent lineages colonized the islands at least four times. Symbiont interactions were investigated using BiSSE to understand if green-algal or cyanobacterial symbiont interactions influenced diversification, only to find that the results were driven almost completely by Type I error. On the other hand, another BiSSE analysis found that an association with Andean tectonic activity increases the speciation rate of species.

Abstract

Brazil is the country with the highest lichen biodiversity on earth, but much of it remains as yet undiscovered. A field trip to rock outcrops in a remnant of Atlantic rain forest in Alagoas, viz. Reserva Biólogica de Pedra Talhada, near the city of Quebrangulo, in the middle of the semiarid region in Northeast Brazil, revealed an unusual fruticose lichen. It forms dense mats on siliceous rock that is influenced by run-off water. It typically grows at the upper ends of gullies that are occupied lower down by cyanophilic lichens such as Peltula and Jenmania. The habitat is extremely poikilohydric; this lichen is occasionally submerged, but usually completely dry. The morphology of this species is enigmatic: it somewhat resembles a Leprocaulon s.lat., but it differs from all known species in that genus by the chemistry; apothecia are not present, but pycnidia are not uncommon. The chemistry is gyrophoric acid, which was not known from any fruticose lichen with chlorococcoid algae. Sequence work places the new species right in the Trapeliaceae, close to or most probably inside Trapeliopsis, with which it indeed shares the chemistry. We propose to describe it in Trapeliopsis, and name the species in honour of the owner of the reserve, Anita Studer, as T. studerae. It represents a new and independent lineage of fruticose lichens.

Abstract

Ascomycota has the highest number of species within the kingdom Fungi, with about one third of these species being lichenized. A particular group of lichenized fungi are those that dwell on leaves of vascular plants, the so-called foliicolous lichens. Gomphillaceae is the most species-rich family of foliicolous lichens, but also contains species growing on other substrata, such as bark, rocks, and soil. Despite that the family features many morphological peculiarities, in particular its conidiomata, the hyphophores, the genus-level classification of Gomphillaceae remained thus far unresolved. This study revised the diversity of foliicolous lichens in the main Brazilian biomes, with emphasis on Gomphillaceae, and made a first attempt at assembling a broad molecular phylogeny using the mitochondrial small subunit and nuclear large subunit rDNA markers (mtSSU, nuLSU), based on new collections from leaves made in areas of Caatinga (in higher altitude forest enclaves), Atlantic Forest, and Amazon forest, completed by collections from other areas in the neotropics (Mexico, Guatemala, Costa Rica, Panama, Cuba). To complete the analysis, we used the method of morphology-based phylogenetic binning, which places species that do not have molecular data into the molecular phylogeny based on phylogeny-weighted assessment of the morphological characters. Four hundred and sixty-seven sequences were obtained for the mtSSU and nuLSU, from three hundred and thirty-seven specimens. As a result, we describe 11 genera: Spinomyces (Aderkomyces albostrigosus group), Roselviria (Aderkomyces purulhensis group), Serusiauxiella (Tricharia farinosa group), Caleniella (Calenia triseptata group), Lumbrispora (Echinoplaca diffluens group), Aptrootidea (Echinoplaca marginata group), Adelphomyces (Gyalideopsis epithallina group), Batistomyces (Tricharia hyalina group), Sipmanidea (Echinoplaca verrucifera group), and Bezerroplaca (Echinoplaca lucernifera group). We also reinstate four genera and propose new combinations in Linhartia (Psorotheciopsis patellarioides group), Microxyphiomyces (Tricharia vainioi group), Psathyromyces (Aderkomyces heterellus group), and Sporocybomyces (Echinoplaca leucotrichoides). This changes the number of genera recognized within the family from twenty-four to approximately forty.

Abstract

Usnea (Parmeliaceae) is a species rich lichen genus with more than 350 species worldwide. It is characterized by a fruticose habit, an elastic central chord inside the branches and the presence of usnic acid in the cortex. Many species are considered to have wide distributions. Usnea in Africa has been insufficiently studied and the availability of molecular data is very limited. The taxonomy of Usnea is difficult due to the high morphological plasticity and chemical variability of the species. Therefore, molecular data is much needed in order to ascertain species recognition. In this study, a dataset of sequences of five nuclear markers (ITS, LSU, SSU, MCM7 and Beta-tubulin) of Usnea samples from Tanzanian mountainous rainforests were generated. These were used for elucidating their phylogenetic relationships in a wider sampling of Usnea, covering many of the Usnea sections. Bayesian analysis was used to infer phylogenetic relationships among the species. The morphology of the samples was assessed and important features such as apothecia and the occurrence and type of soralia, isidiomorphs and fibrils were recorded, along with features such as the cortex and medulla pigmentation, the branch surface characteristics and the branch anatomy. Thin layer chromatography was used to investigate the secondary chemistry. In this study a presentation of the phylogeny of some Tanzanian Usnea from mountainous rainforests is presented, which based on sequence data suggests species identifications and species circumscription.

Abstract

Lichenicolous fungi are pathogens, saprotrophs or commensals which grow obligately on lichens. The lichenicolous habit has evolved many times, with species found in many families across the Ascomycota, the phylum to which the majority of lichenicolous fungi belong. They are, however, often missing from the general consciousness of fungal strategies, and so provide a novel avenue of phylogenetic research regarding fungal diversity and evolution. In this study, a range of lichenicolous fungi from various localities and lichen hosts have been sequenced, including new species, and molecular data from previously sequenced lichenicolous species have been collated from literature. Species were placed in a broad, multilocus phylogeny of the Ascomycota, alongside diverse taxa such as endolichenic, mycoparasitic and rock-inhabiting fungi. An ancestral state reconstruction was performed to show how a lichenicolous perspective influences the big picture of fungal evolution within the Ascomycota.

Abstract

Species circumscription is key to characterize patterns of specificity in symbiotic systems. Here, a phylogenetic framework was used to assess the biodiversity and symbiotic patterns of association among partners of the trimembered lichens in the genus Peltigera. We used a worldwide sampling to explore phylogenetic relationships among taxa included in the section Chloropeltigera. We sequenced six mycobiont loci and performed analyses using species discovery and validation methods to establish species boundaries. Single locus phylogenies were used to establish the identity for both, Nostoc and Coccomyxa photobionts. Distribution and specificity patterns were examined across all Peltigera species from sections Chloropeltigera and Peltidea. The possible role of the reproductive mode (sexual versus asexual) shaping these patterns was explored. Eight fungal species (including five newly delimited) were found in association with nine Nostoc phylogroups and two Coccomyxa species for section Chloropeltigera. In contrast, eight fungal (including three newly delimited) species in section Peltidea were found in association with only four Nostoc phylogroups and the same two Coccomyxa species as for section Chloropeltigera. This difference in cyanobiont biodiversity found between these two sections of the genus Peltigera can potentially be explained by a different type of reproduction driving cyanobiont transmission (vertical versus horizontal) in each section. We found a significantly higher frequency of sexual reproductive structures and higher number of ITS haplotypes in species from section Chloropeltigera compared to species from section Peltidia. This suggests that horizontal transmission might be more prevalent in Chloropeltigera species, while vertical transmission might be more common in Peltidea species. All species in section Chloropeltigera are generalists in their association with Nostoc compared to more specialized Peltigera species in section Peltidea. Constrained distributions of Peltigera species that associate strictly with one species of green algae (Coccomyxa subellipsoidea) demonstrates that the availability of the green alga and the specificity of the interaction might be important factors limiting geographic ranges of trimembered Peltigera, in addition to limitations imposed by their interaction with Nostoc partners.

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Ten editions of the Dictionary of the Fungi were published regularly by the Imperial Mycological Institute and its successor organisations from 1943, providing authoritative information on accepted genera and their diversity (within a much-used higher classification) along with a glossary and biographical entries for mycologists. It is ten years since the last edition of the Dictionary was published. It was avowedly “the last ‘ink-on-paper’ version”, which prompts the question: how can the mycological community build on the ‘marvellously imperfect’ Dictionary and create maximally useful on-line resources? Glossaries and biographies could be implemented via on-line collaborative editing spaces (such as a wiki). Dealing with names and classifications is more challenging; especially providing accepted names within the one classification, which is what most users want. While there are significant existing on-line resources (such as Index/Species Fungorum, MycoBank, GenBank, U.S. National Fungus Collections and NZFungi) there is currently much wasteful effort by users in determining the ‘correct’ name and placement of fungi. There is also misunderstanding of the difference between lists of names (nomenclators, such as Index Fungorum) and lists of taxa (compilations of accepted names with their synonyms, such as Species Fungorum). For names, there should be one version of each name and its associated publication and typification details, compliant with the International Code of Nomenclature for algae, fungi and plants. However, existing nomenclators are not complete, and different databases can have different citation details and even different spellings of names. Full population and checking of nomenclators would benefit from citizen science approaches where volunteers are mobilised, while efficiently utilising scarce nomenclatural and orthographic expertise. For name databases, there is considerable support from hosting institutions and massive voluntary input by key individuals, but a sustainable system is yet to emerge due to lack of clarity around the role and integration of existing databases. For taxa, circumscriptions (such as the delimitation of a species or which genera belong to which families) are opinions by taxonomists, and not governed by rules, but only by peer review (as to whether they appear in the first place) and uptake by mycologists. There are challenges in integrating taxa that lack molecular data into classifications that increasingly do not have morphological synapomorphies. Ideally, knowledgeable mycologists would engage with a simple on-line system that generates a global consensus classification, available at any point in time via web services, with version control. Organisationally, an expanded role of the International Commission on the Taxonomy of Fungi (ICTF) can be envisaged, building on the working groups that have successfully guided the one fungus – one name transition. In addition, existing provisions of the Code could be utilised to create lists of protected names to stabilise taxonomy of important groups. For governance, the International Mycological Association could play an increased role through its ownership of MycoBank. Across both names and taxa, it is vital to trap metadata behind decisions, both nomenclatural (e.g. which spelling is adopted) and taxonomic (which circumscription is adopted) to provide clarity and avoid re-covering old ground in the future.