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1.
An Acad Bras Cienc ; 96(suppl 1): e20230251, 2024.
Article in English | MEDLINE | ID: mdl-39292101

ABSTRACT

The diversity of endophytes and their ecological relationships with the endangered conifer Araucaria angustifolia (a critically endangered species) are unrevealed. This study aimed to characterize the diversity of endophytic fungi associated with A. angustifolia. To this end, we analyzed 90 fragments from five individuals collected from a mixed localized fragment in Guarapuava-PR, Brazil. The total DNA of 61 morphotypes was extracted and the Internal Transcribed Spacer (ITS) region was amplified and sequenced. The sequence analysis allowed the identification of 37 genera belonging to the phylum Ascomycota and the classes Eurotiomycetes, Dothideomycetes, and Sordariomycetes, divided into 11 orders and 13 families. Most of the isolated fungi belonged to the Sordariomycetes class (40%) and to the Xylaria genus (14%), while Eurotiomycetes was the minority class within the community. Our results reveal the high endophytic richness supporting the life cycle of A. angustifolia and reinforce the necessity for the conservation of this conifer, as many genetic resources can be lost owing to its irrational exploration.


Subject(s)
Araucaria , Endophytes , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Brazil , Araucaria/microbiology , DNA, Fungal/genetics , Biodiversity , Ascomycota/classification , Ascomycota/isolation & purification , Ascomycota/genetics , Phylogeny , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , Sequence Analysis, DNA
2.
BMC Microbiol ; 24(1): 356, 2024 Sep 19.
Article in English | MEDLINE | ID: mdl-39300361

ABSTRACT

BACKGROUND: Maple is an important ornamental plant in China. With the increasing use of maple trees in landscaping, a symptom of shoot dieback has been observed in Henan province, China. RESULTS: In this study, 28 Diaporthe isolates were obtained from symptomatic shoots of maple trees between 2020 and 2023. Phylogenetic analyses based on five loci (ITS, TEF, CAL, HIS and TUB) coupled with morphology of 12 representative isolates identified three known species (D. eres, D. pescicola and D. spinosa) and one new species, namely D. pseudoacerina sp. nov. Koch's postulates confirmed that all these species were pathogenic. Additionally, D. pseudoacerina was able to infect China wingnut (Pterocarya stenoptera), pear (Pyrus sp.), and black locust (Robinia pseudoacacia). This study marks the first report of Diaporthe spinosa and D. pescicola pathogens infecting maple trees. CONCLUSIONS: These findings enhance the existing knowledge of the taxonomy and host diversity of Diaporthe species as, while also providing valuable information for managing of maple shoot dieback in Henan Province, China.


Subject(s)
Acer , Ascomycota , Phylogeny , Plant Diseases , Plant Shoots , Acer/microbiology , China , Plant Diseases/microbiology , Plant Shoots/microbiology , Ascomycota/genetics , Ascomycota/classification , Ascomycota/isolation & purification , Ascomycota/physiology , DNA, Fungal/genetics , Sequence Analysis, DNA , Pyrus/microbiology
3.
Ying Yong Sheng Tai Xue Bao ; 35(6): 1689-1694, 2024 Jun.
Article in Chinese | MEDLINE | ID: mdl-39235028

ABSTRACT

We isolated the dark septate endophytic (DSE) fungi from roots of typical plant species in the tundra of Changbai Mountains Nature Reserve, including Rhododendron aureum, R. conferentiatum, Vaccinium uliginosum, and Dryas octopetala, and studied their colonization. We further investigated the DSE community composition and species diversity of the four tundra plant species by using morphological characteristics combined with rDNA ITS sequence analysis. The results showed that DSE formed a typical structure of "microsclerotia" in roots of the four plant species. A total of 69 strains of DSE fungi were isolated from the root samples, belonging to 10 genera, and 12 species. They were Phialocephala fortinii, Alternaria alternata, A. tenuissima, Epicocum nigrum, Canariomyces microsporus, Colletotrichum spaethianum, C. camelliae, Leptophoria sp., Cladosporium cladosporioides, Phoma sp., Cadophora sp., and Discosia italica, respectively. The DSE fungal species diversity was rich, and all these fungal species were firstly reported as DSE fungi in the alpine tundra belt of China. Among them, Phialocephala fortinii was the common and dominant species of all tundra plants. The Simpson, Pielou, and Shannon diversity indices of DSE fungi of the four plant species of tundra differed significantly. Our results showed that tundra plants have rich diversity of DSE fungi, and they can form a good symbiotic relationship, which enhance the adaptability of tundra plants to the harsh environment.


Subject(s)
Biodiversity , Endophytes , Plant Roots , Endophytes/isolation & purification , Endophytes/classification , Endophytes/genetics , China , Plant Roots/microbiology , Rhododendron/microbiology , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , Vaccinium/microbiology , Ascomycota/isolation & purification , Ascomycota/classification , Ascomycota/genetics , Ecosystem , Alternaria/isolation & purification , Alternaria/classification , Alternaria/genetics
4.
Fungal Biol ; 128(6): 2062-2072, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39174241

ABSTRACT

Eucalyptus spp. in plantations are negatively affected by canker and wilt diseases caused by several species of Ceratocystis, particularly those in the Latin American Clade (LAC). Ceratocystis eucalypticola and Ceratocystis manginecans are of particular concern where disease epidemics are reported globally, with recent outbreaks emerging in South African and Indonesian Eucalyptus plantations. Consequently, a rapid screening protocol is required for these pathogens. In this study, a high-resolution melting curve analysis (HRMA) was developed to detect C. eucalypticola and C. manginecans that bypasses time-consuming isolation and post-PCR procedures. Primers targeting a 172 bp region of the cerato-platanin (CP) gene were designed. Using these primers, the accuracy of HRMA to detect and distinguish between these two LAC species was assessed using pure fungal DNA, and DNA extracted directly from Eucalyptus samples naturally infected with C. eucalypticola. The assay accurately detected the presence of C. eucalypticola and C. manginecans and quantifies their DNA, both from cultures, and directly from wood samples. HRMA further differentiated these two species from all other tested LAC individuals. This assay was also able to detect the presence of all the tested LAC species and distinguish seven of these, including C. fimbriata, to species level. Ceratocystis polyconidia was the only non-LAC off-target species detected. Based on these results, the developed assay can be used to rapidly identify C. eucalypticola and C. manginecans directly from infected plant material or fungal cultures, with the potential to also screen for several other LAC species.


Subject(s)
Ascomycota , DNA, Fungal , Eucalyptus , Plant Diseases , Eucalyptus/microbiology , Plant Diseases/microbiology , Ascomycota/genetics , Ascomycota/isolation & purification , Ascomycota/classification , DNA, Fungal/genetics , DNA Primers/genetics , Transition Temperature , Sensitivity and Specificity
5.
Sci Rep ; 14(1): 19817, 2024 08 27.
Article in English | MEDLINE | ID: mdl-39191814

ABSTRACT

Botryosphaeriaceae species are the major causal agents of walnut dieback worldwide, along with Diaporthe species. Botryosphaeria dothidea and Neofusicoccum parvum are the only two Botryosphaeriaceae species associated with this recently emergent disease in France, and little is known about their diversity, structure, origin and dispersion in French walnut orchards. A total of 381 isolates of both species were genetically typed using a sequence-based microsatellite genotyping (SSR-seq) method. This analysis revealed a low genetic diversity and a high clonality of these populations, in agreement with their clonal mode of reproduction. The genetic similarity among populations, regardless of the tissue type and the presence of symptoms, supports the hypothesis that these pathogens can move between fruits and twigs and display latent pathogen lifestyles. Contrasting genetic patterns between N. parvum populations from Californian and Spanish walnut orchards and the French ones suggested no conclusive evidence for pathogen transmission from infected materials. The high genetic similarity with French vineyards populations suggested instead putative transmission between these hosts, which was also observed with B. dothidea populations. Overall, this study provides critical insight into the epidemiology of two important pathogens involved in the emerging dieback of French walnut orchards, including their distribution, potential to mate, putative origin and disease pathways.


Subject(s)
Ascomycota , Genetic Variation , Juglans , Microsatellite Repeats , Plant Diseases , Juglans/microbiology , Ascomycota/genetics , Ascomycota/classification , France , Plant Diseases/microbiology , Microsatellite Repeats/genetics , Genotype
6.
BMC Microbiol ; 24(1): 305, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-39148068

ABSTRACT

BACKGROUND: Phylogeographic studies have gained prominence in linking past geological events to the distribution patterns of biodiversity, primarily in mountainous regions. However, such studies often focus on plant taxa, neglecting the intricate biogeographical patterns of microbes, particularly soil microbial communities. This article explores the spatial distribution of the nematode-trapping fungus Arthrobotrys oligospora, a widespread microorganism, in a tectonically active region at the southeastern edge of the Qinghai-Tibetan Plateau. By analysing the genetic variation of this fungus alongside the historical structure of major river watersheds, we sought to uncover potential connections between the two. Our study involved sampling 149 strains from 116 sites across six major watersheds in the region. RESULTS: The resulting haplotype network revealed five distinct clusters, each corresponding closely to a specific watershed. These clusters exhibited high haplotype diversity and low nucleotide diversity, supporting the notion of watershed-based segregation. Further analysis of haplotypes shared across watersheds provided evidence for three proposed past river connections. In particular, we found numerous shared haplotypes between the Yangtze and Mekong basins, as well as between the Yangtze and the Red basins. Evidence for a Irrawaddy-Salween-Red and a Yangtze-Pearl-Red river connections were also portrayed in our mapping exercise. CONCLUSIONS: These findings emphasize the crucial role of historical geomorphological events in shaping the biogeography of microbial biodiversity, alongside contemporary biotic and abiotic factors. Watershed perimeters emerged as effective predictors of such patterns, suggesting their suitability as analytical units for regional-scale studies. Our study also demonstrates the potential of microorganisms and phylogeographic approaches to complement traditional geological analyses, providing a more comprehensive understanding of past landscape structure and its evolution.


Subject(s)
Genetic Variation , Haplotypes , Phylogeny , Phylogeography , Rivers , Soil Microbiology , China , Rivers/microbiology , Ascomycota/genetics , Ascomycota/classification , Ascomycota/isolation & purification , Biodiversity , DNA, Fungal/genetics
7.
Mycologia ; 116(5): 835-847, 2024.
Article in English | MEDLINE | ID: mdl-38959129

ABSTRACT

Pinibarrenia chlamydospora, sp. nov. isolated from the roots of highbush blueberry in the New Jersey Pine Barrens, is described and illustrated. Based on multigene phylogenetic analysis, as well as morphological and ecological characteristics, Pinibarreniales and Pinibarreniaceae are established to accommodate this novel lineage in Sordariomycetidae, Sordariomycetes. Pinibarreniales, Tracyllalales, and Vermiculariopsiellales are proposed to be included in the subclass Sordariomycetidae. Pinibarreniales likely have a wide distribution and forms association with Ericaceae plants that live in acidic and oligotrophic environments because its DNA barcode matches with environmental sequences from other independent ecological studies. The plant-fungal interaction experiment revealed negative impacts on Arabidopsis, indicating its pathogenicity. This uncovered new fungal lineage will contribute to a better understanding of the diversity and systematics of Sordariomycetes.


Subject(s)
Ascomycota , Ecosystem , Phylogeny , Plant Roots , Plant Roots/microbiology , Ascomycota/classification , Ascomycota/genetics , Ascomycota/isolation & purification , DNA, Fungal/genetics , New Jersey , Arabidopsis/microbiology , Blueberry Plants/microbiology , Plant Diseases/microbiology , Sequence Analysis, DNA , Pinus/microbiology , DNA Barcoding, Taxonomic
8.
Mycologia ; 116(5): 650-658, 2024.
Article in English | MEDLINE | ID: mdl-39024179

ABSTRACT

Fossils can unveil a long-vanished combination of character states that inform inferences about the timing and patterns of diversification of modern fungi. By examining the well-preserved stacked chained vesicular conidiophores developed in clusters from the basal stroma, we describe a new taxon of fossil Zygosporiaceae with a combination of characters unknown among extant taxa on compressed serrated-margined dicot leaf (cf. Fagaceae) recovered from the Siwalik sediments (Late Miocene; ca. 12-8 Ma) of Himachal Pradesh, western Himalaya. Based upon conidiophore morphology, our Siwalik fungal remains, similar to Zygosporium Mont. (Zygosporiaceae: Xylariales: Sordariomycetes), are recognized as a new fossil species, Z. stromaticum Kundu & Khan, sp. nov. Zygosporium stromaticum is the only known fossil anamorphic fungus that occurs on plant cuticles and has a cluster of stacked chained vesicular conidiophores arising from a poorly preserved basal stroma formed by irregular, thick-walled cells. Its combination of morphological characteristics is unknown in extant fungal taxa, so Z. stromaticum likely represents a new anamorphic foliicolous fungus that may now be extinct. This unique evidence may be essential for the calibration of divergence time estimations of fungal lineages.


Subject(s)
Fossils , Fossils/microbiology , India , Plant Leaves/microbiology , Spores, Fungal/cytology , Spores, Fungal/classification , Ascomycota/classification , Ascomycota/genetics , Ascomycota/isolation & purification , Phylogeny
9.
Braz J Microbiol ; 55(3): 2845-2854, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38951477

ABSTRACT

Cotton (Gossypium hirsutum, Malvaceae) is the most important fiber crop in the world. There are published records of many fungal pathogens attacking Gossypium spp., causing numerous diseases, including powdery mildews. Recently, in 2022, non-cultivated spontaneous G. hirsutum plants bearing powdery mildews symptoms were found at roadsides in two municipalities of the state of Minas Gerais (Brazil): Varginha and Ubá. Such localities are situated ca. 260 km apart, suggesting a broader distribution of this fungus-host association in Brazil. Samples were taken to the laboratory, and an Ovulariopsis-like, asexual stage of Phyllactinia, was identified forming amphigenous colonies, that were more evident, white and cottony, abaxially. Morphological and molecular data- of the ITS and LSU regions- have shown that colonies from those two samples were of the same fungus species, belonging to a previously unknown species of Erysiphaceae (Ascomycota). The fungus fits into the Phyllactinia clade and is described herein as the new species Phyllactinia gossypina sp. nov. This new species belongs to the 'basal Phyllactinia group', a lineage that includes species known only from the Americas. This report expands the list of pathogenic fungi on cotton. It is early to anticipate whether this new powdery mildew represents a threat to cultivated cotton, which is a major crop in Brazil. Nevertheless, further studies about its infectivity to commercial cotton varieties are recommended, since all known Erysiphaceae are specialized obligate plant parasites and several species cause major losses to important crops.


Subject(s)
Gossypium , Phylogeny , Plant Diseases , Gossypium/microbiology , Brazil , Plant Diseases/microbiology , Ascomycota/classification , Ascomycota/isolation & purification , Ascomycota/genetics , Ascomycota/physiology , DNA, Fungal/genetics
10.
J Food Sci ; 89(8): 4745-4757, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38955792

ABSTRACT

Food fraud is a problematic yet common phenomenon in the food industry. It impacts numerous sectors, including the market of edible mushrooms. Morel mushrooms are prized worldwide for their culinary and medicinal use. They represent a taxonomically complex group in which food fraud has already been reported. Among the methods to evaluate food fraud, some rely on comparisons of genetic sequences obtained from a sample to existing databases. However, the quality and usefulness of the results are limited by the type of comparison tool and the quality of the database used. The Centroid-based approach is applied by SmartGene in a proprietary artificial intelligence-based method for the generation of automatically curated reference databases that can be further expert curated. In this study, using sequences of the ribosomal internal transcribed spacer (ITS) of the genus Morchella (true morels), we compared this approach to the traditional pairwise alignment tool using two other databases: UNITE and Mycobank (MLST). The Centroid-based approach using an expert-curated database was more performant for the identification of 53 representative ITS sequences corresponding to validated species (83% accuracy, compared to 36% and 47% accuracy for UNITE and MLST, respectively). The Centroid method also revealed an inaccurate taxonomic annotation for sequences of commercial cultivars submitted to public databases. Combined with the web-based commercial software IDNS® available at Smartgene, the Centroid-based approach constitutes a valuable tool to ensure the quality of morel products on the market for actors of the food industry. PRACTICAL APPLICATION: The Centroid-based approach can be used by agri-food actors who need to identify true morels down to the species level without any prior taxonomical knowledge. These include routine laboratories of the food industry, food distributors, and public surveillance agencies. This is a reliable method that requires minimal skills and resources, therefore being particularly adapted for nonspecialists.


Subject(s)
Ascomycota , Ascomycota/genetics , Ascomycota/classification , DNA, Fungal/genetics , Food Contamination/analysis , DNA, Ribosomal Spacer/genetics
11.
Antonie Van Leeuwenhoek ; 117(1): 109, 2024 Jul 31.
Article in English | MEDLINE | ID: mdl-39083124

ABSTRACT

The genetic variety and habitats of Camptophora species, generally known as black yeast, have not been clarified. In this study, we re-evaluated Camptophora based on morphological observations and phylogenetic analyses. Because prior investigations on Camptophora only included a few strains/specimens, 24 Camptophora-related strains were newly obtained from 13 leaf samples of various plant species to redefine the genetic and species concepts of Camptophora. Their molecular phylogenetic relationships were examined using small subunit nuclear ribosomal DNA (nSSU, 18S rDNA), the internal transcribed spacer (ITS) rDNA operon, the large subunit nuclear ribosomal DNA (LSU, 28S rDNA), ß-tubulin, the second largest subunit of RNA polymerase II (rpb2), and mitochondrial small subunit DNA (mtSSU). Single- and multi-locus analyses using nSSU-ITS-LSU-rpb2-mtSSU revealed a robust phylogenetic relationship among Camptophora species within Chaetothyriaceae. Camptophora species could be distinguished from other chaetothyriaceous genera by their snake-shaped conidia with microcyclic conidiation and loosely interwoven mycelial masses. Based on the results of phylogenetic analyses, two undescribed lineages were recognized, and Ca. schimae was excluded from the genus. ITS sequence comparison with environmental DNA sequences revealed that the distribution of the genus is restricted to the Asia-Pacific region. Camptophora has been isolated or detected from abrupt sources, and this was attributed to its microcycle. The mechanisms driving genetic diversity within species are discussed with respect to their phyllosphere habitats.


Subject(s)
DNA, Fungal , Phylogeny , DNA, Fungal/genetics , DNA, Ribosomal/genetics , DNA, Ribosomal Spacer/genetics , Spores, Fungal/genetics , Spores, Fungal/cytology , Spores, Fungal/classification , Sequence Analysis, DNA , Plant Leaves/microbiology , RNA Polymerase II/genetics , Ascomycota/genetics , Ascomycota/classification , Tubulin/genetics
12.
BMC Microbiol ; 24(1): 243, 2024 Jul 04.
Article in English | MEDLINE | ID: mdl-38965478

ABSTRACT

BACKGROUND: Lichens, traditionally considered as a simple partnership primarily between mycobiont and photobiont, are, in reality, complex holobionts comprised of a multitude of microorganisms. Lichen mycobiome represents fungal community residing within lichen thalli. While it is acknowledged that factors like the host lichen species and environmental conditions influence the structure of the lichen mycobiome, the existing research remains insufficient. To investigate which factor, host genus or location, has a greater impact on the lichen mycobiome, we conducted a comparative analysis of mycobiomes within Parmelia and Peltigera collected from both Turkey and South Korea, using high-throughput sequencing based on internal transcribed spacer region amplification. RESULTS: Overall, the lichen mycobiome was dominated by Capnodiales (Dothideomycetes), regardless of host or location. At the order level, the taxonomic composition was not significantly different according to lichen genus host or geographical distance. Hierarchical clustering of the top 100 abundant ASVs did not clearly indicate whether the lichen mycobiome was more influenced by host genus or location. Analyses of community similarity and partitioning variables revealed that the structure of the lichen mycobiome is more significantly influenced by location than by host genus. When analyzing the core mycobiome by host genus, the Peltigera mycobiome contained more ASV members than the Parmelia mycobiome. These two core mycobiomes also share common fungal strains, including basidiomycete yeast. Additionally, we used chi-squared tests to identify host genus-specialists and location-specialists. CONCLUSIONS: By comparing lichen mycobiomes of the same genera across different countries, our study advances our comprehension of these microbial communities. Our study elucidates that, although host species play a contributory role, geographic distance exerts a more pronounced impact on the structure of lichen mycobiome. We have made foundational contributions to understanding the lichen mycobiome occupying ecologically crucial niches. We anticipate that broader global-scale investigations into the fungal community structures will provide more detailed insights into fungal residents within lichens.


Subject(s)
DNA, Fungal , Lichens , Mycobiome , Republic of Korea , Turkey , Lichens/microbiology , Lichens/classification , DNA, Fungal/genetics , Ascomycota/classification , Ascomycota/isolation & purification , Ascomycota/genetics , High-Throughput Nucleotide Sequencing , Phylogeny , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , Parmeliaceae/genetics
13.
PLoS One ; 19(7): e0307747, 2024.
Article in English | MEDLINE | ID: mdl-39052602

ABSTRACT

Field peas are grown by smallholder farmers in Ethiopia for food, fodder, income, and soil fertility. However, leaf diseases such as ascochyta blight, powdery mildew, and leaf spots affect the quantity and quality of this crop as well as crop growth. Experts use visual observation to detect field pea disease. However, this approach is expensive, labor-intensive, and imprecise. Therefore, in this study, we presented a transfer learning approach for the automatic diagnosis of field pea leaf diseases. We classified three field pea leaf diseases: Ascochyta blight, leaf spot, and powdery mildew. A softmax classifier was used to classify the diseases. A total of 1600 images of both healthy and diseased leaves were used to train, validate, and test the pretrained models. According to the experimental results, DenseNet121 achieved 99.73% training accuracy, 99.16% validation accuracy, and 98.33% testing accuracy after 100 epochs. we expect that this research work will offer various benefits for farmers and farm experts. It reduced the cost and time needed for the detection and classification of field pea leaf disease. Thus, a fast, automated, less costly, and accurate detection method is necessary to overcome the detection problem.


Subject(s)
Deep Learning , Pisum sativum , Plant Diseases , Plant Leaves , Plant Diseases/microbiology , Ethiopia , Ascomycota/classification
14.
Fungal Biol ; 128(5): 1968-1981, 2024 Aug.
Article in English | MEDLINE | ID: mdl-39059852

ABSTRACT

Chestnut production is considered one of the most important economic resources of rural mountainous areas in Greece. Lately, producers report a steep rise in the incidence of brown rot disease caused by the fungus Gnomoniopsis smithogilvyi (Gnomoniaceae, Diaporthales), which results in severe chestnut rot. The pathogen is considered an emerging pathogen in many countries worldwide (Italy, France, Switzerland, Australia, New Zealand). This study aimed at (a) exploring the incidence of the brown rot disease in Vria (Regional Unit of Pieria, Region of Central Makedonia, Greece), (b) isolating and identifying the causal agent of the disease, (c) exploring the fungus presence at different phenological stages of the chestnut trees, and (d) implementing species-specific Bar- High Resolution Melting Analysis (HRM) for the early detection of G. smithogilvyi in chestnuts. G. smithogilvyi occurrence in chestnut tissues was more severe in June (59 %), nearly disappeared in July (19 %) and August (7 %) and increased again during harvesting time in September (57 %). This result could be attributed to a sum of different factors, including climate conditions. Moreover, it was demonstrated that G. smithogilvyi can be identified using a Bar-HRM analysis of chestnut tissues (buds, flowers and nuts). Results of this study clearly demonstrate that Bar-HRM can be used for the accurate, rapid and reliable identification of G. smithogilvyi universally on infected samples from different localities.


Subject(s)
Ascomycota , Fagaceae , Flowers , Plant Diseases , Plant Diseases/microbiology , Ascomycota/isolation & purification , Ascomycota/genetics , Ascomycota/classification , Greece , Flowers/microbiology , Fagaceae/microbiology , Incidence
15.
Mycologia ; 116(4): 498-508, 2024.
Article in English | MEDLINE | ID: mdl-38848260

ABSTRACT

Fossil epifoliar fungi are valuable indicators of paleoenvironment and paleoecology. The Meliolaceae, members of which typically inhabit the surface of living plants as biotrophs or pathogens, is one of the largest groups of epifoliar fungi. In this study, we report a novel fossil species of Meliolinites Selkirk (fossil Meliolaceae), Meliolinites tengchongensis, on the lower epidermis of compressed fossil Rhodoleia (Hamamelidaceae) leaves from the Upper Pliocene Mangbang Formation of Tengchong, Yunnan, southwestern China. Meliolinites tengchongensis is characterized by web-like, superficial, brown to dark brown, septate, and branching mycelia bearing 2-celled appressoria and unicellular phialides. The fungal colonies also include ellipsoidal, 5-celled, 4-septate ascospores and dark brown perithecia with suborbicular outline and verrucose surface. The well-preserved vegetative and reproductive organs help us to explore the potential disease process of the new fossil species. Besides, the presence of fungal remains indicates that the fungal taxon might have maintained its host preference since at least the Late Pliocene. Furthermore, the occurrence of both fossil fungi and their host plants in Tengchong indicate a subtropical-tropical, warm, and humid climate during the Late Pliocene, whereas the distribution pattern of the fungi on the host leaves suggests that Rhodoleia may have been a part of the middle-upper canopies in the Tengchong Late Pliocene multilayered forest.


Subject(s)
Fossils , Plant Leaves , Plant Leaves/microbiology , China , Ascomycota/classification , Ascomycota/isolation & purification , Spores, Fungal
16.
Med Mycol ; 62(7)2024 Jul 04.
Article in English | MEDLINE | ID: mdl-38857886

ABSTRACT

Acrophialophora is implicated in superficial and invasive infections, especially in immunosuppressed individuals. The present study was undertaken to provide clinical, microbiological, phylogenetic, and antifungal susceptibility testing (AFST) profile of Acrophialophora isolated from India. All the isolates identified as Acrophialophora species at the National Culture Collection for Pathogenic Fungi, Chandigarh, India were revived. Phenotypic and molecular characterization was performed, followed by temperature studies, scanning electron microscopy (SEM), and AFST. We also performed systematic review of all the cases of Acrophialophora species reported till date. A total of nine isolates identified as Acrophialophora species were identified by molecular method as A. fusispora (n = 8) and A. levis (n = 1), from brain abscess (n = 4), respiratory tract (n = 3), and corneal scraping (n = 2). All patients but two had predisposing factors/co-morbidities. Acrophialophora was identified as mere colonizer in one. Temperature studies and SEM divulged variation between both species. Sequencing of the internal transcribed spacer ribosomal DNA and beta-tubulin loci could distinguish species, while the LSU ribosomal DNA locus could not. AFST showed the lowest minimum inhibitory concentrations (MICs) for triazoles and the highest for echinocandins. Systematic literature review revealed 16 cases (11 studies), with ocular infections, pulmonary and central nervous system infections, and A. fusispora was common species. All the patients except three responded well. High MICs were noted for fluconazole, micafungin, and caspofungin. This is the first study delineating clinical, phenotypic, and genotypic characteristics of Acrophialophora species from India. The study highlights microscopic differences between both species and emphasizes the role of molecular methods in precise identification. Triazoles appear to be the most effective antifungals for managing patients.


We describe clinical, phenotypic, and genotypic characteristics of Acrophialophora species. This species causes mild infection to fatal infection in immunosuppressed individuals. Triazoles are effective in treating such infections.


Subject(s)
Antifungal Agents , Microbial Sensitivity Tests , Mycoses , Phylogeny , India , Humans , Antifungal Agents/pharmacology , Adult , Male , Mycoses/microbiology , Female , Middle Aged , Ascomycota/drug effects , Ascomycota/genetics , Ascomycota/isolation & purification , Ascomycota/classification , DNA, Fungal/genetics , Sequence Analysis, DNA , DNA, Ribosomal Spacer/genetics , Microscopy, Electron, Scanning , Phenotype , Tubulin/genetics , Aged , Young Adult , Child
17.
Appl Environ Microbiol ; 90(7): e0034224, 2024 Jul 24.
Article in English | MEDLINE | ID: mdl-38899884

ABSTRACT

Black apples are the result of late-stage microbial decomposition after falling to the ground. This phenomenon is highly comparable from year to year, with the filamentous fungus Monilinia fructigena most commonly being the first invader, followed by Penicillium expansum. Motivated by the fact that only little chemistry has been reported from apple microbiomes, we set out to investigate the chemical diversity and potential ecological roles of secondary metabolites (SMs) in a total of 38 black apples. Metabolomics analyses were conducted on either whole apples or small excisions of fungal biomass derived from black apples. Annotation of fungal SMs in black apple extracts was aided by the cultivation of 15 recently isolated fungal strains on 9 different substrates in a One Strain Many Compounds (OSMAC) approach, leading to the identification of 3,319 unique chemical features. Only 6.4% were attributable to known compounds based on analysis of high-performance liquid chromatography-high-resolution mass spectrometry (HPLC-HRMS/MS) data using spectral library matching tools. Of the 1,606 features detected in the black apple extracts, 32% could be assigned as fungal-derived, due to their presence in the OSMAC-based training data set. Notably, the detection of several antifungal compounds indicates the importance of such compounds for the invasion of and control of other microbial competitors on apples. In conclusion, the diversity and abundance of microbial SMs on black apples were found to be much higher than that typically observed for other environmental microbiomes. Detection of SMs known to be produced by the six fungal species tested also highlights a succession of fungal growth following the initial invader M. fructigena.IMPORTANCEMicrobial secondary metabolites constitute a significant reservoir of biologically potent and clinically valuable chemical scaffolds. However, their usefulness is hampered by rapidly developing resistance, resulting in reduced profitability of such research endeavors. Hence, the ecological role of such microbial secondary metabolites must be considered to understand how best to utilize such compounds as chemotherapeutics. Here, we explore an under-investigated environmental microbiome in the case of black apples; a veritable "low-hanging fruit," with relatively high abundances and diversity of microbially produced secondary metabolites. Using both a targeted and untargeted metabolomics approach, the interplay between metabolites, other microbes, and the apple host itself was investigated. This study highlights the surprisingly low incidence of known secondary metabolites in such a system, highlighting the need to study the functionality of secondary metabolites in microbial interactions and complex microbiomes.


Subject(s)
Malus , Penicillium , Secondary Metabolism , Malus/microbiology , Penicillium/metabolism , Penicillium/isolation & purification , Penicillium/genetics , Fungi/classification , Fungi/metabolism , Fungi/genetics , Fungi/isolation & purification , Ascomycota/metabolism , Ascomycota/genetics , Ascomycota/classification , Metabolomics , Microbiota , Biodiversity , Mycobiome
18.
Microb Pathog ; 193: 106756, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38901752

ABSTRACT

Eucalyptus spp. are undoubtedly one of the most favored plantation trees globally. Accurately identifying Eucalyptus pathogens is therefore crucial for timely disease prevention and control. Recently, symptoms of a leaf blight disease were observed on Eucalyptus trees in plantations at Jhajjar and Karnal in the state of Haryana, northern India. Asexual morphs resembling the features of the Botryosphaeriaceae were consistently isolated from the symptomatic leaves. Morphological features coupled with DNA sequence analysis confirmed a novel species, which is described and illustrated here as Botryosphaeria eucalypti sp. nov. Conidia of the new taxon are longer and wider than those of its phylogenetic neighbors. A distinct phylogenetic position for the new taxon was established through combined analysis of the internal transcribed spacer (ITS), partial translation elongation factor-1α (tef1) and partial ß-tubulin (tub2) regions. Recombination analysis provided additional support for the new species hypothesis. The pathogenicity of the novel species was proved on Eucalyptus leaves, and Koch's postulates were fulfilled. The discovery of new Botryosphaeria species is important because it will help in understanding the species diversity, host range, possible threats and disease control in the long run.


Subject(s)
Ascomycota , DNA, Fungal , Eucalyptus , Phylogeny , Plant Diseases , Plant Leaves , Sequence Analysis, DNA , Tubulin , Eucalyptus/microbiology , Plant Diseases/microbiology , Ascomycota/genetics , Ascomycota/isolation & purification , Ascomycota/classification , Plant Leaves/microbiology , India , DNA, Fungal/genetics , Tubulin/genetics , Peptide Elongation Factor 1/genetics , Spores, Fungal/genetics , DNA, Ribosomal Spacer/genetics
19.
Fungal Biol ; 128(4): 1815-1826, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38876534

ABSTRACT

Endophytic fungi, pivotal in facilitating plant co-evolution, significantly enhance plant growth, stress resistance, and environmental adaptability. Despite their importance, the spatial distribution of stem endophytic fungi (SEF) within host plants remains poorly characterized. Here, we employed high-throughput sequencing to conduct a comparative analysis of SEF communities in Mussaenda pubescens on a regional scale. Our findings reveal that whole-SEF communities were overwhelmingly dominated by members of the phylum Ascomycota, accounting for 85.9 %, followed by Basidiomycota at 13.9 %, and that alpha diversity within the whole-SEF community of M. pubescens remains relatively consistent across sampling sites. However, significant variation was observed within conditionally abundant taxa (CAT), conditionally rare or abundant taxa (CRAT), and conditionally rare taxa (CRT). Climatic factors emerged as the primary influence on SEF community distribution, followed by spatial distance and stem chemical properties. Neutral community modeling results suggested that both stochastic and deterministic processes play a role in shaping whole-SEF communities, with deterministic processes having a stronger influence on CRT subcommunities. Furthermore, the CRT co-occurrence network exhibited a more complex structure, characterized by higher values of network betweenness and degree relative to CAT and CRAT subcommunities. These findings enhance our understanding of community assembly and ecological interactions between stem fungal endophytes, presenting opportunities for harnessing fungal resources for the benefit of humanity.


Subject(s)
Endophytes , Plant Stems , Endophytes/classification , Endophytes/isolation & purification , Endophytes/genetics , Plant Stems/microbiology , Ascomycota/classification , Ascomycota/genetics , Ascomycota/isolation & purification , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , High-Throughput Nucleotide Sequencing , Basidiomycota/classification , Basidiomycota/genetics , Basidiomycota/isolation & purification , Biodiversity
20.
BMC Microbiol ; 24(1): 195, 2024 Jun 07.
Article in English | MEDLINE | ID: mdl-38849736

ABSTRACT

BACKGROUND: Rhizosphere and endophytic fungi play important roles in plant health and crop productivity. However, their community dynamics during the continuous cropping of Knoxia valerianoides have rarely been reported. K. valerianoides is a perennial herb of the family Rubiaceae and has been used in herbal medicines for ages. Here, we used high-throughput sequencing technology Illumina MiSeq to study the structural and functional dynamics of the rhizosphere and endophytic fungi of K. valerianoides. RESULTS: The findings indicate that continuous planting has led to an increase in the richness and diversity of rhizosphere fungi, while concomitantly resulting in a decrease in the richness and diversity of root fungi. The diversity of endophytic fungal communities in roots was lower than that of the rhizosphere fungi. Ascomycota and Basidiomycota were the dominant phyla detected during the continuous cropping of K. valerianoides. In addition, we found that root rot directly affected the structure and diversity of fungal communities in the rhizosphere and the roots of K. valerianoides. Consequently, both the rhizosphere and endophyte fungal communities of root rot-infected plants showed higher richness than the healthy plants. The relative abundance of Fusarium in two and three years old root rot-infected plants was significantly higher than the control, indicating that continuous planting negatively affected the health of K. valerianoides plants. Decision Curve Analysis showed that soil pH, organic matter (OM), available K, total K, soil sucrase (S_SC), soil catalase (S_CAT), and soil cellulase (S_CL) were significantly related (p < 0.05) to the fungal community dynamics. CONCLUSIONS: The diversity of fungal species in the rhizosphere and root of K. valerianoides was reported for the first time. The fungal diversity of rhizosphere soil was higher than that of root endophytic fungi. The fungal diversity of root rot plants was higher than that of healthy plants. Soil pH, OM, available K, total K, S_CAT, S_SC, and S_CL were significantly related to the fungal diversity. The occurrence of root rot had an effect on the community structure and diversity of rhizosphere and root endophytic fungi.


Subject(s)
Biodiversity , Endophytes , Fungi , Plant Roots , Rhizosphere , Soil Microbiology , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Fungi/classification , Fungi/genetics , Fungi/isolation & purification , Plant Roots/microbiology , DNA, Fungal/genetics , High-Throughput Nucleotide Sequencing , Plant Diseases/microbiology , Ascomycota/genetics , Ascomycota/classification , Ascomycota/growth & development , Ascomycota/isolation & purification , Phylogeny , Mycobiome
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