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1.
Curr Microbiol ; 81(7): 201, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38822823

ABSTRACT

Mucor representatives are mostly rapidly growing cosmopolitan soil saprotrophs of early diverged Mucoromycotina subphylum. Although this is the most speciose genus within the group, some lineages are still understudied. In this study, new species of Mucor was isolated from the post-mining area in southwestern Poland, where soil chemical composition analysis revealed high concentration of hydrocarbons and heavy metals. Phylogenetic analysis based on multigene phylogeny showed that the new isolate clusters distinctly from other Mucor species as a sister group to Mucor microsporus. New species Mucor thermorhizoides Abramczyk (Mucorales, Mucoromycota) is characterized by the extensive rhizoid production in elevated temperatures and formation of two layers of sporangiophores. It also significantly differs from M. microsporus in the shape of spores and the size of sporangia. M. thermorhizoides was shown to be able to grow in oligotrophic conditions at low temperatures. Together with M. microsporus they represent understudied and highly variable lineage of the Mucor genus.


Subject(s)
Mucor , Phylogeny , Soil Microbiology , Mucor/genetics , Mucor/classification , Mucor/isolation & purification , Poland , Mining , DNA, Fungal/genetics , Metals, Heavy
2.
Antonie Van Leeuwenhoek ; 117(1): 77, 2024 May 08.
Article in English | MEDLINE | ID: mdl-38717550

ABSTRACT

The "Shadegan International Wetland" (SIW) is one of the wetlands internationally recognized in the Ramsar convention. The vegetation of this wetland ecosystem consists of mostly grasses and shrubs that host a large number of fungi including endophytes. In this study, Nigrospora isolates were obtained from healthy plants of this wetland and its surrounding salt marshes and identified based on morphological features and multilocus phylogenetic analyses based on three DNA loci, namely the internal transcribed spacer regions 1 and 2 including the intervening 5.8S nuclear ribosomal DNA (ITS), ß-tubulin (tub2), and elongation factor 1-α (tef1-α). Accordingly, the following Nigrospora species were identified: N. lacticolonia, N. oryzae, N. osmanthi, N. pernambucoensis and a novel taxon N. shadeganensis sp. nov., which is described and illustrated. To the best of our knowledge, 10 new hosts for Nigrospora species are here reported, namely Aeluropus lagopoides, Allenrolfea occidentalis, Anthoxanthum monticola, Arthrocnemum macrostachyum, Cressa cretica, Halocnemum strobilaceum, Seidlitzia rosmarinus, Suaeda vermiculata, Tamarix passerinoides, and Typha latifolia. Moreover, the species N. lacticolonia and N. pernambucoensis are new records for the mycobiota of Iran.


Subject(s)
Ascomycota , Endophytes , Phylogeny , Poaceae , Wetlands , Iran , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Poaceae/microbiology , Ascomycota/genetics , Ascomycota/classification , Ascomycota/isolation & purification , DNA, Fungal/genetics , DNA, Ribosomal Spacer/genetics , Tubulin/genetics
3.
Sci Rep ; 14(1): 10601, 2024 05 08.
Article in English | MEDLINE | ID: mdl-38719921

ABSTRACT

A plant parasite associated with the white haze disease in apples, the Basidiomycota Gjaerumia minor, has been found in most samples of the global bathypelagic ocean. An analysis of environmental 18S rDNA sequences on 12 vertical profiles of the Malaspina 2010 expedition shows that the relative abundance of this cultured species increases with depth while its distribution is remarkably different between the deep waters of the Pacific and Atlantic oceans, being present in higher concentrations in the former. This is evident from sequence analysis and a microscopic survey with a species-specific newly designed TSA-FISH probe. Several hints point to the hypothesis that G. minor is transported to the deep ocean attached to particles, and the absence of G. minor in bathypelagic Atlantic waters could then be explained by the absence of this organism in surface waters of the equatorial Atlantic. The good correlation of G. minor biomass with Apparent Oxygen Utilization, recalcitrant carbon and free-living prokaryotic biomass in South Pacific waters, together with the identification of the observed cells as yeasts and not as resting spores (teliospores), point to the possibility that once arrived at deep layers this species keeps on growing and thriving.


Subject(s)
Basidiomycota , Pacific Ocean , Basidiomycota/genetics , Basidiomycota/isolation & purification , Basidiomycota/classification , RNA, Ribosomal, 18S/genetics , Seawater/microbiology , Phylogeny , Atlantic Ocean , DNA, Ribosomal/genetics , DNA, Fungal/genetics
4.
Mycopathologia ; 189(3): 43, 2024 May 06.
Article in English | MEDLINE | ID: mdl-38709328

ABSTRACT

During an epidemiological survey, a potential novel species within the basidiomycetous yeast genus Trichosporon was observed. The clinical strain was obtained from a urine sample taken from a Brazilian kidney transplant recipient. The strain was molecularly identified using the intergenic spacer (IGS1) ribosomal DNA locus and a subsequent phylogenetic analysis showed that multiple strains that were previously reported by other studies shared an identical IGS1-genotype most closely related to that of Trichosporon inkin. However, none of these studies provided an in-depth characterization of the involved strains to describe it as a new taxon. Here, we present the novel clinically relevant yeast for which we propose the name Trichosporon austroamericanum sp. nov. (holotype CBS H-24937). T. austroamericanum can be distinguished from other siblings in the genus Trichosporon using morphological, physiological, and phylogenetic characters.


Subject(s)
DNA, Fungal , DNA, Ribosomal Spacer , Phylogeny , Sequence Analysis, DNA , Transplant Recipients , Trichosporon , Trichosporonosis , Trichosporon/classification , Trichosporon/genetics , Trichosporon/isolation & purification , DNA, Ribosomal Spacer/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Fungal/genetics , Humans , Brazil , Trichosporonosis/microbiology , Cluster Analysis , Mycological Typing Techniques , Kidney Transplantation , Microscopy , Genotype
5.
Front Cell Infect Microbiol ; 14: 1367673, 2024.
Article in English | MEDLINE | ID: mdl-38707512

ABSTRACT

Most species of Dothiora are known from the dead parts of various host plants as saprobic fungi in terrestrial habitats occurring in tropical and temperate regions. In the present study, samples of Dothiora were collected from dead twigs and branches of Capparis spinosa, Rhaponticum repens, and an unknown angiosperm plant from the Tashkent and Jizzakh regions of Uzbekistan. Multi-gene phylogenetic analyses based on a combined ITS, LSU, SSU, TEF1, and TUB2 sequence data revealed their taxonomic positions within the Dothideaceae. Three new species of Dothiora, namely, Dothiora capparis, Dothiora rhapontici, and Dothiora uzbekistanica were proposed by molecular and morphological data. Likewise, the phylogenetic relationship and morphology of Dothiora are discussed. In addition, we provide a list of accepted Dothiora species, including host information, distribution, morphology descriptions, and availability of sequence data, to enhance the current knowledge of the diversity within Dothiora.


Subject(s)
Ascomycota , DNA, Fungal , Phylogeny , Sequence Analysis, DNA , DNA, Fungal/genetics , Ascomycota/genetics , Ascomycota/classification , Ascomycota/isolation & purification , Uzbekistan , DNA, Ribosomal/genetics , Plant Diseases/microbiology
6.
Article in English | MEDLINE | ID: mdl-38713196

ABSTRACT

The genus Exophiala is polymorphic, able to transition between yeast, hyphal and pseudohyphal forms. Species of the genus Exophiala are ubiquitous fungi that are distributed in various environments around the world. During a survey of fungal diversity in the gut of amphipods (Floresorchestia amphawaensis and undescribed Dogielinotid amphipods) from the Amphawa estuary, Samut Songkhram province, Thailand, five black yeast strains (DMKU-MG01, DMKU-MG07, DMKU-MG08, DMKU-HG10 and DMKU-FG04) were identified as representing a novel taxon on the basis of a combination of morphological and molecular phylogenetic features. The five strains did not produce filamentous hyphae or pseudohyphae. Only budding yeast cells were observed. On the basis of the phenotypic characteristics and the results of molecular analyses of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region, the five strains were identified as representing a novel species via applied nucleotide pairwise analysis. They differed from the most closely related species Exophiala alcalophiala by 3.54 % nucleotide substitutions (20 nucleotide substitutions in 572 bp) in the D1/D2 domains of the LSU rRNA gene. Moreover, the sequences of the ITS region of the five strains differed from those of the most closely related species E. alcalophiala, by 7.44-9.62 % nucleotide substitutions, and Exophiala halophiala, by 7.2-7.53 % nucleotide substitutions. The results of phylogenetic analyses based on the concatenated sequences of the ITS regions and the D1/D2 domains of the LSU rRNA gene confirmed that the five black yeast strains represented a single novel species of the genus Exophiala. In this study, Exophiala amphawaensis sp. nov. is proposed to accommodate these strains. The holotype is TBRC 15626T and the isotype is PYCC9020. The MycoBank accession number of the novel species is MB 851477.


Subject(s)
Amphipoda , DNA, Fungal , DNA, Ribosomal Spacer , Exophiala , Phylogeny , Sequence Analysis, DNA , Animals , Thailand , Amphipoda/microbiology , DNA, Fungal/genetics , DNA, Ribosomal Spacer/genetics , Exophiala/genetics , Exophiala/isolation & purification , Exophiala/classification , Mycological Typing Techniques , Gastrointestinal Tract/microbiology
7.
Article in English | MEDLINE | ID: mdl-38713197

ABSTRACT

Two isolates representing a novel species of the genus Wickerhamiella were obtained in India from nectar of flowers of Lantana camara, an ornamental exotic species native to Central and South America. Phylogenetic analyses of the D1/D2 domain of the 26S large subunit (LSU) rRNA gene, internal transcribed spacer (ITS) region, and physiological characteristics, supported the recognition of the novel species, that we designate Wickerhamiella lachancei sp. nov (MycoBank no. MB851709), with MCC 9929T as the holotype and PYCC 10003T as the isotype. Considering pairwise sequence similarity, the type strain of the novel species differs from the type strain of the most closely related species, Wickerhamiella drosophilae CBS 8459T, by 16 nucleotide substitutions and two gaps (3.9 % sequence variation) in the D1/D2 region (560 bp compared) and 28 nucleotide substitutions and five gaps (7.22 % sequence variation) in the ITS region (444 bp compared).


Subject(s)
DNA, Fungal , DNA, Ribosomal Spacer , Flowers , Lantana , Phylogeny , Sequence Analysis, DNA , India , Flowers/microbiology , DNA, Fungal/genetics , Lantana/microbiology , DNA, Ribosomal Spacer/genetics , Molecular Sequence Data , Mycological Typing Techniques , RNA, Ribosomal/genetics , Saccharomycetales/genetics , Saccharomycetales/isolation & purification , Saccharomycetales/classification
8.
J Appl Microbiol ; 135(5)2024 May 01.
Article in English | MEDLINE | ID: mdl-38724454

ABSTRACT

AIMS: Neocosmospora species are saprobes, endophytes, and pathogens belonging to the family Nectriaceae. This study aims to investigate the taxonomy, biosynthetic potential, and application of three newly isolated Neocosmospora species from mangrove habitats in the southern part of Thailand using phylogeny, bioactivity screening, genome sequencing, and bioinformatics analysis. METHODS AND RESULTS: Detailed descriptions, illustrations, and a multi-locus phylogenetic tree with large subunit ribosomal DNA (LSU), internal transcribed spacer (ITS), translation elongation factor 1-alpha (ef1-α), and RNA polymerase II second largest subunit (RPB2) regions showing the placement of three fungal strains, MFLUCC 17-0253, MFLUCC 17-0257, and MFLUCC 17-0259 clustered within the Neocosmospora clade with strong statistical support. Fungal crude extracts of the new species N. mangrovei MFLUCC 17-0253 exhibited strong antifungal activity to control Colletotrichum truncatum CG-0064, while N. ferruginea MFLUCC 17-0259 exhibited only moderate antifungal activity toward C. acutatum CC-0036. Thus, N. mangrovei MFLUCC 17-0253 was sequenced by Oxford nanopore technology. The bioinformatics analysis revealed that 49.17 Mb genome of this fungus harbors 41 potential biosynthetic gene clusters. CONCLUSION: Two fungal isolates of Neocosmospora and a new species of N. mangrovei were reported in this study. These fungal strains showed activity against pathogenic fungi causing anthracnose in chili. In addition, full genome sequencing and bioinformatics analysis of N. mangrovei MFLUCC 17-0253 were obtained.


Subject(s)
Colletotrichum , Phylogeny , Colletotrichum/genetics , Thailand , Ascomycota/genetics , Antifungal Agents/pharmacology , Plant Diseases/microbiology , Plant Diseases/prevention & control , Biological Control Agents , DNA, Fungal/genetics , Genome, Fungal , Southeast Asian People
9.
Article in English | MEDLINE | ID: mdl-38747851

ABSTRACT

Microsporum canis, one of the most widespread dermatophytes worldwide, is a zoonotic microorganism that transmits infection from reservoirs such as cats and dogs to humans. This microorganism is associated with Tinea corporis and other clinical manifestations; however, few studies have used genetic surveillance to determine and characterize the process of zoonotic transmission. In this study, we show a clear example of zoonotic transmission from a cat to an intrafamilial environment, where it caused Tinea corporis by infection with M. canis. Molecular characterization using the b-tubulin gene and Random Amplified Polymorphic DNA analysis made it possible to determine that the six isolates of M. canis obtained in this study belonged to the same genetic variant or clone responsible for reservoir-reservoir or reservoir-human transmission.


Subject(s)
Cat Diseases , Microsporum , Tinea , Zoonoses , Microsporum/isolation & purification , Microsporum/genetics , Microsporum/classification , Cats/microbiology , Animals , Tinea/microbiology , Tinea/transmission , Tinea/veterinary , Cat Diseases/microbiology , Cat Diseases/transmission , Zoonoses/microbiology , Zoonoses/transmission , Pets/microbiology , Humans , Dogs , Random Amplified Polymorphic DNA Technique , Male , Female , Dog Diseases/microbiology , Dog Diseases/transmission , DNA, Fungal/genetics
10.
Mycopathologia ; 189(3): 44, 2024 May 11.
Article in English | MEDLINE | ID: mdl-38734862

ABSTRACT

A 50-year-old man, previously diagnosed with pulmonary tuberculosis and lung cavities, presented with symptoms including fever, shortness of breath, and cough. A pulmonary CT scan revealed multiple cavities, consolidation and tree-in-bud in the upper lungs. Further investigation through direct examination of bronchoalveolar lavage fluid showed septate hyphae with dichotomous acute branching. Subsequent isolation and morphological analysis identified the fungus as belonging to Aspergillus section Nigri. The patient was diagnosed with probable invasive pulmonary aspergillosis and successfully treated with a three-month oral voriconazole therapy. Phylogenetic analysis based on partial ß-tubulin, calmodulin and RNA polymerase second largest subunit sequences revealed that the isolate represents a putative new species related to Aspergillus brasiliensis, and is named Aspergillus hubkae here. Antifungal susceptibility testing demonstrated that the isolate is resistant to itraconazole but susceptible to voriconazole. This phenotypic and genetic characterization of A. hubkae, along with the associated case report, will serve as a valuable resource for future diagnoses of infections caused by this species. It will also contribute to more precise and effective patient management strategies in similar clinical scenarios.


Subject(s)
Antifungal Agents , Aspergillus , Invasive Pulmonary Aspergillosis , Microbial Sensitivity Tests , Phylogeny , Sequence Analysis, DNA , Voriconazole , Humans , Male , Middle Aged , Antifungal Agents/therapeutic use , Antifungal Agents/pharmacology , Aspergillus/isolation & purification , Aspergillus/genetics , Aspergillus/classification , Aspergillus/drug effects , Bronchoalveolar Lavage Fluid/microbiology , Cluster Analysis , DNA, Fungal/genetics , DNA, Fungal/chemistry , Invasive Pulmonary Aspergillosis/microbiology , Invasive Pulmonary Aspergillosis/drug therapy , Invasive Pulmonary Aspergillosis/diagnosis , Itraconazole/pharmacology , Microscopy , Tomography, X-Ray Computed , Treatment Outcome , Tubulin/genetics , Voriconazole/therapeutic use , Voriconazole/pharmacology
11.
Fungal Biol ; 128(3): 1751-1757, 2024 May.
Article in English | MEDLINE | ID: mdl-38796259

ABSTRACT

This study explores the fungal diversity associated with tarballs, weathered crude oil deposits, on Goa's tourist beaches. Despite tarball pollution being a longstanding issue in Goa state in India, comprehensive studies on associated fungi are scarce. Our research based on amplicon sequence analysis of fungal ITS region fills this gap, revealing a dominance of Aspergillus, particularly Aspergillus penicillioides, associated with tarballs from Vagator and Morjim beaches. Other notable species, including Aspergillus sydowii, Aspergillus carbonarius, and Trichoderma species, were identified, all with potential public health and ecosystem implications. A FUNGuild analysis was conducted to investigate the potential ecological roles of these fungi, revealing a diverse range of roles, including nutrient cycling, disease propagation, and symbiotic relationships. The study underscores the need for further research and monitoring, given the potential health risks and contribution of tarball-associated fungi to the bioremediation of crude oil-contaminated beaches.


Subject(s)
Biodiversity , DNA, Fungal , Fungi , India , DNA, Fungal/genetics , Fungi/genetics , Fungi/classification , Fungi/isolation & purification , Sequence Analysis, DNA , DNA, Ribosomal Spacer/genetics , Phylogeny , Petroleum/microbiology
12.
Fungal Biol ; 128(3): 1758-1770, 2024 May.
Article in English | MEDLINE | ID: mdl-38796260

ABSTRACT

Starting in the fall of 2019, mortality, blight symptoms, and signs of white fungal mycelia were observed on external host tissues of non-native landscape trees as well as numerous native trees, understory shrubs, and vines throughout northern and central Florida, USA. We determined that the fungus is an undescribed species of Basidiomycota based on morphological characteristics and DNA sequence analysis. Phylogenetic analyses of the internal transcribed spacer (ITS), large subunit (LSU), and translation elongation factor 1-alpha (tef1) regions revealed that this novel plant pathogen is an undescribed taxon of the genus Parvodontia (Cystostereaceae, Agaricales). We propose the name Parvodontia relampaga sp. nov. which describes its unique morphological features and phylogenetic placement. We confirmed the pathogenicity of P. relampaga in greenhouse inoculations on host plants from which strains of this novel pathogen were isolated, including the non-native gymnosperm Afrocarpus falcatus, the non-native and commercially important Ligustrum japonicum, and the native tree Quercus hemisphaerica. P. relampaga was also detected on a total of 27 different species of woody host plants, including such economically and ecologically important hosts as Fraxinus, Ilex, Magnolia, Persea, Prunus, Salix, Vitis, and Vaccinium. For this new plant disease, we propose the name "relampago blight," which refers to the lightning-like rhizomorph growth (relámpago means 'lightning' in Spanish). This study presents a newly discovered fungal taxon with a wide host range on both angiosperms and gymnosperms that may be an emerging pathogen of concern in Florida and the Gulf Coast region.


Subject(s)
DNA, Fungal , Phylogeny , Plant Diseases , Plant Diseases/microbiology , Florida , DNA, Fungal/genetics , Agaricales/genetics , Agaricales/classification , Agaricales/isolation & purification , Agaricales/physiology , Agaricales/pathogenicity , Sequence Analysis, DNA , DNA, Ribosomal Spacer/genetics , DNA, Ribosomal Spacer/chemistry
13.
Fungal Biol ; 128(3): 1780-1789, 2024 May.
Article in English | MEDLINE | ID: mdl-38796262

ABSTRACT

Anthracnose caused by Colletotrichum is the most severe and widely occurring cashew disease in Brazil. Colletotrichum species are commonly found as pathogens, endophytes and occasionally as saprophytes in a wide range of hosts. The endophytic species associated with cashew trees are poorly studied. In this study, we report the Colletotrichum endophytic species associated with cashew trees in two locations in the state of Pernambuco, their prevalence in different plant organs (leaves, veins, branches and inflorescences), and compare the species in terms of pathogenicity and aggressiveness using different inoculation methods (wounded × unwounded). Six species of Colletotrichum were identified according to multilocus phylogenetic analyses, including Colletotrichum asianum, Colletotrichum chrysophilum, Colletotrichum karsti, Colletotrichum siamense, Colletotrichum theobromicola, and Colletotrichum tropicale. There were differences in the percentage of isolation in relation to the prevalence of colonized tissues and collection locations. C. tropicale was the prevalent species in both geographic areas and plant tissues collected, with no pattern of distribution of species between areas and plant tissues. All isolates were pathogenic in injured tissues of cashew plants. The best method to test the pathogenicity of Colletotrichum species was utilizing the combination of leaves + presence of wounds + conidial suspension, as it better represents the natural infection process. C. siamense was the most aggressive species.


Subject(s)
Anacardium , Colletotrichum , Endophytes , Phylogeny , Plant Diseases , Colletotrichum/genetics , Colletotrichum/classification , Colletotrichum/isolation & purification , Brazil , Anacardium/microbiology , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Plant Diseases/microbiology , DNA, Fungal/genetics , Multilocus Sequence Typing
14.
Fungal Biol ; 128(3): 1790-1799, 2024 May.
Article in English | MEDLINE | ID: mdl-38796263

ABSTRACT

Species in the Melastomataceae (Myrtales) include trees and woody shrubs that are amongst the most common hosts of Chrysoporthe and related fungi. These fungi cause stem cankers, branch death and in extreme cases, kill their hosts. Chrysoporthe-like fungi were observed on Miconia spp. and Rhynchanthera grandiflora (Melastomataceae) plants during tree disease surveys in south-eastern Brazil including the states of Minas Gerais and Rio de Janeiro. The aims of this study were to isolate and identify the fungi utilising morphological characteristics and phylogenetic analyses. This led to the identification of a new species of Chrysoporthe described here as Chrysoporthe brasilensis sp.nov. Inoculations were conducted on R. grandiflora and M. theaezans, showing that C. brasiliensis is an aggressive pathogen. This study adds to a growing number of reports of new and pathogenic species of Chrysoporthe that potentially threaten native Myrtales globally, including important trees such as Eucalyptus, both in natural ecosystems and in planted forests.


Subject(s)
Melastomataceae , Phylogeny , Plant Diseases , Brazil , Melastomataceae/microbiology , Plant Diseases/microbiology , DNA, Fungal/genetics , Ascomycota/classification , Ascomycota/genetics , Ascomycota/isolation & purification , DNA, Ribosomal/genetics , Sequence Analysis, DNA , DNA, Ribosomal Spacer/genetics , DNA, Ribosomal Spacer/chemistry , Cluster Analysis
15.
Fungal Biol ; 128(3): 1800-1805, 2024 May.
Article in English | MEDLINE | ID: mdl-38796264

ABSTRACT

It has been the aim of this study to molecular-taxonomically identify 15 Beauveria isolates collected from different geographical regions and insect hosts in Argentina and to investigate the levels of inter- and intra-specific diversity across this set of isolates. Based on phylogenetic analyses of EF1A-RPB1-RPB2 concatenated genes and BLOC markers, all Beauveria strains were identify as Beauveria bassiana. Within the B. bassiana clades of both phylogenies, isolates from Argentina were not clustered according to geographic origin or host. The 15 fungal isolates were further analyzed by PCR amplification of the intron insertion hot spot region of the nuclear 28S rRNA encoding sequence. By intron sequence and position, seven different group-I intron combinations termed variants A, B1, B2, C, D, E and F were found in the 15 isolates under study. Variants B1/B2 consisting of a single 28Si2 intron were found in ten isolates, whereas variant A occurred twice and variants C through F were unique across the set of isolates under study. The determination of the different introns and intron combinations in the 28S rRNA gene is a powerful tool for achieving infraspecific differentiation of B. bassiana isolates from Argentina.


Subject(s)
Beauveria , Genetic Variation , Phylogeny , RNA, Ribosomal, 28S , Beauveria/genetics , Beauveria/classification , Beauveria/isolation & purification , Argentina , RNA, Ribosomal, 28S/genetics , Animals , DNA, Fungal/genetics , Insecta/microbiology , Sequence Analysis, DNA , Molecular Sequence Data , Introns , DNA, Ribosomal/genetics , Cluster Analysis
16.
Mol Biol Rep ; 51(1): 687, 2024 May 25.
Article in English | MEDLINE | ID: mdl-38796652

ABSTRACT

BACKGROUND: Alternaria blotch disease in Himachal Pradesh, India, caused by Alternaria spp., adversely affects apple cultivars, resulting in reduced fruit size and quality accompanied by premature leaf fall. METHODS AND RESULTS: Sixteen Alternaria isolates from apple growing regions underwent comprehensive analysis including morphology, pathogenicity, and molecular characterization. Variations in conidiophore and conidia dimensions, shapes, and divisions were observed among isolates. Pathogenicity assays revealed differences in incubation periods, latent phases, and disease responses. Molecular characterization via nuclear ITS rDNA and RAPD analysis indicated 99-100% homology with Alternaria alternata, Alternaria mali, and other Alternaria spp., with a close phylogenetic relationship to Chinese isolates. Differentiation of isolates based on origin, cultural characteristics, and morphology was achieved using RAPD markers. CONCLUSIONS: The study identifies diverse genotypes and morphotypes of Alternaria contributing to apple blotch disease in Himachal Pradesh. These findings highlight the complexity of the pathogenic environment and hold significant implications for disease management in apple orchards.


Subject(s)
Alternaria , Malus , Phylogeny , Plant Diseases , Alternaria/pathogenicity , Alternaria/genetics , Malus/microbiology , India , Plant Diseases/microbiology , Random Amplified Polymorphic DNA Technique , DNA, Fungal/genetics , Spores, Fungal/genetics
17.
BMC Microbiol ; 24(1): 180, 2024 May 24.
Article in English | MEDLINE | ID: mdl-38789974

ABSTRACT

BACKGROUND: Cobweb disease is a fungal disease that commonly affects the cultivation and production of edible mushrooms, leading to serious yield and economic losses. It is considered a major fungal disease in the realm of edible mushrooms. The symptoms of cobweb disease were found during the cultivation of Lyophyllum decastes. This study aimed to identify the causative pathogen of cobweb disease and evaluate effective fungicides, providing valuable insights for field control and management of L. decastes cobweb disease. RESULTS: The causal agent of cobweb disease was isolated from samples infected and identified as Cladobotryum mycophilum based on morphological and cultural characteristics, as well as multi-locus phylogeny analysis (ITS, RPB1, RPB2, and TEF1-α). Pathogenicity tests further confirmed C. mycophilum as the responsible pathogen for this condition. Among the selected fungicides, Prochloraz-manganese chloride complex, Trifloxystrobin, tebuconazole, and Difenoconazole exhibited significant inhibitory effects on the pathogen's mycelium, with EC50 values of 0.076 µg/mL, 0.173 µg/mL, and 0.364 µg/mL, respectively. These fungicides can serve as references for future field control of cobweb disease in L. decastes. CONCLUSION: This study is the first report of C. mycophilum as the causing agent of cobweb disease in L. decastes in China. Notably, Prochloraz-manganese chloride complex demonstrated the strongest inhibitory efficacy against C. mycophilum.


Subject(s)
Fungicides, Industrial , Phylogeny , China , Fungicides, Industrial/pharmacology , Agaricales/genetics , Agaricales/drug effects , Agaricales/classification , Ascomycota/drug effects , Ascomycota/genetics , Ascomycota/isolation & purification , Ascomycota/classification , DNA, Fungal/genetics , Triazoles/pharmacology , Microbial Sensitivity Tests , Strobilurins , Acetates , Dioxolanes , Imines
18.
Int J Mol Sci ; 25(10)2024 May 10.
Article in English | MEDLINE | ID: mdl-38791224

ABSTRACT

Cotton Verticillium wilt is mainly caused by the fungus Verticillium dahliae, which threatens the production of cotton. Its pathogen can survive in the soil for several years in the form of microsclerotia, making it a destructive soil-borne disease. The accurate, sensitive, and rapid detection of V. dahliae from complex soil samples is of great significance for the early warning and management of cotton Verticillium wilt. In this study, we combined the loop-mediated isothermal amplification (LAMP) with CRISPR/Cas12a technology to develop an accurate, sensitive, and rapid detection method for V. dahliae. Initially, LAMP primers and CRISPR RNA (crRNA) were designed based on a specific DNA sequence of V. dahliae, which was validated using several closely related Verticillium spp. The lower detection limit of the LAMP-CRISPR/Cas12a combined with the fluorescent visualization detection system is approximately ~10 fg/µL genomic DNA per reaction. When combined with crude DNA-extraction methods, it is possible to detect as few as two microsclerotia per gram of soil, with the total detection process taking less than 90 min. Furthermore, to improve the method's user and field friendliness, the field detection results were visualized using lateral flow strips (LFS). The LAMP-CRISPR/Cas12a-LFS system has a lower detection limit of ~1 fg/µL genomic DNA of the V. dahliae, and when combined with the field crude DNA-extraction method, it can detect as few as six microsclerotia per gram of soil, with the total detection process taking less than 2 h. In summary, this study expands the application of LAMP-CRISPR/Cas12a nucleic acid detection in V. dahliae and will contribute to the development of field-deployable diagnostic productions.


Subject(s)
CRISPR-Cas Systems , Nucleic Acid Amplification Techniques , Plant Diseases , Soil Microbiology , Nucleic Acid Amplification Techniques/methods , Plant Diseases/microbiology , Ascomycota/genetics , Ascomycota/isolation & purification , Molecular Diagnostic Techniques/methods , Gossypium/microbiology , DNA, Fungal/genetics , DNA, Fungal/isolation & purification , Verticillium/genetics
19.
Int J Mol Sci ; 25(10)2024 May 14.
Article in English | MEDLINE | ID: mdl-38791411

ABSTRACT

Melon (Cucumis melo L.) is a global commercial crop that is sensitive to seed-borne wilt infections caused by Fusarium oxysporum f. sp. melonis (Fom). To address the challenge of detecting Fom contamination, we designed a probe-based real-time PCR method, TDCP2, in combination with rapid or column-based DNA extraction protocols to develop reliable molecular detection methods. Utilizing TDCP2, the detection rate reached 100% for both artificially Fom-inoculated (0.25-25%) and pod-inoculated melon seeds in conjunction with DNA samples from either the rapid or column-based extraction protocol. We performed analyses of precision, recall, and F1 scores, achieving a maximum F1 score of 1 with TDCP2, which highlights the robustness of the method. Additionally, intraday and interday assays were performed, which revealed the high reproducibility and stability of column-based DNA extraction protocols combined with TDCP2. These metrics confirm the reliability of our developed protocols, setting a foundation for future enhancements in seed pathology diagnostics and potentially broadening their applicability across various Fom infection levels. In the future, we hope that these methods will reduce food loss by improving the control and management of melon diseases.


Subject(s)
Fusarium , Plant Diseases , Real-Time Polymerase Chain Reaction , Seeds , Fusarium/genetics , Fusarium/isolation & purification , Seeds/microbiology , Plant Diseases/microbiology , Real-Time Polymerase Chain Reaction/methods , Cucurbitaceae/microbiology , DNA, Fungal/genetics , DNA, Fungal/isolation & purification , Cucumis melo/microbiology , Reproducibility of Results
20.
Diagn Microbiol Infect Dis ; 109(3): 116337, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38718662

ABSTRACT

Lecanicillium dimorphum and Lecanicillium psalliotae are fungi that exist naturally in plants or insects, and are generally considered non-pathogenic to humans. However, in this case, we cultured Lecanicillium from the synovial fluid of a patient, and identified it through genome sequencing and sequence alignment as Lecanicillium dimorphum or Lecanicillium psalliotae. Due to the conservation of sequences, we can only identify the genus and not the species. There are very few reports on the human infection and pathogenicity of these two fungi, and this case also cannot completely prove that the pathogenic agent is this fungus. But this case also holds clinical significance, as the discovery of Lecanicillium in a human sample can alert the clinician to the presence of an uncommon mold with unclear clinical significance.


Subject(s)
Hypocreales , Mycoses , Humans , Hypocreales/isolation & purification , Hypocreales/genetics , Hypocreales/classification , Mycoses/microbiology , Mycoses/diagnosis , Synovial Fluid/microbiology , Male , Phylogeny , Sequence Analysis, DNA , DNA, Fungal/genetics
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