Biocontrol Potential of Cladosporium sphaerospermum Against the Wheat Powdery Mildew Fungus Blumeria graminis f. sp. tritici.

Cladosporium spp. are known to be mycoparasites and inhibit phytopathogenic fungi. However, so far, little information is available on the impact of Cladosporium spp. on powdery mildews. Based on the morphological characteristics and molecular analysis, C. sphaerospermum was identified as a mycoparasite on the wheat powdery mildew fungus Blumeria graminis f. sp. tritici (Bgt), recently named B. graminis s. str. C. sphaerospermum was capable of preventing colony formation and conidial distribution of Bgt. The biomasses of Bgt notably decreased by 1.3, 2.2, 3.6, and 3.8 times at 2, 4, 6, and 8 days postinoculation (dpi), respectively. In addition, biomasses of C. sphaerospermum at 2, 4, 6, and 8 dpi significantly increased to 5.6, 13.9, 18.2, and 67.3 times, respectively. In vitro, C. sphaerospermum exudates significantly impaired appressorial formation of Bgt. Thus, C. sphaerospermum acts as a potential biological control agent by suppressing the formation, distribution, and development of Bgt conidia and is a viable alternative for managing the wheat powdery mildew. These results suggest that C. sphaerospermum is an antagonistic parasite of the wheat powdery mildew fungus and, hence, provide new knowledge about the biological control of phytopathogenic fungi.

Gene expression changes in human bronchial epithelial cells (BEAS-2B) and human pulmonary alveolar epithelial cells (HPAEpiC) after interaction with Cladosporium sphaerospermum.

Cladosporium is one of the most abundant spore. Fungi of this genus can cause respiratory allergy and intrabronchial lesion. We studied the differential expression of host genes after the interaction of Cladosporium sphaerospermum conidia with Human Bronchial Epithelial Cells (BEAS-2B) and Human Pulmonary Alveolar Epithelial Cells (HPAEpiC). C. sphaerospermum conidia were harvested and co-cultured with BEAS-2B cells or HPAEpiC cells for 48 hours respectively. This culture duration was chosen as it was associated with high germination rate. RNA was extracted from two biological replicates per treatment. RNA of BEAS-2B cells was used to assess changes in gene expression using AffymetrixGeneChip® Human Transcriptome Array 2.0. After co-culture with Cladosporium spores, 68 individual genes were found differentially expressed (P ≤ 0.05) and up-regulated ≥ 1.5 folds while 75 genes were found differentially expressed at ≤ -1.5 folds compared with controls. Reverse transcription and qPCR were performed on the RNA collected from both BEAS-2B cells and HPAEpiC cells to validate the microarray results with 7 genes. Based on the findings, infected pulmonary epithelial cells exhibited an increase in cell death-related genes and genes associated with innate immunity.

Hybrid Polyketides from a Hydractinia-Associated Cladosporium sphaerospermum SW67 and Their Putative Biosynthetic Origin.

Five hybrid polyketides (1a, 1b, and 2-4) containing tetramic acid core including a new hybrid polyketide, cladosin L (1), were isolated from the marine fungus Cladosporium sphaerospermum SW67, which was isolated from the marine hydroid polyp of Hydractinia echinata. The hybrid polyketides were isolated as a pair of interconverting geometric isomers. The structure of 1 was determined based on 1D and 2D NMR spectroscopic and HR-ESIMS analyses. Its absolute configuration was established by quantum chemical electronic circular dichroism (ECD) calculations and modified Mosher's method. Tetramic acid-containing compounds are reported to be derived from a hybrid PKS-NRPS, which was also proved by analyzing our 13C-labeling data. We investigated whether compounds 1-4 could prevent cell damage induced by cisplatin, a platinum-based anticancer drug, in LLC-PK1 cells. Co-treatment with 2 and 3 ameliorated the damage of LLC-PK1 cells induced by 25 µM of cisplatin. In particular, the effect of compound 2 at 100 µM (cell viability, 90.68 ± 0.81%) was similar to the recovered cell viability of 88.23 ± 0.25% with 500 µM N-acetylcysteine (NAC), a positive control.

Unstable Tetramic Acid Derivatives from the Deep-Sea-Derived Fungus Cladosporium sphaerospermum EIODSF 008.

Seven new unstable tetramic acid derivatives, cladosporiumins I-O (1⁻7), together with the known analogue cladodionen (8) were isolated from the extract of the deep-sea-derived fungus Cladosporium sphaerospermum EIODSF 008. Their structures were elucidated by spectroscopic analysis, quantum chemical calculations and ECD spectra. Compound 4 was a Mg complex of tetramic acid derivative. In acidic solvent, 4 could change to 1 and 6, and 7 could change to 5. In addition, 1, 5 and 8 existed as two exchangeable isomers, respectively. The structures of cladosporiumins E-H were reassigned as their Na complexes. The antibacterial and cytotoxic activities of 1⁻8 were also evaluated. However, because of their instability, all of the isolated compounds did not show significant antibacterial activity as the preliminary EtOAc extracts of the fungal strain.

Cladosins F and G, two new hybrid polyketides from the deep-sea-derived Cladosporium sphaerospermum 2005-01-E3.

Two new fungal hybrid polyketides, cladosins F (1) and G (2), with rare 6(3)-enamino-8,10-dihydroxy-tetraketide system were discovered from the deep-sea-derived fungus Cladosporium sphaerospermum 2005-01-E3 guided by OSMAC approach. Their structures were elucidated on the basis of comprehensive spectroscopic analyses, and cytotoxicity, antitubercular, anti-influenza A H1N1 virus, and NF-κB inhibitory activities were evaluated.

Mycodegradation of low-density polyethylene by Cladosporium sphaerospermum, isolated from platisphere.

Plastic accumulation is a severe threat to the environment due to its resistivity to thermal, mechanical and biological processes. In recent years, microbial degradation of plastic waste disposal is of interest because of its eco-friendly nature. In this study, a total of 33 fungi were isolated from the plastisphere and out of which 28 fungal species showed halo zone of clearance in agarized LDPE media. The fungus showing highest zone of clearance was further used to evaluate its degradation potential. Based on morphological and molecular technique, the fungus was identified as Cladosporium sphaerospermum. The biodegradation of LDPE by C. sphaerospermum was evaluated by various methods. The exposure of LDPE with C. sphaerospermum resulted in weight loss (15.23%) in seven days, higher reduction rate (0.0224/day) and lower half-life (30.93 days). FTIR analysis showed changes in functional group and increased carbonyl index in LDPE treated with C. sphaerospermum. SEMimages evidenced the formation of pits, surface aberrations and grooves on the LDPE film treated with the fungus whereas the untreated control LDPE film showed no change. AFM analysis confirmed the surface changes and roughness in fungus treated LDPE film. This might be due to the extracellular lignolytic enzymes secreted by C. sphaerospermum grown on LDPE. The degradation of polyethylene by Short chain alkanes such as dodecane, hexasiloxane and silane were identified in the extract of fungus incubated with LDPE film through GC-MS analysis which might be due to the degradation of LDPE film by C. sphaerospermum. This was the first report on the LDPE degradation by C. sphaerospermum in very short duration which enables green scavenging of plastic wastes.

Truncation of a novel C-terminal domain of a ß-glucanase improves its thermal stability and specific activity.

Enzymes that degrade ß-glucan play important roles in various industries, including those related to brewing, animal feed, and health care. Csph16A, an endo-ß-1,3(4)-glucanase encoded by a gene from the halotolerant, xerotolerant, and radiotrophic black fungus Cladosporium sphaerospermum, was cloned and expressed in Pichia pastoris. Two isoforms (Csph16A.1 and Csph16A.2) are produced, arising from differential glycosylation. The proteins were predicted to contain a catalytic Lam16A domain, along with a C-terminal domain (CTD) of unknown function which exhibits minimal secondary structure. Employing PCR-mediated gene truncation, the CTD of Csph16A was excised to assess its functional impact on the enzyme and determine potential alterations in biotechnologically relevant characteristics. The truncated mutant, Csph16A-ΔC, exhibited significantly enhanced thermal stability at 50°C, with D-values 14.8 and 23.5 times greater than those of Csph16A.1 and Csph16A.2, respectively. Moreover, Csph16A-ΔC demonstrated a 20%-25% increase in halotolerance at 1.25 and 1.5 M NaCl, respectively, compared to the full-length enzymes. Notably, specific activity against cereal ß-glucan, lichenan, and curdlan was increased by up to 238%. This study represents the first characterization of a glucanase from the stress-tolerant fungus C. sphaerospermum and the first report of a halotolerant and engineered endo-ß-1,3(4)-glucanase. Additionally, it sheds light on a group of endo-ß-1,3(4)-glucanases from Antarctic rock-inhabiting black fungi harboring a Lam16A catalytic domain and a novel CTD of unknown function.

Cladosporium sphaerospermum extract inhibits quorum sensing associated virulence factors of Serratia marcescens.

Serratia marcescens is now becoming a propensity for its highly antimicrobial-resistant clinical infections. Currently, it provides a novel strategy to prevent and control microbial infection by regulating S. marcescens quorum sensing (QS). Deep-sea-derived fungi are rich in QS bioactive constituents. In this work, the extracts from Cladosporium sphaerospermum SCSGAF0054 showed potent QS-related virulence factors and biofilm-inhibiting activities against S. marcescens NJ01. The swimming motility and multiple virulence factors such as prodigiosin, exopolysaccharide (EPS), lipase, protease and hemolysin were moderately inhibited by the extracts at varied concentrations. The confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) images revealed that C. sphaerospermum extracts moderately arrested biofilm formation and cell viability. Further, real-time quantitative PCR (RT-qPCR) analysis revealed that expressions of genes associated with virulence factors, flhD, fimA, fimC, bsmA, bsmB, pigA, pigC, and shlA, were significantly down-regulated compared with control. In addition, the extracts combined with imipenem inhibited the QS system of S. marcescens NJ01, disrupted its preformed biofilm, released the intra-biofilm bacteria and killed the bacteria gradually. Therefore, the extracts combined with imipenem can partially restore bacterial drug sensitivity. These results suggest that the extracts from SCSGAF0054 effectively interfere with the QS system to treat S. marcescens infection alone or combining with classical antimicrobial drugs.

Cultivation of the Dematiaceous Fungus Cladosporium sphaerospermum Aboard the International Space Station and Effects of Ionizing Radiation.

In Space, cosmic radiation is a strong, ubiquitous form of energy with constant flux, and the ability to harness it could greatly enhance the energy-autonomy of expeditions across the solar system. At the same time, radiation is the greatest permanent health risk for humans venturing into deep space. To protect astronauts beyond Earth's magnetosphere, advanced shielding against ionizing as well as non-ionizing radiation is highly sought after. In search of innovative solutions to these challenges, biotechnology appeals with suitability for in situ resource utilization (ISRU), self-regeneration, and adaptability. Where other organisms fail, certain microscopic fungi thrive in high-radiation environments on Earth, showing high radioresistance. The adaptation of some of these molds to areas, such as the Chernobyl Exclusion Zone has coined the terms positive "radiotropism" and "radiotrophy", reflecting the affinity to and stimulation by radiation, and sometimes even enhanced growth under ionizing conditions. These abilities may be mediated by the pigment melanin, many forms of which also have radioprotective properties. The expectation is that these capabilities are extendable to radiation in space. To study its growth in space, an experiment cultivating Cladosporium sphaerospermum Penzig ATCC® 11289™ aboard the International Space Station (ISS) was conducted while monitoring radiation beneath the formed biomass in comparison to a no-growth negative control. A qualitative growth advantage in space was observable. Quantitatively, a 1.21 ± 0.37-times higher growth rate than in the ground control was determined, which might indicate a radioadaptive response to space radiation. In addition, a reduction in radiation compared to the negative control was discernable, which is potentially attributable to the fungal biomass.

Cladosporium sphaerospermum as a Rare Cause of Pneumonia.

Cladosporium sphaerospermum isa radiotrophic dematiaceous fungus that can rarely cause disease in humans such as infections of the skin, eye, upper airways, and brain. To the best of our knowledge, we present the first reported case of Cladosporium sphaerospermum-induced invasive lung infection. This case presents a 51-year-old male with a medical history significant for heavy smoking and severe alcohol abuse who was admitted for acute hypoxic respiratory failure secondary to a large exudative right pleural effusion compounded by hydropneumothorax. Despite an initial positive clinical response, appropriate medical treatment, and eradication of the infection, which was confirmed by repeat negative culture studies, the patient had a complicated hospital course. It is suspected that the patient's medical history played a role in the acquisition of the Cladosporium sphaerospermum infection as smoking and alcohol use are known risk factors for aspiration of pathogens into the pulmonary tract. We believe it is important to bring to attention this less known organism as a potential differential diagnosis for a complicated lung infection.

Cladosins L-O, new hybrid polyketides from the endophytic fungus Cladosporium sphaerospermum WBS017.

The endophytic fungus Cladosporium sphaerospermum WBS017 was obtained from healthy bulbs of Fritillaria unibracteata var. wabuensis. Fermentation of C. sphaerospermum on solid rice medium yielded three new hybrid polyketides, cladosins L-N (1-3), and a known derivative cladodionen (4). Further cultivation of this fungus on white bean medium afforded an additional new hybrid polyketide, cladosin O (5) along with three known analogues (6-8). The structures of the new compounds were elucidated using a combination of NMR and HRESIMS data. The absolute configurations of compounds 2 and 3 were determined by Mosher's method and TDDFT-ECD calculations. All isolated compounds were evaluated for their cytotoxic and antimicrobial activities. Cladodionen (4) exhibited cytotoxicity against the mouse lymphoma cell line L5178Y with an IC50 value of 3.7 µM, and also exhibited antifungal activity against Ustilago maydis and Saccharomyces cerevisiae, while cladosin L (1) displayed week antibacterial activity against Staphylococcus aureus ATCC 29213 and S. aureus ATCC 700699 with MIC values of 50 and 25 µM, respectively.

Cladosporium sphaerospermum causing brain abscess, a saprophyte turning pathogen: Case and review of published reports.

Emergence of saprophytic fungi thriving in dead plant material and soil as opportunistic human pathogens is of great concern. Cladosporium species are environmental saprophytes reported to cause various superficial and invasive fungal infections worldwide. C. sphaerospermum, a predominantly indoor fungus has been reported from cases of meningitis, subcutaneous and pulmonary fungal infections in the past. Herein we report the first case of cerebral abscess due to C. sphaerospermum in an immunocompetent host who was successfully managed by combined medical and surgical therapy.

Novel resources of Taxol from endophytic and entomopathogenic fungi: Isolation, characterization and LC-Triple mass spectrometric quantification.

This work presents a second-to-none method for Taxol isolation from the Endophytic fungus Cladosporium sphaerospermum (AUMC 6896) and the Entomopathogenic fungus Metarizium anisopliae (AUMC 5130). The extracts were analyzed by high performance liquid chromatography (HPLC) and Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) using positive electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode. This is rapid, consistent, reproducible, accurate, and sensitive for quantifying Taxol across multiple samples. The yield of crude Taxol product obtained from Potato Dextrose broth (PDB) medium inoculated with Cladosporium sphaerospermu and Metarizium anisopliae was found to be 3.732, and 0.0023 µg L-1 respectively. The yield can be improved by adding ammonium acetate or salicylic acid to the culture broth. Addition of ammonium acetate (AA) (20 mg L-1) to culture media resulted in an increase of Taxol yield to 30.365 and 27.289 µg L-1 respectively. Production of Taxol was 29.844 and 67.254 µg L-1 for the two fungus species when ammonium acetate was substituted by 90 mg L-1 salicylic acid (SA). Adding both AA (20 mg L-1) and SA (90 mg L-1) to the culture media resulted in an increase of the Taxol yield to 4.054 and 116.373 µg L-1 respectively. Our proposed analytical method offers very fast (3 min) quantitation of Taxol in comparison with other published methods. These findings represent a new bioprospecting of the endophytic fungi that may serve as a potential material for the production of Taxol for anticancer treatment.

A peculiar case of onychomycosis caused by Cladosporium halotolerans

ABSTRACT A 49-year-old patient with changes in the nails of the hallux for 10 years was diagnosed with onychomycosis. The identity of the causative agent was confirmed as Cladosporium halotolerans from the Cladosporium sphaerospermum species complex using molecular techniques. MALDI-TOF identified the agent as C. sphaerospermum complex species. Overall, species such as onychomycosis agents should attract special attention to avoid mistakes in the identification process while considering a probable contaminant as responsible for the disease. These species deserve attention since there are rare descriptions of them as causes of onychomycosis. It is important to recognize them as causes of disease and not just as a probable contaminant.

Exposure in vitro to an Environmentally Isolated Strain TC09 of Cladosporium sphaerospermum Triggers Plant Growth Promotion, Early Flowering, and Fruit Yield Increase.

A growing number of bacteria and fungi have been found to promote plant growth through mutualistic interactions involving elements such as volatile organic compounds (VOCs). Here, we report the identification of an environmentally isolated strain of Cladosporium sphaerospermum (herein named TC09), that substantially enhances plant growth after exposure in vitro beyond what has previously been reported. When cultured on Murashige and Skoog (MS) medium under in vitro conditions, tobacco seedlings (Nicotiana tabacum) exposed to TC09 cultures for 20 days increased stem height and whole plant biomass up to 25- and 15-fold, respectively, over controls without exposure. TC09-mediated growth promotion required >5 g/L sucrose in the plant culture medium and was influenced by the duration of exposure ranging from one to 10 days, beyond which no differences were detected. When transplanted to soil under greenhouse conditions, TC09-exposed tobacco plants retained higher rates of growth. Comparative transcriptome analyses using tobacco seedlings exposed to TC09 for 10 days uncovered differentially expressed genes (DEGs) associated with diverse biological processes including cell expansion and cell cycle, photosynthesis, phytohormone homeostasis and defense responses. To test the potential efficacy of TC09-mediated growth promotion on agricultural productivity, pepper plants (Capsicum annuum L.) of two different varieties, Cayenne and Minisweet, were pre-exposed to TC09 and planted in the greenhouse to monitor growth, flowering, and fruit production. Results showed that treated pepper plants flowered 20 days earlier and yielded up to 213% more fruit than untreated controls. Altogether the data suggest that exposure of young plants to C. sphaerospermum produced VOCs may provide a useful tool to improve crop productivity.

Morphological and molecular identification of Cladosporium sphaerospermum isolates collected from tomato plant residues / Identificação morfológica e molecular de isolados de Cladosporium sphaerospermum coletados de resíduos de plantas de tomate

This study was conducted at the Agriculture College University of Karbala, Iraq to isolate and morphologically and molecularly diagnose thirteen Cladosporium isolates collected from tomato plant residues present in desert regions of Najaf and Karbala provinces, Iraq. We diagnosed the obtained isolates by PCR amplification using the ITS1 and ITS4 universal primer pair followed by sequencing. PCR amplification and analysis of nucleotide sequences using the BLAST program showed that all isolated fungi belong to Cladosporium sphaerospermum. Analysis of the nucleotide sequences of the identified C. sphaerospermum isolates 2, 6, 9, and 10 showed a genetic similarity reached 99%, 98%, 99%, and 99%, respectively, with those previously registered at the National Center for Biotechnology Information (NCBl). By comparing the nucleotide sequences of the identified C. sphaerospermum isolates with the sequences belong to the same fungi and available at NCBI, it was revealed that the identified C. sphaerospermum isolates 2, 6, 9, and 10 have a genetic variation with those previously recorded at the National Center for Biotechnology Information (NCBl); therefore, the identified sequences of C. sphaerospermum isolates have been registered in GenBank database (NCBI) under the accession numbers MN896004, MN896107, MN896963, and MN896971, respectively.

Este estudo foi conduzido na Agriculture College University of Karbala, Iraque, para isolar e diagnosticar morfológica e molecularmente treze isolados de Cladosporium coletados de resíduos de plantas de tomate presentes nas regiões desérticas das províncias de Najaf e Karbala, no Iraque. Diagnosticamos os isolados obtidos por amplificação por PCR usando o par de primers universais ITS1 e ITS4 seguido de sequenciamento. A amplificação por PCR e a análise de sequências de nucleotídeos usando o programa BLAST mostraram que todos os fungos isolados pertencem a Cladosporium sphaerospermum. A análise das sequências de nucleotídeos dos isolados 2, 6, 9 e 10 de C. sphaerospermum identificados mostrou similaridade genética de 99%, 98%, 99% e 99%, respectivamente, com aqueles previamente registrados no National Center for Biotechnology Informações (NCBl). Ao comparar as sequências de nucleotídeos dos isolados de C. sphaerospermum identificados com as sequências pertencentes aos mesmos fungos e disponíveis no NCBI, foi revelado que os isolados 2, 6, 9 e 10 de C. sphaerospermum identificados têm variação genética com aqueles anteriormente registrados no National Center for Biotechnology Information (NCBl). Portanto, as sequências identificadas de isolados de C. sphaerospermum foram registradas no banco de dados GenBank (NCBI) sob os números de acesso MN896004, MN896107, MN896963 e MN896971, respectivamente.

Morphological and molecular identification of Cladosporium sphaerospermum isolates collected from tomato plant residues

Abstract This study was conducted at the Agriculture College University of Karbala, Iraq to isolate and morphologically and molecularly diagnose thirteen Cladosporium isolates collected from tomato plant residues present in desert regions of Najaf and Karbala provinces, Iraq. We diagnosed the obtained isolates by PCR amplification using the ITS1 and ITS4 universal primer pair followed by sequencing. PCR amplification and analysis of nucleotide sequences using the BLAST program showed that all isolated fungi belong to Cladosporium sphaerospermum. Analysis of the nucleotide sequences of the identified C. sphaerospermum isolates 2, 6, 9, and 10 showed a genetic similarity reached 99%, 98%, 99%, and 99%, respectively, with those previously registered at the National Center for Biotechnology Information (NCBl). By comparing the nucleotide sequences of the identified C. sphaerospermum isolates with the sequences belong to the same fungi and available at NCBI, it was revealed that the identified C. sphaerospermum isolates 2, 6, 9, and 10 have a genetic variation with those previously recorded at the National Center for Biotechnology Information (NCBl); therefore, the identified sequences of C. sphaerospermum isolates have been registered in GenBank database (NCBI) under the accession numbers MN896004, MN896107, MN896963, and MN896971, respectively.

Resumo Este estudo foi conduzido na Agriculture College University of Karbala, Iraque, para isolar e diagnosticar morfológica e molecularmente treze isolados de Cladosporium coletados de resíduos de plantas de tomate presentes nas regiões desérticas das províncias de Najaf e Karbala, no Iraque. Diagnosticamos os isolados obtidos por amplificação por PCR usando o par de primers universais ITS1 e ITS4 seguido de sequenciamento. A amplificação por PCR e a análise de sequências de nucleotídeos usando o programa BLAST mostraram que todos os fungos isolados pertencem a Cladosporium sphaerospermum. A análise das sequências de nucleotídeos dos isolados 2, 6, 9 e 10 de C. sphaerospermum identificados mostrou similaridade genética de 99%, 98%, 99% e 99%, respectivamente, com aqueles previamente registrados no National Center for Biotechnology Informações (NCBl). Ao comparar as sequências de nucleotídeos dos isolados de C. sphaerospermum identificados com as sequências pertencentes aos mesmos fungos e disponíveis no NCBI, foi revelado que os isolados 2, 6, 9 e 10 de C. sphaerospermum identificados têm variação genética com aqueles anteriormente registrados no National Center for Biotechnology Information (NCBl). Portanto, as sequências identificadas de isolados de C. sphaerospermum foram registradas no banco de dados GenBank (NCBI) sob os números de acesso MN896004, MN896107, MN896963 e MN896971, respectivamente.

Morphological and molecular identification of Cladosporium sphaerospermum isolates collected from tomato plant residues / Identificação morfológica e molecular de isolados de Cladosporium sphaerospermum coletados de resíduos de plantas de tomate

This study was conducted at the Agriculture College University of Karbala, Iraq to isolate and morphologically and molecularly diagnose thirteen Cladosporium isolates collected from tomato plant residues present in desert regions of Najaf and Karbala provinces, Iraq. We diagnosed the obtained isolates by PCR amplification using the ITS1 and ITS4 universal primer pair followed by sequencing. PCR amplification and analysis of nucleotide sequences using the BLAST program showed that all isolated fungi belong to Cladosporium sphaerospermum. Analysis of the nucleotide sequences of the identified C. sphaerospermum isolates 2, 6, 9, and 10 showed a genetic similarity reached 99%, 98%, 99%, and 99%, respectively, with those previously registered at the National Center for Biotechnology Information (NCBl). By comparing the nucleotide sequences of the identified C. sphaerospermum isolates with the sequences belong to the same fungi and available at NCBI, it was revealed that the identified C. sphaerospermum isolates 2, 6, 9, and 10 have a genetic variation with those previously recorded at the National Center for Biotechnology Information (NCBl); therefore, the identified sequences of C. sphaerospermum isolates have been registered in GenBank database (NCBI) under the accession numbers MN896004, MN896107, MN896963, and MN896971, respectively.

Este estudo foi conduzido na Agriculture College University of Karbala, Iraque, para isolar e diagnosticar morfológica e molecularmente treze isolados de Cladosporium coletados de resíduos de plantas de tomate presentes nas regiões desérticas das províncias de Najaf e Karbala, no Iraque. Diagnosticamos os isolados obtidos por amplificação por PCR usando o par de primers universais ITS1 e ITS4 seguido de sequenciamento. A amplificação por PCR e a análise de sequências de nucleotídeos usando o programa BLAST mostraram que todos os fungos isolados pertencem a Cladosporium sphaerospermum. A análise das sequências de nucleotídeos dos isolados 2, 6, 9 e 10 de C. sphaerospermum identificados mostrou similaridade genética de 99%, 98%, 99% e 99%, respectivamente, com aqueles previamente registrados no National Center for Biotechnology Informações (NCBl). Ao comparar as sequências de nucleotídeos dos isolados de C. sphaerospermum identificados com as sequências pertencentes aos mesmos fungos e disponíveis no NCBI, foi revelado que os isolados 2, 6, 9 e 10 de C. sphaerospermum identificados têm variação genética com aqueles anteriormente registrados no National Center for Biotechnology Information (NCBl). Portanto, as sequências identificadas de isolados de C. sphaerospermum foram registradas no banco de dados GenBank (NCBI) sob os números de acesso MN896004, MN896107, MN896963 e MN896971, respectivamente.

Cladosporium lebrasiae, a new fungal species isolated from milk bread rolls in France.

The fungal genus Cladosporium (Cladosporiaceae, Dothideomycetes) is composed of a large number of species, which can roughly be divided into three main species complexes: Cladosporium cladosporioides, Cladosporium herbarum, and Cladosporium sphaerospermum. The aim of this study was to characterize strains isolated from contaminated milk bread rolls by phenotypic and genotypic analyses. Using multilocus data from the internal transcribed spacer ribosomal DNA (rDNA), partial translation elongation factor 1-α, actin, and beta-tubulin gene sequences along with Fourier-transform infrared (FTIR) spectroscopy and morphological observations, three isolates were identified as a new species in the C. sphaerospermum species complex. This novel species, described here as Cladosporium lebrasiae, is phylogenetically and morphologically distinct from other species in this complex.