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Scientific Reports Feb 2022Mangrove endophytic fungi are tolerant to numerous stresses and are inevitably capable of exhibiting excellent biological activity by producing impressive numbers of...
Mangrove endophytic fungi are tolerant to numerous stresses and are inevitably capable of exhibiting excellent biological activity by producing impressive numbers of metabolites with special biological functions, based on previous work on the biological potential of mangrove-derived endophytic fungi. To obtain marked antimicrobial and cytotoxic fermentation products of culturable endophytic fungi from mangrove forests, our research evaluated the antimicrobial and cytotoxic activities of crude extracts of endophytic fungi from Rhizophora stylosa and Rhizophora mucronata. Forty-six fungal isolates were cultured on four different media, namely, dextrose agar (PDA), Czapek's agar (CZA), rice medium (RM) and grain medium (GM) and harvested by ethyl acetate solvent at 40 days. The extracts were tested for antimicrobial activity by the microdilution method against the gram-negative bacteria Pseudomonas adaceae (PA), gram-positive bacteria Enterococcus faecalis (EF), methicillin-resistant Staphylococcus aureus (MRSA) and pathogenic fungus Monilia albicans (MA). The cytotoxic activity of the extracts was evaluated by MTT assay using A549 human lung cancer cells, HeLa human cervical carcinoma cells, and HepG2 human hepatocellular cells. The results showed that rice medium could promote the secretion of antimicrobial and antitumour secondary metabolites of endophytic fungi in comparison with other cultivation media. Seventeen strains (68%) from R. stylosa exhibited inhibitory effects on indicators, especially N. protearum HHL46, which could inhibit the growth of four microbes with MIC values reaching 0.0625 mg/mL. Fifteen strains (71.4%) from R. mucronata displayed activities against human pathogenic microbes; in particular, Pestalotiopsis sp. HQD6 and N. protearum HQD5 could resist the growth of four microbes with MIC values ranging from 0.015 to 1 mg/mL. In the cytotoxicity assay, the extracts of 10 strains (40%), 9 strains (40%) and 13 strains (52%) of R. stylosa and 13 strains (61.9%), 10 strains (47.6%) and 10 strains (47.6%) of R. mucronata displayed cytotoxicity against A549, HeLa and HepG2 cancer cells with cell viability values ≤ 50%. Neopestalotiopsis protearum HHL46, Phomopsis longicolla HHL50, Botryosphaeria fusispora HQD83, Fusarium verticillioides HQD48 and Pestalotiopsis sp. HQD6 displayed significant antitumour activity with IC values below 20 μg/mL. These results highlighted the antimicrobial and antitumour potential of endophytic fungi from R. stylosa and R. mucronata and the possibility of exploiting their antimicrobial and cytotoxic agents.
Topics: A549 Cells; Anti-Infective Agents; Bacteria; Complex Mixtures; Cytotoxins; Drug Evaluation, Preclinical; Endophytes; Fungi; HeLa Cells; Hep G2 Cells; Humans; Rhizophoraceae
PubMed: 35177749
DOI: 10.1038/s41598-022-06711-9 -
PloS One 2021Loquat [Eriobotrya japonica (Thunb.) Lindl.] is an important fruit crop in Pakistan; however, a constant decline in its production is noted due biotic and abiotic...
Loquat [Eriobotrya japonica (Thunb.) Lindl.] is an important fruit crop in Pakistan; however, a constant decline in its production is noted due biotic and abiotic stresses, particularly disease infestation. Fungal pathogens are the major disease-causing agents; therefore, their identification is necessary for devising management options. This study explored Taxila, Wah-Cantt, Tret, Chatar, Murree, Kalar-Kahar, Choa-Saidan-Shah and Khan-Pur districts in the Punjab and Khyber Paktoon Khawa (KPK) provinces of Pakistan to explore the diversity of fungal pathogens associated with loquat. The samples were collected from these districts and their microscopic characterizations were accomplished for reliable identification. Alternaria alternata, Curvularia lunata, Lasiodiplodia theobromae, Aspergilus flavis, Botrytis cinerea, Chaetomium globosum, Pestalotiopsis mangiferae and Phomopsis sp. were the fungal pathogens infesting loquat in the study area. The isolates of A. alternata and C. lunata were isolated from leaf spots and fruit rot, while the isolates of L. theobromae were associated with twig dieback. The remaining pathogens were allied with fruit rot. The nucleotide evidence of internal transcribed spacer (ITS) regions (ITS1, 5.8S, and ITS2) were computed from all the pathogens and submitted in the database of National Center for Biotechnology Information (NCBI). For multigene analysis, beta-tubulin (BT) gene and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) regions were explored for A. alternata and C. lunata isolates, respectively. The virulence scales of leaf spots, fruit rot, and twig dieback diseases of loquat were developed for the first time through this study. It is the first comprehensive study with morpho-molecular identification, and newly developed virulence scales of the fungal pathogens associated with loquat, which improves the understanding of these destructive diseases.
Topics: Ascomycota; Biodiversity; Crops, Agricultural; DNA, Fungal; Eriobotrya; Fruit; Mitosporic Fungi; Pakistan; Phylogeny; Plant Diseases; Plant Leaves; Virulence
PubMed: 34648523
DOI: 10.1371/journal.pone.0257951 -
Molecules (Basel, Switzerland) Feb 2022In Mexico, the mango crop is affected by anthracnose caused by species. In the search for environmentally friendly fungicides, chitosan has shown antifungal activity....
In Mexico, the mango crop is affected by anthracnose caused by species. In the search for environmentally friendly fungicides, chitosan has shown antifungal activity. Therefore, fungal isolates were obtained from plant tissue with anthracnose symptoms from the state of Guerrero in Mexico and identified with the ITS and β-Tub genetic markers. Isolates of the complex were again identified with the markers ITS, Act, β-Tub, GADPH, CHS-1, CaM, and ApMat. Commercial chitosan (Aldrich, lot # STBF3282V) was characterized, and its antifungal activity was evaluated on the radial growth of the fungal isolates. The isolated anthracnose-causing species were , , , and . Other fungi found were sp., , sp., sp., , , and . Chitosan showed 78% deacetylation degree and a molecular weight of 32 kDa. Most of the species and the other identified fungi were susceptible to 1 g L chitosan. However, two isolates were less susceptible to chitosan. Although chitosan has antifungal activity, the interactions between species of the complex and their effect on chitosan susceptibility should be studied based on genomic changes with molecular evidence.
Topics: Antifungal Agents; Chitosan; Colletotrichum; Mangifera
PubMed: 35209032
DOI: 10.3390/molecules27041244 -
International Journal of Molecular... Jul 2023sp. is an important class of plant pathogenic fungi that can infect a variety of crops. We have proved the pathogenicity of on bayberry leaves and caused bayberry...
sp. is an important class of plant pathogenic fungi that can infect a variety of crops. We have proved the pathogenicity of on bayberry leaves and caused bayberry blight. Phenazine-1-carboxylic acid (PCA) has the characteristics of high efficiency, low toxicity, and environmental friendliness, which can prevent fungal diseases on a variety of crops. In this study, the effect of PCA on the morphological, physiological, and molecular characteristics of has been investigated, and the potential antifungal mechanism of PCA against was also explored. We applied PCA on in vitro and in vivo to determine its inhibitory effect on PCA. It was found that PCA was highly efficient against with EC around 2.32 μg/mL, and the in vivo effect was 57% at 14 μg/mL. The mechanism of PCA was preliminarily explored by transcriptomics technology. The results showed that after the treatment of PCA, 3613 differential genes were found, focusing on redox processes and various metabolic pathways. In addition, it can also cause mycelial development malformation, damage cell membranes, reduce mitochondrial membrane potential, and increase ROS levels. This result expanded the potential agricultural application of PCA and revealed the possible mechanism against .
Topics: Antifungal Agents; Pestalotiopsis; Fungi
PubMed: 37511033
DOI: 10.3390/ijms241411274 -
Archives of Microbiology Oct 2021Endophytes associated with plants have the property to produce active biomolecules with their possible applications in agro-industrial sectors. This study provides a...
Endophytes associated with plants have the property to produce active biomolecules with their possible applications in agro-industrial sectors. This study provides a project work on analyzing various activities of fungal endophytes isolated from Swertia chirayita of Sikkim Himalayan region. Among several fungal endophytes screened, isolate UTCRF6 was found most active with the secretion of enzymes protease, cellulase, amylase and chitinase, as well as other metabolites Indoleacetic acid and siderophores. This endophyte was found active in restricting the growth of phyto-pathogens, including strains of Fusarium solani, Colletotrichum gloeosporioides, Alternaria alternata, Pestalotiopsis theae and Sclerotinia sclerotiorum. Morphological and molecular studies of this endophytic fungus showed similarity with Penicillium citrinum.
Topics: Alternaria; Ascomycota; Biological Control Agents; Colletotrichum; Endophytes; Fusarium; Penicillium; Plants, Medicinal; Swertia
PubMed: 34338823
DOI: 10.1007/s00203-021-02498-x -
Pathogens (Basel, Switzerland) Dec 2019This study contributes new knowledge on the diversity of conidial fungi in edible pine () seeds found in Yunnan Province, China and emphasizes the importance of edible...
This study contributes new knowledge on the diversity of conidial fungi in edible pine () seeds found in Yunnan Province, China and emphasizes the importance of edible seed products to ensure food safety standards. We isolated two fungal species, one on the pine seed coat and the other on the endosperm of the pine seed. The two fungal species were identified as sp. nov. and a new host record . Characteristic morphological features of were used alongside results from multi-gene phylogenetic analysis to distinguish it from currently known species within the genus. was identified as a new host record based on morphological features and phylogenetic analysis. In addition, detailed descriptions, scanned electron microscopy morphology, illustrations, and phylogenetic trees are provided to show the placement of these species.
PubMed: 31817121
DOI: 10.3390/pathogens8040285 -
Plant Disease Jul 2023Photinia bodinieri Lévl. is an evergreen broadleaf species widely cultivated in subtropical China as an ornamental value (Zhang et al. 2018). In July 2021, leaf spot...
Photinia bodinieri Lévl. is an evergreen broadleaf species widely cultivated in subtropical China as an ornamental value (Zhang et al. 2018). In July 2021, leaf spot symptoms were observed on the campus of Jiangxi Agricultural University (28°45'56″N, 115°50'21″E), Jiangxi province, China. The spots were circular to irregular, gray in the center, and dark brown on the lesion margin. The disease incidence was estimated 15%. Leaf pieces (5 × 5 mm) from the lesion borders were surface-sterilized in 70% ethanol for 30 s, followed by 2% NaOCl for 1 min, and then rinsed three times with sterile water. Tissues were placed on potato dextrose agar (PDA) and incubated at 25°C in the dark. Pure cultures were obtained by monosporic isolation, and the representative isolates, SN-3, SN-7, and SN-11 were used for morphological studies and phylogenetic analyses. The colonies of three isolates grown on PDA were white, cottony, and exhibited flocculent, contained undulate edges with dense aerial mycelium on the surface. Conidia were 5-celled, clavate to fusiform, smooth, 18.2-24.3 × 5.5-8.4 μm (n = 100). The 3 median cells were dark brown to olivaceous, central cell was darker than other 2 cells, and the basal and apical cells were hyaline. Conidia developed filiform appendages; one basal appendage (3.3-8.2 μm long; n = 100), and 2-3 apical appendages (16-29 μm long; n = 100). Morphological features were similar to Neopestalotiopsis sp. (Maharachchikumbura et al. 2014). Portions of internal transcribed spacer (ITS) regions, β-tubulin 2 (TUB2) and translation elongation factor 1-alpha (TEF1-α) genes were amplified from genomic DNA for the three isolates using primers ITS1/ITS4, T1/Bt-2b, EF1-728F/EF-2 (Maharachchikumbura et al. 2014), respectively. All sequences were deposited into GenBank (ITS, OQ572345 - OQ572347; TUB2, OQ597847 - OQ597849; TEF1-α, OQ597844 - OQ597846). A maximum likelihood and Bayesian posterior probability analyses using IQtree v. 1.6.8 and Mr. Bayes v. 3.2.6 with the concatenated sequences placed SN-3, SN-7, and SN-11 in the clade of N. clavispora. Based on the multi-locus phylogeny and morphology, three isolates were identified as N. clavispora. Pathogenicity of the three isolates was verified on nine disease-free 7-year-old Photinia bodinieri plants, which were grown in the field. Two healthy leaves per plant were wounded with two pricks using a sterile needle (Φ=0.5 mm) and inoculated with 20 μL conidial suspension per leaf (106 conidia/mL). Another nine control plants were inoculated with sterile water. 36 leaves were used for the pathogenicity test of three isolates. All leaves were covered with plastic bags to maintain a humid environment for 2 days. The inoculated leaves showed similar symptoms to those observed in the field, whereas control leaves were asymptomatic after 10 days. The fungi were consistently reisolated only from the inoculated and symptomatic leaves, fulfilling Koch's postulates. N. clavispora can cause leaf diseases in a variety of hosts, including Kadsura coccinea (Xie et al. 2018), Photinia serratifolia (Yang et al. 2018), Camellia chrysantha (Zhao et al. 2020). Photinia spp. is an excellent landscape gardening plant, threatened with grey blight (Pestalotiopsis microspore) (Ye et al. 2022), anthracnose (Colletotrichum sp.) (Guan et al. 2013). However, this is the first report of N. clavispora infecting Photinia bodinieri in China. This work provided crucial information for epidemiologic studies and appropriate control strategies for this newly emerging disease.
PubMed: 37467128
DOI: 10.1094/PDIS-06-23-1108-PDN -
Journal of Fungi (Basel, Switzerland) May 2023(J. U. Santos) is an endemic plant species in Morro do Urucum in the Pantanal wetland (Brazil). is used for the restoration of areas impacted by iron mining...
(J. U. Santos) is an endemic plant species in Morro do Urucum in the Pantanal wetland (Brazil). is used for the restoration of areas impacted by iron mining activities. This study evaluates the diversity (composition, value and abundance) of endophytic fungal communities, considering parts of the plant and soil condition. The leaves and roots of were collected from native vegetation areas (NVA) and recovery areas (RCA) in Morro do Urucum. Illumina sequencing technology was used to investigate variation in endophytic fungal biodiversity. The operational taxonomic units detected in NVA ranged from 183 to 263 (leaf) and 115 to 285 (root), while RCA samples ranged from 200 to 282 (leaf) and 156 to 348 (root). Ascomycota phylum was the most common species among all plant samples. The most significant classes identified were Lecanoromycetes and Dothideomycetes that differed significantly ( ≤ 0.05) according to their plant hosts and soil stress. The relative abundance of (Sordariomycetes class) and (Lecanoromycetes class) genera was influenced by the iron mining activities according to the leaf samples analysed. However, the abundance and wealth of endophytic fungal communities in from RCA were evidence that could explain their high resilience to environmental disturbances and the source-sink dynamics of fungal propagules.
PubMed: 37367568
DOI: 10.3390/jof9060632 -
Pathogens (Basel, Switzerland) Mar 2021Bayberry is an important fruit tree native to the subtropical regions of China. However, a systematic twig blight disease caused by and , resulted in the death of the...
Bayberry is an important fruit tree native to the subtropical regions of China. However, a systematic twig blight disease caused by and , resulted in the death of the whole tree of bayberry. The main variety Dongkui is highly sensitive to the twig blight disease, but the variety Zaojia is very highly resistant to the disease. Therefore, it is very necessary to clear the difference between resistant and susceptible varieties in response to the fungal infection. In this paper, we investigated the response of resistant and susceptible bayberry cultivars to infection of twig blight pathogen by histological observation and gibberellin signaling pathway-related genes expression. Microscopic observation revealed the difference in the infection process between resistant and susceptible varieties. The results of frozen scanning electron microscopy showed that the conidia were shrunk, the mycelium was shriveled and did not extend into the cells of resistant cultivars, while the conidia were full and the top was extended, the mycelia was normal and continued to extend to the cells of a susceptible cultivar. Indeed, the medulla cells were almost intact in resistant cultivar, but obviously damaged in susceptible cultivar after inoculation of the main fungal pathogen conidia, which is earlier germinated on sterile glass slide than that of a hard plastic slide. The quantitative real-time PCR results showed a significant difference between resistant and susceptible cultivars in the expression of gibberellin signaling pathway-related genes in leaves and stems of bayberry, which is closely related to infection time, the type of genes and varieties. Overall, this study provides a clue for our understanding of the resistance mechanism of bayberry against the twig blight disease.
PubMed: 33805451
DOI: 10.3390/pathogens10040402 -
PloS One 2018Pestalopezia brunneopruinosa, the type species of Pestalopezia in Leotiomycetes, produces typical cup-shaped ascomata. Because its asexual morph has conidia comprised of...
Pestalopezia brunneopruinosa, the type species of Pestalopezia in Leotiomycetes, produces typical cup-shaped ascomata. Because its asexual morph has conidia comprised of five cells including apical and basal appendages and three pigmented median cells, it was first described as Pestalotia gibbosa, which belongs to Sordariomycetes. This contradiction has not been resolved due to the difficulty in isolating this fungus in culture. In this study, we isolated separate strains from the sexual morph and the asexual morph for molecular analysis. Phylogenetic trees of Sporocadaceae based on internal transcribed spacer, partial β-tubulin, and partial translation elongation factor 1-alpha sequence datasets revealed that both strains fall into the same taxon, in a clade in Pestalotiopsis sensu stricto alongside P. gaultheriae and P. spathulata. We provide the first evidence that fungi producing cup-shaped ascomata in Pestalotiopsis belong to Sordariomycetes, and we have proposed the transfer of Pestalopezia brunneopruinosa to Pestalotiopsis gibbosa.
Topics: Ascomycota; Fungal Proteins; Phylogeny; Tubulin
PubMed: 29949577
DOI: 10.1371/journal.pone.0197025