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Plant Disease Nov 2022Garcinia mangostana L. is a famous tropical fruit in Asia. In April 2021, a leaf disease on G. mangostana cv. Huazhu was observed in Zhanjiang (21.17° N, 110.18° E),...
Garcinia mangostana L. is a famous tropical fruit in Asia. In April 2021, a leaf disease on G. mangostana cv. Huazhu was observed in Zhanjiang (21.17° N, 110.18° E), Guangdong province, China. Symptoms was on new leaves of 2 year old plants. The spots were circular to irregular, gray in the center, and brown on the lesion margin. The disease incidence was estimated 25% (n = 500 investigated plants from about 50-ha). Twenty diseased leaves were collected from the orchard. The margin of the diseased tissues was cut into 2 mm × 2 mm pieces; surface disinfected with 75% ethanol and 2% sodium hypochlorite for 30 and 60 s, respectively; and rinsed thrice with sterile water. The tissues were plated onto potato dextrose agar (PDA) medium and incubated at 28 ℃. Twenty-eight isolates were obtained (isolation frequency = 28/4×20 = 35%). Single-spore isolation method was used to recover pure cultures for three isolates (GMN-1, GMN-2, and GMN-3) (Liu et al. 2021). The colonies were initially white with cottony aerial mycelium at 7 days on PDA. Then, they developed black acervular conidiomata at 10 days. Conidia were clavate to fusiform, four-septate, straight or slightly curved, and measured 16.5 to 21.4 µm long (average 19.5 µm; n = 40) × 4.5 to 6.5 µm wide (average 5.2 µm; n = 40). The three median cells were versicolored, whereas the basal and apical cells were hyaline. Conidia had a single basal appendage (4.5 to 5.5 µm long; n = 40) and three apical appendages (19.2 to 24.5 µm long; n = 40). The morphological characteristics of the isolates are comparable with those of the genus Neopestalotiopsis (Sajeewa et al. 2012). Molecular identification was performed using the colony polymerase chain reaction method with MightyAmp DNA Polymerase (Takara-Bio, Dalian, China) (Lu et al. 2012). Sequences were generated from the isolates using primers for the rDNA ITS (ITS1/ITS4), TEF1-α (EF1-728F/EF1-986R), and β-tubulin (T1/βt2b) loci (Sajeewa et al. 2012). The sequences of the isolates were submitted to GenBank (ITS, MZ026535-MZ026537; TEF, MZ032203-MZ032205; β-tubulin, MZ032206-MZ032208). The sequences of the isolates were 100% identical to the type strain MFLUCC12-0281 (accession nos. JX398979, JX399014, and JX399045) through BLAST analysis. The isolates clustered with N. clavispora (MFLUCC12-0280 and MFLUCC12-0281). The pathogenicity was tested in vivo. Individual plants (cv. Huazhu) were grown (n = 2, 1-1.5 year old) in a greenhouse at 24 ℃-30 ℃ with 80% relative humidity. Wounded leaflets were inoculated with 5-mm-diameter mycelial plugs or agar plugs (as control). Besides, sterile cotton balls were immersed in the spore suspension (1 × 105 per mL) and sterile distilled water (control) for about 15 s before they were fixed on the leaves for 3 days. One plant employed for each isolate with nine leaves. The test was performed thrice. Disease symptoms were found on the leaflets after 10 days, whereas the controls remained healthy. The pathogen was re-isolated from infected leaves and phenotypically identical to the original isolates to fulfill Koch's postulates. Neopestalotiopsis clavispora and Pestalotiopsis clavispora are synonyms. The fungus appeared to have a wide host range and distribution including in Thailand, Malaysia, North Queensland, and Australia (Sajeewa et al. 2012;Shahriar et al. 2022). Thus, this is the first report of N. clavispora causing leaf spot on G. mangostana in China. This finding will help improve management strategies against the leaf spots on G. mangostana in China.
PubMed: 36350731
DOI: 10.1094/PDIS-05-22-1120-PDN -
Plant Disease Oct 2022Machilus pauhoi Kaneh. is an excellent evergreen broad-leaved tree species widely grown in China for its ornamental and economic value (He et al. 2022). In September...
Machilus pauhoi Kaneh. is an excellent evergreen broad-leaved tree species widely grown in China for its ornamental and economic value (He et al. 2022). In September 2021, a leaf spot was observed on M. pauhoi plants on Guantian forest farm (27°06'15.6″N, 114°34'20.72″E) in ji' an city, Jiangxi province, China. The disease incidence was estimated to be above 20%. The symptoms began as brown irregular spots, then the spots gradually expand over time, with a gray-to-brown center and dark brown-to-black edges. Small infected tissues (3 to 5 mm2) were surface-sterilized in 70% ethanol for 30 s and 2% NaClO for 60 s, and rinsed three times with sterile water (Ju et al. 2021). Tissues were placed on potato dextrose agar (PDA) and incubated at 25°C. Pure cultures were obtained by transferring hyphal tips to new PDA plates. Twenty-two isolates of Colletotrichum ssp. were obtained (isolation frequency about 78%). Three representative single-spore isolates (PN-1, PN-4, and PN-9) were used for morphological studies and phylogenetic analyses. Colonies on the PDA of the three isolates were white to gray with cottony mycelia and grayish-white on the undersides of the culture. Conidia were single-celled, straight, hyaline, cylindrical, clavate, and measured 11.4-16.8 ×4.1-5.5 µm (13.2 ± 1.0 × 4.4 ± 0.3 µm, n = 100). Appressoria were brown to dark brown, ovoid to clavate, slightly irregular to irregular, and ranged from 5.2-8.8 × 4.1-6.2 µm (6.7 ± 0.2 × 5.1 ± 0.3 µm, n=100). Morphological features were similar to Colletotrichum gloeosporioides species complex (Weir et al. 2012). The internal transcribed spacer (ITS) regions, actin (ACT), calmodulin (CAL), beta-tubulin 2 (TUB2), chitin synthase (CHS-1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were amplified from genomic DNA for the three isolates using primers ITS1/ITS4, ACT-512F/ACT-783R, CL1/CL2, T1/Bt2b, CHS-79F/CHS-354R and GDF/GDR (Weir et al. 2012), respectively. All sequences were deposited into GenBank (ITS, ON176154 - ON176156; ACT, ON185554 - ON185556; GAPDH, ON185563 - ON185565; TUB2, ON185566 - ON185568; CHS-1, ON185560 - ON185562; CAL, ON185557 - ON185559). 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 PN-1, PN-4, and PN-9 in the clade of C. siamense. Based on the multi-locus phylogeny and morphology, three isolates were identified as C. siamense. The pathogenicity of three isolates was tested on nine M. pauhoi plants, which were grown in the field. Healthy leaves were wounded with a sterile needle and inoculated with 10 µL of spore suspension (106 conidia/mL). The spore suspension of each isolate was inoculated onto six leaves. Another three plants inoculated with ddH2O served as the control (Wan et al. 2022). All the inoculated leaves were covered with plastic bags to keep them moist for 2 days (relative humidity > 80%). All the inoculated leaves showed similar symptoms to those observed in the field, whereas control leaves were asymptomatic for 7 days. C. siamense was reisolated from the lesions, whereas no fungus was isolated from control leaves. Up to now, Pestalotiopsis chamaeropis, Corynespora cassiicola and Arthrinium arundinis could infect M. pauhoi plants (Zhang et al. 2021), and cause leaf spots in China. To our knowledge, this is the first report of C. siamense causing leaf spots on M. pauhoi. This work provided crucial information for epidemiologic studies and appropriate control strategies for this newly emerging disease.
PubMed: 36210334
DOI: 10.1094/PDIS-08-22-2005-PDN -
Plant Disease Oct 2021Late-season bunch rots cause major losses in grape production every year in the Mid-Atlantic United States, but the causal agents are not well characterized. In this...
Late-season bunch rots cause major losses in grape production every year in the Mid-Atlantic United States, but the causal agents are not well characterized. In this study, 265 fungal isolates were collected from rotten grapes from 2014 to 2020 and identified to the genus level according to internal transcribed spacer sequences. The most prevalent of the 15 genera were , , , , , and Of these, isolates within three prevalent, yet understudied, genera were identified to be , , and . The pathogenicity of these three fungal species was evaluated in two field trials by artificially inoculating wounded and nonwounded grapes () of four cultivars at the phenological stages of bloom, véraison, and preharvest. Upon ripening, fruit were weighed and assessed for severity of multiple diseases. On nonwounded fruit, caused significantly higher disease severity than the control in both seasons. On wounded fruit, each inocula caused significantly higher disease than the respective controls in the first season, but only and caused this in the second season. Also, wounding was found to have a detrimental effect on cluster weight, which was significantly influenced by inoculation timing and cultivar. Lastly, and were tested for sensitivity to azoxystrobin, boscalid, and difenoconazole. The isolates were found to be more sensitive to boscalid and difenoconazole in general, with varying sensitivity to azoxystrobin. isolates were resistant to boscalid and azoxystrobin but displayed much higher sensitivity to difenoconazole. Evidence from the isolate collection and field trials demonstrates that could be a significant pathogen of wine grapes in the Mid-Atlantic United States. Results from this study will be useful for the identification and management of the understudied Alternaria, Aspergillus, and Neopestalotiopsis fruit rots of wine grapes.
Topics: Fungicides, Industrial; Seasons; United States; Virulence; Vitis; Wine
PubMed: 33656367
DOI: 10.1094/PDIS-01-21-0006-RE -
International Journal of Food... Apr 2018The aim of this study was to find alternatives to conventional synthetic fungicides to control postharvest decay of longan fruit. The antifungal potential of thymol,...
The aim of this study was to find alternatives to conventional synthetic fungicides to control postharvest decay of longan fruit. The antifungal potential of thymol, carvacrol and trans-cinnamaldehyde was evaluated against four major longan pathogens, Lasiodiplodia spp., Phomopsis spp., Pestalotiopsis spp. and Geotrichum candidum, using vapor phase and direct contact methods. The vapor phase of all active compounds was more effective on fungal growth than direct contact. A volatile vapor of thymol and carvacrol had strong antifungal activity against the tested fungi, exhibiting minimum inhibitory concentration (MIC) in the range of 40 to 80 mg/L air; trans-cinnamaldehyde showed the least efficiency, with MIC ranging from 80 to 160 mg/L air for G. candidum and Phomopsis spp., while it could not inhibit Lasiodiplodia spp. and Pestalotiopsis spp. at 160 mg/L air. The minimum fungicidal concentration (MFC) of thymol and carvacrol varied from 40 to 80 mg/L air, while trans-cinnamaldehyde completely inhibited the mycelial growth of the tested fungi at higher concentrations. Mycelial growth of all tested fungi decreased with increasing active compound concentration, except for trans-cinnamaldehyde. Thymol proved to be the most effective compound against the four tested fungi, with effective concentration 50 (EC) of 5.68 ± 0.59, 6.86 ± 0.52, 8.27 ± 0.22 and 9.99 ± 1.28 mg/L air for Lasiodiplodia spp., Phomopsis spp., Pestalotiopsis spp. and G. candidum, respectively. Fungal growth curves were adequately fitted (0.958 < R < 0.996) by a modified Gompertz model. For all tested fungi, the lag phase (λ) of fungal mycelia exposed to thymol and carvacrol increased, while the maximum colony diameter (A) and maximum growth rate (v) decreased. A combination of thymol and carvacrol exhibited an antagonistic effect against G. candidum but an indifferent effect against Lasiodiplodia spp., Phomopsis spp. and Pestalotiopsis spp.
Topics: Acrolein; Antifungal Agents; Ascomycota; Cymenes; Fruit; Fungicides, Industrial; Microbial Sensitivity Tests; Monoterpenes; Oils, Volatile; Phytochemicals; Sapindaceae; Thymol
PubMed: 29459244
DOI: 10.1016/j.ijfoodmicro.2018.02.009 -
The Journal of Antibiotics Apr 2022Four new pestalone-type benzophenones, pestalotinones A-D (1-4), along with six known congeners, pestalone, pestalone E-F, SB87-Cl, SB87-H, and pestalachloride B, were...
Four new pestalone-type benzophenones, pestalotinones A-D (1-4), along with six known congeners, pestalone, pestalone E-F, SB87-Cl, SB87-H, and pestalachloride B, were isolated from the endophytic fungus Pestalotiopsis trachicarpicola SC-J551 cultivated on rice grains. Their structures were established by extensive spectroscopic analysis. Compounds 1-3 exhibited potent activity against Staphylococcus aureus and MRSA (MIC: 1.25-2.5 μg ml) while no cytotoxicity against Vero cells (IC > 50 μM). The activity profile of this group of compounds suggested that replacement of the C-14 aldehyde with an oxymethyl greatly increases their activity and selectivity towards the bacteria and chlorine substitutions result in the increase of antibacterial activity and slight decrease of cytotoxicity against the mammalian cells.
Topics: Animals; Anti-Bacterial Agents; Benzophenones; Chlorocebus aethiops; Fungi; Molecular Structure; Pestalotiopsis; Vero Cells
PubMed: 35115699
DOI: 10.1038/s41429-022-00510-0 -
Current Medical Mycology Sep 2023Onychomycosis caused by dematiaceous fungi is rarely reported and the identification is also quite tricky due to poor sporulation. Recent emergence of dematiaceous fungi...
BACKGROUND AND PURPOSE
Onychomycosis caused by dematiaceous fungi is rarely reported and the identification is also quite tricky due to poor sporulation. Recent emergence of dematiaceous fungi as a major cause of onychomycosis is a matter of concern in the field of mycology. Therefore, this study aimed to understand the dematiaceous fungi as a possible cause of onychomycosis, especially among agricultural workers. In addition, the evaluation of the antifungal susceptibility patterns led to the idea of an accurate drug that will help to treat and prevent antifungal resistance.
MATERIALS AND METHODS
The standard procedure was followed for direct microscopic examination and fungi isolation. Furthermore, antifungal susceptibility testing was conducted in accordance with the Clinical and Laboratory Standards Institute M-38-A2 protocol.
RESULTS
Both potassium hydroxide and fungal positivity were found in 275 out of 356 suspected cases, 52%, 4.3%, 28.7%, and 14.9% of which were non-dermatophytic molds (NDMs), yeast, dermatophytes, and sterile hyphae, respectively. Among NDMs (52%, n=143), 45.5% (n=65) were hyaline hyphomycetes and 54.5% (n=78) were dematiaceous hyphomycetes. Among dematiaceous fungi, spp. and spp. were the commonly isolated ones. Additionally, azoles, amphotericin-B, and anidulafungin showed excellent antifungal activity against tested isolates.
CONCLUSION
Dematiaceous fungi are now becoming a potential cause of onychomycosis. A more detailed study is needed on the identification of these emerging isolates and the mode of action of antifungal drugs for a better treatment strategy.
PubMed: 38361959
DOI: 10.22034/cmm.2023.345077.1428 -
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 -
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