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Plant Disease Jun 2024Psidium guajava L. is widely cultivated in southern China. In May 2021, guava scab on cv. Zhenzhu was observed in Zhanjiang (21.18° N, 110.21° E), Guangdong province,...
Psidium guajava L. is widely cultivated in southern China. In May 2021, guava scab on cv. Zhenzhu was observed in Zhanjiang (21.18° N, 110.21° E), Guangdong province, China. Guava scab was corky with ovoid or round lesions on the surfaces of green fruits. Gradually the lesions sunk. Disease incidence was estimated as 85% in 500 investigated plants in about 50 ha. Twenty diseased fruits were collected from twenty trees in the field. From each fruit 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, successively; and rinsed thrice with sterile water. The tissues were plated onto potato dextrose agar (PDA) medium and incubated at 28 ℃. Thirty-four isolates were obtained. Single-spore isolation method (Liu et al. 2021) was used to recover pure cultures of three isolates (PGNC-1, PGNC-2, and PGNC-3) . The colonies were initially white with cottony aerial mycelium at 7 days on PDA. Then, these colonies form black acervular conidiomata at 10 days. Conidia were clavate to fusiform, four-septate, straight or slightly curved, and measured 15.8 to 21.2 µm × 4.5 to 6.5 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 were consistent with the description of Neopestalotiopsis clavispora (Maharachchikumbura et al. 2012). Molecular identification was performed using PCR 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 (Maharachchikumbura et al. 2012). The sequences of the isolates were submitted to GenBank (ITS, OQ996557 to OQ996559; TEF, OR101037 to OR101039; β-tubulin, OR100971 to OR100973). 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). N. clavispora and Pestalotiopsis clavispora are synonyms. The pathogenicity was tested in vivo. Plants (cv. Zhenzhu) were grown ( 3 years old) in a quarantine orchard at 25 ℃ to 32 ℃ with 60 to 80% relative humidity in May 2022. Disease-free green fruits were inoculated. 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 wounded fruits with transparent tape. Five fruits on one plant per isolate were inoculated. Five fruits on one plant severed as control. The test was performed thrice. Disease symptoms were found on the inoculated fruits after 20 days, whereas the controls remained healthy. The pathogen was re-isolated from infected fruits and was phenotypically identical to the original isolates thus fulfilling Koch's postulates. Neopestalotiopsis or Pestalotiopsis spp. were reported to be the causal agents of guava scab in Colombia and in Hawaii (Keith et al. 2006; Solarte et al. 2018). N. clavispora has been reported to cause disease in a broad range of hosts (Ge et al. 2009; Chen et al. 2018), but not in guava. This is the first report of N. clavispora causing guava scab in China. There would be no harvest if this disease is left unmanaged.
PubMed: 38885025
DOI: 10.1094/PDIS-11-23-2357-PDN -
Marine Drugs Nov 2022Five undescribed polyketide derivatives, pestaloketides A-E (-), along with eleven known analogues (-), were isolated from the sponge-derived fungus sp. Their...
Five undescribed polyketide derivatives, pestaloketides A-E (-), along with eleven known analogues (-), were isolated from the sponge-derived fungus sp. Their structures, including absolute configurations, were elucidated by analyses of NMR spectroscopic HRESIMS data and electronic circular dichroism (ECD) calculations. Compounds , , , and exhibited weak cytotoxicities against four human cancer cell lines, with IC values ranging from 22.1 to 100 μM. Pestaloketide A () is an unusual polyketide, featuring a rare 5/10/5-fused ring system. Pestaloketides A () and B () exhibited moderately inhibited LPS-induced NO production activity, with IC values of 23.6 and 14.5 μM, respectively, without cytotoxicity observed. Preliminary bioactivity evaluations and molecular docking analysis indicated that pestaloketides A () and B () had the potential to be developed into anti-inflammatory activity drug leads.
Topics: Humans; Polyketides; Pestalotiopsis; Molecular Docking Simulation; Fungi; Anti-Inflammatory Agents
PubMed: 36421989
DOI: 10.3390/md20110711 -
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 -
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 -
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 -
Frontiers in Microbiology 2022Tea () is an important crop that is mainly used in the food industry. This study using the metabolome and microbiome investigates the resistance factors of wild tea...
Tea () is an important crop that is mainly used in the food industry. This study using the metabolome and microbiome investigates the resistance factors of wild tea plant resources against tea gray blight disease, which is caused by (Sawada) Steyaert. According to the interaction analysis of tea leaves and pathogenic fungus, the resistance of wild tea plant resource "R1" (Resistance 1) to tea gray blight disease was significantly higher than that of wild tea plant resource "S1" (Susceptibility 1). The difference between "R1" and "S1" in the metabolome was obvious. There were 145 metabolites that significantly changed. The phenolic acids and flavonoids were the major increased categories in "R1," and it included 4-O-glucosyl-sinapate and petunidin-3-o-(6"-o-p-coumaroyl) rutinoside. Six metabolic pathways were significantly enriched, including aminoacyl-tRNA biosynthesis, flavone, and flavonol biosynthesis. In terms of bacteria, there was no significant difference between "S1" and "R1" in the principal component analysis (PCA). was the major bacterial genus in "S1" and "R1." In addition, each of the two resources had its own predominant genus: was a predominant bacterial genus in "S1" and was a predominant bacterial genus in "R1." In terms of fungi, the fungal diversity and the abundance of the two tea plant resource samples could be distinguished clearly. The fungal component of "S1" was more abundant than that of "R1" at the genus level. x was the predominant fungal genus of "S1," and was the predominant fungal genus of "R1." The relative abundance of and were significantly different between "S1" and "R1." was identified as a potential biomarker. They correlated with some metabolites enriched in "S1" or "R1," such as L-arginine and quercetin-3-o-(2"-o-rhamnosyl) rutinoside-7-o-glucoside. Overall, phenolic acids, flavonoids, and could be functional metabolites or microorganisms that contributed to improving the resistance of wild tea plant resources to tea gray blight disease.
PubMed: 35910661
DOI: 10.3389/fmicb.2022.907962 -
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