Stefan Hobi, Wing Yan Jacqueline Tam, May Tse, Omid Nekouei, Yingfei Chai, Fraser I Hill, Edmund Cheung, Wietz Botes, Francois Saulnier-Troff, Colin T McDermott, Vanessa R Barrs
Dermatophytic pseudomycetoma (DPM) is a rarely reported invasive fungal infection of humans and animals, especially cats. This study aimed to identify dermatophytes, breed associations, and the frequency of extracutaneous (EC) involvement in feline DPM. Electronic records and formalin-fixed paraffin-embedded tissue (FFPET) from 32 suspected DPM cases in 30 cats were retrieved from a diagnostic laboratory between 2018 and 2024. To confirm DPM and molecular identity, DNA was extracted from FFPET for ITS2 sequencing, and immunohistochemistry was performed on PCR-negative cases. All cases were confirmed as DPM. Microsporum canis was the only dermatophyte identified. The sensitivity and specificity of ITS2 sequencing for M. canis identification in FFPET were 22/32 (68.8%) and 21/22 (95.5%), respectively. Exotic (36.7%) and Persian (23.3%) but not British breeds (26.3%) were over-represented compared to feline admissions at an affiliated veterinary hospital (8.5%, p < 0.001; 3%, p < 0.001; 21.6%, p = 0.817, respectively). Five cases (16.7%) had EC lesions; two had intra-abdominal masses; two had oral cavity masses, including one which extended into the cranial vault; and one had superficial cervical lymph node invasion. Exotic and Persian breeds are over-represented for DPM and M. canis is the primary cause. EC lesions of DPM may occur more commonly than previously thought.
{"title":"<i>Microsporum canis</i> Causes Cutaneous and Extracutaneous Feline Dermatophytic Pseudomycetomas: Molecular Identification and Clinicopathological Characteristics.","authors":"Stefan Hobi, Wing Yan Jacqueline Tam, May Tse, Omid Nekouei, Yingfei Chai, Fraser I Hill, Edmund Cheung, Wietz Botes, Francois Saulnier-Troff, Colin T McDermott, Vanessa R Barrs","doi":"10.3390/jof10080576","DOIUrl":"10.3390/jof10080576","url":null,"abstract":"<p><p>Dermatophytic pseudomycetoma (DPM) is a rarely reported invasive fungal infection of humans and animals, especially cats. This study aimed to identify dermatophytes, breed associations, and the frequency of extracutaneous (EC) involvement in feline DPM. Electronic records and formalin-fixed paraffin-embedded tissue (FFPET) from 32 suspected DPM cases in 30 cats were retrieved from a diagnostic laboratory between 2018 and 2024. To confirm DPM and molecular identity, DNA was extracted from FFPET for ITS2 sequencing, and immunohistochemistry was performed on PCR-negative cases. All cases were confirmed as DPM. <i>Microsporum canis</i> was the only dermatophyte identified. The sensitivity and specificity of ITS2 sequencing for <i>M. canis</i> identification in FFPET were 22/32 (68.8%) and 21/22 (95.5%), respectively. Exotic (36.7%) and Persian (23.3%) but not British breeds (26.3%) were over-represented compared to feline admissions at an affiliated veterinary hospital (8.5%, <i>p</i> < 0.001; 3%, <i>p</i> < 0.001; 21.6%, <i>p</i> = 0.817, respectively). Five cases (16.7%) had EC lesions; two had intra-abdominal masses; two had oral cavity masses, including one which extended into the cranial vault; and one had superficial cervical lymph node invasion. Exotic and Persian breeds are over-represented for DPM and <i>M. canis</i> is the primary cause. EC lesions of DPM may occur more commonly than previously thought.</p>","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.2,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11355761/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142080563","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Serpula himantioides is a globally distributed wood decay fungus that causes heartwood decay in several tree species. We investigated the occurrence of S. himantioides fruiting bodies in Japan for two years and six months to characterize their biology. The fruiting bodies matured in autumn and occurred on living Chamaecyparis pisifera, Chamaecyparis obtusa, Larix kaempferi, and Cryptomeria japonica trees, as well as on dead trees and soil. Assessing three circular plots, the incidence of living trees with S. himantioides fruiting bodies was lowest in the plot with the most advanced heartwood decay. Furthermore, fruiting bodies occurred more frequently in the lower slope direction of the trunk. Analysis using the pair correlation function suggested that the spatial distribution pattern of living trees with fruiting bodies may change from intensive to random with heartwood decay progress. Finally, according to generalized linear and generalized linear mixed models, which were used to investigate the factors affecting the development of fruiting bodies in C. pisifera, C. obtusa, and L. kaempferi, no clear relationship was found between the presence or absence of fruiting bodies and heartwood decay. Thus, we suggest that fruiting bodies can occur in healthy living trees as well as in living trees in the early stages of heartwood decay.
Serpula himantioides是一种分布于全球的木材腐朽真菌,会导致多个树种的心材腐朽。我们对日本两年零六个月来发生的S. himantioides子实体进行了调查,以了解其生物学特性。子实体在秋季成熟,出现在活的 Chamaecyparis pisifera、Chamaecyparis obtusa、Larix kaempferi 和 Cryptomeria japonica 树木上,也出现在死树和土壤上。在对三个环形地块进行评估后发现,在心材腐烂程度最严重的地块,活树上出现 S. himantioides 子实体的几率最低。此外,在树干下坡方向出现子实体的频率更高。利用成对相关函数进行的分析表明,随着心材腐烂程度的加深,带有子实体的活树的空间分布模式可能会从密集型变为随机型。最后,利用广义线性模型和广义线性混合模型研究了影响 C.pisifera、C.obtusa 和 L. kaempferi 子实体发育的因素,发现子实体的有无与心材腐朽之间没有明显的关系。因此,我们认为健康的活树和处于心材腐烂早期的活树都可能出现子实体。
{"title":"Occurrence and Characteristics of <i>Serpula himantioides</i> Fruiting Bodies on Living Trees in Japan.","authors":"Ryusei Haraguchi, Toshihide Hirao, Toshihiro Yamada","doi":"10.3390/jof10080572","DOIUrl":"10.3390/jof10080572","url":null,"abstract":"<p><p><i>Serpula himantioides</i> is a globally distributed wood decay fungus that causes heartwood decay in several tree species. We investigated the occurrence of <i>S. himantioides</i> fruiting bodies in Japan for two years and six months to characterize their biology. The fruiting bodies matured in autumn and occurred on living <i>Chamaecyparis pisifera</i>, <i>Chamaecyparis obtusa</i>, <i>Larix kaempferi</i>, and <i>Cryptomeria japonica</i> trees, as well as on dead trees and soil. Assessing three circular plots, the incidence of living trees with <i>S. himantioides</i> fruiting bodies was lowest in the plot with the most advanced heartwood decay. Furthermore, fruiting bodies occurred more frequently in the lower slope direction of the trunk. Analysis using the pair correlation function suggested that the spatial distribution pattern of living trees with fruiting bodies may change from intensive to random with heartwood decay progress. Finally, according to generalized linear and generalized linear mixed models, which were used to investigate the factors affecting the development of fruiting bodies in <i>C. pisifera</i>, <i>C. obtusa</i>, and <i>L. kaempferi</i>, no clear relationship was found between the presence or absence of fruiting bodies and heartwood decay. Thus, we suggest that fruiting bodies can occur in healthy living trees as well as in living trees in the early stages of heartwood decay.</p>","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.2,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11355443/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142080615","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yunmin Wen, Meng Li, Shuzhen Yang, Litao Peng, Gang Fan, Huilin Kang
In this study, antagonistic endophytic fungi were isolated from postharvest chestnut fruits; endophytic antagonistic fungi and their combination of inhibitory effects on the fungal pathogen Neofusicoccum parvum were evaluated. A total of 612 endophytic fungi were isolated from 300 healthy chestnut kernels, and 6 strains out of them including NS-3, NS-11, NS-38, NS-43, NS-56, and NS-58 were confirmed as antagonistic endophytic fungi against Neofusicoccum parvum; these were separately identified as Penicillium chermesinum, Penicillium italicum, Penicillium decaturense, Penicillium oxalicum, Talarmyces siamensis, and Penicillium guanacastense. Some mixed antagonistic endophytic fungi, such as NS-3-38, NS-11-38, NS-43-56, and NS-56-58-38, exhibited a much stronger antifungal activity against N. parvum than that applied individually. Among them, the mixture of NS-3-38 showed the highest antifungal activity, and the inhibition rate was up to 86.67%. The fermentation broth of NS-3, NS-38, and their combinations exhibited an obvious antifungal activity against N. parvum, and the ethyl acetate phase extract of NS-3-38 had the strongest antifungal activity, for which the inhibitory rate was up to 90.19%. The NS-3-38 fermentation broth combined with a chitosan coating significantly reduced N. parvum incidence in chestnuts from 100% to 19%. Furthermore, the fruit decay and weight loss of chestnuts during storage were significantly decreased by the NS-3-38 fermentation broth mixture along with a chitosan coating. Therefore, a mixture of P. chermesinum and P. decaturense could be used as a potential complex biocontrol agent to control postharvest fruit decay in chestnuts.
{"title":"Isolation of Antagonistic Endophytic Fungi from Postharvest Chestnuts and Their Biocontrol on Host Fungal Pathogens.","authors":"Yunmin Wen, Meng Li, Shuzhen Yang, Litao Peng, Gang Fan, Huilin Kang","doi":"10.3390/jof10080573","DOIUrl":"10.3390/jof10080573","url":null,"abstract":"<p><p>In this study, antagonistic endophytic fungi were isolated from postharvest chestnut fruits; endophytic antagonistic fungi and their combination of inhibitory effects on the fungal pathogen <i>Neofusicoccum parvum</i> were evaluated. A total of 612 endophytic fungi were isolated from 300 healthy chestnut kernels, and 6 strains out of them including NS-3, NS-11, NS-38, NS-43, NS-56, and NS-58 were confirmed as antagonistic endophytic fungi against <i>Neofusicoccum parvum</i>; these were separately identified as <i>Penicillium chermesinum</i>, <i>Penicillium italicum</i>, <i>Penicillium decaturense</i>, <i>Penicillium oxalicum, Talarmyces siamensis</i>, and <i>Penicillium guanacastense</i>. Some mixed antagonistic endophytic fungi, such as NS-3-38, NS-11-38, NS-43-56, and NS-56-58-38, exhibited a much stronger antifungal activity against <i>N. parvum</i> than that applied individually. Among them, the mixture of NS-3-38 showed the highest antifungal activity, and the inhibition rate was up to 86.67%. The fermentation broth of NS-3, NS-38, and their combinations exhibited an obvious antifungal activity against <i>N. parvum</i>, and the ethyl acetate phase extract of NS-3-38 had the strongest antifungal activity, for which the inhibitory rate was up to 90.19%. The NS-3-38 fermentation broth combined with a chitosan coating significantly reduced <i>N. parvum</i> incidence in chestnuts from 100% to 19%. Furthermore, the fruit decay and weight loss of chestnuts during storage were significantly decreased by the NS-3-38 fermentation broth mixture along with a chitosan coating. Therefore, a mixture of <i>P. chermesinum</i> and <i>P. decaturense</i> could be used as a potential complex biocontrol agent to control postharvest fruit decay in chestnuts.</p>","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.2,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11355363/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142080612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Diaporthe longicolla (syn. Phomopsis longicolla) is an important seed-borne fungal pathogen and the primary cause of Phomopsis seed decay (PSD) in soybean. PSD is one of the most devastating seed diseases, reducing soybean seed quality and yield worldwide. As part of a genome sequencing project on the fungal Diaporthe-Phomopsis complex, draft genomes of eight D. longicolla isolates were sequenced and assembled. Sequences of mitochondrial genomes were extracted and analyzed. The circular mitochondrial genomes ranged from 52,534 bp to 58,280 bp long, with a mean GC content of 34%. A total of 14 core protein-coding genes, 23 tRNA, and 2 rRNA genes were identified. Introns were detected in the genes of atp6, cob, cox1, cox2, cox3, nad1, nad2, nad5, and rnl. Three isolates (PL7, PL10, and PL185E) had more introns than other isolates. Approximately 6.4% of the mitochondrial genomes consist of repetitive elements. Moreover, 48 single-nucleotide polymorphisms (SNPs) and were identified. The mitochondrial genome sequences of D. longicolla will be useful to further study the molecular basis of seed-borne pathogens causing seed diseases, investigate genetic variation among isolates, and develop improved control strategies for Phomopsis seed decay of soybean.
{"title":"Comparative Analysis of the Mitochondrial Genome Sequences of <i>Diaporthe longicolla</i> (syn. <i>Phomopsis longicolla</i>) Isolates Causing Phomopsis Seed Decay in Soybean.","authors":"Shuxian Li, Xiaojun Hu, Qijian Song","doi":"10.3390/jof10080570","DOIUrl":"10.3390/jof10080570","url":null,"abstract":"<p><p><i>Diaporthe longicolla</i> (syn. <i>Phomopsis longicolla</i>) is an important seed-borne fungal pathogen and the primary cause of Phomopsis seed decay (PSD) in soybean. PSD is one of the most devastating seed diseases, reducing soybean seed quality and yield worldwide. As part of a genome sequencing project on the fungal <i>Diaporthe-Phomopsis</i> complex, draft genomes of eight <i>D. longicolla</i> isolates were sequenced and assembled. Sequences of mitochondrial genomes were extracted and analyzed. The circular mitochondrial genomes ranged from 52,534 bp to 58,280 bp long, with a mean GC content of 34%. A total of 14 core protein-coding genes, 23 tRNA, and 2 rRNA genes were identified. Introns were detected in the genes of <i>atp6</i>, <i>cob</i>, <i>cox1</i>, <i>cox2</i>, <i>cox3</i>, <i>nad1</i>, <i>nad2</i>, <i>nad5</i>, and <i>rnl</i>. Three isolates (PL7, PL10, and PL185E) had more introns than other isolates. Approximately 6.4% of the mitochondrial genomes consist of repetitive elements. Moreover, 48 single-nucleotide polymorphisms (SNPs) and were identified. The mitochondrial genome sequences of <i>D. longicolla</i> will be useful to further study the molecular basis of seed-borne pathogens causing seed diseases, investigate genetic variation among isolates, and develop improved control strategies for Phomopsis seed decay of soybean.</p>","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.2,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11355892/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142080605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Serine protease is an extracellular protease secreted by biocontrol fungi that can effectively control nematode diseases by degrading nematode eggshells and enhancing plant resistance. Trichoderma longibrachiatum T6, an important biocontrol fungus, has been demonstrated to effectively parasitize and degrade Heterodera avenae cysts, eggs, and second-stage juveniles (J2s). However, the genes that encoding serine protease and their functions in T. longibrachiatum T6 have not been thoroughly investigated. In this study, we successfully cloned and sequenced the serine protease gene TlSP1 in T. longibrachiatum T6. Our results revealed that the expression level of the TlSP1 gene was induced and significantly increased in T. longibrachiatum T6 after inoculation with H. avenae cysts. The full-length sequence of the coding region (CDS) of TlSP1 gene was 1230 bp and encoded a protein consisting of 409 amino acids. Upon the transformation of the TlSP1 gene into Pichia pastoris X33, the purified recombinant TlSP1 protein exhibited optimal activity at a temperature of 50 °C and pH 8.0. Following 4–10-day of treatment with the purified recombinant TlSP1 protein, the eggshells and content were dissolved and exuded. The number of nematodes invading wheat roots was reduced by 38.43% in the group treated with both TlSP1 and eggs on one side (P1+N) compared to the control group, while the number of nematodes invading wheat roots was reduced by 30.4% in the TlSP1 and eggs two-sided treatment group (P1/N). Furthermore, both the P1+N and P1/N treatments significantly upregulated genes associated with defense enzymes (TaPAL, TaCAT, TaSOD, and TaPOD), genes involved in the lignin synthesis pathway (TaC4H, Ta4CL2, TaCAD1, and TaCAD12), and salicylic acid (SA)-responsive genes (TaNPR1, TaPR1, and TaPR2) and led to the high expression of jasmonic acid (JA)-responsive genes (TaPR4, TaOPR3, and TaAOS2). This study has highlighted the significant role of the TlSP1 gene in facilitating H. avenae eggshells’ dissolution, preventing nematode invasion in the host plant, and boosting plant resistance in wheat.
{"title":"Characterization of the Serine Protease TlSP1 from Trichoderma longibrachiatum T6 and Its Function in the Control of Heterodera avenae in Wheat","authors":"Xiujuan Wang, Shuwu Zhang, Bingliang Xu","doi":"10.3390/jof10080569","DOIUrl":"https://doi.org/10.3390/jof10080569","url":null,"abstract":"Serine protease is an extracellular protease secreted by biocontrol fungi that can effectively control nematode diseases by degrading nematode eggshells and enhancing plant resistance. Trichoderma longibrachiatum T6, an important biocontrol fungus, has been demonstrated to effectively parasitize and degrade Heterodera avenae cysts, eggs, and second-stage juveniles (J2s). However, the genes that encoding serine protease and their functions in T. longibrachiatum T6 have not been thoroughly investigated. In this study, we successfully cloned and sequenced the serine protease gene TlSP1 in T. longibrachiatum T6. Our results revealed that the expression level of the TlSP1 gene was induced and significantly increased in T. longibrachiatum T6 after inoculation with H. avenae cysts. The full-length sequence of the coding region (CDS) of TlSP1 gene was 1230 bp and encoded a protein consisting of 409 amino acids. Upon the transformation of the TlSP1 gene into Pichia pastoris X33, the purified recombinant TlSP1 protein exhibited optimal activity at a temperature of 50 °C and pH 8.0. Following 4–10-day of treatment with the purified recombinant TlSP1 protein, the eggshells and content were dissolved and exuded. The number of nematodes invading wheat roots was reduced by 38.43% in the group treated with both TlSP1 and eggs on one side (P1+N) compared to the control group, while the number of nematodes invading wheat roots was reduced by 30.4% in the TlSP1 and eggs two-sided treatment group (P1/N). Furthermore, both the P1+N and P1/N treatments significantly upregulated genes associated with defense enzymes (TaPAL, TaCAT, TaSOD, and TaPOD), genes involved in the lignin synthesis pathway (TaC4H, Ta4CL2, TaCAD1, and TaCAD12), and salicylic acid (SA)-responsive genes (TaNPR1, TaPR1, and TaPR2) and led to the high expression of jasmonic acid (JA)-responsive genes (TaPR4, TaOPR3, and TaAOS2). This study has highlighted the significant role of the TlSP1 gene in facilitating H. avenae eggshells’ dissolution, preventing nematode invasion in the host plant, and boosting plant resistance in wheat.","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141949058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To investigate the effect and mechanism of plasma-activated water (PAW) on Aspergillus niger, PAW was prepared using a needle array–plate dielectric barrier discharge plasma system. The concentrations of long-lived reactive oxygen and nitrogen species (RONS), namely, H2O2, NO2−, and NO3−, in the PAW were 48.76 mg/L, 0.046 mg/L, and 172.36 mg/L, respectively. Chemically activated water (CAW) with the same concentration of long-lived RONS was also prepared for comparison. A. niger A32 was treated with PAW and CAW. After treatment, the treated strains were observed and analyzed with scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to screen probable mutants. The results indicated that the pH, conductivity, and ORP values of PAW were 2.42, 1935 μS/cm, and 517.07 mV, respectively. In contrast, the pH and ORP values of CAW were 6.15 and 301.73 mV, respectively, which differed significantly from those of PAW. In addition, the conductivity of CAW showed no change. SEM and TEM analyses revealed that A. niger A32 treated with CAW exhibited less damage compared with the control. In contrast, A. niger A32 treated with PAW showed significant shrinkage, deformation, and exudate attachment over time. Following PAW treatment, after four passages, a high cellulase-producing stable mutant strain A-WW5 was screened, exhibiting a filter paper enzyme activity of 29.66 U/mL, a cellulose endonuclease activity of 13.79 U/mL, and a β-glucosidase activity of 27.13 U/mL. These values were found to be 33%, 38%, and 2.1% higher than those of the original fungus sample, respectively. In total, 116 SNPs and 61 InDels were present in the genome of the mutant strain A-WW5. The above findings indicate that the impact of PAW on A. niger is not only attributed to long-lasting H2O2, NO2−, and NO3− particles but also to other short-lived active particles; PAW is expected to become a new microbial breeding mutagen.
{"title":"Effect of Plasma-Activated Water on the Cellulase-Producing Strain Aspergillus niger A32","authors":"Zhiqing Song, Yingwei Jiang, Chan Chen, Changjiang Ding, Hao Chen","doi":"10.3390/jof10080568","DOIUrl":"https://doi.org/10.3390/jof10080568","url":null,"abstract":"To investigate the effect and mechanism of plasma-activated water (PAW) on Aspergillus niger, PAW was prepared using a needle array–plate dielectric barrier discharge plasma system. The concentrations of long-lived reactive oxygen and nitrogen species (RONS), namely, H2O2, NO2−, and NO3−, in the PAW were 48.76 mg/L, 0.046 mg/L, and 172.36 mg/L, respectively. Chemically activated water (CAW) with the same concentration of long-lived RONS was also prepared for comparison. A. niger A32 was treated with PAW and CAW. After treatment, the treated strains were observed and analyzed with scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to screen probable mutants. The results indicated that the pH, conductivity, and ORP values of PAW were 2.42, 1935 μS/cm, and 517.07 mV, respectively. In contrast, the pH and ORP values of CAW were 6.15 and 301.73 mV, respectively, which differed significantly from those of PAW. In addition, the conductivity of CAW showed no change. SEM and TEM analyses revealed that A. niger A32 treated with CAW exhibited less damage compared with the control. In contrast, A. niger A32 treated with PAW showed significant shrinkage, deformation, and exudate attachment over time. Following PAW treatment, after four passages, a high cellulase-producing stable mutant strain A-WW5 was screened, exhibiting a filter paper enzyme activity of 29.66 U/mL, a cellulose endonuclease activity of 13.79 U/mL, and a β-glucosidase activity of 27.13 U/mL. These values were found to be 33%, 38%, and 2.1% higher than those of the original fungus sample, respectively. In total, 116 SNPs and 61 InDels were present in the genome of the mutant strain A-WW5. The above findings indicate that the impact of PAW on A. niger is not only attributed to long-lasting H2O2, NO2−, and NO3− particles but also to other short-lived active particles; PAW is expected to become a new microbial breeding mutagen.","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141949057","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Louis S. Phillips-Rose, Chendi K. Yu, Nicholas P. West, James A. Fraser
The plethora of genome sequences produced in the postgenomic age has not resolved many of our most pressing biological questions. Correlating gene expression with an interrogatable and easily observable characteristic such as the surrogate phenotype conferred by a reporter gene is a valuable approach to gaining insight into gene function. Many reporters including lacZ, amdS, and the fluorescent proteins mRuby3 and mNeonGreen have been used across all manners of organisms. Described here is an investigation into the creation of a robust, synthetic, fusion reporter system for Cryptococcus neoformans that combines some of the most useful fluorophores available in this system with the versatility of the counter-selectable nature of amdS. The reporters generated include multiple composition and orientation variants, all of which were investigated for differences in expression. Evaluation of known promoters from the TEF1 and GAL7 genes was undertaken, elucidating novel expression tendencies of these biologically relevant C. neoformans regulators of transcription. Smaller than lacZ but providing multiple useful surrogate phenotypes for interrogation, the fusion ORF serves as a superior whole-cell assay compared to traditional systems. Ultimately, the work described here bolsters the array of relevant genetic tools that may be employed in furthering manipulation and understanding of the WHO fungal priority group pathogen C. neoformans.
{"title":"A Chimeric ORF Fusion Phenotypic Reporter for Cryptococcus neoformans","authors":"Louis S. Phillips-Rose, Chendi K. Yu, Nicholas P. West, James A. Fraser","doi":"10.3390/jof10080567","DOIUrl":"https://doi.org/10.3390/jof10080567","url":null,"abstract":"The plethora of genome sequences produced in the postgenomic age has not resolved many of our most pressing biological questions. Correlating gene expression with an interrogatable and easily observable characteristic such as the surrogate phenotype conferred by a reporter gene is a valuable approach to gaining insight into gene function. Many reporters including lacZ, amdS, and the fluorescent proteins mRuby3 and mNeonGreen have been used across all manners of organisms. Described here is an investigation into the creation of a robust, synthetic, fusion reporter system for Cryptococcus neoformans that combines some of the most useful fluorophores available in this system with the versatility of the counter-selectable nature of amdS. The reporters generated include multiple composition and orientation variants, all of which were investigated for differences in expression. Evaluation of known promoters from the TEF1 and GAL7 genes was undertaken, elucidating novel expression tendencies of these biologically relevant C. neoformans regulators of transcription. Smaller than lacZ but providing multiple useful surrogate phenotypes for interrogation, the fusion ORF serves as a superior whole-cell assay compared to traditional systems. Ultimately, the work described here bolsters the array of relevant genetic tools that may be employed in furthering manipulation and understanding of the WHO fungal priority group pathogen C. neoformans.","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141949061","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Amira Yacoub, David Renault, Rana Haidar, Florian Boulisset, Patricia Letousey, Rémy Guyoneaud, Eleonore Attard, Patrice Rey
Grapevine trunk diseases (GTDs) are currently limiting grapevine productivity in many vineyards worldwide. As no chemical treatments are registered to control GTDs, biocontrol agents are being tested against these diseases. Esquive® WP, based on the fungus Trichoderma atroviride I-1237 strain, is the first biocontrol product registered in France to control GTDs. In this study, we determine whether, following grapevine pruning wound treatments with Esquive® WP, changes occurred or not in the indigenous microbial communities that are colonizing grapevine wood. Over a 6-year period, Esquive® WP was applied annually to pruning wounds on three grapevine cultivars located in three different regions. Wood samples were collected at 2 and 10 months after the Esquive® WP treatments. Based on MiSeq high-throughput sequencing analyses, the results showed that specific microbial communities were linked to each ‘region/cultivar’ pairing. In certain cases, a significant modification of alpha diversity indexes and the relative abundance of some microbial taxa were observed between treated and non-treated grapevines 2 months after Esquive® WP treatment. However, these modifications disappeared over time, i.e., 10 months post-treatment. This result clearly showed that Esquive® WP pruning wood treatment did not induce significant changes in the grapevine wood’s microbiome, even after 6 years of recurrent applications on the plants.
{"title":"Impact of the Biocontrol Product, Esquive® WP, on the Indigenous Grapevine Wood Microbiome after a 6-Year Application Period","authors":"Amira Yacoub, David Renault, Rana Haidar, Florian Boulisset, Patricia Letousey, Rémy Guyoneaud, Eleonore Attard, Patrice Rey","doi":"10.3390/jof10080566","DOIUrl":"https://doi.org/10.3390/jof10080566","url":null,"abstract":"Grapevine trunk diseases (GTDs) are currently limiting grapevine productivity in many vineyards worldwide. As no chemical treatments are registered to control GTDs, biocontrol agents are being tested against these diseases. Esquive® WP, based on the fungus Trichoderma atroviride I-1237 strain, is the first biocontrol product registered in France to control GTDs. In this study, we determine whether, following grapevine pruning wound treatments with Esquive® WP, changes occurred or not in the indigenous microbial communities that are colonizing grapevine wood. Over a 6-year period, Esquive® WP was applied annually to pruning wounds on three grapevine cultivars located in three different regions. Wood samples were collected at 2 and 10 months after the Esquive® WP treatments. Based on MiSeq high-throughput sequencing analyses, the results showed that specific microbial communities were linked to each ‘region/cultivar’ pairing. In certain cases, a significant modification of alpha diversity indexes and the relative abundance of some microbial taxa were observed between treated and non-treated grapevines 2 months after Esquive® WP treatment. However, these modifications disappeared over time, i.e., 10 months post-treatment. This result clearly showed that Esquive® WP pruning wood treatment did not induce significant changes in the grapevine wood’s microbiome, even after 6 years of recurrent applications on the plants.","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141949059","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yunxiang Tan, Yunhang Lv, Mengyu Xv, Laiye Qu, Wenjuan Wang
Populus euphratica, Tamarix ramosissima, and Sophora alopecuroides are, respectively, typical arboreal, shrubby, and herbaceous species in oases of arid regions. It is important to study the difference in metabolic characteristics of the rhizosphere fungal community of these plant species and their relationships with soil factors for the preservation of delicate arid oasis ecosystems with future environmental changes. In this study, we, respectively, collected 18 rhizosphere soil samples of P. euphratica, T. ramosissima, and S. alopecuroides to explore the difference in rhizosphere fungal metabolic characteristics of different plant life forms and their underlying driving factors. The results showed that (1) soil physicochemical properties (including soil water content, pH, etc.) were significantly different among different plant species (p < 0.05). (2) Rhizosphere fungal metabolic characteristics were significantly different between S. alopecuroides and T. ramosissima (ANOSIM, p < 0.05), which was mainly caused by the different utilization of carboxylic carbon. (3) The RDA showed that the main driving factors of the variations in rhizosphere fungal metabolic characteristics were different among different plant species. The main explanatory variables of the variations in the metabolic characteristics of the rhizosphere fungal community were carbon to nitrogen ratio (23%) and available potassium (17.4%) for P. euphratica, while soil organic carbon (23.1%), pH (8.6%), and total nitrogen (8.2%) for T. ramosissima, and soil clay content (36.6%) and soil organic carbon (12.6%) for S. alopecuroides. In conclusion, the variations in rhizosphere fungal metabolic characteristics in arid oases are dominantly affected by soil factors rather than plant life forms.
胡杨、柽柳和槐分别是干旱地区绿洲中典型的乔木、灌木和草本物种。研究这些植物物种根圈真菌群落代谢特征的差异及其与土壤因子的关系,对于在未来环境变化中保护脆弱的干旱绿洲生态系统具有重要意义。在本研究中,我们分别采集了 P. euphratica、T. ramosissima 和 S. alopecuroides 的 18 个根圈土壤样本,探讨了不同植物生命形式根圈真菌代谢特征的差异及其背后的驱动因素。结果表明:(1)土壤理化性质(包括土壤含水量、pH 值等)在不同植物物种之间存在显著差异(p < 0.05)。(2)根瘤菌代谢特征在 S. alopecuroides 和 T. ramosissima 之间存在显著差异(ANOSIM,p < 0.05),这主要是由于对羧基碳的利用率不同造成的。(3)RDA 显示,不同植物物种根瘤菌代谢特征变化的主要驱动因素不同。对 P. euphratica 而言,根圈真菌群落代谢特征变化的主要解释变量是碳氮比(23%)和可利用钾(17.4%);对 T. ramosissima 而言,主要解释变量是土壤有机碳(23.1%)、pH 值(8.6%)和全氮(8.2%);对 S. alopecuroides 而言,主要解释变量是土壤粘土含量(36.6%)和土壤有机碳(12.6%)。总之,干旱绿洲根圈真菌代谢特征的变化主要受土壤因素而非植物生命形式的影响。
{"title":"Differences in Metabolic Characteristics of Rhizosphere Fungal Community of Typical Arboreal, Shrubby and Herbaceous Species in Oasis of Arid Region","authors":"Yunxiang Tan, Yunhang Lv, Mengyu Xv, Laiye Qu, Wenjuan Wang","doi":"10.3390/jof10080565","DOIUrl":"https://doi.org/10.3390/jof10080565","url":null,"abstract":"Populus euphratica, Tamarix ramosissima, and Sophora alopecuroides are, respectively, typical arboreal, shrubby, and herbaceous species in oases of arid regions. It is important to study the difference in metabolic characteristics of the rhizosphere fungal community of these plant species and their relationships with soil factors for the preservation of delicate arid oasis ecosystems with future environmental changes. In this study, we, respectively, collected 18 rhizosphere soil samples of P. euphratica, T. ramosissima, and S. alopecuroides to explore the difference in rhizosphere fungal metabolic characteristics of different plant life forms and their underlying driving factors. The results showed that (1) soil physicochemical properties (including soil water content, pH, etc.) were significantly different among different plant species (p < 0.05). (2) Rhizosphere fungal metabolic characteristics were significantly different between S. alopecuroides and T. ramosissima (ANOSIM, p < 0.05), which was mainly caused by the different utilization of carboxylic carbon. (3) The RDA showed that the main driving factors of the variations in rhizosphere fungal metabolic characteristics were different among different plant species. The main explanatory variables of the variations in the metabolic characteristics of the rhizosphere fungal community were carbon to nitrogen ratio (23%) and available potassium (17.4%) for P. euphratica, while soil organic carbon (23.1%), pH (8.6%), and total nitrogen (8.2%) for T. ramosissima, and soil clay content (36.6%) and soil organic carbon (12.6%) for S. alopecuroides. In conclusion, the variations in rhizosphere fungal metabolic characteristics in arid oases are dominantly affected by soil factors rather than plant life forms.","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141949060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The entomopathogenic fungus (EPF) Metarhizium acridum is a typical filamentous fungus and has been used to control migratory locusts (Locusta migratoria manilensis). This study examines the impact of the Zn(II)2Cys6 transcription factor, MaAzaR, in the virulence of M. acridum. Disruption of MaAzaR (ΔMaAzaR) diminished the fungus’s ability to penetrate the insect cuticle, thereby decreasing its virulence. The median lethal time (LT50) for the ΔMaAzaR strain increased by approximately 1.5 d compared to the wild-type (WT) strain when topically inoculated, simulating natural infection conditions. ΔMaAzaR compromises the formation, turgor pressure, and secretion of extracellular hydrolytic enzymes in appressoria. However, the growth ability of ΔMaAzaR within the hemolymph is not impaired; in fact, it grows better than the WT strain. Moreover, RNA-sequencing (RNA-Seq) analysis of ΔMaAzaR and WT strains grown for 20 h on locust hindwings revealed 87 upregulated and 37 downregulated differentially expressed genes (DEGs) in the mutant strain. Pathogen–host interaction database (PHI) analysis showed that about 40% of the total DEGs were associated with virulence, suggesting that MaAzaR is a crucial transcription factor that directly regulates the expression of downstream genes. This study identifies a new transcription factor involved in EPF cuticle penetration, providing theoretical support and genetic resources for the developing highly virulent strains.
{"title":"MaAzaR Influences Virulence of Metarhizium acridum against Locusta migratoria manilensis by Affecting Cuticle Penetration","authors":"Geng Hong, Siqing Wang, Yuxian Xia, Guoxiong Peng","doi":"10.3390/jof10080564","DOIUrl":"https://doi.org/10.3390/jof10080564","url":null,"abstract":"The entomopathogenic fungus (EPF) Metarhizium acridum is a typical filamentous fungus and has been used to control migratory locusts (Locusta migratoria manilensis). This study examines the impact of the Zn(II)2Cys6 transcription factor, MaAzaR, in the virulence of M. acridum. Disruption of MaAzaR (ΔMaAzaR) diminished the fungus’s ability to penetrate the insect cuticle, thereby decreasing its virulence. The median lethal time (LT50) for the ΔMaAzaR strain increased by approximately 1.5 d compared to the wild-type (WT) strain when topically inoculated, simulating natural infection conditions. ΔMaAzaR compromises the formation, turgor pressure, and secretion of extracellular hydrolytic enzymes in appressoria. However, the growth ability of ΔMaAzaR within the hemolymph is not impaired; in fact, it grows better than the WT strain. Moreover, RNA-sequencing (RNA-Seq) analysis of ΔMaAzaR and WT strains grown for 20 h on locust hindwings revealed 87 upregulated and 37 downregulated differentially expressed genes (DEGs) in the mutant strain. Pathogen–host interaction database (PHI) analysis showed that about 40% of the total DEGs were associated with virulence, suggesting that MaAzaR is a crucial transcription factor that directly regulates the expression of downstream genes. This study identifies a new transcription factor involved in EPF cuticle penetration, providing theoretical support and genetic resources for the developing highly virulent strains.","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":null,"pages":null},"PeriodicalIF":4.2,"publicationDate":"2024-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141925401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}