Pub Date : 2025-06-27DOI: 10.1016/j.funbio.2025.101619
Christina M. Kelliher , Jay C. Dunlap
In many model organisms, the circadian system has been proposed to comprise multiple oscillators that interact to promote accuracy of the clock as well as intricacies of rhythmic outputs. In Neurospora crassa, the circadian transcriptional/translational loop comprising of the FRQ (Frequency) and WCC (White Collar Complex) proteins has been instrumental in explaining many attributes of the clock including entrainment and rhythms in development and gene expression; in addition, some non-circadian oscillations can be unmasked when the FRQ-WCC feedback loop is eliminated. These rhythms have often lost defining circadian characteristics and are potentially controlled by other oscillators, termed FRQ-less oscillators (FLOs) in Neurospora. Understanding the biology of these oscillators and their hierarchical relationship with the FRQ-WCC oscillator (FWO) are salient questions in rhythms research. In this study, we examined candidate FLO effector pathways involving peroxiredoxins (Prxs) and mTOR. We find that independent gene knockouts compromising each pathway do not alter circadian period length or decrease the amplitude of the core circadian FWO rhythm in any meaningful way in Neurospora. Our findings suggest that molecular rhythms in Prx oxidation and in mTOR activity on the chol-1 FLO oscillator are neither required for nor strongly regulate FWO components during a normal circadian day.
{"title":"Individual peroxiredoxin or Tor pathway components are not required for circadian clock function in Neurospora crassa","authors":"Christina M. Kelliher , Jay C. Dunlap","doi":"10.1016/j.funbio.2025.101619","DOIUrl":"10.1016/j.funbio.2025.101619","url":null,"abstract":"<div><div>In many model organisms, the circadian system has been proposed to comprise multiple oscillators that interact to promote accuracy of the clock as well as intricacies of rhythmic outputs. In <em>Neurospora crassa</em>, the circadian transcriptional/translational loop comprising of the FRQ (Frequency) and WCC (White Collar Complex) proteins has been instrumental in explaining many attributes of the clock including entrainment and rhythms in development and gene expression; in addition, some non-circadian oscillations can be unmasked when the FRQ-WCC feedback loop is eliminated. These rhythms have often lost defining circadian characteristics and are potentially controlled by other oscillators, termed FRQ-less oscillators (FLOs) in <em>Neurospora</em>. Understanding the biology of these oscillators and their hierarchical relationship with the FRQ-WCC oscillator (FWO) are salient questions in rhythms research. In this study, we examined candidate FLO effector pathways involving peroxiredoxins (Prxs) and mTOR. We find that independent gene knockouts compromising each pathway do not alter circadian period length or decrease the amplitude of the core circadian FWO rhythm in any meaningful way in <em>Neurospora</em>. Our findings suggest that molecular rhythms in Prx oxidation and in mTOR activity on the <em>chol-1</em> FLO oscillator are neither required for nor strongly regulate FWO components during a normal circadian day.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 6","pages":"Article 101619"},"PeriodicalIF":2.9,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144535417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-25DOI: 10.1016/j.funbio.2025.101620
Katarzyna Stojek , Barbara Bobrowska-Korczak , Justyna Frączek , Marcin Piotrowski , Mirosław Krośniak , Bogdan Jaroszewicz
Wild mushrooms can be an important source of protein and essential amino acids, however very little is known about the environmental factors affecting the content of these compounds. In our study, we investigated the influence of soil properties (soil type, C/N ratio, pH) and tree stand characteristics (tree diversity, canopy cover, understory cover, and the proportion of deciduous trees) on total protein and essential amino acids (Valine, Leucine, Isoleucine, Phenylalanine, Lysine, Methionine, Arginine, Histidine) contents in seven wild-growing mushroom species (Macrolepiota procera, Rhodocolybia butyracea, Russula cyanoxantha, R.heterophylla, Lactifluus vellereus, Armillaria mellea s.l., and Xerocomellus chrysenteron). Our study showed that the identity of mushroom species determines, to a large extent, the protein content and the amino acid profile of the mushrooms. The highest protein content was revealed in X. chrysenteron, M. procera and R. butyracea. Effects of environmental factors were weaker and species specific. The protein content in X. chrysenteron was mainly influenced by soil type (Cambisols vs. Luvisols) and soil characteristics (positively by C/N ratio and negatively by soil pH). In L. vellereus the protein content was negatively influenced by stand characteristics (canopy cover, understory cover, and tree diversity). In M. procera the protein content decreased with decreasing understory cover, while for all the other studied species, the effects of environmental factors were negligible. Similarly, the amino acid profiles were not affected by any environmental factors, however, they seem to be consistent with ecological roles of the species (ectomycorrhizal, saprotrophic, parasitic fungi). This last result requires further investigation.
{"title":"Protein content and amino acid profile of wild mushrooms depend on environmental conditions","authors":"Katarzyna Stojek , Barbara Bobrowska-Korczak , Justyna Frączek , Marcin Piotrowski , Mirosław Krośniak , Bogdan Jaroszewicz","doi":"10.1016/j.funbio.2025.101620","DOIUrl":"10.1016/j.funbio.2025.101620","url":null,"abstract":"<div><div>Wild mushrooms can be an important source of protein and essential amino acids, however very little is known about the environmental factors affecting the content of these compounds. In our study, we investigated the influence of soil properties (soil type, C/N ratio, pH) and tree stand characteristics (tree diversity, canopy cover, understory cover, and the proportion of deciduous trees) on total protein and essential amino acids (Valine, Leucine, Isoleucine, Phenylalanine, Lysine, Methionine, Arginine, Histidine) contents in seven wild-growing mushroom species (<em>Macrolepiota procera</em>, <em>Rhodocolybia butyracea</em>, <em>Russula cyanoxantha</em>, <em>R.heterophylla</em>, <em>Lactifluus vellereus</em>, <em>Armillaria mellea</em> s.l., and <em>Xerocomellus chrysenteron).</em> Our study showed that the identity of mushroom species determines, to a large extent, the protein content and the amino acid profile of the mushrooms. The highest protein content was revealed in <em>X. chrysenteron</em>, <em>M. procera</em> and <em>R. butyracea</em>. Effects of environmental factors were weaker and species specific. The protein content in <em>X. chrysenteron</em> was mainly influenced by soil type (Cambisols vs. Luvisols) and soil characteristics (positively by C/N ratio and negatively by soil pH). In <em>L. vellereus</em> the protein content was negatively influenced by stand characteristics (canopy cover, understory cover, and tree diversity). In <em>M. procera</em> the protein content decreased with decreasing understory cover, while for all the other studied species, the effects of environmental factors were negligible. Similarly, the amino acid profiles were not affected by any environmental factors, however, they seem to be consistent with ecological roles of the species (ectomycorrhizal, saprotrophic, parasitic fungi). This last result requires further investigation.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 6","pages":"Article 101620"},"PeriodicalIF":2.9,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144501023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-24DOI: 10.1016/j.funbio.2025.101621
Gustavo Hernán Ramírez , María Virginia Bianchinotti , Freda Elizabeth Anderson
Tritrophic interactions involving host plants, fungal pathogens and mycoparasites play an important role in the dynamics of natural ecosystems. In this work, we investigate the impact of the rust fungus Puccinia araujiae on the growth of Araujia hortorum plants in the presence/absence of a mycoparasitic Cladosporium species identified here as Cladosporium sphaerospermum, supported by both morphological and molecular studies. The capacity of the latter to grow and reproduce at the expense of teliospores of the rust was confirmed through microscopic observations. P. araujiae is added to the list of hosts of C. sphaerospermum. An experiment was carried out to assess the impact of rust infection on host plant biomass and whether C. sphaerospermum affected the outcome of the interaction. Plants were subjected to three treatments: inoculation with the rust alone, inoculation with both the rust and the mycoparasite, and uninoculated controls. Rust-infected plants (both with and without the mycoparasite) exhibited 50–60 % reductions in biomass, primarily in root tissues, with premature senescence and leaf abscission contributing to overall decline. However, rust-infected plants in the absence of the mycoparasite produced 10 % less biomass than those in the treatment where it was present, showing that C. sphaerospermum is capable of exerting a detrimental effect on the rust which in turn reflects on a better performance of the plant host. It is argued that this fact does not preclude the potential of P. araujiae as a biological control agent.
{"title":"Exploring the tritrophic interactions between Araujia hortorum, Puccinia araujiae, and a mycoparasitic Cladosporium: implications for the biological control of moth plant","authors":"Gustavo Hernán Ramírez , María Virginia Bianchinotti , Freda Elizabeth Anderson","doi":"10.1016/j.funbio.2025.101621","DOIUrl":"10.1016/j.funbio.2025.101621","url":null,"abstract":"<div><div>Tritrophic interactions involving host plants, fungal pathogens and mycoparasites play an important role in the dynamics of natural ecosystems. In this work, we investigate the impact of the rust fungus <em>Puccinia araujiae</em> on the growth of <em>Araujia hortorum</em> plants in the presence/absence of a mycoparasitic <em>Cladosporium</em> species identified here as <em>Cladosporium sphaerospermum</em>, supported by both morphological and molecular studies. The capacity of the latter to grow and reproduce at the expense of teliospores of the rust was confirmed through microscopic observations. <em>P. araujiae</em> is added to the list of hosts of <em>C. sphaerospermum</em>. An experiment was carried out to assess the impact of rust infection on host plant biomass and whether <em>C. sphaerospermum</em> affected the outcome of the interaction. Plants were subjected to three treatments: inoculation with the rust alone, inoculation with both the rust and the mycoparasite, and uninoculated controls. Rust-infected plants (both with and without the mycoparasite) exhibited 50–60 % reductions in biomass, primarily in root tissues, with premature senescence and leaf abscission contributing to overall decline. However, rust-infected plants in the absence of the mycoparasite produced 10 % less biomass than those in the treatment where it was present, showing that <em>C. sphaerospermum</em> is capable of exerting a detrimental effect on the rust which in turn reflects on a better performance of the plant host. It is argued that this fact does not preclude the potential of <em>P. araujiae</em> as a biological control agent.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 6","pages":"Article 101621"},"PeriodicalIF":2.9,"publicationDate":"2025-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144501131","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-14DOI: 10.1016/j.funbio.2025.101618
Flavia de Fatima Costa , Bruno Paulo Rodrigues Lustosa , Camila Pereira Perico , Ricardo Belmonte-Lopes , João Lucas Vitório Ribeiro Carvalho , Emanuel L. Razzolini , Germana Davila dos Santos , Bruna Jacomel Favoreto de Souza Lima , Cristina Maria de Souza-Motta , Roberto Tadeu Raittz , Yinggai Song , Laura Selbmann , G. Sybren de Hoog , Jacques Meis , Vania Aparecida Vicente
Lichens exemplify a unique symbiotic relationship between fungi and algae or cyanobacteria, where fungi (mycobionts) provide structural support, while algae or cyanobacteria (photobionts) provide nutrients. Recent discoveries in the order Chaetothyriales have led to the description of several lichenicolous species, underscoring an intricate relationship of some black yeast-like fungi with lichens. The present study aims to investigate public metagenomic data of lichens available in the SRA database, covering a total of 2888 samples. The analysis incorporated 122 molecular marker sequences (barcodes and padlock probes) previously documented in the literature for species classified within Chaetothyriales. Additionally, 11 novel barcodes for species recently identified in lichens of the genera Cladophialophora and Paracladophialophora are described. The selected metagenomes were then compared with molecular marker sequences using local BLASTn (v2.6.0+), considering only alignments with a coverage cut-off and 100 % identity (perfect match). Reads from each sample were retrieved from the SRA as a multifasta file and analyzed with the SWeeP method for vector-based, alignment-free sequence analysis. The analysis identified fungi that are known as environmental inhabitants and, occasionally, opportunistic pathogens of vertebrates, including species in the genera Cladophialophora, Cyphellophora, and Exophiala. These species were distributed across 11 BioProjects from various locations around the world. The findings of this study corroborate extant knowledge concerning fungal colonization in diverse extremophilic environments, including deserts, tundra, and rocky surfaces.
{"title":"In silico search reveals the association of lichens with black yeast-like fungi in the order Chaetothyriales","authors":"Flavia de Fatima Costa , Bruno Paulo Rodrigues Lustosa , Camila Pereira Perico , Ricardo Belmonte-Lopes , João Lucas Vitório Ribeiro Carvalho , Emanuel L. Razzolini , Germana Davila dos Santos , Bruna Jacomel Favoreto de Souza Lima , Cristina Maria de Souza-Motta , Roberto Tadeu Raittz , Yinggai Song , Laura Selbmann , G. Sybren de Hoog , Jacques Meis , Vania Aparecida Vicente","doi":"10.1016/j.funbio.2025.101618","DOIUrl":"10.1016/j.funbio.2025.101618","url":null,"abstract":"<div><div>Lichens exemplify a unique symbiotic relationship between fungi and algae or cyanobacteria, where fungi (mycobionts) provide structural support, while algae or cyanobacteria (photobionts) provide nutrients. Recent discoveries in the order Chaetothyriales have led to the description of several lichenicolous species, underscoring an intricate relationship of some black yeast-like fungi with lichens. The present study aims to investigate public metagenomic data of lichens available in the SRA database, covering a total of 2888 samples. The analysis incorporated 122 molecular marker sequences (barcodes and padlock probes) previously documented in the literature for species classified within Chaetothyriales. Additionally, 11 novel barcodes for species recently identified in lichens of the genera <em>Cladophialophora</em> and <em>Paracladophialophora</em> are described. The selected metagenomes were then compared with molecular marker sequences using local BLASTn (v2.6.0+), considering only alignments with a coverage cut-off and 100 % identity (perfect match). Reads from each sample were retrieved from the SRA as a multifasta file and analyzed with the SWeeP method for vector-based, alignment-free sequence analysis. The analysis identified fungi that are known as environmental inhabitants and, occasionally, opportunistic pathogens of vertebrates, including species in the genera <em>Cladophialophora</em>, <em>Cyphellophora</em>, and <em>Exophiala</em>. These species were distributed across 11 BioProjects from various locations around the world. The findings of this study corroborate extant knowledge concerning fungal colonization in diverse extremophilic environments, including deserts, tundra, and rocky surfaces.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 6","pages":"Article 101618"},"PeriodicalIF":2.9,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144312750","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-14DOI: 10.1016/j.funbio.2025.101617
Mara Edith Martin , Juan Santiago Guidobono , Eliana Melignani , Laura Gasoni , Mario Carlos Nazareno Saparrat , Viviana Andrea Barrera
Cladorrhinum, a genus of soil fungi belonging to the Podosporaceae family within the Sordariales order of the Ascomycota phylum, comprises several species with diverse physiological capabilities. While there are existing data on the cellulolytic activity of Cladorrhinum bulbillosum and Cladorrhinum foecundissimum, no reports are available on the potential of other species in this genus. In this study, different strains of Cladorrhinum were evaluated for carboxymethyl cellulase (CMCase) activity on solid medium, and the strains exhibiting the highest activity levels were selected. In liquid culture, Cladorrhinum samala INTA-AR 59 and C. samala INTA-AR 7 revealed maximum levels of endo-1,4-β-glucanase activity at 14 incubation d (27.55 ± 3.39 IU/L and 17.52 ± 0.87 IU/L, respectively), while C. samala INTA-AR 156 exhibited it at 31 incubation d (13.32 ± 3.59 IU/L). Concerning β-glucosidase activity, C. samala INTA-AR 59 and C. samala INTA-AR 156 displayed maximum levels of activity at 38 d (17.52 ± 0.87 IU/L and 0.5 ± 0.21 IU/L, respectively), and INTA-AR 7 at 31 d (9.44 ± 1.02 IU/L). Additionally, we present data on CMCase activity obtained through zymogram analysis of the C. samala INTA-AR 59 strain.
{"title":"Comparative analysis of the cellulolytic enzymatic ability of strains belonging to the genus Cladorrhinum (Podosporaceae, Sordariales, Ascomycota) and preliminary data for Cladorrhinum samala INTA-AR 59","authors":"Mara Edith Martin , Juan Santiago Guidobono , Eliana Melignani , Laura Gasoni , Mario Carlos Nazareno Saparrat , Viviana Andrea Barrera","doi":"10.1016/j.funbio.2025.101617","DOIUrl":"10.1016/j.funbio.2025.101617","url":null,"abstract":"<div><div><em>Cladorrhinum</em>, a genus of soil fungi belonging to the Podosporaceae family within the Sordariales order of the Ascomycota phylum, comprises several species with diverse physiological capabilities. While there are existing data on the cellulolytic activity of <em>Cladorrhinum bulbillosum</em> and <em>Cladorrhinum foecundissimum</em>, no reports are available on the potential of other species in this genus. In this study, different strains of <em>Cladorrhinum</em> were evaluated for carboxymethyl cellulase (CMCase) activity on solid medium, and the strains exhibiting the highest activity levels were selected. In liquid culture, <em>Cladorrhinum samala</em> INTA-AR 59 and <em>C. samala</em> INTA-AR 7 revealed maximum levels of endo-1,4-β-glucanase activity at 14 incubation d (27.55 ± 3.39 IU/L and 17.52 ± 0.87 IU/L, respectively), while <em>C. samala</em> INTA-AR 156 exhibited it at 31 incubation d (13.32 ± 3.59 IU/L). Concerning β-glucosidase activity, <em>C. samala</em> INTA-AR 59 and <em>C. samala</em> INTA-AR 156 displayed maximum levels of activity at 38 d (17.52 ± 0.87 IU/L and 0.5 ± 0.21 IU/L, respectively), and INTA-AR 7 at 31 d (9.44 ± 1.02 IU/L). Additionally, we present data on CMCase activity obtained through zymogram analysis of the <em>C. samala</em> INTA-AR 59 strain.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 6","pages":"Article 101617"},"PeriodicalIF":2.9,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144338637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-11DOI: 10.1016/j.funbio.2025.101616
Xiuwen Wang , Hassan Mohamed , Qing Liu , Zhaosen Fan , Yuanda Song
Oleaginous filamentous fungus Mucor circinelloides harbors a GATA transcription activator AreA, which regulates nitrogen metabolism. In our previous study, deletion of AreA resulted in increased lipid production, while its overexpression reduced lipid synthesis. Although it is not a direct lipogenesis regulator, AreA influences metabolic flux by modulating nitrogen utilization pathways, which in turn affects carbon distribution. One such pathway is the GABA shunt, an alternative route that bypasses two steps of TCA cycle and contributes to replenishing TCA intermediates under nitrogen-limited conditions. The uga2 gene, encoding succinate semialdehyde dehydrogenase, plays a key role in this shunt. We constructed UGA2-knockout strain in wild-type, AreA-knockout and AreA-overexpression strains, respectively. Results showed that single knockout of uga2 increased fatty acid content from 24.5 % to 32.7 %, while double knockout of areA and uga2 exhibited the highest fatty acid content at 36.1 %. The areA overexpression combined with uga2 knockout also resulted in increased lipid accumulation up to 32.4 %. The gene expression levels and enzyme activities related to fatty acid synthesis showed that deletion of uga2 may lead to carbon-nitrogen metabolism disequilibrium, shifting carbon flux towards lipogenesis. This indicated that AreA might regulate lipid metabolism through the modulation of uga2 in M. circinelloides.
{"title":"The nitrogen regulator AreA modulates lipid metabolism through uga2 in Mucor circinelloides","authors":"Xiuwen Wang , Hassan Mohamed , Qing Liu , Zhaosen Fan , Yuanda Song","doi":"10.1016/j.funbio.2025.101616","DOIUrl":"10.1016/j.funbio.2025.101616","url":null,"abstract":"<div><div>Oleaginous filamentous fungus <em>Mucor circinelloides</em> harbors a GATA transcription activator AreA, which regulates nitrogen metabolism. In our previous study, deletion of AreA resulted in increased lipid production, while its overexpression reduced lipid synthesis. Although it is not a direct lipogenesis regulator, AreA influences metabolic flux by modulating nitrogen utilization pathways, which in turn affects carbon distribution. One such pathway is the GABA shunt, an alternative route that bypasses two steps of TCA cycle and contributes to replenishing TCA intermediates under nitrogen-limited conditions. The <em>uga2</em> gene, encoding succinate semialdehyde dehydrogenase, plays a key role in this shunt. We constructed UGA2-knockout strain in wild-type, AreA-knockout and AreA-overexpression strains, respectively. Results showed that single knockout of <em>uga2</em> increased fatty acid content from 24.5 % to 32.7 %, while double knockout of <em>areA</em> and <em>uga2</em> exhibited the highest fatty acid content at 36.1 %. The <em>areA</em> overexpression combined with <em>uga2</em> knockout also resulted in increased lipid accumulation up to 32.4 %. The gene expression levels and enzyme activities related to fatty acid synthesis showed that deletion of <em>uga2</em> may lead to carbon-nitrogen metabolism disequilibrium, shifting carbon flux towards lipogenesis. This indicated that AreA might regulate lipid metabolism through the modulation of <em>uga2</em> in <em>M. circinelloides</em>.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 6","pages":"Article 101616"},"PeriodicalIF":2.9,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144279426","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-04DOI: 10.1016/j.funbio.2025.101615
Johannes W. Debler, Robert C. Lee, Karam B. Singh, Lars G. Kamphuis, Bernadette M. Henares
Ascochyta lentis and Ascochyta rabiei are fungal pathogens affecting lentil and chickpea crops, respectively. They employ effector proteins to facilitate infection, and understanding the role of effector genes is crucial for unravelling host–pathogen interactions and developing disease-resistant crops. Traditional methods for studying effectors in lentil and chickpea face challenges, such as the ability to perform gene overexpression or knockout studies, due to the difficulty of effector protein infiltration and the limitations of using non-host plants for expression studies. Here, we introduce an alternative tool to enhance the genetic modification toolkit for A. lentis and A. rabiei by developing boscalid-resistant mutants using targeted mutations in the succinate dehydrogenase subunit B (SdhB). This allows for the generation of multiple gene knockouts and gene complementation in A. lentis, where previously only one selectable marker was available. By using the SdhB H277L mutation, we transformed both pathogens and successfully selected transformants using the fungicide boscalid as the selective agent. The method was validated through gene complementation studies of AlScd1 in A. lentis and ArPks1 in A. rabiei, restoring wild type melanin production phenotypes and demonstrating the utility of the new marker system. Additionally, we generated double knockouts in both pathogens, highlighting the potential for more sophisticated genetic studies. The boscalid resistance marker system described here represents a significant advancement in the functional genomics of Ascochyta species, providing a new tool for dissecting the molecular mechanisms underlying pathogenicity and host–pathogen interactions. This approach opens new avenues for research on disease management strategies for lentil and chickpea.
{"title":"Boscalid – a new selectable marker for Ascochyta lentis and Ascochyta rabiei","authors":"Johannes W. Debler, Robert C. Lee, Karam B. Singh, Lars G. Kamphuis, Bernadette M. Henares","doi":"10.1016/j.funbio.2025.101615","DOIUrl":"10.1016/j.funbio.2025.101615","url":null,"abstract":"<div><div><em>Ascochyta lentis</em> and <em>Ascochyta rabiei</em> are fungal pathogens affecting lentil and chickpea crops, respectively. They employ effector proteins to facilitate infection, and understanding the role of effector genes is crucial for unravelling host–pathogen interactions and developing disease-resistant crops. Traditional methods for studying effectors in lentil and chickpea face challenges, such as the ability to perform gene overexpression or knockout studies, due to the difficulty of effector protein infiltration and the limitations of using non-host plants for expression studies. Here, we introduce an alternative tool to enhance the genetic modification toolkit for <em>A. lentis</em> and <em>A. rabiei</em> by developing boscalid-resistant mutants using targeted mutations in the succinate dehydrogenase subunit B (SdhB). This allows for the generation of multiple gene knockouts and gene complementation in <em>A. lentis</em>, where previously only one selectable marker was available. By using the SdhB H277L mutation, we transformed both pathogens and successfully selected transformants using the fungicide boscalid as the selective agent. The method was validated through gene complementation studies of <em>AlScd1</em> in <em>A. lentis</em> and <em>ArPks1</em> in <em>A. rabiei</em>, restoring wild type melanin production phenotypes and demonstrating the utility of the new marker system. Additionally, we generated double knockouts in both pathogens, highlighting the potential for more sophisticated genetic studies. The boscalid resistance marker system described here represents a significant advancement in the functional genomics of Ascochyta species, providing a new tool for dissecting the molecular mechanisms underlying pathogenicity and host–pathogen interactions. This approach opens new avenues for research on disease management strategies for lentil and chickpea.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 5","pages":"Article 101615"},"PeriodicalIF":2.9,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144262471","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-04DOI: 10.1016/j.funbio.2025.101610
Susmita Jana , Dona Das , Sankar Bhattacharyya , Subrata Raha
Nanoparticle research is currently a topic of significant scientific interest, due to its vast array of application in biological field. An effort was made to produce silver nanoparticles (AgNPs) from two endophytic fungi; Fusarium oxysporum (FoAgNPs) and Fusarium proliferatum (FpAgNPs), which were isolated from a Pteridophyte Pyrrosia lanceolata (L.) Farw. The AgNPs were characterised using UV–Vis spectroscopy, exhibiting sizes ranging from 3 nm to 27 nm and displaying a polycrystalline nature as determined by scanning and transmission microscopy, along with SAED pattern analysis. Additionally, we identify phenolic groups at 1067.12 cm−1 as the capping agent that facilitates the reduction of silver ions and stabilizes the nanoparticles evaluated via FTIR. The in vitro antibacterial potency AgNPs had the maximum activity against Escherichia fergusonii, followed by Proteus mirabilis; for both organisms, the minimum inhibitory concentration (MIC) value was 10 μg/ml. AgNPs also demonstrated strong antifungal activity against various plant pathogens, MIC was 15 μg/ml. Additionally, SEM analysis revealed that AgNPs caused pathogen hypha shrinkage and deformation, indicating structural deterioration in cellular and organelle structures due to ROS production. Further, the antifungal efficacy of manufactured AgNPs was investigated against F. oxysporum f.sp. lycopersici in Solanum lycopersicum, a plant pathogen affecting tomato growth and yield, and nano-formulation (150 ppm) completely prevented infection in the greenhouse settings. Biogenic AgNPs at 40 ppm enhanced root-shoot length in Vigna radiata seeds compared to untreated seeds, suggesting phyto-stimulatory action. The cytotoxicity assessment indicated that the synthesized AgNPs are safe for a variety of bio-applications. Furthermore, we chose NADPH Oxidase (F. oxysporum f.sp. lycopersici) as the protein to perform the molecular docking and results indicates that the active site of the selected protein serves as a critical region for inhibiting disease propagation. The study findings enhanced our knowledge of the antifungal properties and mechanisms of AgNPs, providing a novel perspective on utilising this antifungal alternative for the treatment of plant diseases.
{"title":"Mycogenic synthesis of silver nanoparticles using endophytic fungi and their characterization, biological activities, including in-silico studies with special reference to Fusarium wilt of tomato","authors":"Susmita Jana , Dona Das , Sankar Bhattacharyya , Subrata Raha","doi":"10.1016/j.funbio.2025.101610","DOIUrl":"10.1016/j.funbio.2025.101610","url":null,"abstract":"<div><div>Nanoparticle research is currently a topic of significant scientific interest, due to its vast array of application in biological field. An effort was made to produce silver nanoparticles (AgNPs) from two endophytic fungi; <em>Fusarium oxysporum</em> (FoAgNPs) and <em>Fusarium proliferatum</em> (FpAgNPs), which were isolated from a Pteridophyte <em>Pyrrosia lanceolata</em> (L.) Farw. The AgNPs were characterised using UV–Vis spectroscopy, exhibiting sizes ranging from 3 nm to 27 nm and displaying a polycrystalline nature as determined by scanning and transmission microscopy, along with SAED pattern analysis. Additionally, we identify phenolic groups at 1067.12 cm<sup>−1</sup> as the capping agent that facilitates the reduction of silver ions and stabilizes the nanoparticles evaluated via FTIR. The <em>in vitro</em> antibacterial potency AgNPs had the maximum activity against <em>Escherichia fergusonii</em>, followed by <em>Proteus mirabilis</em>; for both organisms, the minimum inhibitory concentration (MIC) value was 10 μg/ml. AgNPs also demonstrated strong antifungal activity against various plant pathogens, MIC was 15 μg/ml. Additionally, SEM analysis revealed that AgNPs caused pathogen hypha shrinkage and deformation, indicating structural deterioration in cellular and organelle structures due to ROS production. Further, the antifungal efficacy of manufactured AgNPs was investigated against <em>F. oxysporum</em> f.sp. <em>lycopersici</em> in <em>Solanum lycopersicum</em>, a plant pathogen affecting tomato growth and yield, and nano-formulation (150 ppm) completely prevented infection in the greenhouse settings. Biogenic AgNPs at 40 ppm enhanced root-shoot length in <em>Vigna radiata</em> seeds compared to untreated seeds, suggesting phyto-stimulatory action. The cytotoxicity assessment indicated that the synthesized AgNPs are safe for a variety of bio-applications. Furthermore, we chose NADPH Oxidase (<em>F. oxysporum</em> f.sp. <em>lycopersici</em>) as the protein to perform the molecular docking and results indicates that the active site of the selected protein serves as a critical region for inhibiting disease propagation. The study findings enhanced our knowledge of the antifungal properties and mechanisms of AgNPs, providing a novel perspective on utilising this antifungal alternative for the treatment of plant diseases.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 5","pages":"Article 101610"},"PeriodicalIF":2.9,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144272486","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-04DOI: 10.1016/j.funbio.2025.101614
Lilla Szendrei , Annamária Tóth , Mátyás Szépligeti , László Palkovics , János Ágoston
Round-leaved sundew (Drosera rotundifolia L.) is a protected glacial relict plant inhabiting Sphagnum bogs, which are endangered habitats in Hungary. In 2020 and 2021 greyish mycelium growth was observed on the hibernacula of D. rotundifolia in Czech Republic, Germany and Hungary. Samples have been collected in possession of the required permits. The fungus was isolated and identified with classical and molecular methods. Koch's postulates were fulfilled. The novel pathogen was identified as the highly polyphagous Botrytis cinerea in each sample. Simultaneously, field assessments of wild Hungarian populations were carried out. Throughout the survey of three different Hungarian collection sites, altogether 207 hibernacula were carefully examined for gray mold symptoms. Interestingly, only plants grown on milled peat substrate were affected by the pathogen. The antifungal and antimicrobial properties of Sphagnum mosses have been reported by other researchers, which could aid in the protection of D. rotundifolia hibernacula. These results indicate that live Sphagnum moss is a better substrate for this species than milled peat, both for commercial production and for in situ conservation. This information can be vital to the survival and conservation of this species. Sphagnum bogs may protect and allow the expansion and re-establishment of D. rotundifolia.
{"title":"Identification of a novel pathogen of the glacial relict Drosera rotundifolia and the impact of the fungus on the conservation of the plant and its habitat","authors":"Lilla Szendrei , Annamária Tóth , Mátyás Szépligeti , László Palkovics , János Ágoston","doi":"10.1016/j.funbio.2025.101614","DOIUrl":"10.1016/j.funbio.2025.101614","url":null,"abstract":"<div><div>Round-leaved sundew (<em>Drosera rotundifolia</em> L.) is a protected glacial relict plant inhabiting <em>Sphagnum</em> bogs, which are endangered habitats in Hungary. In 2020 and 2021 greyish mycelium growth was observed on the hibernacula of <em>D. rotundifolia</em> in Czech Republic, Germany and Hungary. Samples have been collected in possession of the required permits. The fungus was isolated and identified with classical and molecular methods. Koch's postulates were fulfilled. The novel pathogen was identified as the highly polyphagous <em>Botrytis cinerea</em> in each sample. Simultaneously, field assessments of wild Hungarian populations were carried out. Throughout the survey of three different Hungarian collection sites, altogether 207 hibernacula were carefully examined for gray mold symptoms. Interestingly, only plants grown on milled peat substrate were affected by the pathogen. The antifungal and antimicrobial properties of <em>Sphagnum</em> mosses have been reported by other researchers, which could aid in the protection of <em>D. rotundifolia</em> hibernacula. These results indicate that live <em>Sphagnum</em> moss is a better substrate for this species than milled peat, both for commercial production and for <em>in situ</em> conservation. This information can be vital to the survival and conservation of this species. <em>Sphagnum</em> bogs may protect and allow the expansion and re-establishment of <em>D. rotundifolia</em>.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 6","pages":"Article 101614"},"PeriodicalIF":2.9,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144366373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-03DOI: 10.1016/j.funbio.2025.101611
Gabrielle Felizardo , Adrián Adolfo Álvarez Padilla , Amanda Teixeira de Melo , Ricardo Ferreira Lima , Grasielle Pereira Jannuzzi , Kevin Felipe Cruz Martho , Sandro Rogério de Almeida , Karen Spadari Ferreira , Renata Castiglioni Pascon , Marcelo Afonso Vallim
Cryptococcosis is a systemic mycosis caused by Cryptococcus neoformans with significant clinical importance, mainly affecting immunodeficient patients. The treatment options are limited to a few drugs, and resistance to them has been reported. Therefore, research is essential to broaden knowledge regarding the biology of this yeast, aiming to identify traits that could serve as new targets for antifungal drugs. This study aims to expand the current understanding of the autophagy process in this pathogenic yeast. Autophagy is a conserved intracellular degradation and recycling process among eukaryotes, indispensable in cellular homeostasis. In Saccharomyces cerevisiae, the PEP4 gene encodes a protease required during the final stages of autophagy, playing a role in the maturation and activation of vacuolar hydrolases, which contributes to cell survival under conditions of nutritional deprivation and stress. However, PEP4 has never been studied in C. neoformans. Thus, we evaluated the impact of PEP4 deletion on the expression of virulence factors and the cell response to multiple stress conditions. Our results demonstrated that the pep4Δ mutant exhibited attenuated virulence in Galleria mellonella and a decreased fungal burden in macrophages. Notably, we observed the accumulation of autophagic bodies in the pep4Δ strain under nutrient starvation, suggesting a defect in the final steps of autophagic degradation. These findings suggest that the Pep4 protein of C. neoformans plays a crucial role in vacuolar function and the adaptation and survival of yeast cells under stressful conditions, as well as in the host–pathogen interaction.
{"title":"The role of Pep4 protease in Cryptococcus neoformans cell survival and virulence factors","authors":"Gabrielle Felizardo , Adrián Adolfo Álvarez Padilla , Amanda Teixeira de Melo , Ricardo Ferreira Lima , Grasielle Pereira Jannuzzi , Kevin Felipe Cruz Martho , Sandro Rogério de Almeida , Karen Spadari Ferreira , Renata Castiglioni Pascon , Marcelo Afonso Vallim","doi":"10.1016/j.funbio.2025.101611","DOIUrl":"10.1016/j.funbio.2025.101611","url":null,"abstract":"<div><div>Cryptococcosis is a systemic mycosis caused by <em>Cryptococcus neoformans</em> with significant clinical importance, mainly affecting immunodeficient patients. The treatment options are limited to a few drugs, and resistance to them has been reported. Therefore, research is essential to broaden knowledge regarding the biology of this yeast, aiming to identify traits that could serve as new targets for antifungal drugs. This study aims to expand the current understanding of the autophagy process in this pathogenic yeast. Autophagy is a conserved intracellular degradation and recycling process among eukaryotes, indispensable in cellular homeostasis. In <em>Saccharomyces cerevisiae</em>, the <em>PEP4</em> gene encodes a protease required during the final stages of autophagy, playing a role in the maturation and activation of vacuolar hydrolases, which contributes to cell survival under conditions of nutritional deprivation and stress. However, <em>PEP4</em> has never been studied in <em>C. neoformans</em>. Thus, we evaluated the impact of <em>PEP4</em> deletion on the expression of virulence factors and the cell response to multiple stress conditions. Our results demonstrated that the <em>pep4</em>Δ mutant exhibited attenuated virulence in <em>Galleria mellonella</em> and a decreased fungal burden in macrophages. Notably, we observed the accumulation of autophagic bodies in the <em>pep4</em>Δ strain under nutrient starvation, suggesting a defect in the final steps of autophagic degradation. These findings suggest that the Pep4 protein of <em>C. neoformans</em> plays a crucial role in vacuolar function and the adaptation and survival of yeast cells under stressful conditions, as well as in the host–pathogen interaction.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 5","pages":"Article 101611"},"PeriodicalIF":2.9,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144223220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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