Fungal biology最新文献

{"title":"Diversity of tremellalean Pertusaria-associated fungi in Norway and the role of secondary metabolites in host specificity","authors":"Mika H. Kirkhus ,&nbsp;Andreas Frisch ,&nbsp;Ann M. Evankow ,&nbsp;Rakel Blaalid ,&nbsp;Raffaele Zane ,&nbsp;Mika Bendiksby ,&nbsp;Marie L. Davey","doi":"10.1016/j.funbio.2025.101563","DOIUrl":"10.1016/j.funbio.2025.101563","url":null,"abstract":"<div><div>The diversity and host relationships of lichen-inhabiting tremellalean fungi remain understudied, partly due to their asymptomatic yeast phase. This study employs ITS2 metabarcoding to investigate the diversity and colonization patterns of these fungi within 13 <em>Pertusaria</em> lichen species collected in Norway. We discovered a high prevalence, with tremellalean fungi present in 57 % of the examined specimens, representing 12 of the 13 species included, 10 of which were not previously recognized as hosts. Our findings suggest a broad host range for the presumed yeast phase of these fungi, though occurrences of the five most frequent OTUs appeared non-random across different host species. Additionally, negative-binomial general linear models indicated a negative correlation between the presence of tremellalean fungi and the occurrence of specific secondary metabolites, such as thiophaninic acid, fumarprotocetraric acid, and gyrophoric acid, in <em>Pertusaria</em> hosts. These results highlight previously undocumented genetic diversity within tremellalean fungi and suggest that host chemotype significantly influences colonization patterns. Our findings demonstrate a higher frequency of asymptomatic colonization than previously reported, emphasizing the need for further research into their ecological roles and interactions.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 3","pages":"Article 101563"},"PeriodicalIF":2.9,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143561951","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combined metabolome and transcriptome analyses reveal the pivotal role of mycorrhizal fungi Tulasnella sp. BJ1 in the growth and accumulation of secondary metabolites in Bletilla striata (Thunb.) Reiehb.f.","authors":"Yueyu Ye ,&nbsp;Shiqing Li ,&nbsp;Xiaomei Li ,&nbsp;Man Chen ,&nbsp;Haimin Chen ,&nbsp;Qiang Yuan ,&nbsp;Dongfeng Yang ,&nbsp;Meiya Li ,&nbsp;Fusheng Jiang ,&nbsp;Chunchun Zhang","doi":"10.1016/j.funbio.2025.101553","DOIUrl":"10.1016/j.funbio.2025.101553","url":null,"abstract":"<div><div>The orchids usually coexist with mycorrhizal fungi during their growth and development. Numerous studies have substantiated the pivotal regulatory role of <em>Tulasnella</em> sp. mycorrhizal fungi in the germination and growth of orchid seeds. However, there remains a dearth of research elucidating the effects and underlying mechanisms of <em>Tulasnella</em> sp. on the growth, development, and metabolite accumulation in <em>Bletilla striata</em> seedlings. In the current study, metabolomics and transcriptomic analysis were used to reveal the key role of the mycorrhizal fungus <em>Tulasnella</em> sp.BJ1 in the growth and accumulation of secondary metabolites in <em>B. striata</em>. The results demonstrated that the application of BJ1 significantly enhanced the growth and development of <em>B. striata</em> seedlings. In September, the plant weight, tuber diameter, and tuber weight in the BJ1 treatment group reached 44.27 ± 6.79 g, 6.13 ± 0.53 cm, and 23.35 ± 3.06 g, respectively, surpassing those in the control group. The polysaccharide content in the BJ1 treatment group and control group peaked in June, reaching 14.91 ± 2.26 % and 14.38 ± 0.25 %, respectively. Total phenol content in both groups decreased in May and June, and the total phenol content in BJ1 treatment group was significantly lower than that in control group. The significant decrease observed in total phenol content during May and June may be attributed to an increase in proportion of polysaccharides promoted by BJ1. The transcriptome results showed that BJ1 upregulated polysaccharide biosynthesis-related genes, such as mannose phosphatase, transferase, mannose 6-phosphate isomerase, hexokinase, fructose kinase, and glucose 6-phosphate isomerase, as well as genes involved in stilbenes biosynthesis, including hydroxycinnamyltransferase and transcinnamate 4-monooxygenase. Metabolomics data indicated that the content of mannose and seven stilbene compounds in the tubers increased significantly after BJ1 treatment. Interestingly, the accumulation of these compounds corresponds to the pathway of upregulated genes. These findings suggest that an upregulation in mannose synthesis may facilitate the accumulation of polysaccharides in <em>B. striata</em>. Therefore, the current study uncovered that the mycorrhizal fungus <em>Tulasnella</em> sp. BJ1 can not only promote the growth and development of <em>B</em>. <em>striata</em> seedlings and increase tuber yield but also promote the accumulation of polysaccharides and stilbenes.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 3","pages":"Article 101553"},"PeriodicalIF":2.9,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143549030","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}

{"title":"Pleurotus ostreatus mek1 is essential for meiosis and basidiospore production","authors":"Takehito Nakazawa ,&nbsp;Takeshi Kobukata ,&nbsp;Fuga Yamasaki ,&nbsp;Junko Sugano ,&nbsp;Minji Oh ,&nbsp;Moriyuki Kawauchi ,&nbsp;Yoichi Honda","doi":"10.1016/j.funbio.2025.101562","DOIUrl":"10.1016/j.funbio.2025.101562","url":null,"abstract":"<div><div>Breeding strains without basidiospores is important for the mushroom industry. However, target genes for sporeless breeding remain limited. To identify a new gene essential for basidiospore production in <em>Pleurotus ostreatus</em>, the RNA sequencing data obtained in our previous study were re-analyzed. Among the 36 <em>P. ostreatus</em> genes that are exclusively expressed in the gills where basidiospores are formed and produced, candidate genes were narrowed down using transcriptome data during the fruiting stage of <em>Coprinopsis cinerea</em>, in which the meiotic steps progress synchronously. Three <em>C. cinerea</em> genes homologous to the gill-specific <em>P. ostreatus</em> genes were upregulated during meiosis. One of these three genes encodes a protein homologous to <em>Saccharomyces cerevisiae</em> Mek1p, a protein kinase important in the meiotic recombination checkpoint. Plasmids containing expression cassettes for hygromycin B-resistance screening, Cas9, and single-guide RNA targeting <em>mek1</em> were introduced into the protoplasts of the dikaryotic <em>P. ostreatus</em> strain PC9×#64, which showed that the obtained dikaryotic gene disruptant produced no basidiospores. Microscopic analysis suggests that meiosis is suspended during telophase I. These results suggested that <em>P. ostreatus</em> Mek1 is essential for meiosis II progression and basidiospore production. In addition, disturbed orientation and loss of negative gravitropism during fruiting were observed.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 3","pages":"Article 101562"},"PeriodicalIF":2.9,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143534758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolomic profiles of the infection pathways of Calcarisporium cordycipiticola on the cultivated and medicinal mushroom, Cordyceps militaris","authors":"Li Lu ,&nbsp;Mahesh C.A. Galappaththi ,&nbsp;Nimesha M. Patabendige ,&nbsp;Yu-Zhe Feng ,&nbsp;Tian Yang ,&nbsp;Samantha C. Karunarathna ,&nbsp;Jiang-Tao Xie ,&nbsp;Eleni Gentekaki ,&nbsp;Sylvie Rapior ,&nbsp;Esteban Charria-Girón ,&nbsp;Marc Stadler ,&nbsp;Wei-Feng Ding ,&nbsp;Feng-Hua Tian ,&nbsp;Xiang-Yu Zeng","doi":"10.1016/j.funbio.2025.101561","DOIUrl":"10.1016/j.funbio.2025.101561","url":null,"abstract":"<div><div><em>Cordyceps militaris</em> is a widely cultivated mushroom with multiple medicinal properties. However, the emergence of white mildew disease caused by <em>Calcarisporium cordycipiticola</em> has become a serious dilemma, leading to economic losses in its industrial production. The genome of <em>Ca</em>. <em>cordycipiticola</em> possesses more secondary metabolite biosynthetic gene clusters and a smaller number of genes encoding for carbohydrate-active enzyme secretion than other mycoparasites. To uncover those functional metabolites correlated with the infection process, metabolomic profiles between healthy <em>C. militaris</em> fruit bodies (CK) and healthy (HFB) and diseased (DFB) parts of infected <em>C. militaris</em> fruit bodies by <em>Ca. cordicipiticola</em> were compared based on untargeted metabolomic analyses. The function of different metabolites during the pathogen infection and host response processes were further analyzed according to their respective metabolic pathways. Results of key metabolic pathway analyses suggested that a sterigmatocystin-like metabolite functions as one of the virulence factors of white mildew disease on <em>C. militaris</em>, whereas <em>S</em>-adenosyl-L-methionine represents a hub intermediate in both processes of pathogen infection and host response, highlighting the relevance of methyl group turnovers in this battle. More importantly, the detection of toxic metabolites in diseased <em>C. militaris</em> fruiting bodies suggests that this macrofungus contaminated by <em>Ca. cordycipiticola</em> should not be consumed due to the risk that it may contain related instead toxins. This study hypothesizes on the scenario of key metabolic biosynthesis in the battle between <em>Ca</em>. <em>cordycipiticola</em> and <em>C. militaris</em>. Our instead findings not only shed light on the interaction between the pathogen and the host but also provide crucial insights for the development of effective prevention and control strategies in the future.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 3","pages":"Article 101561"},"PeriodicalIF":2.9,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143549029","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}

{"title":"Mango Microcosm: Unveiling the species diversity and pathogenic potential of fungi isolated from mango (Mangifera indica) in northern Thailand","authors":"A. Armand ,&nbsp;M. Fallahi ,&nbsp;S. Absalan ,&nbsp;A.R. Rathnayaka ,&nbsp;U. Braun ,&nbsp;P.W.J. Taylor ,&nbsp;K. Khongphinitbunjong ,&nbsp;E.H.C. McKenzie ,&nbsp;K.D. Hyde ,&nbsp;F. Al-Otibi ,&nbsp;R.S. Jayawardena","doi":"10.1016/j.funbio.2025.101552","DOIUrl":"10.1016/j.funbio.2025.101552","url":null,"abstract":"<div><div>This study aimed to identify fungal pathogens responsible for diseases affecting mango orchards and fruits in the primary mango-growing regions of northern Thailand, and to evaluate their pathogenicity against mango. Mango leaves, branches, and fruits showing fruit and leaf anthracnose, leaf spots, fruit rot, stem-end rot, fruit decline, leaf and stem blight, and shoot dieback were collected and examined from Chiang Rai, Kalasin, Nan, and Sakon Nakhon provinces located in northern Thailand. The isolated fungi were identified based on morphological characteristics and multi-locus phylogenetic analysis based on the most reliable markers and loci combinations suggested for each genus. This study illustrates and describes nine pathogenic genera, including eight known genera and one new genus, <em>Pseudopallidocercospora</em>. <em>Botryosphaeria fabicerciana</em>, <em>Botryosphaeria scharifii</em>, <em>Colletotrichum asianum</em>, <em>Colletotrichum gigasporum</em>, <em>Colletotrichum guajavae</em>, <em>Colletotrichum musae</em>, <em>Colletotrichum plurivorum</em>, <em>Colletotrichum siamense</em>, <em>Diaporthe rosae</em>, <em>Diaporthe siamensis</em>, <em>Diaporthe subellipicola</em>, <em>Neofusicoccum sinoeucalypti</em>, <em>Neopestalotiopsis chrysea</em>, <em>Neopestalotiopsis cubana</em>, <em>Neopestalotiopsis dendrobii</em>, <em>Neopestalotiopsis psidii</em>, <em>Neopestalotiopsis rhododendri</em>, <em>Neoscytalidium dimidiatum</em>, <em>Pestalotiopsis humicola</em> and <em>Pestalotiopsis menhaiensis</em> were identified as known species. Five new species, including <em>Neopestalotiopsis mangiferae</em>, <em>Neopestalotiopsis mangifericola</em>, <em>Neopestalotiopsis mangiferigena</em>, <em>Pseudopallidocercospora mangiferae</em>, and <em>Zasmidium mangiferae</em> were fully illustrated and justified. Moreover, this study reports twelve globally new host-fungus occurrences and six new geographical records. Furthermore, pathogenicity assessment revealed that the majority of the isolates (91.6 %) were pathogenic against mango, while their virulence was significantly different. <em>C. asianum</em> was found to be the predominant pathogenic species with a frequency of 31.6 %. Haplotype networks generated using ITS sequence data for 220 <em>C. asianum</em> strains isolated from mango across 14 countries revealed eight haplotypes, indicating high genetic diversity (Hd = 0.5936) within the species. However, the analysis did not reveal a clear grouping of isolates based on their country of origin.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 3","pages":"Article 101552"},"PeriodicalIF":2.9,"publicationDate":"2025-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143519221","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}

{"title":"Isolate and plant host specificity of rhizosphere competence in Trichoderma species","authors":"N. Cripps-Guazzone ,&nbsp;H.J. Ridgway ,&nbsp;L.M. Condron ,&nbsp;K.L. McLean ,&nbsp;A. Stewart ,&nbsp;E.E. Jones","doi":"10.1016/j.funbio.2025.101554","DOIUrl":"10.1016/j.funbio.2025.101554","url":null,"abstract":"<div><div>Rhizosphere competence, the ability of a microorganism to colonise and proliferate in the rhizosphere of developing roots, is often studied when mechanisms of individual <em>Trichoderma</em> biocontrol agents are investigated. However, the extent of rhizosphere competence of <em>Trichoderma</em> species and isolates within species has not been widely studied. The rhizosphere competence of 22 <em>Trichoderma</em> isolates from a range of species was assessed using <em>Trichoderma</em> coated sweet corn (<em>Zea mays</em>) seeds grown in non-sterile soil. Results showed that 82 % of the <em>Trichoderma</em> isolates inoculated onto the seeds produced rhizosphere populations significantly greater than the control, indicating rhizosphere competence was widespread within the species that were tested. The least and most rhizosphere-competent isolates belonged to the same species indicating that rhizosphere competence was not species specific. The three least (<em>T. crassum</em> LU555, <em>Trichoderma harzianum</em> LU672, and <em>T. virens</em> LU556) and most (<em>Trichoderma atroviride</em> LU132, <em>T. harzianum</em> LU151, and LU673) rhizosphere-competent isolates were assessed on six plant species (sweet corn, ryegrass, cauliflower, carrot, onion, and white clover). Ryegrass and cauliflower were the most receptive plants to colonisation of the rhizosphere by <em>Trichoderma</em> species, and clover the least. Preferential rhizosphere colonisation was observed between some <em>Trichoderma</em> isolates and the plant species indicating that overall rhizosphere competence was dependent on specific interactions between the <em>Trichoderma</em> isolate and the plant species. However, some isolates were more broadly rhizosphere-competent than others and may have greater potential as plant protection agents. Since only one time point was sampled, future work is required to determine the temporal dynamics of rhizosphere colonisation as well as the spatial colonisation along the length of the root to determine whether different isolates preferentially colonise different regions of the root over different time periods.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 3","pages":"Article 101554"},"PeriodicalIF":2.9,"publicationDate":"2025-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143512285","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}

{"title":"Phytopathogenic fungi and oomycetes causing diseases in Theobroma cacao: Chemical and genetic features","authors":"Luis A. Ramírez-Camejo ,&nbsp;Candelario Rodríguez ,&nbsp;Ximena Florez-Buitrago","doi":"10.1016/j.funbio.2025.101551","DOIUrl":"10.1016/j.funbio.2025.101551","url":null,"abstract":"<div><div>Phytopathogenic fungi (PF) and oomycetes (Oo) represent some of the most significant plant pathogens globally, causing extensive damage and economic losses in the chocolate tree, <em>Theobroma cacao.</em> This review aims to elucidate the molecular mechanisms behind cacao-PF/Oo interactions, with a particular emphasis on virulence factors. Despite their importance, the secondary metabolites (SMs) produced during controlled interactions between PF, Oo, and <em>T. cacao</em> remain underexplored. We have conducted a comprehensive review of the most critical PF and Oo species that infect <em>T. cacao</em> and highlighted the agricultural relevance of their SM chemistry. This investigation analyzes peer-reviewed papers from electronic databases PubMed, MDPI, ScienceDirect, Google Scholar, and SCOPUS. Through this analysis, we identify gaps in the current understanding and propose potential directions for future research. This includes a deeper investigation into the role of SMs in pathogen virulence, which could inform the development of more effective disease management strategies.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 3","pages":"Article 101551"},"PeriodicalIF":2.9,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143464228","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}

{"title":"Revealing robust antioxidant defences of a mycoparasitic Trichoderma species","authors":"Pierson Siebe ,&nbsp;Fricker Mark ,&nbsp;Lichius Alexander ,&nbsp;Sandbichler Adolf Michael ,&nbsp;Zeilinger Susanne","doi":"10.1016/j.funbio.2025.101549","DOIUrl":"10.1016/j.funbio.2025.101549","url":null,"abstract":"<div><div>The fungal genus <em>Trichoderma</em> contains a vast array of species well known for their high opportunistic potential and adaptability to various ecological niches. The ability of many <em>Trichoderma</em> species to both colonize the rhizosphere and parasitize plant pathogenic fungi has led to their use in biological pathogen control for several decades. Reactive oxygen species (ROS) are linked to both the antagonism imposed by the mycoparasite <em>Trichoderma</em> and the elicited defence reaction by its fungal hosts during the mycoparasitic interaction. <em>Trichoderma</em> spp. likely tolerate higher levels of ROS compared with some of their host species, thereby giving them an advantage during the mycoparasitic interaction.</div><div>In the present study, we investigated glutathione redox dynamics using the fluorescent reporter Grx1-roGFP2 stably expressed in <em>Trichoderma asperellum</em> following electrotransformation. Grx1-roGFP2 undergoes reversible changes in its excitation spectrum in response to variations in the cellular glutathione redox potential, providing a real-time indication of intracellular oxidative load. Considering the putative importance of ROS in mycoparasitic interactions, we performed live-cell imaging of the <em>T. asperellum</em> reporter strain interacting with the cereal pathogen <em>Fusarium graminearum.</em> Surprisingly, the glutathione redox potential did not change during this mycoparasitic interaction. We found no evidence that host-induced tip growth arrest within <em>T. asperellum</em> hyphae is induced by intracellular ROS accumulation. Furthermore, we show that the <em>F. graminearum</em> mycotoxins deoxynivalenol and zearalenone do not induce detectable changes in glutathione redox potential, even at very high concentrations. We infer that <em>T. asperellum</em> has a robust anti-oxidant defence system, supported by the observation that high concentrations of H₂O₂ are required to fully oxidize the reporter during <em>in vivo</em> calibration. We cannot rule out a role for ROS as a signal during mycoparasitic interactions, but, if present, this does not appear to be mediated by glutathione redox potential.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 3","pages":"Article 101549"},"PeriodicalIF":2.9,"publicationDate":"2025-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143454716","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Establishment of Agrobacterium tumefaciens-mediated genetic transformation of the entomopathogenic fungus Hirsutella satumaensis","authors":"Yongli Feng ,&nbsp;Jiaojiao Qu ,&nbsp;Xiao Zou ,&nbsp;Wei Cao ,&nbsp;Yeming Zhou","doi":"10.1016/j.funbio.2025.101548","DOIUrl":"10.1016/j.funbio.2025.101548","url":null,"abstract":"<div><div><em>Hirsutella satumaensis</em>, an endoparasitic fungus that targets Lepidoptera insects, holds significant potential for biocontrol applications. However, its molecular study has been limited due to the absence of an efficient genetic transformation system. In this study, an optimized <em>Agrobacterium tumefaciens</em>-mediated transformation protocol was developed for <em>H. satumaensis</em> using binary vectors pBARGPE1-GFP and pK2-bar, which carry the green fluorescent protein (<em>eGFP</em>) and phosphinothricin resistance (<em>bar</em>) genes, respectively. The optimal transformation conditions included a conidial concentration of 10⁵ conidia/mL, an <em>A. tumefaciens</em> (strain AGL-1) concentration of OD₆₆₀ = 0.6, and a 3-day co-cultivation period with 200 μM acetosyringone, resulting in an average of 121 ± 5.07 transformants. Successful integration was confirmed by green fluorescence in the transformants. Furthermore, the ribotoxin gene <em>hirsutellin A</em> (<em>HtA</em>), specific to the genus <em>Hirsutella</em>, was successfully overexpressed using this system. Insect bioassays demonstrated that the gpdA promoter effectively drives <em>HtA</em> expression in <em>H. satumaensis</em>. The transformation system exhibited stable gene integration, strong fluorescence, and bioactivity. This study establishes the first genetic transformation protocol for <em>H. satumaensis</em>, providing a valuable tool for exploring insect-pathogen interactions and the functional roles of key genes in this entomopathogenic fungus.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 2","pages":"Article 101548"},"PeriodicalIF":2.9,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143436588","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}

{"title":"Functional analysis of two genes coding for distinct cation diffusion facilitators of the cadmium-accumulating fungus Agaricus crocodilinus","authors":"Jan Sácký ,&nbsp;Veronika Liščáková ,&nbsp;Jan Šnábl ,&nbsp;Jaroslav Zelenka ,&nbsp;Jan Borovička ,&nbsp;Tereza Leonhardt ,&nbsp;Pavel Kotrba","doi":"10.1016/j.funbio.2025.101550","DOIUrl":"10.1016/j.funbio.2025.101550","url":null,"abstract":"<div><div>The gilled mushroom <em>Agaricus crocodilinus</em> (<em>Agaricaceae</em>) analyzed in this study hyperaccumulated Cd and showed common Zn and very low Mn concentrations. To gain an insight into the handling of heavy metals in this saprotrophic species, its two genes of the cation diffusion facilitator (CDF) protein family were isolated, Ac<em>CDF1</em> and Ac<em>CDF2</em>, encoding the membrane transporters of the Zn-CDF and Mn-CDF subfamilies, respectively. When expressed in the model, metal-sensitive yeast, Ac<em>CDF1</em> conferred marked Zn tolerance and promoted the intracellular accumulation of Zn. Green fluorescent protein (GFP) tagging of AcCDF1 visualized the functional protein predominantly in the tonoplast, indicating that AcCDF1 can mediate the transport of Zn into vacuoles, which are used for deposition of excess Zn in most fungi. Ac<em>CDF2</em> conferred a high degree of Mn tolerance to model yeast, in which the transport-active AcCDF2:GFP fusion was localized to the plasma membrane, suggesting a role in Mn export and thus reduced Mn accumulation. Furthermore, the Ac<em>CDF2</em> gene appeared to be Mn-inducible in <em>A. crocodilinus</em>, suggesting an Mn efflux function of AcCDF2. Neither AcCDFs nor the mutant AcCDF1 variants constructed to mimic transmembrane tetrahedral Cd transport sites manifested appreciable Cd-related phenotypes in yeast models, and further efforts are needed to elucidate the mechanism underlying Cd hyperaccumulation in <em>A. crocodilinus</em>.</div></div>","PeriodicalId":12683,"journal":{"name":"Fungal biology","volume":"129 2","pages":"Article 101550"},"PeriodicalIF":2.9,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143420172","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}