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Characterization of a High-Affinity Copper Transporter CTR1a in the White-Nose Syndrome Causing Fungal Pathogen Pseudogymnoascus destructans. 白鼻综合征致病真菌假革囊菌中高亲和性铜转运体 CTR1a 的特征。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-21 DOI: 10.3390/jof10100729
Saika Anne, Alyssa D Friudenberg, Ryan L Peterson

Copper is an essential micronutrient and the ability to scavenge tightly bound or trace levels of copper ions at the host-pathogen interface is vital for fungal proliferation in animal hosts. Recent studies suggest that trace metal ion acquisition is critical for the establishment and propagation of Pseudogymnoascus destructans, the fungal pathogen responsible for white-nose syndrome (WNS), on their bat host. However, little is known about these metal acquisition pathways in P. destructans. In this study, we report the characterization of the P. destructans high-affinity copper transporter VC83_00191 (PdCTR1a), which is implicated as a virulence factor associated with the WNS disease state. Using Saccharomyces cerevisiae as a recombinant expression host, we find that PdCTR1a can efficiently traffic Cu ions into the yeast cytoplasm. Complementary studies in the native P. destructans fungus provide evidence that PdCTR1a transcripts and protein levels are dictated by Cu-bioavailability in the growth media. Our study demonstrates that PdCTR1a is a functional high-affinity copper transporter and is relevant to Cu homeostasis pathways in P. destructans.

铜是一种必需的微量营养元素,在宿主-病原体界面清除紧密结合或痕量水平的铜离子的能力对于真菌在动物宿主中的繁殖至关重要。最近的研究表明,痕量金属离子的获取对于导致白鼻综合症(WNS)的真菌病原体--破坏性假丝酵母菌(Pseudogymnoascus destructans)在蝙蝠宿主上的建立和繁殖至关重要。然而,人们对毁灭假丝酵母的这些金属获取途径知之甚少。在本研究中,我们报告了毁坏真菌高亲和性铜转运体 VC83_00191 (PdCTR1a)的特征,它被认为是与 WNS 疾病状态相关的毒力因子。利用酿酒酵母作为重组表达宿主,我们发现 PdCTR1a 能有效地将铜离子转运到酵母细胞质中。在原生毁灭酵母菌中进行的补充研究证明,PdCTR1a 的转录本和蛋白质水平受生长介质中铜生物可用性的影响。我们的研究证明,PdCTR1a 是一种功能性高亲和性铜转运体,与去壳酵母中的铜平衡途径有关。
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引用次数: 0
Diversity and Antifungal Susceptibilities of Yeasts from Mangroves in Hong Kong, China-A One Health Aspect. 中国香港红树林中酵母菌的多样性和抗真菌敏感性--一个健康角度。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-20 DOI: 10.3390/jof10100728
Pak-Ting Hau, Anson Shiu, Emily Wan-Ting Tam, Eddie Chung-Ting Chau, Michaela Murillo, Eva Humer, Wai-Wai Po, Ray Chun-Wai Yu, Joshua Fung, Sai-Wang Seto, Chi-Ching Tsang, Franklin Wang-Ngai Chow

While mangrove ecosystems are rich in biodiversity, they are increasingly impacted by climate change and urban pollutants. The current study provides first insights into the emergence of potentially pathogenic yeasts in Hong Kong's mangroves. Sediment and water samples were collected from ten urban and rural mangroves sites. Initial CHROMagarTM Candida Plus screening, representing the first application of this differential medium for water and soil samples collected from a non-clinical environment, enabled the rapid, preliminary phenotypic identification of yeast isolates from mangroves. Subsequent molecular profiling (ITS and/or 28S nrDNA sequencing) and antifungal drug susceptibility tests were conducted to further elucidate yeast diversity and drug resistance. A diversity of yeasts, including 45 isolates of 18 distinct species across 13 genera/clades, was isolated from sediments and waters from Hong Kong mangroves. Molecular profiling revealed a dominance of the Candida/Lodderomyces clade (44.4%), a group of notorious opportunistic pathogens. The findings also reveal a rich biodiversity of non-Candida/Lodderomyces yeasts in mangroves, including the first reported presence of Apiotrichum domesticum and Crinitomyces flavificans. A potentially novel Yamadazyma species was also discovered. Remarkably, 14.3% of the ubiquitous Candida parapsilosis isolates displayed resistance to multiple antifungal drugs, suggesting that mangroves may be reservoirs of multi-drug resistance. Wildlife, especially migratory birds, may disseminate these hidden threats. With significant knowledge gaps regarding the environmental origins, drug resistance, and public health impacts of pathogenic yeasts, urgent surveillance is needed from a One Health perspective. This study provides an early warning that unrestrained urbanization can unleash resistant pathogens from coastal ecosystems globally. It underscores the necessity for enhanced surveillance studies and interdisciplinary collaboration between clinicians, ornithologists, and environmental microbiologists to effectively monitor and manage this environmental health risk, ensuring the maintenance of 'One Health'.

虽然红树林生态系统具有丰富的生物多样性,但它们受到气候变化和城市污染物的影响日益严重。本研究首次揭示了香港红树林中可能出现的致病酵母菌。研究人员从十个城市和农村红树林地点采集了沉积物和水样本。初步的 CHROMagarTM Candida Plus 筛选(这是首次将这种差异培养基用于从非临床环境中采集的水和土壤样本)使从红树林中分离的酵母菌得到了快速、初步的表型鉴定。随后进行了分子剖析(ITS 和/或 28S nrDNA 测序)和抗真菌药敏试验,以进一步阐明酵母菌的多样性和耐药性。从香港红树林的沉积物和水体中分离出了多种酵母菌,包括 13 个属/支系 18 个不同物种的 45 个分离物。分子分析表明,念珠菌/巴豆菌支系(44.4%)占主导地位,这是一组臭名昭著的机会致病菌。研究结果还揭示了红树林中丰富的非念珠菌/巴豆酵母菌生物多样性,包括首次报道的 Apiotrichum domesticum 和 Crinitomyces flavificans。此外,还发现了一个潜在的新物种 Yamadazyma。值得注意的是,在无处不在的副丝状念珠菌分离物中,有 14.3% 显示出对多种抗真菌药物的耐药性,这表明红树林可能是多种药物耐药性的储存库。野生动物,尤其是候鸟,可能会传播这些隐藏的威胁。由于对病原酵母菌的环境起源、耐药性和对公共卫生的影响存在巨大的知识差距,因此需要从 "一体健康 "的角度进行紧急监测。这项研究提供了一个预警,即无节制的城市化可能会从全球沿海生态系统中释放出具有抗药性的病原体。它强调了加强监测研究以及临床医生、鸟类学家和环境微生物学家之间跨学科合作的必要性,以有效监测和管理这一环境健康风险,确保维护 "一体健康"。
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引用次数: 0
Exogenous Nitric Oxide Induces Pathogenicity of Alternaria alternata on Huangguan Pear Fruit by Regulating Reactive Oxygen Species Metabolism and Cell Wall Modification. 外源一氧化氮通过调节活性氧代谢和细胞壁修饰诱导黄冠梨果上的交替孢属致病性
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-19 DOI: 10.3390/jof10100726
Di Wang, Haijue Zhang, Lingkui Meng, Xinyu Tan, Rong Liu, Qingchao Gao, Yan Wu, Yuhan Zhu, Xueyan Ren, Yongcai Li, Qingjun Kong

Black spot caused by Alternaria alternata is one of the most common postharvest diseases in fruit and vegetables. A comprehensive investigation into its pathogenicity mechanism is imperative in order to propose a targeted and effective control strategy. The effect of nitric oxide (NO) on the pathogenicity of A. alternata and its underlying mechanism was studied. The results showed that treatment with 0.5 mM L-1 of sodium nitroprusside (SNP) (NO donor) increased the lesion diameter of A. alternata in vivo and in vitro, which was 22.8% and 13.2% higher than that of the control, respectively. Exogenous NO treatment also induced endogenous NO accumulation by activating nitric oxide synthase (NOS). In addition, NO triggered an increase in reactive oxygen species (ROS) levels. NO enhanced activities and gene expression levels of NADPH oxidase (NOX), superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione peroxidase (GPX), and glutathione reductase (GR). Moreover, NO stimulated cell wall degrading enzymes by activating the corresponding gene expression in vivo and in vitro. These results suggested that exogenous NO promoted the pathogenicity of A. alternata by inducing ROS accumulation and activating antioxidants and cell wall degrading enzymes. The present results could establish a theoretical foundation for the targeted control of the black spot disease in pear fruit.

由交替花叶病毒(Alternaria alternata)引起的黑斑病是水果和蔬菜收获后最常见的病害之一。为了提出有针对性的有效防治策略,对其致病机理进行全面研究势在必行。研究了一氧化氮(NO)对交替孢霉致病性的影响及其内在机制。结果表明,用 0.5 mM L-1 的硝普钠(SNP)(一氧化氮供体)处理交替蚁,其体内和体外病变直径分别比对照组增加了 22.8%和 13.2%。外源性 NO 处理还能通过激活一氧化氮合酶(NOS)诱导内源性 NO 的积累。此外,NO 还会引发活性氧(ROS)水平的升高。NO 提高了 NADPH 氧化酶(NOX)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、谷胱甘肽过氧化物酶(GPX)和谷胱甘肽还原酶(GR)的活性和基因表达水平。此外,氮氧化物还通过激活体内和体外相应基因的表达来刺激细胞壁降解酶。这些结果表明,外源 NO 通过诱导 ROS 积累、激活抗氧化剂和细胞壁降解酶促进了交替穗霉的致病性。这些结果为有针对性地防治梨果黑斑病奠定了理论基础。
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引用次数: 0
Recent Advances in Diagnostic Approaches for Mucormycosis. 粘孢子菌病诊断方法的最新进展。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-19 DOI: 10.3390/jof10100727
Jawad Safiia, Marco Aurelio Díaz, Hassan Alshaker, Christine J Atallah, Paul Sakr, Dimitrios G Moshovitis, Ahmad Nawlo, Andres E Franceschi, Alexis Liakos, Sophia Koo

Mucormycosis, an invasive fungal infection caused by members of the order Mucorales, often progresses fulminantly if not recognized in a timely manner. This comprehensive review discusses the latest developments in diagnostic approaches for mucormycosis, from traditional histopathology and culture-based methods to advanced and emerging techniques such as molecular assays, imaging, serology, and metabolomics. We discuss challenges in the diagnosis of mucormycosis and emphasize the importance of rapid and accurate identification of this life-threatening infection.

粘孢子菌病是由粘孢子菌目成员引起的一种侵袭性真菌感染,如果不能及时发现,病情往往会急剧恶化。本综述讨论了粘孢子菌病诊断方法的最新进展,从传统的组织病理学和培养方法到先进的新兴技术,如分子检测、成像、血清学和代谢组学。我们讨论了粘孢子虫病诊断中的挑战,并强调了快速准确地识别这种威胁生命的感染的重要性。
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引用次数: 0
Overview of the Current Challenges in Pulmonary Coccidioidomycosis. 概述肺球孢子菌病目前面临的挑战。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-18 DOI: 10.3390/jof10100724
Mohamed A Fayed, Timothy M Evans, Eyad Almasri, Kathryn L Bilello, Robert Libke, Michael W Peterson

Coccidioidomycosis is a disease caused by soil fungi of the genus Coccidioides, divided genetically into Coccidioides immitis (California isolates) and Coccidioides posadasii (isolates outside California). Coccidioidomycosis is transmitted through the inhalation of fungal spores, arthroconidia, which can cause disease in susceptible mammalian hosts, including humans. Coccidioidomycosis is endemic to the western part of the United States of America, including the central valley of California, Arizona, New Mexico, and parts of western Texas. Cases have been reported in other regions in different states, and endemic pockets are present in these states. The incidence of reported cases of coccidioidomycosis has notably increased since it became reportable in 1995. Clinically, the infection ranges from asymptomatic to fatal disease due to pneumonia or disseminated states. The recognition of coccidioidomycosis can be challenging, as it frequently mimics bacterial community-acquired pneumonia. The diagnosis of coccidioidomycosis is frequently dependent on serologic testing, the results of which can take several days or longer to obtain. Coccidioidomycosis continues to present challenges for clinicians, and suspected cases can be easily missed. The challenges of coccidioidomycosis disease, from presentation to diagnosis to treatment, remain a hurdle for clinicians, and further research is needed to address these challenges.

球孢子菌病是由球孢子菌属的土壤真菌引起的一种疾病,从基因上分为 Coccidioides immitis(加利福尼亚州分离物)和 Coccidioides posadasii(加利福尼亚州以外分离物)。球孢子菌病通过吸入真菌孢子、节孢子传播,可导致包括人类在内的易感哺乳动物宿主发病。球孢子菌病流行于美国西部,包括加利福尼亚州中部山谷、亚利桑那州、新墨西哥州和得克萨斯州西部部分地区。不同州的其他地区也有病例报告,这些州也有地方病流行区。自 1995 年可报告球孢子菌病以来,报告病例的发病率明显上升。在临床上,从无症状感染到因肺炎或播散而致命的疾病都有。由于球孢子菌病经常与细菌性社区获得性肺炎相似,因此识别球孢子菌病具有挑战性。球孢子菌病的诊断通常取决于血清学检测,而检测结果可能需要几天或更长时间才能获得。球孢子菌病一直是临床医生面临的难题,疑似病例很容易被漏诊。球孢子菌病从发病到诊断再到治疗的整个过程都给临床医生带来了挑战,需要进一步的研究来应对这些挑战。
{"title":"Overview of the Current Challenges in Pulmonary Coccidioidomycosis.","authors":"Mohamed A Fayed, Timothy M Evans, Eyad Almasri, Kathryn L Bilello, Robert Libke, Michael W Peterson","doi":"10.3390/jof10100724","DOIUrl":"https://doi.org/10.3390/jof10100724","url":null,"abstract":"<p><p>Coccidioidomycosis is a disease caused by soil fungi of the genus <i>Coccidioides</i>, divided genetically into <i>Coccidioides immitis</i> (California isolates) and <i>Coccidioides posadasii</i> (isolates outside California). Coccidioidomycosis is transmitted through the inhalation of fungal spores, arthroconidia, which can cause disease in susceptible mammalian hosts, including humans. Coccidioidomycosis is endemic to the western part of the United States of America, including the central valley of California, Arizona, New Mexico, and parts of western Texas. Cases have been reported in other regions in different states, and endemic pockets are present in these states. The incidence of reported cases of coccidioidomycosis has notably increased since it became reportable in 1995. Clinically, the infection ranges from asymptomatic to fatal disease due to pneumonia or disseminated states. The recognition of coccidioidomycosis can be challenging, as it frequently mimics bacterial community-acquired pneumonia. The diagnosis of coccidioidomycosis is frequently dependent on serologic testing, the results of which can take several days or longer to obtain. Coccidioidomycosis continues to present challenges for clinicians, and suspected cases can be easily missed. The challenges of coccidioidomycosis disease, from presentation to diagnosis to treatment, remain a hurdle for clinicians, and further research is needed to address these challenges.</p>","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":"10 10","pages":""},"PeriodicalIF":4.2,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11508864/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142516477","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}
引用次数: 0
Distributions of Lanostene-Derived Triterpenoids and Glucan Content in the Fruiting Bodies of the Australian Ganoderma Species. 澳大利亚灵芝菌种果实体内的兰氏三萜类化合物和葡聚糖含量的分布。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-18 DOI: 10.3390/jof10100723
Aline De Oliveira Campos, Mark D Harrison, David L Marshall, Peter James Strong

Lanostene-derived triterpenoids and β-glucans are important metabolites in Ganoderma mushrooms associated with benefits to human health. The medicinal value of the Australian Ganoderma species remains unclear, with no data on triterpenoid distribution or glucan content. In the present study, 22 Australian Ganoderma specimens were analyzed for triterpenoid and glucan contents. Thirty-two triterpenoids were identified in the fruiting bodies of 19 of the specimens. Distinct patterns in triterpenoid distribution between laccate and matte fruiting bodies were observed, leading to the classification of four groups of Ganoderma. Most of the glucans in the Ganoderma fruiting bodies were β-glucans (~99%), with a nominal α-glucan content (~1%). The β-glucan content ranged from 19.5 to 43.5% (w/w). A range of antioxidant activities was observed for methanol extracts using the ABTS (1.8 to 8.4 mg GAE.g-1), DPPH (1.7 to 9.4 mg GAE/g-1) and FRAP (24.7 to 111.6 mmol FeSO4.g-1) assays, with four specimens presenting relatively high radical scavenging and reducing activities. For the first time, we demonstrated that Australian Ganoderma mushrooms contain medicinal triterpenoids, including ganoderic acid A, and we established a link between its distribution and the fruiting body morphology. However, further research is required to isolate diploid clones and determine factors that impact triterpenoid and glucan synthesis in these strains.

兰芝三萜类化合物和β-葡聚糖是灵芝中的重要代谢产物,对人类健康有益。澳大利亚灵芝物种的药用价值尚不清楚,也没有关于三萜类化合物分布或葡聚糖含量的数据。本研究分析了 22 个澳大利亚灵芝标本的三萜类化合物和葡聚糖含量。在其中 19 个标本的子实体中鉴定出 32 种三萜类化合物。观察到三萜类化合物在长条形子实体和无光泽子实体之间的不同分布模式,从而将灵芝分为四类。灵芝子实体中的大部分葡聚糖都是β-葡聚糖(约占 99%),α-葡聚糖含量很少(约占 1%)。β-葡聚糖含量介于 19.5%至 43.5%(重量/重量)之间。使用 ABTS(1.8 至 8.4 毫克 GAE.g-1)、DPPH(1.7 至 9.4 毫克 GAE/g-1)和 FRAP(24.7 至 111.6 毫摩尔 FeSO4.g-1)测定法观察到甲醇提取物具有一定范围的抗氧化活性,其中四个样本具有相对较高的自由基清除和还原活性。我们首次证明澳大利亚灵芝含有包括灵芝酸 A 在内的药用三萜类化合物,并建立了灵芝酸 A 的分布与子实体形态之间的联系。不过,还需要进一步研究分离二倍体克隆,并确定影响这些菌株三萜类化合物和葡聚糖合成的因素。
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引用次数: 0
Three New Species of Mytilinidioid Fungi (Dothideomycetes, Ascomycota) from Mexico. 来自墨西哥的三个 Mytilinidioid 真菌新种(Dothideomycetes, Ascomycota)。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-18 DOI: 10.3390/jof10100725
Tania Raymundo, César R Martínez-González, Michelle Martínez-Pineda, Aurora Cobos-Villagrán, Isabel Ramírez-Rosales, Ricardo Valenzuela

Mytilinidioid fungi are conchiform in nature, with the appearance of bivalve shells or wedge-shaped, rigid, brittle, and carbonaceous hysterothecia growing on the bark of gymnosperms or angiosperms. Based on their morphological characteristics and molecular markers (ITS and LSU), this study describes three new species of mytilinidioid fungi: Ericboehmia mexicana of the family Hysteriaceae of the order Hysteriales and Lophium pinicola and Mytilinidion mexicanum of the family Mytilinidiaceae of the order Mytilinidiales. The first species grows on Liquidambar styracyphlua, the second species grows on Pinus patula, and the third species grows on Acacia californica subsp. pringlei. The specimens studied were deposited in the ENCB Herbarium.

栉孔真菌(Mytilinidioid fungi)呈贝壳状或楔形,生长在裸子植物或被子植物的树皮上,质地坚硬、脆硬、碳质。根据它们的形态特征和分子标记(ITS 和 LSU),本研究描述了三种蕈菌类真菌新种:这三个新种分别是:Hysteriales 目 Hysteriaceae 科的 Ericboehmia mexicana,以及 Mytilinidiales 目 Mytilinidiaceae 科的 Lophium pinicola 和 Mytilinidion mexicanum。第一个物种生长在枫香树上,第二个物种生长在松树上,第三个物种生长在加州金合欢亚种上。所研究的标本保存在 ENCB 标本馆。
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引用次数: 0
Stratocorticium sinensis gen. et sp. nov. and Cericium gloeocystidiatum sp. nov. (Cyphellaceae, Agaricales) from East Asia. 来自东亚的 Stratocorticium sinensis gen.
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-17 DOI: 10.3390/jof10100722
Yu-Peng Zhang, Yue Li, Karen K Nakasone, Shuang-Hui He

Cyphellaceae, a small and under-studied family of Agaricales, includes mostly saprophytic taxa with varied basidiomes. In this study, we focus on wood-decay species with corticioid or stereoid basidiomes. Phylogenetic analyses of concatenated ITS-nrLSU sequences uncovered seven generic lineages of corticioid or stereoid fungi-Acanthocorticium, Cericium, Chondrostereum, Cunninghammyces, Gloeostereum, Granulobasidium, and Stratocorticium gen. nov. The genus Cericium is shown to be in the Cyphellaceae family, and two new species, Cericium gloeocystidiatum and Stratocorticium sinensis, are described from East Asia. Morphologically, Ce. gloeocystidiatum is characterized by resupinate basidiomes with smooth hymenophores, a dimitic hyphal system with clamped generative hyphae and micro-binding hyphae, cystidia with resinous-like or golden yellow contents, and ellipsoid basidiospores. Stratocorticium is monotypic, differing from Cericium by a trimitic hyphal system of clamped generative, micro-binding, and brown, thick-walled skeletal-like hyphae, clavate to cylindrical cystidia with homogenous, colorless contents, and hyphidia. Descriptions and illustrations are provided for the new taxa and Cericium luteoincrustatum, and a key to corticioid or stereoid genera in Cyphellaceae is included.

黄柏科(Cyphellaceae)是姬松属(Agaricales)中一个规模较小、研究较少的科,主要包括基部形态各异的吸浆类群。在本研究中,我们主要研究具有皮质基部或立体基部的木腐物种。ITS-nrLSU 序列的系统发育分析发现了皮层真菌或立体真菌的 7 个属系--Acanthocorticium、Cericium、Chondrostereum、Cunninghammyces、Gloeostereum、Granulobasidium 和 Stratocorticium gen.结果表明 Cericium 属属于黄柏科,并从东亚描述了两个新种:Cericium gloeocystidiatum 和 Stratocorticium sinensis。从形态上看,Ce. gloeocystidiatum 的特征是复杯状基生体,具有光滑的膜柄;二纤细的菌丝系统,具有夹持的生成菌丝和微结合菌丝;子囊腔具有树脂状或金黄色内容物;基生孢子椭圆形。Stratocorticium 为单型,与 Cericium 的不同之处在于其三生菌丝系统,包括夹生的生成菌丝、微结合菌丝和棕色的厚壁骨架状菌丝,棍棒状至圆柱状的子囊菌,内含物均匀无色,以及菌褶。该研究为新分类群和 Cericium luteoincrustatum 提供了描述和插图,并包含了黄柏科皮质类属或立体类属的检索表。
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引用次数: 0
Molecular Mechanism of Mok I Gene Overexpression in Enhancing Monacolin K Production in Monascus pilosus. 莫克 I 基因过表达促进莫纳可林 K 生产的分子机制
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-16 DOI: 10.3390/jof10100721
Zhiwei Huang, Lishi Xiao, Wenlan Mo, Yaru Zhang, Yiyang Cai, Simei Huang, Zhiting Chen, Chuannan Long

Monascus species are capable of producing various active metabolites, including monacolin K (MK) and pigments. Studies have shown that the overexpression of the mok I gene from the MK synthesis gene cluster in Monascus species can significantly increase MK production; however, the molecular mechanism has not yet been fully elucidated. Therefore, this study focused on the mok I gene of Monascus pilosus to construct overexpression strains of the mok I gene, resulting in high-yield MK production. Sixteen positive transformants were obtained, seven of which produced 9.63% to 41.39% more MK than the original strain, with no citrinin detected in any of the transformants. The qRT-PCR results revealed that the expression levels of mok I in the transformed strains TI-13, TI-24, and TI-25 increased by more than 50% compared to the original strain at various fermentation times, with the highest increase being 10.9-fold. Furthermore, multi-omics techniques were used to analyze the molecular mechanisms underlying enhanced MK production in transformed strains. The results indicated that mok I overexpression may enhance MK synthesis in M. pilosus by regulating the expression of key genes (such as MAO, HPD, ACX, and PLC) and the synthesis levels of key metabolites (such as delta-tocopherol and alpha-linolenic acid) in pathways linked to the biosynthesis of cofactors, the biosynthesis of unsaturated fatty acids, tyrosine metabolism, ubiquinone and other terpenoid-quinone biosynthesis, alpha-linolenic acid metabolism, and glycerophospholipid metabolism. These findings provide a theoretical basis for further study of the metabolic regulation of MK in Monascus species and for effectively enhancing their MK production.

莫纳科菌类能够产生各种活性代谢产物,包括莫纳可林 K(MK)和色素。研究表明,过表达莫纳可林合成基因簇中的 mok I 基因可显著提高莫纳可林的产量,但其分子机制尚未完全阐明。因此,本研究聚焦于Monascus pilosus的mok I基因,构建mok I基因的过表达菌株,从而获得高产的MK产量。研究获得了 16 个阳性转化株,其中 7 个转化株的 MK 产量比原始菌株高出 9.63% 至 41.39%,所有转化株均未检测到柠檬蛋白。qRT-PCR 结果显示,在不同的发酵时间,转化菌株 TI-13、TI-24 和 TI-25 中 mok I 的表达水平比原菌株提高了 50%以上,最高提高了 10.9 倍。此外,研究人员还利用多组学技术分析了转化菌株MK产量增加的分子机制。结果表明,mok I 的过表达可通过调控 M. pilosus 菌株的表达来提高 MK 的合成。结果表明,mok I 的过表达可通过调节关键基因(如 MAO、HPD、ACX 和 PLC)的表达和关键代谢产物(如 delta-生育酚和α-亚麻酸)在辅助因子生物合成、不饱和脂肪酸生物合成、酪氨酸代谢、泛醌和其他萜类-醌类生物合成、α-亚麻酸代谢和甘油磷脂代谢等相关途径中的合成水平来提高 MK 的合成。这些发现为进一步研究莫纳树属物种的 MK 代谢调节和有效提高其 MK 产量提供了理论依据。
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引用次数: 0
The Aspergillus flavus hacA Gene in the Unfolded Protein Response Pathway Is a Candidate Target for Host-Induced Gene Silencing. 未折叠蛋白反应途径中的黄曲霉 hacA 基因是宿主诱导基因沉默的候选靶标
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-16 DOI: 10.3390/jof10100719
Perng-Kuang Chang

Fungal HacA/Hac1 transcription factors play a crucial role in regulating the unfolded protein response (UPR). The UPR helps cells to maintain endoplasmic reticulum (ER) protein homeostasis, which is critical for growth, development, and virulence. The Aspergillus flavus hacA gene encodes a domain rich in basic and acidic amino acids (Bsc) and a basic leucine zipper (bZip) domain, and features a non-conventional intron (Nt20). In this study, CRISPR/Cas9 was utilized to dissect the Bsc-coding, bZip-coding, and Nt20 sequences to elucidate the relationship between genotype and phenotype. In the Bsc and bZip experimental sets, all observed mutations in both coding sequences were in frame, suggesting that out-of-frame mutations are lethal. The survival rate of transformants in the Nt20 experiment set was low, at approximately 7%. Mutations in the intron primarily consisted of out-of-frame insertions and deletions. In addition to the wild-type-like conidial morphology, the mutants exhibited varied colony morphologies, including sclerotial, mixed (conidial and sclerotial), and mycelial morphologies. An ER stress test using dithiothreitol revealed that the sclerotial and mycelial mutants were much more sensitive than the conidial mutants. Additionally, the mycelial mutants were unable to produce aflatoxin but still produced aspergillic acid and kojic acid. RNAi experiments targeting the region encompassing Bsc and bZip indicated that transformant survival rates generally decreased, with a small number of transformants displaying phenotypic changes. Defects in the hacA gene at the DNA and transcript levels affected the survival, growth, and development of A. flavus. Thus, this gene may serve as a promising target for future host-induced gene-silencing strategies aimed at controlling infection and reducing aflatoxin contamination in crops.

真菌 HacA/Hac1 转录因子在调节未折叠蛋白反应(UPR)中发挥着至关重要的作用。UPR 帮助细胞维持内质网(ER)蛋白质的平衡,这对生长、发育和毒力至关重要。黄曲霉的 hacA 基因编码一个富含碱性和酸性氨基酸(Bsc)的结构域和一个碱性亮氨酸拉链(bZip)结构域,并具有一个非常规内含子(Nt20)。本研究利用 CRISPR/Cas9 对 Bsc 编码、bZip 编码和 Nt20 序列进行了剖析,以阐明基因型与表型之间的关系。在 Bsc 和 bZip 实验集中,所有观察到的编码序列突变都在框内,这表明框外突变是致死的。Nt20 实验集中转化子的存活率很低,约为 7%。内含子中的突变主要包括框架外插入和缺失。除了类似野生型的分生孢子形态外,突变体还表现出不同的菌落形态,包括硬核、混合(分生孢子和硬核)和菌丝形态。使用二硫苏糖醇进行的ER压力测试表明,硬核突变体和菌丝突变体比分生孢子突变体敏感得多。此外,菌丝突变体不能产生黄曲霉毒素,但仍能产生曲霉酸和曲霉酸。针对包括 Bsc 和 bZip 的区域进行的 RNAi 实验表明,转化体的存活率普遍下降,少数转化体出现表型变化。hacA 基因在 DNA 和转录本水平上的缺陷影响了黄曲霉的存活、生长和发育。因此,该基因有可能成为未来宿主诱导基因沉默策略的目标,以控制感染并减少农作物中的黄曲霉毒素污染。
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引用次数: 0
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Journal of Fungi
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