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New saga in Finland: The rise of Diplodia sapinea in Scots pine 芬兰的新传奇:苏格兰松中sapinea的崛起。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2025-02-01 DOI: 10.1016/j.fgb.2024.103955
Eeva Terhonen , Tiina Ylioja , Tuija Hytönen , Katri Leino , Linda Mutanen , Markus Melin , Eeva Vaahtera , Suvi Sutela
The intensity of fungal virulence is likely to increase in northern forests as climate change alters environmental conditions, favoring pathogen proliferation in existing ecosystems while also facilitating their expansion into new geographic areas. In Finland, Diplodia sapinea, the causal agent of disease called “Diplodia tip blight”, has emerged as a new pathogen within the past few years. To reveal the current distribution of the novel fungal pathogen, and the effect of temperature and rainfall on its distribution, we utilized citizen science for the detection and collection of symptomatic Scots pine (Pinus sylvestris) shoots. The Finnish culture collection of D. sapinea was initiated using in vitro cultured symptomatic samples, and selected strains were studied for their virulence and disease cycle. Furthermore, the mycobiome of selected symptomatic and asymptomatic Scots pine shoots was studied using amplicon sequencing and the presence of D. sapinea was confirmed with culturing, qPCR, and species-specific PCR. Based on over 500 Scots pine shoots testing positive for D. sapinea, the distribution of this fungal pathogen is concentrated along the coastal areas of Finland, extending up to 200 km inland from the coastline. The observed presence of D. sapinea followed the period of highest average temperatures recorded in Finland in 2023 and was also found to be related to less precipitation. The amplicon sequencing showed that abundance of D. sapinea was higher in the healthy tissues of symptomatic shoots compared to visually healthy shoots. Similarly, the abundance was higher in samples collected from coastal areas in Southwestern Finland, which are the most heavily impacted by this disease. Here, we show that the presence of D. sapinea is more extensive than previously assumed, and lastly illustrate the hypothesized disease cycle of the fungal pathogen in Finland based on observations made in the field from 2021 to 2024 and in vivo and in vitro studies.
随着气候变化改变环境条件,北方森林真菌毒力的强度可能会增加,有利于病原体在现有生态系统中的增殖,同时也促进它们向新的地理区域扩张。在芬兰,被称为“尖叶枯病”的疾病的致病因子——sapinea在过去几年中作为一种新的病原体出现。为了揭示该新型真菌病原体的分布现状,以及温度和降雨对其分布的影响,我们利用公民科学方法对有症状的苏格兰松(Pinus sylvestris)枝条进行了检测和采集。采用体外培养的有症状的样品,开始了芬兰的sapinea培养收集,并对所选菌株的毒力和疾病周期进行了研究。此外,我们还利用扩增子测序技术对选定的有症状和无症状的苏格兰松枝条进行了真菌组研究,并通过培养、qPCR和种特异性PCR证实了D. sapinea的存在。根据500多根检测呈sapinea阳性的苏格兰松芽,这种真菌病原体的分布集中在芬兰沿海地区,从海岸线向内陆延伸200 公里。在芬兰2023年平均气温最高的时期观测到的D. sapinea的存在也被发现与降水减少有关。扩增子测序结果表明,与视觉健康的幼苗相比,有症状的幼苗健康组织中sapinea的丰度更高。同样,在芬兰西南部沿海地区收集的样本中,丰度更高,这些地区受这种疾病的影响最严重。在这里,我们证明了D. sapinea的存在比以前假设的更广泛,最后说明了基于2021年至2024年在芬兰进行的实地观察以及体内和体外研究的真菌病原体的假设疾病周期。
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引用次数: 0
The sensor protein VdSLN1 is involved in regulating melanin biosynthesis and pathogenicity via MAPK pathway in Verticillium dahliae 传感器蛋白VdSLN1在大丽花黄萎病中通过MAPK通路参与调节黑色素生物合成和致病性。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2025-02-01 DOI: 10.1016/j.fgb.2025.103960
XiaYu Wang , JunJiao Li , XiaoBin Ji , Dan Wang , ZhiQiang Kong , XiaoFeng Dai , JieYin Chen , DanDan Zhang
The vascular wilt fungus Verticillium dahliae is a destructive soil-borne pathogen that causes yield loss on various economically important crops. Membrane-spanning sensor protein SLN1 have been demonstrated to contribute to virulence in varying degrees among numerous devastating fungal pathogens. However, the biological function of SLN1 in V. dahliae remains unclear. In this study, we identified the membrane-spanning sensor protein encoding gene VdSLN1 and it interacts physically with Vst50 and regulates the expression of MAPK module Vst50-Vst11-Vst7. The expression of VdSLN1 was also positively regulated by the MAPK signaling pathways transmembrane-associated members VdSho1 and VdMsb2, suggesting that the expression of VdSLN1 is associated with VdSho1 and VdMsb2. In addition, we found that VdSLN1, similar to VdSho1 and VdMsb2, is not required for V. dahliae vegetative growth and response to various abiotic stresses. While, ΔVdSLN1 mutant exhibited slightly reduced ability to penetrate a cellophane membrane and melanin synthesis compared with the wild type strain. Further experiments indicate that VdSLN1, VdSho1 and VdMsb2 has an additive effect on the virulence, cellophane penetration and melanin biosynthesis and of V. dahliae. In short, VdSLN1, though not essential, plays a role in cellophane penetration, melanin biosynthesis, also contributes to the virulence, as the downstream factor of VdSho1 and VdMsb2.
大丽花黄萎病(Verticillium dahliae)是一种破坏性的土传病原菌,对多种重要经济作物造成产量损失。跨膜传感器蛋白SLN1已被证明在不同程度上对许多破坏性真菌病原体的毒力有贡献。然而,SLN1在大丽花中的生物学功能尚不清楚。在本研究中,我们发现了跨膜传感器蛋白编码基因VdSLN1,它与Vst50物理相互作用,调控MAPK模块Vst50- vst11 - vst7的表达。VdSLN1的表达也受到MAPK信号通路跨膜相关成员VdSho1和VdMsb2的正调控,提示VdSLN1的表达与VdSho1和VdMsb2相关。此外,我们发现VdSLN1与VdSho1和VdMsb2类似,不是大丽花营养生长和应对各种非生物胁迫所必需的。与野生型菌株相比,ΔVdSLN1突变体穿透玻璃膜和合成黑色素的能力略有降低。进一步的实验表明,VdSLN1、VdSho1和VdMsb2对大丽花病原菌的毒力、透光性和黑色素合成具有加性作用。总之,VdSLN1虽然不是必需的,但作为VdSho1和VdMsb2的下游因子,在玻璃膜渗透、黑色素生物合成中发挥作用,也有助于毒力。
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引用次数: 0
Exploring endophytic fungi from Cynodon dactylon: GC–MS profiling and biological activity 短爪蟹内生真菌的GC-MS分析及生物活性研究。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2025-02-01 DOI: 10.1016/j.fgb.2024.103959
Aishwarya K. Kamunkar , R. Nischitha
The rapid decline of significant plant species due to deforestation and slow regrowth has endangered many trees that are crucial for producing life-saving medications. This dual crisis of conserving plant biodiversity while meeting pharmaceutical demands necessitates innovative solutions. Endophytic fungi, naturally occurring symbionts within plants, present an eco-friendly and economically viable alternative. These fungi can produce a wide range of bioactive compounds, offering a sustainable source of pharmaceuticals. This study investigated endophytic fungi isolated from the inflorescence, leaf, and culm of Cynodon dactylon, a perennial medicinal grass. The research involved the isolation of endophytic fungi on potato dextrose agar (PDA) and water agar (WA), extracting secondary metabolites, and performing antimicrobial and antioxidant assays and gas chromatography-mass spectroscopy (GC–MS) profiling. A total of 21 endophytic fungi were isolated, with species of Alternaria, Aspergillus, and Cladosporium being predominant. These fungi were identified through morphological and molecular (internal transcribed sequences-ITS) characterization. Based on factors such as fungal dominance and specificity, five fungi (Aspergillus chevalieri, Aspergillus stellatus, Hypoxylon sp., and Xylaria apiculate) were selected and they exhibited significant activity against plant pathogens (Sclerotium rolfsii and Aspergillus niger) and radical scavenging properties in DPPH assays. GC–MS analysis revealed over twenty bioactive compounds in each fungal extract. These findings underscore the potential of endophytic fungi as sustainable sources of novel pharmaceuticals and effective biocontrol agents, offering a promising approach to address the current ecological and medicinal challenges.
由于森林砍伐和再生缓慢,重要植物物种的迅速减少已经危及许多对生产救命药物至关重要的树木。在满足制药需求的同时保护植物生物多样性的双重危机需要创新的解决方案。内生真菌是植物内天然存在的共生体,是一种生态友好且经济可行的替代品。这些真菌可以产生广泛的生物活性化合物,提供可持续的药物来源。研究了从多年生药用禾本科植物Cynodon dactylon的花序、叶片和茎中分离得到的内生真菌。本研究包括在马铃薯葡萄糖琼脂(PDA)和水琼脂(WA)上分离内生真菌,提取次生代谢产物,进行抗菌和抗氧化检测以及气相色谱-质谱(GC-MS)分析。共分离到内生真菌21种,以Alternaria、Aspergillus和Cladosporium为优势种。这些真菌通过形态学和分子(内部转录序列- its)鉴定。基于真菌优势度和特异性等因素,选择了5种真菌(chevaliaspergillus, Aspergillus stellatus, Hypoxylon sp.和Xylaria apiculate),它们在DPPH实验中对植物病原菌(Sclerotium rolfsii和Aspergillus niger)表现出显著的活性和自由基清除能力。GC-MS分析显示,每种真菌提取物中含有20多种生物活性化合物。这些发现强调了内生真菌作为新型药物和有效生物防治剂的可持续来源的潜力,为解决当前的生态和医学挑战提供了有希望的方法。
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引用次数: 0
FungANI, a BLAST-based program for analyzing average nucleotide identity (ANI) between two fungal genomes, enables easy fungal species delimitation FungANI是一个基于blast的程序,用于分析两个真菌基因组之间的平均核苷酸同源性(ANI),使真菌物种划分变得容易。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2025-01-31 DOI: 10.1016/j.fgb.2025.103969
Christophe Lalanne , Philippe Silar
Fungal species delimitation and phylogeny will likely rely in the future upon whole genome sequence comparison, as the costs of such sequences are rapidly decreasing. Average Nucleotide Identity (ANI) between genomes is a convenient metric that can be rapidly calculated for species delimitation. However, there is presently no easy-to-use program calculating the ANI between two fungal genomes and providing easy-to interpret results that can be help mycologists having limited access to bioinformatic facilities. Here, we present FungANI, a customizable BLAST-based program that calculate ANI between genomes. The program primarily targets Linux workstations or servers but it can be run on the latest Windows, macOS and Linux 64-Bit operating systems as a standalone desktop application. It was tested with various publicly-available genomes from species belonging to the Sordariales order. It proved efficient to differentiate closely related species and retrace their possible phylogenetic relationships. However, FungANI did not perform well for phylogenetic reconstruction on a broader evolutionary scale such as inferring relationships between distant genera. The program is freely available at https://github.com/podo-gec/fungani.
真菌物种的划分和系统发育在未来很可能依赖于全基因组序列的比较,因为这种序列的成本正在迅速下降。基因组之间的平均核苷酸同一性(ANI)是一种方便的度量,可以快速计算物种划分。然而,目前还没有易于使用的程序来计算两个真菌基因组之间的ANI,并提供易于解释的结果,可以帮助有限的生物信息设施的真菌学家。在这里,我们提出了FungANI,一个可定制的基于blast的程序,计算基因组之间的ANI。该程序主要针对Linux工作站或服务器,但它可以作为一个独立的桌面应用程序在最新的Windows、macOS和64位Linux操作系统上运行。研究人员用各种公开的Sordariales目物种的基因组进行了测试。事实证明,它能有效地区分近亲物种,并追溯它们可能的系统发育关系。然而,FungANI在更广泛的进化尺度上的系统发育重建(如推断远缘属之间的关系)表现不佳。该程序可在https://github.com/podo-gec/fungani免费获得。
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引用次数: 0
Physical forces supporting hyphal growth 支持菌丝生长的物理力量。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2025-01-27 DOI: 10.1016/j.fgb.2025.103961
Nicholas P. Money
Hyphae are viscoelastic tubes whose internal pressure pushes the cell membrane against the inner surface of the cell wall. Catalytic yielding of the wall allows this turgor to force its polymers apart as new materials are added to the surface of the growing tip. Turgor drops slightly as the wall expands, creating a pressure gradient that causes the cytoplasm to flow toward the tip. These physiological processes affect the rate of extension of the hypha and determine the magnitude of the force that it uses for invasive growth. This paper provides an overview of the experimental basis for this description of hyphal mechanics and explains the wider significance of biophysical studies on fungi and water molds.
菌丝是粘弹性管,其内部压力将细胞膜推向细胞壁的内表面。当新材料被添加到生长尖端的表面时,壁的催化生成允许这种膨胀迫使其聚合物分离。当细胞壁膨胀时,膨压会轻微下降,产生压力梯度,导致细胞质向尖端流动。这些生理过程影响菌丝的延伸速度,并决定其用于侵入性生长的力的大小。本文概述了菌丝力学描述的实验基础,并解释了真菌和水霉菌生物物理研究的更广泛意义。
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引用次数: 0
Efficient genome editing using CRISPR/Cas9 technology and its application for identifying Sesquiterpene synthases involved in the biosynthesis of Steperoxides in Steccherinum ochraceum 利用 CRISPR/Cas9 技术进行高效基因组编辑并将其应用于鉴定参与 Steccherinum ochraceum 中 Steperoxides 生物合成的倍半萜合成酶。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2024-12-01 DOI: 10.1016/j.fgb.2024.103944
Jia-Yu Shen , Fei-Hong Mao , Qiwen Wang , Pei-Pei Ou , Ji-Kai Liu , Qunfei Zhao , Qing-Li He
CRISPR technology has been widely used for gene editing in various species,but the genetic manipulation in basidiomycete mushrooms is still notoriously difficult for unknown endogenous promoters and inefficient DNA delivery. Steccherinum ochraceum is a white rot basidiomycete fungus with abundant secondary metabolites and plays an important ecological role worldwide. To facilitate the study of gene function in S. ochraceum, an effective CRISPR/Cas9 system was successfully developed by identifying highly efficient endogenous promoters, and utilizing the Agrobacterium-transformation method. Two efficient endogenous RNA polymerase II promoters (Psogpd and Psotef1) and one efficient RNA polymerase III promoter (Pu6-d) were identified and characterized, with an editing efficiency of 61.5 % at the ura3 locus. Using this optimized system, the sesquiterpene gene A0064, which could produce 10 possible sesquiterpenes in the heterologous expression system of A. oryzae, was knocked out to obtain A0064 knockout strain S. ochraceum (∆A0064). Steperoxide A could not be detected in S. ochraceum (∆A0064), demonstrating that A0064 was the only enzyme responsible for the biosynthesis of β-chamigrene (the sesquiterpene skeleton of steperoxide A) in S. ochraceum. This efficient system will enable precise targeting and multiplex editing of S. ochraceum genes, facilitating functional studies of genes involved in lignin degradation and natural product biosynthesis in S. ochraceum, and providing some valuable guidance for gene editing in tens of thousands of macrofungi.
CRISPR技术已被广泛应用于各种物种的基因编辑,但由于内源启动子未知和DNA传递效率低下,基枝菌的基因操作仍是众所周知的难题。Steccherinum ochraceum 是一种白腐基生真菌,具有丰富的次生代谢产物,在世界范围内发挥着重要的生态作用。为了便于研究赭色链格孢的基因功能,研究人员通过识别高效的内源启动子,并利用农杆菌转化法,成功开发了一套有效的 CRISPR/Cas9 系统。该系统识别并鉴定了两个高效的内源 RNA 聚合酶 II 启动子(Psogpd 和 Psotef1)和一个高效的 RNA 聚合酶 III 启动子(Pu6-d),在 ura3 基因座上的编辑效率为 61.5%。利用这一优化系统,敲除了在 A. oryzae 异源表达系统中可产生 10 种倍半萜的倍半萜基因 A0064,得到了 A0064 基因敲除株 S. ochraceum(ΔA0064)。在 S. ochraceum(ΔA0064)中检测不到菊酯氧化物 A,这表明 A0064 是 S. ochraceum 中负责β-chamigrene(菊酯氧化物 A 的倍半萜骨架)生物合成的唯一酶。这一高效系统将实现对赭色真菌基因的精确靶向和多重编辑,有助于对赭色真菌中参与木质素降解和天然产物生物合成的基因进行功能研究,并为数以万计的大型真菌的基因编辑提供一些有价值的指导。
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引用次数: 0
Physiological function of hydrophobin Hydph16 in cell wall formation in agaricomycete Pleurotus ostreatus 疏水蛋白hyph16在平菇细胞壁形成中的生理功能。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2024-11-28 DOI: 10.1016/j.fgb.2024.103943
Junxian Han , Moriyuki Kawauchi , Yuki Terauchi , Kenya Tsuji , Akira Yoshimi , Chihiro Tanaka , Takehito Nakazawa , Yoichi Honda
Hydrophobins are small-secreted proteins with both hydrophobic and hydrophilic regions, enabling the mycelium to break through the air-medium interface by reducing the medium surface tension. Over 20 putative hydrophobin-encoding genes have been predicted in the agaricomycete Pleurotus ostreatus. Three hydrophobin-encoding genes, vmh2, vmh3, and hydph16, were predominantly expressed in the vegetative mycelium. Despite these common properties, we have previously demonstrated the distinct functions of Vmh2 and Vmh3 in environmental stress resistance. In this study, we focused on hydph16 and found that Δhydph16 strains had sparser aerial mycelium than control strains. The cell wall thickness of Δhydph16 strains reduced by 40 % compared to that of control strains, but no significant differences were found in the relative chitin and glucan percentages or relative putative cell wall synthesis-related gene expression levels. Furthermore, unlike vmh2 and vmh3, hydph16 deletion did not change the hydrophobicity of the aerial mycelium. This study is the first to report that the lack of hydrophobin can lead to a significant change in aerial hyphae cell wall formation without altering the major cell wall polysaccharide composition. Additionally, this study revealed multiple roles for Hydph16, distinct from those of other highly expressed hydrophobins, Vmh2 and Vmh3. These results suggested that agaricomycetes, including P. ostreatus, have evolved to possess multiple hydrophobins with different functions.
疏水蛋白是一种小分泌蛋白,同时具有疏水和亲水区域,使菌丝通过降低介质表面张力突破空气-介质界面。在平菇真菌中已经发现了20多个推测的疏水性编码基因。3个疏水蛋白编码基因vmh2、vmh3和hyph16主要在营养菌丝中表达。尽管有这些共同的特性,我们之前已经证明了Vmh2和Vmh3在环境胁迫抗性中的不同功能。在本研究中,我们将重点放在hydph16上,发现Δhydph16菌株的气相菌丝比对照菌株少。与对照菌株相比,Δhydph16菌株的细胞壁厚度减少了40 %,但几丁质和葡聚糖的相对百分比和细胞壁合成相关基因的相对推定表达量无显著差异。此外,与vmh2和vmh3不同,hyph16的缺失没有改变气生菌丝的疏水性。本研究首次报道了疏水蛋白的缺乏可导致气生菌丝细胞壁形成的显著变化,而不改变主要细胞壁多糖的组成。此外,本研究揭示了Hydph16的多种作用,不同于其他高表达的疏水蛋白Vmh2和Vmh3。这些结果表明,包括P. ostreatus在内的木链菌已经进化出具有多种不同功能的疏水酶。
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引用次数: 0
The impact of glutamine synthetase PbgsA on the growth, conidiation and mycophenolic acid production of Penicillium brevicompactum 谷氨酰胺合成酶 PbgsA 对破伤风青霉的生长、分生孢子和产生霉酚酸的影响。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2024-11-16 DOI: 10.1016/j.fgb.2024.103941
TingTing Hu , Jishou Wu , Zixuan Lin , Yi Lin , Lin Lin , Wei Wei , Dongzhi Wei
Glutamine synthetase (GS) is a critical enzyme in nitrogen metabolism regulation and plays an essential role in the metabolic pathways involved in microbial growth and development. Penicillium brevicompactum, known for its rich repertoire of secondary metabolites, including mycophenolic acid (MPA), lacks research on the regulatory mechanisms of GS within this species. This study aimed to investigate the influence of GS on the growth, sporulation, and secondary metabolism of P. brevicompactum to elucidate the biological function of GS in this organism. We identified the glutamine synthetase gene (PbgsA) from P. brevicompactum and constructed PbgsA gene-overexpression and gene-silencing transformants. The impact of PbgsA on growth and sporulation was evaluated, revealing that PbgsA gene-overexpression transformants exhibited enhanced growth and significantly increased the expression levels of sporulation pathway genes (brlA, abaA, and wetA). Additionally, PbgsA gene-overexpression transformants produced higher MPA yields, with a maximum of 4.78 g/L, representing a 54.19 % increase compared to the wild type (WT). Conversely, PbgsA gene-silencing transformants showed reduced MPA production, with a minimum yield of 1.13 g/L, a 63.55 % decrease relative to the WT. Transcriptional analysis of the MPA biosynthetic gene cluster indicated that PbgsA exerted regulatory effects on certain biosynthetic pathway genes, such as mpaA and mpaB. This study demostrated the potential positive regulatory role of glutamine synthetase PbgsA in the growth, spore development, and secondary metabolism of P. brevicompactum, which provided a new strategy for genetic regulation in filamentous fungal.
谷氨酰胺合成酶(GS)是氮代谢调节中的一种关键酶,在微生物生长发育的代谢途径中发挥着重要作用。布氏青霉(Penicillium brevicompactum)以其丰富的次生代谢产物(包括霉酚酸(MPA))而闻名,但对该菌种内 GS 的调控机制缺乏研究。本研究旨在调查 GS 对 P. brevicompactum 的生长、孢子和次生代谢的影响,以阐明 GS 在该生物体内的生物学功能。我们确定了 P. brevicompactum 的谷氨酰胺合成酶基因(PbgsA),并构建了 PbgsA 基因外表达和基因沉默转化子。结果表明,PbgsA 基因缺失表达转化子的生长能力增强,孢子途径基因(brlA、abaA 和 wetA)的表达水平显著提高。此外,PbgsA 基因缺失转化体产生了更高的 MPA 产量,最高达 4.78 克/升,与野生型(WT)相比增加了 54.19%。相反,PbgsA 基因沉默转化体的 MPA 产量降低,最低产量为 1.13 克/升,比 WT 降低了 63.55%。对 MPA 生物合成基因簇的转录分析表明,PbgsA 对某些生物合成途径基因(如 mpaA 和 mpaB)具有调控作用。该研究证明了谷氨酰胺合成酶PbgsA在P. brevicompactum的生长、孢子发育和次生代谢中的潜在正向调控作用,为丝状真菌的遗传调控提供了一种新策略。
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引用次数: 0
Screens for mutants defective in UapA trafficking highlight the importance of ER-exit as a primary control point in transporter biogenesis 对 UapA 转运缺陷突变体的筛选突显了 ER 出口作为转运体生物发生过程中主要控制点的重要性。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2024-11-07 DOI: 10.1016/j.fgb.2024.103940
Effie Demos , Sofia Dimou , Claudio Scazzocchio , George Diallinas
Most transmembrane membrane proteins are thought to traffic to the plasma membrane (PM) via the conventional secretory pathway through sorting from the Golgi. However, our recent work has shown that in the filamentous fungus Aspergillus nidulans several nutrient transporters and other major membrane proteins traffic to the PM via Golgi-bypass and independently of known post-Golgi secretory mechanisms. Here in an effort to dissect the molecular mechanism underlying membrane cargo trafficking via Golgi-bypass we design and use unbiased genetic screens, based on the UapA uric acid-xanthine transporter, which allowed the isolation of mutants defective in UapA translocation to the plasma membrane. Analyses of these mutants highlight the importance of ER-exit as the primary control point in transporter trafficking via Golgi-bypass. Most mutants isolated concerned mutations within the uapA gene, albeit we also obtained uapA extragenetic mutants affecting secretion and growth pleiotropically or leading on apparent activation of an efflux transporter related to purine-detoxification. Our work paves the way to use genetic approaches targeting specifically trafficking mutations affecting Golgi-bypass.
大多数跨膜蛋白被认为是通过传统的分泌途径,从高尔基体分拣进入质膜(PM)的。然而,我们最近的研究表明,在丝状真菌黑曲霉(Aspergillus nidulans)中,几种营养物质转运体和其他主要膜蛋白通过高尔基体旁路(Golgi-bypass)转运到质膜(PM),与已知的高尔基体后分泌机制无关。在这里,为了剖析通过高尔基体旁路进行膜货物运输的分子机制,我们以 UapA 尿酸-黄嘌呤转运体为基础,设计并使用了无偏见的遗传筛选,从而分离出了 UapA 向质膜转运缺陷的突变体。对这些突变体的分析凸显了ER-出口作为通过高尔基体旁路进行转运的主要控制点的重要性。大多数分离出的突变体涉及 uapA 基因内部的突变,不过我们也获得了 uapA 基因外突变体,这些突变体会影响分泌和多向生长,或导致与嘌呤解毒有关的外排转运体的明显激活。我们的工作为使用遗传方法专门针对影响高尔基体旁路的转运突变铺平了道路。
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引用次数: 0
Candida albicans strains adapted to the mouse gut are resistant to bile salts via a Flo8-dependent mechanism 适应小鼠肠道的白色念珠菌菌株通过 Flo8 依赖性机制对胆汁盐产生抗性。
IF 2.4 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-31 DOI: 10.1016/j.fgb.2024.103939
Susana Hidalgo-Vico , Daniel Prieto , Rebeca Alonso-Monge , Elvira Román , Corinne Maufrais , Christophe d’Enfert , Jesús Pla
Candida albicans normally colonizes the human gastrointestinal tract as a commensal. Studying fungal factors involved in colonizing the mammalian gastrointestinal tract requires mouse models with altered microbiota. We have obtained strains of C. albicans through microevolution in the mouse gut for a prolonged period (one year) that display a substantial increase in fitness in this niche. These strains show resistance to bile salts, an increase in their adhesion to the intestinal mucosa, and are unable to filament in response to serum. Genetic analysis revealed some alterations, mainly a triploidy of chr7, a whole chr6 homozygosis, and an SNP in the FLO8 gene (located in the chr6), resulting in a truncated protein version. A wild type FLO8 gene complemented filamentation and bile salt sensitivity but showed an intermediate fitness phenotype in colonization. Alterations in bile salt sensitivity were also evident in bmt mutants, defective in β-mannosylation, and transcriptional targets of Flo8, suggesting a link between the fungal cell wall and mammalian gut colonization via the Flo8 transcriptional regulator.
白色念珠菌通常作为共生菌定植于人类胃肠道。要研究真菌在哺乳动物胃肠道定植的相关因素,需要使用微生物群发生变化的小鼠模型。我们通过在小鼠肠道中长期(一年)的微进化获得了白僵菌菌株,这些菌株在这一生态位中的适应性大大提高。这些菌株表现出对胆汁盐的抵抗力,对肠粘膜的粘附力增强,并且无法对血清产生丝状反应。遗传分析发现了一些改变,主要是 chr7 的三倍体、整个 chr6 的同源染色体以及 FLO8 基因(位于 chr6 上)的 SNP,从而导致蛋白截短。野生型 FLO8 基因可补充丝状化和胆盐敏感性,但在定殖过程中表现出中等的适应性表型。胆盐敏感性的改变在 bmt 突变体中也很明显,该突变体在 β-甘露糖基化和 Flo8 的转录靶标方面存在缺陷,这表明真菌细胞壁与哺乳动物肠道定殖之间通过 Flo8 转录调节因子存在联系。
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Fungal Genetics and Biology
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