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Corrigendum: The Sordariomycetes: an expanding resource with Big Data for mining in evolutionary genomics and transcriptomics. 更正:鞭毛霉菌纲:用于挖掘进化基因组学和转录组学大数据的不断扩大的资源。
IF 2.1 Q3 MYCOLOGY Pub Date : 2024-08-20 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1478516
Zheng Wang, Wonyong Kim, Yen-Wen Wang, Elizabeta Yakubovich, Caihong Dong, Frances Trail, Jeffrey P Townsend, Oded Yarden

[This corrects the article DOI: 10.3389/ffunb.2023.1214537.].

[此处更正了文章 DOI:10.3389/ffunb.2023.1214537]。
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引用次数: 0
Dual-RNA-sequencing to elucidate the interactions between sorghum and Colletotrichum sublineola. 通过双 RNA 测序阐明高粱与 Colletotrichum sublineola 之间的相互作用。
IF 2.1 Q3 MYCOLOGY Pub Date : 2024-08-16 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1437344
Saddie Vela, Emily S A Wolf, Jeffrey A Rollins, Hugo E Cuevas, Wilfred Vermerris

In warm and humid regions, the productivity of sorghum is significantly limited by the fungal hemibiotrophic pathogen Colletotrichum sublineola, the causal agent of anthracnose, a problematic disease of sorghum (Sorghum bicolor (L.) Moench) that can result in grain and biomass yield losses of up to 50%. Despite available genomic resources of both the host and fungal pathogen, the molecular basis of sorghum-C. sublineola interactions are poorly understood. By employing a dual-RNA sequencing approach, the molecular crosstalk between sorghum and C. sublineola can be elucidated. In this study, we examined the transcriptomes of four resistant sorghum accessions from the sorghum association panel (SAP) at varying time points post-infection with C. sublineola. Approximately 0.3% and 93% of the reads mapped to the genomes of C. sublineola and Sorghum bicolor, respectively. Expression profiling of in vitro versus in planta C. sublineola at 1-, 3-, and 5-days post-infection (dpi) indicated that genes encoding secreted candidate effectors, carbohydrate-active enzymes (CAZymes), and membrane transporters increased in expression during the transition from the biotrophic to the necrotrophic phase (3 dpi). The hallmark of the pathogen-associated molecular pattern (PAMP)-triggered immunity in sorghum includes the production of reactive oxygen species (ROS) and phytoalexins. The majority of effector candidates secreted by C. sublineola were predicted to be localized in the host apoplast, where they could interfere with the PAMP-triggered immunity response, specifically in the host ROS signaling pathway. The genes encoding critical molecular factors influencing pathogenicity identified in this study are a useful resource for subsequent genetic experiments aimed at validating their contributions to pathogen virulence. This comprehensive study not only provides a better understanding of the biology of C. sublineola but also supports the long-term goal of developing resistant sorghum cultivars.

炭疽病是高粱(Sorghum bicolor (L.) Moench)的一种严重病害,可导致粮食和生物量减产高达 50%。尽管宿主和真菌病原体都有基因组资源,但人们对高粱-C. sublineola 相互作用的分子基础知之甚少。通过采用双 RNA 测序方法,可以阐明高粱与亚线虫之间的分子串扰。在本研究中,我们研究了高粱协会(SAP)中四个抗性高粱品种在感染亚线虫后不同时间点的转录组。大约 0.3% 和 93% 的读数分别映射到亚线虫和双色高粱的基因组。感染后 1 天、3 天和 5 天(dpi)亚线虫体外与植物体内的表达谱分析表明,在从生物营养阶段向坏死阶段过渡期间(3 dpi),编码分泌型候选效应物、碳水化合物活性酶(CAZymes)和膜转运体的基因表达量增加。病原体相关分子模式(PAMP)触发的高粱免疫特征包括活性氧(ROS)和植物毒素的产生。据预测,亚线虫分泌的候选效应物大多定位于宿主的细胞外质,它们可能干扰 PAMP 触发的免疫反应,特别是宿主的 ROS 信号通路。本研究中发现的编码影响致病性的关键分子因子的基因为后续遗传实验提供了有用的资源,这些实验旨在验证这些基因对病原体毒力的贡献。这项全面的研究不仅有助于更好地了解亚线虫的生物学特性,还有助于实现开发抗性高粱栽培品种的长期目标。
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引用次数: 0
Phosphate availability conditions caspofungin tolerance, capsule attachment and titan cell formation in Cryptococcus neoformans. 磷酸盐的可用性决定了新生隐球菌对卡泊芬净的耐受性、菌盖附着和泰坦细胞的形成。
IF 2.1 Q3 MYCOLOGY Pub Date : 2024-08-14 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1447588
Xianya Qu, Kabir Bhalla, Linda C Horianopoulos, Guanggan Hu, Armando Alcázar Magaña, Leonard J Foster, Leandro Buffoni Roque da Silva, Matthias Kretschmer, James W Kronstad

There is an urgent need for new antifungal drugs to treat invasive fungal diseases. Unfortunately, the echinocandin drugs that are fungicidal against other important fungal pathogens are ineffective against Cryptococcus neoformans, the causative agent of life-threatening meningoencephalitis in immunocompromised people. Contributing mechanisms for echinocandin tolerance are emerging with connections to calcineurin signaling, the cell wall, and membrane composition. In this context, we discovered that a defect in phosphate uptake impairs the tolerance of C. neoformans to the echinocandin caspofungin. Our previous analysis of mutants lacking three high affinity phosphate transporters revealed reduced elaboration of the polysaccharide capsule and attenuated virulence in mice. We investigated the underlying mechanisms and found that loss of the transporters and altered phosphate availability influences the cell wall and membrane composition. These changes contribute to the shedding of capsule polysaccharide thus explaining the reduced size of capsules on mutants lacking the phosphate transporters. We also found an influence of the calcineurin pathway including calcium sensitivity and an involvement of the endoplasmic reticulum in the response to phosphate limitation. Furthermore, we identified membrane and lipid composition changes consistent with the role of phosphate in phospholipid biosynthesis and with previous studies implicating membrane integrity in caspofungin tolerance. Finally, we discovered a contribution of phosphate to titan cell formation, a cell type that displays modified cell wall and capsule composition. Overall, our analysis reinforces the importance of phosphate as a regulator of cell wall and membrane composition with implications for capsule attachment and antifungal drug susceptibility.

治疗侵袭性真菌疾病急需新的抗真菌药物。遗憾的是,对其他重要真菌病原体具有杀真菌作用的棘白菌素类药物对新隐球菌无效,而新隐球菌是导致免疫力低下人群患上脑膜脑炎并危及生命的病原体。导致棘球蚴耐受性的机制正在出现,这些机制与钙调素信号、细胞壁和膜组成有关。在这种情况下,我们发现磷酸盐摄取的缺陷会影响新变形杆菌对棘白菌素类药物卡泊芬净的耐受性。我们之前对缺乏三种高亲和力磷酸盐转运体的突变体进行了分析,结果发现小鼠体内多糖胶囊的合成减少,毒力减弱。我们对其潜在机制进行了研究,发现转运体的缺失和磷酸盐供应的改变会影响细胞壁和膜的组成。这些变化有助于胶囊多糖的脱落,从而解释了缺乏磷酸盐转运体的突变体胶囊大小减小的原因。我们还发现了钙调磷酸酶通路的影响,包括钙敏感性和内质网对磷酸盐限制反应的参与。此外,我们还发现了膜和脂质成分的变化,这与磷酸盐在磷脂生物合成中的作用以及之前关于膜完整性与卡泊芬净耐受性有关的研究相一致。最后,我们发现了磷酸盐对钛细胞形成的作用,这种细胞类型的细胞壁和囊的组成发生了改变。总之,我们的分析加强了磷酸盐作为细胞壁和膜组成调节剂的重要性,并对胶囊附着和抗真菌药物敏感性产生了影响。
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引用次数: 0
Antagonistic interactions between maize seeds microbiome species and the late wilt disease agent, Magnaporthiopsis maydis. 玉米种子微生物组物种与晚疫病病原体 Magnaporthiopsis maydis 之间的拮抗相互作用。
IF 2.1 Q3 MYCOLOGY Pub Date : 2024-08-07 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1436759
Ofir Degani, Aseel Ayoub, Elhanan Dimant, Asaf Gordani

Magnaporthiopsis maydis is a maize pathogen that causes severe damage to commercial corn fields in the late growth stages. Late wilt disease (LWD) has spread since its discovery in the 1960s in Egypt and is now reported in about 10 countries. The pathogen has a hidden endophytic lifecycle in resistant corn plants and secondary hosts such as green foxtail, watermelon lupin and cotton. At the same time, it could be an opportunist and hinder the host development under the right conditions. This study uncovered M. maydis interactions with newly identified maize endophytes. To this end, six fungi were isolated from the seeds of three sweet corn cultivars having varying susceptibility to LWD. These isolates were identified using colony morphology and microscopic characterization, universal internal transcribed spacer (ITS) molecular targeting and phylogenetic analysis. Most of them belonged to pathogenic species. Compared to three previously identified bioprotective microorganisms, the new species were tested for their ability to secrete metabolites that repress M. maydis in vitro and to antagonize it in a solid media confront test and a seedlings pathogenicity assay. The opportunistic fungal species Aspergillus flavus (ME1), Aspergillus terreus (PE3) and the reference biocontrol bacteria Bacillus subtilis (R2) achieved the highest M. maydis inhibition degree in the plates tests (74-100% inhibition). The seedlings' pathogenicity assay that predicts the seeds' microflora resistance to M. maydis highlighted the bio-shielding potential of most species (23% or more epicotyl elongation over the infected control). Fusarium sp. (ME2) was the leading species in this measure (43% enhancement), and B. subtilis gave the best protection in terms of seeds' germination (50%) and sprouts' biomass (34%). The results of this study could enhance our understanding of the pathobiome's role in the context of LWD and represent a first step in using the seeds' natural protective microflora to develop novel management strategies.

Magnaporthiopsis maydis 是一种玉米病原体,会在玉米生长后期对商品玉米田造成严重破坏。自 20 世纪 60 年代在埃及发现以来,晚疫病(LWD)已经蔓延开来,现在大约有 10 个国家都有报道。病原体在抗性玉米植株和次要寄主(如绿狐尾、西瓜羽扇豆和棉花)中具有隐蔽的内生生命周期。同时,它也可能是机会主义者,在适当的条件下阻碍寄主的发展。本研究揭示了 M. maydis 与新发现的玉米内生菌之间的相互作用。为此,我们从对 LWD 敏感性不同的三个甜玉米栽培品种的种子中分离出了六种真菌。利用菌落形态和显微特征、通用内部转录间隔(ITS)分子定位和系统发育分析对这些分离物进行了鉴定。其中大部分属于致病物种。与之前发现的三种生物保护性微生物相比,新菌种在体外分泌代谢物抑制 M. maydis 的能力以及在固体培养基对抗试验和幼苗致病性试验中拮抗 M. maydis 的能力得到了检验。在平板试验中,机会真菌黄曲霉(ME1)、赤曲霉(PE3)和参考生防菌枯草芽孢杆菌(R2)对麦地那龙线虫的抑制率最高(74%-100%)。秧苗致病性检测可预测种子微生物菌群对麦地那龙线虫的抗性,该检测强调了大多数物种的生物屏蔽潜力(外胚轴伸长率比受感染的对照高 23% 或更多)。镰刀菌(ME2)在这一指标中居首位(提高 43%),而枯草芽孢杆菌则在种子发芽率(50%)和萌芽生物量(34%)方面提供了最佳保护。这项研究的结果可以加深我们对病原生物群在LWD中的作用的理解,并为利用种子的天然保护性微生物群来制定新型管理策略迈出了第一步。
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引用次数: 0
Opportunities for optimizing fungal biological control agents for long-term and effective management of insect pests of orchards and vineyards: a review. 优化真菌生物防治制剂以长期有效治理果园和葡萄园害虫的机会:综述。
IF 2.1 Q3 MYCOLOGY Pub Date : 2024-08-01 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1443343
Christopher M Wallis, Mark S Sisterson

Novel tactics for controlling insect pests in perennial fruit and nut crops are needed because target pests often display decreased susceptibility to chemical controls due to overreliance on a handful of active ingredients and regulatory issues. As an alternative to chemical controls, entomopathogenic fungi could be utilized as biological control agents to manage insect pest populations. However, development of field ready products is hampered by a lack of basic knowledge. Development of field ready products requires collecting, screening, and characterizing a greater variety of potential entomopathogenic fungal species and strains. Creation of a standardized research framework to study entomopathogenic fungi will aid in identifying the potential mechanisms of biological control activity that fungi could possess, including antibiotic metabolite production; strains and species best suited to survive in different climates and agroecosystems; and optimized combinations of entomopathogenic fungi and novel formulations. This mini review therefore discusses strategies to collect and characterize new entomopathogenic strains, test different potential mechanisms of biocontrol activity, examine ability of different species and strains to tolerate different climates, and lastly how to utilize this information to develop strains into products for growers.

由于过度依赖少数活性成分和监管问题,目标害虫往往对化学防治的敏感性降低,因此需要采用新的策略来控制多年生水果和坚果作物中的害虫。作为化学防治的替代品,昆虫病原真菌可用作生物防治剂来控制害虫数量。然而,由于缺乏基础知识,田间现成产品的开发受到阻碍。开发可在田间使用的产品需要收集、筛选和鉴定更多潜在的昆虫病原真菌种类和菌株。建立研究昆虫病原真菌的标准化研究框架将有助于确定真菌可能具有的生物防治活性机制,包括抗生素代谢物的产生;最适合在不同气候和农业生态系统中生存的菌株和物种;以及昆虫病原真菌和新型制剂的优化组合。因此,这篇小型综述讨论了收集和鉴定新的昆虫病原菌菌株、测试生物防治活动的不同潜在机制、研究不同物种和菌株耐受不同气候的能力,以及如何利用这些信息将菌株开发成产品供种植者使用的策略。
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引用次数: 0
Editorial: Insights into fungal biology with emphasis on pathogenesis in humans. 社论:对真菌生物学的见解,重点是人类的致病机理。
IF 2.1 Q3 MYCOLOGY Pub Date : 2024-07-02 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1438060
Angel Gonzalez, Marilene Henning Vainstein, Patrícia Albuquerque, Ildinete Silva-Pereira
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引用次数: 0
Submerged cultivation and phytochemical analysis of medicinal mushrooms (Trametes sp.) 药用蘑菇(Trametes sp.)
Pub Date : 2024-06-11 DOI: 10.3389/ffunb.2024.1414349
Malay Srivastava, Moni Kumari, S. K. Karn, A. Bhambri, Vivek G. Mahale, Sushma Mahale
Mushrooms are widely available around the world and have various nutritional as well as therapeutic values. Many Asian cultures believe that medicinal mushrooms can prolong life and improve vitality. This study aims to characterize the phytochemical and polysaccharide content, mainly β-glucan content, of mycelial biomass and fruiting bodies collected from the Himalayan region, particularly Uttarakhand. Through molecular analysis of the LSU F/R-rDNA fragment sequence and phylogenetic analysis, the strain was identified as Trametes sp. We performed screening of phytochemicals and polysaccharides in mushroom and biomass extracts using high-performance liquid chromatography (HPLC) and a PC-based UV-Vis spectrophotometer. The macrofungal biomass was found to be high in saponin, anthraquinone, total phenolic, flavonoid, and β-glucan content. In biomass extract, we observed a high level of saponin (70.6µg/mL), anthraquinone (14.5µg/mL), total phenolic (12.45 µg/mL), and flavonoid (9.500 µg/mL) content. Furthermore, we examined the contents of alkaloids, tannins, terpenoids, and sterols in the biomass and mushroom extracts; the concentration of these compounds in the ethanol extract tested was minimal. We also looked for antioxidant activity, which is determined in terms of the IC50 value. Trametes sp. mushroom extract exhibits higher DPPH radical scavenging activity (62.9% at 0.5 mg/mL) than biomass extract (59.19% at 0.5 mg/mL). We also analyzed β-glucan in Trametes sp. from both mushroom and biomass extracts. The biomass extract showed a higher β-glucan content of 1.713 mg/mL than the mushroom extract, which is 1.671 mg/mL. Furthermore, β-glucan analysis was confirmed by the Megazyme β-glucan assay kit from both biomass and mushroom extract of Trametes sp. β-glucans have a promising future in cancer treatment as adjuncts to conventional medicines. Producing pure β-glucans for the market is challenging because 90–95% of β glucan sold nowadays is thought to be manipulated or counterfeit. The present study supports the recommendation of Trametes sp. as rich in β-glucan, protein, phytochemicals, and antioxidant activities that help individuals with cancer, diabetes, obesity, etc.
蘑菇在世界各地广泛存在,具有各种营养和治疗价值。许多亚洲文化都认为药用蘑菇可以延年益寿。本研究旨在分析喜马拉雅地区(尤其是北阿坎德邦)采集的蘑菇菌丝体和子实体的植物化学成分和多糖含量(主要是β-葡聚糖含量)。通过对 LSU F/R-rDNA 片段序列的分子分析和系统进化分析,确定该菌株为金针菇属。 我们使用高效液相色谱法(HPLC)和基于 PC 的紫外可见分光光度计对蘑菇和生物质提取物中的植物化学物质和多糖进行了筛选。研究发现,大型真菌生物质中的皂苷、蒽醌、总酚、类黄酮和β-葡聚糖含量较高。在生物质提取物中,我们观察到皂苷(70.6 微克/毫升)、蒽醌(14.5 微克/毫升)、总酚(12.45 微克/毫升)和黄酮(9.500 微克/毫升)含量较高。此外,我们还检测了生物质和蘑菇提取物中生物碱、单宁酸、萜类化合物和甾醇的含量;这些化合物在乙醇提取物中的浓度极低。我们还研究了抗氧化活性,这是以 IC50 值来确定的。金针菇提取物的 DPPH 自由基清除活性(0.5 毫克/毫升时为 62.9%)高于生物质提取物(0.5 毫克/毫升时为 59.19%)。我们还分析了从蘑菇和生物质提取物中提取的曲霉中的β-葡聚糖。生物质提取物中的β-葡聚糖含量为 1.713 毫克/毫升,高于蘑菇提取物中的 1.671 毫克/毫升。此外,用 Megazyme β-葡聚糖检测试剂盒对金针菇生物质和蘑菇提取物中的β-葡聚糖进行了分析,结果表明,β-葡聚糖作为传统药物的辅助药物,在癌症治疗中具有广阔的前景。为市场生产纯净的 β-葡聚糖具有挑战性,因为目前销售的 β-葡聚糖有 90-95% 被认为是经过处理或假冒的。本研究支持推荐使用富含β-葡聚糖、蛋白质、植物化学物质和抗氧化活性的 Trametes sp.
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引用次数: 0
Molecular cloning, expression, and purification, along with in silico epitope analysis of recombinant enolase proteins (a potential vaccine candidate) from Candida albicans and Candida auris. 对来自白色念珠菌和白色念珠菌的重组烯醇化酶蛋白(潜在候选疫苗)进行分子克隆、表达、纯化和表位分析。
Pub Date : 2024-05-31 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1399546
Manisha Shukla, Rohit Singh, Pankaj Chandley, Soma Rohatgi

Candida albicans is the predominant cause of systemic candidiasis, although other non albicans Candida species are progressively becoming more widespread nowadays. Candida auris has emerged as a deadly multidrug-resistant fungal pathogen, posing a significant threat to global public health. In the absence of effective antifungal therapies, the development of a vaccine against C. auris infections is imperative. Enolase, a key glycolytic enzyme, has emerged as a promising vaccine candidate due to its immunogenic properties and essential role in fungal virulence. Herein, full-length Enolase gene sequences from C. albicans and C. auris were cloned into suitable expression vector and transformed into Escherichia coli expression hosts. Recombinant Enolase proteins were successfully expressed and purified using affinity chromatography under native conditions, followed by SDS-PAGE characterization and Western blot analysis. CD spectroscopy verified the existence of expressed proteins in soluble native conformation. Preliminary in silico studies verified the immunogenicity of recombinant Enolase proteins isolated from both C. albicans and C. auris. Furthermore, bioinformatics analysis revealed conserved B-cell and T-cell epitopes across C. albicans and C. auris Enolase proteins, suggesting potential cross-reactivity and broad-spectrum vaccine efficacy. Our findings are anticipated to play a role in advancing therapeutic as well as diagnostic strategies against systemic candidiasis.

白念珠菌是全身性念珠菌病的主要致病菌,但如今其他非白念珠菌属的念珠菌也逐渐广泛传播。白色念珠菌已成为一种致命的耐多药真菌病原体,对全球公共卫生构成重大威胁。在缺乏有效抗真菌疗法的情况下,开发针对念珠菌感染的疫苗势在必行。烯醇化酶是一种关键的糖酵解酶,由于其免疫原性和在真菌毒力中的重要作用,它已成为一种有希望的候选疫苗。在此,我们将白僵菌和法氏囊菌的全长烯醇化酶基因序列克隆到合适的表达载体中,并将其转化到大肠杆菌表达宿主中。重组烯醇化酶蛋白被成功表达,并在原生条件下使用亲和层析法纯化,随后进行了SDS-PAGE表征和Western印迹分析。CD 光谱分析验证了表达的蛋白质以可溶性原生构象存在。初步的硅学研究验证了从白喉杆菌和白喉杆菌中分离出来的重组烯醇化酶蛋白的免疫原性。此外,生物信息学分析表明,白僵菌酵素酶蛋白和阿氏杆菌酵素酶蛋白的B细胞和T细胞表位是一致的,这表明它们可能具有交叉反应性和广谱疫苗功效。预计我们的研究结果将在推进系统性念珠菌病的治疗和诊断策略方面发挥作用。
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引用次数: 0
Fungal biomass and ectomycorrhizal community assessment of phosphorus responsive Pinus taeda plantations. 对磷响应性尾叶松种植园进行真菌生物量和外生菌根群落评估。
Pub Date : 2024-05-28 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1401427
Jacob Hackman, Alex Woodley, David Carter, Brian Strahm, Collin Averill, Rytas Vilgalys, Kevin Garcia, Rachel Cook

Ectomycorrhizal fungi and non-ectomycorrhizal fungi are responsive to changes in environmental and nutrient availabilities. Although many species of ectomycorrhizas are known to enhance the uptake of phosphorus and other nutrients for Pinus taeda, it is not understood how to optimize these communities to have tangible effects on plantation silviculture and P use efficiency. The first step of this process is the identification of native fungi present in the system that are associated with P. taeda and influence P uptake efficiency. We used sand-filled mesh bags baited with finely ground apatite to sample ectomycorrhizal and non-ectomycorrhizal fungi associated with the rhizosphere of P-responsive P. taeda under several field conditions. Mesh bags were assessed for biomass accumulation over three years using a single three-month burial period pre-harvest and three six-month burial periods post-planting. Amplicon sequencing assessed ectomycorrhizal and non-ectomycorrhizal communities between phosphorus treatments, sites, mesh bags, and the rhizosphere of actively growing P. taeda in the field. We found biomass accumulation within the mesh bags was inversely related to increasing phosphorus fertilization (carryover) rates from pre-harvest to post-planting. Up to 25% increases in total biomass within the bags were observed for bags baited with P. Taxonomic richness was highest in Alfisol soils treated with phosphorus from the previous rotation and lowest in the Spodosol regardless of phosphorus treatment.

外生菌根真菌和非外生菌根真菌对环境和养分供应的变化反应灵敏。虽然已知许多种外生菌根真菌能提高太田松对磷和其他养分的吸收,但如何优化这些群落,使其对人工林造林和钾利用效率产生切实影响,目前尚不清楚。这一过程的第一步是确定系统中存在的与泰达松相关并影响钾吸收效率的本地真菌。我们使用装有细磨磷灰石饵料的装沙网袋,在几种田间条件下对与对钾有反应的泰达红豆杉根圈相关的外生菌根真菌和非外生菌根真菌进行采样。通过采收前一次为期三个月的掩埋和种植后三次为期六个月的掩埋,对网袋三年的生物量积累进行了评估。扩增子测序评估了磷处理、地点、网袋和田间生长旺盛的泰达红豆杉根圈之间的外生菌根和非外生菌根群落。我们发现,从收获前到种植后,网袋内生物量的积累与磷肥施用量(结转)的增加成反比。无论磷肥处理方式如何,上一轮轮作中施用磷肥的 Alfisol 土壤中的分类丰富度最高,而 Spodosol 土壤中的分类丰富度最低。
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引用次数: 0
First-class – biosynthesis of 6-MSA and bostrycoidin type I polyketides in Yarrowia lipolytica Yarrowia lipolytica 中 6-MSA 和 Bostrycoidin I 型多酮类化合物的一级生物合成
Pub Date : 2024-03-22 DOI: 10.3389/ffunb.2024.1327777
Mihaela Bejenari, Eva Mie Lang Spedtsberg, Julie Mathiesen, Alexandra Claire Jeppesen, Lucia Cernat, Aouregane Toussaint, Cristina Apostol, Victor Stoianov, T. B. Pedersen, M. R. Nielsen, Jens Laurids Sørensen
Fungal polyketides are a large group of secondary metabolites, valuable due to their diverse spectrum of pharmacological activities. Polyketide biosynthesis in filamentous fungi presents some challenges: small yield and low-purity titers. To tackle these issues, we switched to the yeast Yarrowia lipolytica, an easily cultivable heterologous host. As an oleaginous yeast, Y. lipolytica displays a high flux of acetyl- and malonyl-CoA precursors used in lipid synthesis. Likewise, acetyl- and malonyl-CoA are the building blocks of many natural polyketides, and we explored the possibility of redirecting this flux toward polyketide production. Despite its promising prospect, Y. lipolytica has so far only been used for heterologous expression of simple type III polyketide synthases (PKSs) from plants. Therefore, we decided to evaluate the potential of Y. lipolytica by targeting the more complex fungal polyketides synthesized by type I PKSs. We employed a CRISPR-Cas9-mediated genome editing method to achieve markerless gene integration of the genes responsible for bostrycoidin biosynthesis in Fusarium solani (fsr1, fsr2, and fsr3) and 6-methylsalicylic acid (6-MSA) biosynthesis in Aspergillus hancockii (6MSAS). Moreover, we attempted titer optimization through metabolic engineering by overexpressing two enzymes, TGL4 and AOX2, involved in lipid β-oxidation, but we did not observe an effect on polyketide production. With maximum titers of 403 mg/L 6-MSA and 35 mg/L bostrycoidin, the latter being substantially higher than our previous results in Saccharomyces cerevisiae (2.2 mg/L), this work demonstrates the potential of Y. lipolytica as a platform for heterologous production of complex fungal polyketides.
真菌多酮苷是一大类次级代谢产物,因其具有多种药理活性而弥足珍贵。丝状真菌中的多酮生物合成面临一些挑战:产量小、纯度低。为了解决这些问题,我们转而使用容易培养的异源宿主--脂肪分解酵母菌(Yarrowia lipolytica)。作为一种含油酵母,脂溶性酵母显示出用于脂质合成的乙酰基和丙二酰-CoA 前体的高通量。同样,乙酰-CoA 和丙二酰-CoA 也是许多天然多酮的组成成分,因此我们探索了将这种通量重新导向多酮生产的可能性。尽管Y.lipolytica具有广阔的前景,但迄今为止它仅被用于异源表达来自植物的简单III型多酮合成酶(PKSs)。因此,我们决定以 I 型 PKSs 合成的更复杂的真菌多酮化合物为目标,评估 Y. lipolytica 的潜力。我们采用了 CRISPR-Cas9 介导的基因组编辑方法,对茄属镰刀菌(fsr1、fsr2 和fsr3)中负责梭菌毒素生物合成的基因和汉氏曲霉(Aspergillus hancockii)中负责 6-甲基水杨酸(6-MSA)生物合成的基因(6MSAS)进行了无标记基因整合。此外,我们还尝试通过代谢工程优化滴度,过量表达两种参与脂质β氧化的酶 TGL4 和 AOX2,但我们没有观察到它们对多酮生产的影响。这项工作证明了溶脂酵母作为异源生产复杂真菌多酮的平台的潜力。
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Frontiers in fungal biology
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