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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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引用次数: 0
A Bacillus velezensis strain shows antimicrobial activity against soilborne and foliar fungi and oomycetes. 一种 Velezensis 杆菌菌株对土传真菌、叶面真菌和卵菌具有抗菌活性。
IF 2.1 Q3 MYCOLOGY Pub Date : 2024-02-23 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1332755
Anna Wockenfuss, Kevin Chan, Jessica G Cooper, Timothy Chaya, Megan A Mauriello, Sarah M Yannarell, Julia A Maresca, Nicole M Donofrio

Biological control uses naturally occurring antagonists such as bacteria or fungi for environmentally friendly control of plant pathogens. Bacillus spp. have been used for biocontrol of numerous plant and insect pests and are well-known to synthesize a variety of bioactive secondary metabolites. We hypothesized that bacteria isolated from agricultural soil would be effective antagonists of soilborne fungal pathogens. Here, we show that the Delaware soil isolate Bacillus velezensis strain S4 has in vitro activity against soilborne and foliar plant pathogenic fungi, including two with a large host range, and one oomycete. Further, this strain shows putative protease and cellulase activity, consistent with our prior finding that the genome of this organism is highly enriched in antifungal and antimicrobial biosynthetic gene clusters. We demonstrate that this bacterium causes changes to the fungal and oomycete hyphae at the inhibition zone, with some of the hyphae forming bubble-like structures and irregular branching. We tested strain S4 against Magnaporthe oryzae spores, which typically form germ tubes and penetration structures called appressoria, on the surface of the leaf. Our results suggest that after 12 hours of incubation with the bacterium, fungal spores form germ tubes, but instead of producing appressoria, they appear to form rounded, bubble-like structures. Future work will investigate whether a single antifungal molecule induces all these effects, or if they are the result of a combination of bacterially produced antimicrobials.

生物防治利用细菌或真菌等天然拮抗剂对植物病原体进行环境友好型控制。芽孢杆菌属已被用于对多种植物和昆虫害虫进行生物防治,而且众所周知,它们能合成多种具有生物活性的次级代谢产物。我们假设,从农业土壤中分离出来的细菌将成为土传真菌病原体的有效拮抗剂。在这里,我们发现特拉华州土壤中分离出的芽孢杆菌 S4 菌株对土传和叶传植物致病真菌具有体外活性,其中包括两种寄主范围较大的真菌和一种卵菌。此外,该菌株还显示出潜在的蛋白酶和纤维素酶活性,这与我们之前发现的该生物体基因组中抗真菌和抗微生物生物合成基因簇的高度富集是一致的。我们证明,这种细菌会导致抑制区的真菌和卵菌菌丝发生变化,其中一些菌丝会形成气泡状结构和不规则分枝。我们对菌株 S4 进行了针对木格诺氏菌孢子的测试,这种孢子通常会在叶片表面形成芽管和称为附着体的穿透结构。我们的结果表明,真菌孢子在与该细菌培养 12 小时后,会形成芽管,但不会产生附着体,而是形成圆形的气泡状结构。未来的工作将研究是单一的抗真菌分子诱导了所有这些效应,还是细菌产生的抗菌剂共同作用的结果。
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引用次数: 0
Healthcare-associated fungal infections and emerging pathogens during the COVID-19 pandemic. COVID-19 大流行期间与医疗保健相关的真菌感染和新出现的病原体。
Pub Date : 2024-02-23 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1339911
Krish Shah, Mukund Deshpande, P Shah

Historically, fungi were mainly identified as plant and insect pathogens since they grow at 28°C. At the same time, bacteria are known to be the most common human pathogens as they are compatible with the host body temperature of 37°C. Because of immunocompromised hosts, cancer therapy, and malnutrition, fungi are rapidly gaining attention as human pathogens. Over 150 million people have severe fungal infections, which lead to approximately more than one million deaths per year. Moreover, diseases like cancer involving long-term therapy and prophylactic use of antifungal drugs in high-risk patients have increased the emergence of drug-resistant fungi, including highly virulent strains such as Candida auris. This clinical spectrum of fungal diseases ranges from superficial mucocutaneous lesions to more severe and life-threatening infections. This review article summarizes the effect of hospital environments, especially during the COVID-19 pandemic, on fungal infections and emerging pathogens. The review also provides insights into the various antifungal drugs and their existing challenges, thereby driving the need to search for novel antifungal agents.

一直以来,真菌主要被认为是植物和昆虫的病原体,因为它们在 28 摄氏度下生长。与此同时,众所周知,细菌是最常见的人类病原体,因为它们与 37°C 的宿主体温相适应。由于宿主免疫力低下、癌症治疗和营养不良等原因,真菌作为人类病原体正迅速受到关注。超过 1.5 亿人患有严重的真菌感染,每年导致约 100 多万人死亡。此外,癌症等疾病需要长期治疗,高危患者需要预防性使用抗真菌药物,这也增加了耐药性真菌的出现,其中包括白色念珠菌等高致病性菌株。真菌疾病的临床范围从浅表皮肤粘膜病变到更严重和危及生命的感染。这篇综述文章总结了医院环境对真菌感染和新病原体的影响,尤其是在 COVID-19 大流行期间。该综述还深入探讨了各种抗真菌药物及其现有挑战,从而推动了寻找新型抗真菌药物的需求。
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引用次数: 0
Editorial: Endophytic fungi producers of biomolecules of interest to human health. 社论:内生真菌生产对人类健康有益的生物大分子。
Pub Date : 2024-01-30 eCollection Date: 2024-01-01 DOI: 10.3389/ffunb.2024.1239470
Pérola O Magalhães, João Vicente Braga de Souza, Danilo Batista Pinho
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引用次数: 0
Editorial: Nematophagous fungi as nematode control agents 社论:作为线虫控制剂的食线虫真菌
Pub Date : 2024-01-17 DOI: 10.3389/ffunb.2023.1353132
F. E. Soares, Liliana Aguilar-Marcelino, Fabio Ribeiro Braga
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引用次数: 0
Utilization of Metarhizium as an insect biocontrol agent and a plant bioinoculant with special reference to Brazil 利用 Metarhizium 作为昆虫生物控制剂和植物生物接种剂,特别是在巴西
Pub Date : 2023-12-21 DOI: 10.3389/ffunb.2023.1276287
E. Mesquita, Shasha Hu, Tais B. Lima, P. Gôlo, M. Bidochka
Brazil has a long history of using biological control and has the largest program in sugarcane agriculture to which a biocontrol program has been applied. This achievement is at least partly due to the utilization of the entomopathogenic fungus Metarhizium. This well-known fungal genus exhibits pathogenicity against a broad range of arthropod hosts and has been used globally as a biocontrol agent. This fungus is also a root symbiont, and in this capacity, it is a plant growth promoter. However, this feature (i.e., as a plant symbiont) has yet to be fully explored and implemented in Brazil, although the number of reports demonstrating Metarhizium’s utility as a plant bioinoculant is increasing. The Brazilian bioproduct industry targets agricultural pests, and is limited to two Metarhizium species represented by four fungal isolates as active ingredients. Entomopathogenic fungi have also been successful in controlling arthropods of public health concern, as shown in their control of mosquitoes, which are vectors of diseases. The isolation of new indigenous Metarhizium isolates from a variety of substrates such as soil, insects, and plants shows the wide genetic diversity within this fungal genus. In this review, we emphasize the significance of Metarhizium spp. for the biological control of insects in Brazil. We also suggest that the experience and success of biological control with fungi in Brazil is an important resource for developing integrated pest management and sustainable strategies for pest control worldwide. Moreover, the future implementation prospects of species of Metarhizium being used as bioinoculants and possible new advances in the utility of this fungus are discussed.
巴西使用生物防治的历史悠久,是甘蔗农业中使用生物防治计划最多的国家。这一成就至少部分归功于对昆虫病原真菌 Metarhizium 的利用。这种著名的真菌属对多种节肢动物宿主具有致病性,已被全球用作生物控制剂。这种真菌也是根系的共生体,因此能促进植物生长。不过,尽管证明 Metarhizium 可用作植物生物接种剂的报告数量正在增加,但巴西尚未充分探索和实施这一特性(即作为植物共生体)。巴西的生物产品产业以农业害虫为目标,其有效成分仅限于以四种真菌分离物为代表的两种 Metarhizium。昆虫病原真菌也成功地控制了与公共卫生有关的节肢动物,如对疾病传播媒介蚊子的控制。从土壤、昆虫和植物等多种基质中分离出新的本地 Metarhizium 分离物,显示了该真菌属内部广泛的遗传多样性。在这篇综述中,我们强调了 Metarhizium 对巴西昆虫生物防治的重要意义。我们还认为,巴西在利用真菌进行生物防治方面的经验和成功,是全球虫害综合治理和可持续虫害防治战略发展的重要资源。此外,我们还讨论了将 Metarhizium 菌种用作生物接种剂的未来实施前景,以及这种真菌的用途可能取得的新进展。
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引用次数: 0
Associated bacterial communities, confrontation studies, and comparative genomics reveal important interactions between Morchella with Pseudomonas spp. 相关细菌群落、对抗研究和比较基因组学揭示了莫切莱氏菌与假单胞菌属之间的重要相互作用。
Pub Date : 2023-12-13 DOI: 10.3389/ffunb.2023.1285531
G. Cailleau, Buck T. Hanson, Melissa Cravero, Sami Zhioua, Patrick Hilpish, Celia Ruiz, Aaron J. Robinson, Julia M. Kelliher, Demosthenes P Morales, La Verne Gallegos-Graves, Gregory Bonito, P. S. Chain, S. Bindschedler, Pilar Junier
Members of the fungal genus Morchella are widely known for their important ecological roles and significant economic value. In this study, we used amplicon and genome sequencing to characterize bacterial communities associated with sexual fruiting bodies from wild specimens, as well as vegetative mycelium and sclerotia obtained from Morchella isolates grown in vitro. These investigations included diverse representatives from both Elata and Esculenta Morchella clades. Unique bacterial community compositions were observed across the various structures examined, both within and across individual Morchella isolates or specimens. However, specific bacterial taxa were frequently detected in association with certain structures, providing support for an associated core bacterial community. Bacteria from the genus Pseudomonas and Ralstonia constituted the core bacterial associates of Morchella mycelia and sclerotia, while other genera (e.g., Pedobacter spp., Deviosa spp., and Bradyrhizobium spp.) constituted the core bacterial community of fruiting bodies. Furthermore, the importance of Pseudomonas as a key member of the bacteriome was supported by the isolation of several Pseudomonas strains from mycelia during in vitro cultivation. Four of the six mycelial-derived Pseudomonas isolates shared 16S rDNA sequence identity with amplicon sequences recovered directly from the examined fungal structures. Distinct interaction phenotypes (antagonistic or neutral) were observed in confrontation assays between these bacteria and various Morchella isolates. Genome sequences obtained from these Pseudomonas isolates revealed intriguing differences in gene content and annotated functions, specifically with respect to toxin-antitoxin systems, cell adhesion, chitinases, and insecticidal toxins. These genetic differences correlated with the interaction phenotypes. This study provides evidence that Pseudomonas spp. are frequently associated with Morchella and these associations may greatly impact fungal physiology.
真菌莫氏菌属的成员因其重要的生态作用和巨大的经济价值而广为人知。在这项研究中,我们使用扩增子和基因组测序技术,分析了与野生标本有性子实体相关的细菌群落,以及从离体生长的莫氏菌分离物中获得的无性菌丝和硬菌。这些研究涵盖了莫切拉氏菌 Elata 支系和 Esculenta 支系的各种代表菌。在所研究的各种结构中,都观察到了独特的细菌群落组成,包括在单个莫氏菌分离物或标本内部和之间。不过,在某些结构中经常检测到特定的细菌类群,这为相关的核心细菌群落提供了支持。假单胞菌属(Pseudomonas)和雷氏菌属(Ralstonia)的细菌构成了莫氏菌菌丝体和硬核菌的核心细菌群落,而其他菌属(如Pedobacter属、Deviosa属和Bradyrhizobium属)则构成了子实体的核心细菌群落。此外,在离体培养过程中从菌丝体中分离出的几株假单胞菌也证明了假单胞菌作为细菌群落关键成员的重要性。从菌丝体中分离出的六株假单胞菌中,有四株的 16S rDNA 序列与直接从受检真菌结构中回收的扩增子序列具有相同性。在这些细菌与各种莫切拉氏菌分离物的对抗试验中观察到了不同的相互作用表型(拮抗或中性)。从这些假单胞菌分离物中获得的基因组序列揭示了基因内容和注释功能方面的惊人差异,特别是在毒素-抗毒素系统、细胞粘附性、几丁质酶和杀虫毒素方面。这些基因差异与相互作用表型相关。这项研究提供的证据表明,假单胞菌属经常与莫切拉菌发生关联,而这些关联可能会对真菌的生理学产生重大影响。
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引用次数: 0
Prevalence of groundnut dry root rot (Macrophomina phaseolina (Tassi) Goid.) and its pathogenic variability in Southern India 印度南部花生干根腐病(Macrophomina phaseolina (Tassi) Goid.)的流行及其病原变异性
Pub Date : 2023-12-04 DOI: 10.3389/ffunb.2023.1189043
Prince Jayasimha Pamala, R. S. Jayalakshmi, K. Vemana, G. M. Naidu, R. Varshney, Hari Kishan Sudini
Macrophomina phaseolina is the most devastating and emerging threat to groundnut production in India. An increase in average temperature and inconsistent rainfalls resulting from changing climatic conditions are strongly believed to aggravate the disease and cause severe yield losses. The present study aims to conduct a holistic survey to assess the prevalence and incidence of dry root rot of groundnut in major groundnut growing regions of Southern India, viz., Andhra Pradesh, Telangana, Karnataka, and Tamil Nadu. Furthermore, the pathogenic variability was determined using different assays such as morphological, cultural, pathogenic, and molecular assays. Results indicate that disease incidence in surveyed locations ranged from 8.06 to 20.61%. Both temperature and rainfall played a major role in increasing the disease incidence. The pathogenic variability of M. phaseolina isolates differed significantly, based on the percent disease incidence induced on cultivars of JL-24 groundnut and K-6 groundnut. Morphological variations in terms of growth pattern, culture color, sclerotia number, and sclerotia size were observed. The molecular characterization of M. phaseolina isolates done by ITS rDNA region using ITS1 and ITS4 primers yielded approximately 600 bp PCR amplicons, sequenced and deposited in GenBank (NCBI). Molecular variability analysis using SSR primers indicated the genetic variation among the isolates collected from different states. The present investigation revealed significant variations in pathogenic variability among isolates of M. phaseolina and these may be considered important in disease management and the development of resistant cultivars against groundnut dry root rot disease.
在印度,菜豆大蠹是对花生生产最具破坏性和新出现的威胁。人们强烈认为,气候条件变化导致的平均气温升高和降雨不稳定会加剧这种疾病,造成严重的产量损失。本研究旨在进行一项全面调查,以评估印度南部主要花生种植区,即安得拉邦、特伦甘纳邦、卡纳塔克邦和泰米尔纳德邦花生干根腐病的患病率和发病率。此外,采用不同的检测方法,如形态学、文化、致病性和分子检测来确定致病变异性。结果调查地点的发病率为8.06 ~ 20.61%。气温和降雨对发病率的增加起主要作用。从JL-24花生和K-6花生品种的致病率来看,菜豆分枝杆菌分离株的致病变异性存在显著差异。在生长模式、培养颜色、菌核数量和菌核大小方面观察到形态学变化。利用ITS1和ITS4引物对菜绿分枝杆菌的ITS rDNA区进行分子鉴定,得到约600 bp的PCR扩增片段,测序并存入GenBank (NCBI)。利用SSR引物进行分子变异分析,发现不同产地分离物间存在遗传变异。本研究揭示了花生干根腐病分离株间致病变异的显著差异,这可能对疾病管理和开发抗花生干根腐病品种具有重要意义。
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引用次数: 0
Corrigendum: Fusarium verticillioides of maize plant: Potentials of propitious phytomicrobiome as biocontrol agents. 更正:玉米植株上的疣状镰刀菌:植物微生物群作为生物控制剂的潜力。
Pub Date : 2023-11-29 eCollection Date: 2023-01-01 DOI: 10.3389/ffunb.2023.1298350
Oluwadara Pelumi Omotayo, Olubukola Oluranti Babalola

[This corrects the article DOI: 10.3389/ffunb.2023.1095765.].

[此处更正了文章 DOI:10.3389/ffunb.2023.1095765]。
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引用次数: 0
Exploring Psilocybe spp. mycelium and fruiting body chemistry for potential therapeutic compounds. 探索 Psilocybe spp.菌丝体和子实体化学的潜在治疗化合物。
Pub Date : 2023-11-29 eCollection Date: 2023-01-01 DOI: 10.3389/ffunb.2023.1295223
Adam Waldbillig, Maria Baranova, Sarah Neumann, Jonathan Andrade, Sharan Sidhu

Psilocybe mushrooms, otherwise known as "magic" mushrooms, owe their psychedelic effect to psilocin, a serotonin subtype 2A (5-HT2A) receptor agonist and metabolite of psilocybin, the primary indole alkaloid found in Psilocybe species. Metabolomics is an advanced fingerprinting tool that can be utilized to identify the differences among fungal life stages that may otherwise be unaccounted for. In this study, by using targeted and untargeted (metabolomic) multivariate analysis, we demonstrate that the chemical composition of Psilocybe differs among mycelia, grain mycelia, and fruiting bodies. The preferential accumulation of psilocybin, baeocystin, tryptophan, ergothioneine, and phenylethylamine in fruiting bodies differentiated them from mycelia; however, the levels of alpha-glycerylphosphorylcholine (α-GPC), N-acetylglucosamine, and trimethylglycine were found to be proportionally higher in mycelia than in fruiting bodies based on Pareto-scaled data. Considering the wealth of compounds with therapeutic potential that have been isolated from various fungal genera, it would be pertinent to study the compounds found in Psilocybe mycelia as potential naturally derived therapeutic targets.

被称为 "神奇 "蘑菇的银环蛇蘑菇之所以具有迷幻效果,是因为银环蛇毒素是一种血清素亚型 2A(5-HT2A)受体激动剂,也是银环蛇毒素的代谢物,银环蛇毒素是银环蛇属真菌中的主要吲哚生物碱。代谢组学是一种先进的指纹识别工具,可用于识别真菌生命阶段之间的差异,否则这些差异可能无法解释。在这项研究中,通过使用有针对性和无针对性(代谢组学)的多元分析,我们证明了银莲花菌丝体、子实体和谷粒菌丝体之间的化学成分差异。在子实体中,茜草素、芍药苷、色氨酸、麦角硫因和苯乙胺的优先积累将它们与菌丝体区分开来;然而,根据帕累托标度数据,我们发现菌丝体中的α-甘油酰磷酰胆碱(α-GPC)、N-乙酰葡糖胺和三甲基甘氨酸的含量在比例上高于子实体。考虑到已从各种真菌属中分离出大量具有治疗潜力的化合物,将研究在银铃草菌丝体中发现的化合物作为潜在的自然衍生治疗靶标将是很有意义的。
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引用次数: 0
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Frontiers in fungal biology
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