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Whole genomes of deep-sea sponge-associated bacteria exhibit high novel natural product potential. 深海海绵相关细菌的全基因组显示出很高的新型天然产物潜力。
Pub Date : 2023-02-22 eCollection Date: 2023-01-01 DOI: 10.1093/femsmc/xtad005
Poppy J Hesketh-Best, Grant G January, Matthew J Koch, Philip J Warburton, Kerry L Howell, Mathew Upton

Global antimicrobial resistance is a health crisis that can change the face of modern medicine. Exploring diverse natural habitats for bacterially-derived novel antimicrobial compounds has historically been a successful strategy. The deep-sea presents an exciting opportunity for the cultivation of taxonomically novel organisms and exploring potentially chemically novel spaces. In this study, the draft genomes of 12 bacteria previously isolated from the deep-sea sponges Phenomena carpenteri and Hertwigia sp. are investigated for the diversity of specialized secondary metabolites. In addition, early data support the production of antibacterial inhibitory substances produced from a number of these strains, including activity against clinically relevant pathogens Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. Draft whole-genomes are presented of 12 deep-sea isolates, which include four potentially novel strains: Psychrobacter sp. PP-21, Streptomyces sp. DK15, Dietzia sp. PP-33, and Micrococcus sp. M4NT. Across the 12 draft genomes, 138 biosynthetic gene clusters were detected, of which over half displayed less than 50% similarity to known BGCs, suggesting that these genomes present an exciting opportunity to elucidate novel secondary metabolites. Exploring bacterial isolates belonging to the phylum Actinomycetota, Pseudomonadota, and Bacillota from understudied deep-sea sponges provided opportunities to search for new chemical diversity of interest to those working in antibiotic discovery.

全球抗菌素耐药性是一场可以改变现代医学面貌的健康危机。为细菌衍生的新型抗菌化合物探索不同的自然栖息地历来是一种成功的策略。深海为培育分类上的新生物和探索潜在的化学新空间提供了一个令人兴奋的机会。在这项研究中,研究了以前从深海海绵中分离出来的12种细菌的基因组草案,研究了专门的次级代谢物的多样性。此外,早期数据支持从这些菌株中生产抗菌抑制物质,包括抗临床相关病原体鲍曼不动杆菌、大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌和金黄色葡萄球菌的活性。本文报道了12个深海分离株的全基因组草图,其中包括4个潜在的新菌株:Psychrobacter sp. PP-21、Streptomyces sp. DK15、Dietzia sp. PP-33和Micrococcus sp. M4NT。在12个基因组草案中,检测到138个生物合成基因簇,其中一半以上与已知bgc的相似性低于50%,这表明这些基因组为阐明新的次级代谢物提供了令人兴奋的机会。从深海海绵中探索属于放线菌门、假单胞菌门和芽孢杆菌门的细菌分离物,为那些从事抗生素发现工作的人提供了寻找新的化学多样性的机会。
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引用次数: 0
Editorial: Thematic issue on bacterial-viral co-infections. 社论:细菌病毒共同感染专题。
Pub Date : 2023-01-19 eCollection Date: 2023-01-01 DOI: 10.1093/femsmc/xtac031
Sam Manna, Daniel M Weinberger, Catherine Satzke
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引用次数: 0
Direct wastewater extraction as a simple and effective method for SARS-CoV-2 surveillance and COVID-19 community-level monitoring. 直接废水提取是SARS-CoV-2监测和新冠肺炎社区级监测的一种简单有效的方法。
Pub Date : 2023-01-12 eCollection Date: 2023-01-01 DOI: 10.1093/femsmc/xtad004
Megan E J Lott, William A Norfolk, Cody A Dailey, Amelia M Foley, Carolina Melendez-Declet, Megan J Robertson, Stephen L Rathbun, Erin K Lipp

Wastewater surveillance has proven to be an effective tool to monitor the transmission and emergence of infectious agents at a community scale. Workflows for wastewater surveillance generally rely on concentration steps to increase the probability of detection of low-abundance targets, but preconcentration can substantially increase the time and cost of analyses while also introducing additional loss of target during processing. To address some of these issues, we conducted a longitudinal study implementing a simplified workflow for SARS-CoV-2 detection from wastewater, using a direct column-based extraction approach. Composite influent wastewater samples were collected weekly for 1 year between June 2020 and June 2021 in Athens-Clarke County, Georgia, USA. Bypassing any concentration step, low volumes (280 µl) of influent wastewater were extracted using a commercial kit, and immediately analyzed by RT-qPCR for the SARS-CoV-2 N1 and N2 gene targets. SARS-CoV-2 viral RNA was detected in 76% (193/254) of influent samples, and the recovery of the surrogate bovine coronavirus was 42% (IQR: 28%, 59%). N1 and N2 assay positivity, viral concentration, and flow-adjusted daily viral load correlated significantly with per-capita case reports of COVID-19 at the county-level (ρ = 0.69-0.82). To compensate for the method's high limit of detection (approximately 106-107 copies l-1 in wastewater), we extracted multiple small-volume replicates of each wastewater sample. With this approach, we detected as few as five cases of COVID-19 per 100 000 individuals. These results indicate that a direct-extraction-based workflow for SARS-CoV-2 wastewater surveillance can provide informative and actionable results.

废水监测已被证明是在社区范围内监测传染源传播和出现的有效工具。废水监测的工作流程通常依赖于浓缩步骤来增加检测低丰度目标的概率,但预浓缩可以显著增加分析的时间和成本,同时在处理过程中还会引入额外的目标损失。为了解决其中的一些问题,我们进行了一项纵向研究,使用基于直接柱的提取方法,实现了从废水中检测严重急性呼吸系统综合征冠状病毒2型的简化工作流程。2020年6月至2021年6月,在美国佐治亚州雅典克拉克县每周采集一次复合进水废水样本,为期一年。通过任何浓缩步骤,使用商业试剂盒提取低体积(280µl)的进水废水,并立即通过RT-qPCR分析严重急性呼吸系统综合征冠状病毒2型N1和N2基因靶标。在76%(193/254)的进水样本中检测到严重急性呼吸系统综合征冠状病毒2型病毒RNA,替代牛冠状病毒的回收率为42%(IQR:28%,59%)。N1和N2检测阳性率、病毒浓度和流量调整后的每日病毒载量与县级新冠肺炎的每个病例报告显著相关(ρ=0.69-0.82)。为了补偿该方法的高检测限(废水中约106-107个拷贝l-1),我们提取了每个废水样本的多个小容量复制品。通过这种方法,我们每10万人中只检测到5例新冠肺炎病例。这些结果表明,基于直接提取的严重急性呼吸系统综合征冠状病毒2型废水监测工作流程可以提供信息和可操作的结果。
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引用次数: 0
Show us the data: global COVID-19 wastewater monitoring efforts, equity, and gaps. 向我们展示数据:全球COVID-19废水监测工作、公平性和差距。
Pub Date : 2023-01-12 eCollection Date: 2023-01-01 DOI: 10.1093/femsmc/xtad003
Colleen C Naughton, Fernando A Roman, Ana Grace F Alvarado, Arianna Q Tariqi, Matthew A Deeming, Krystin F Kadonsky, Kyle Bibby, Aaron Bivins, Gertjan Medema, Warish Ahmed, Panagis Katsivelis, Vajra Allan, Ryan Sinclair, Joan B Rose

A year since the declaration of the global coronavirus disease 2019 (COVID-19) pandemic, there were over 110 million cases and 2.5 million deaths. Learning from methods to track community spread of other viruses such as poliovirus, environmental virologists and those in the wastewater-based epidemiology (WBE) field quickly adapted their existing methods to detect SARS-CoV-2 RNA in wastewater. Unlike COVID-19 case and mortality data, there was not a global dashboard to track wastewater monitoring of SARS-CoV-2 RNA worldwide. This study provides a 1-year review of the "COVIDPoops19" global dashboard of universities, sites, and countries monitoring SARS-CoV-2 RNA in wastewater. Methods to assemble the dashboard combined standard literature review, Google Form submissions, and daily, social media keyword searches. Over 200 universities, 1400 sites, and 55 countries with 59 dashboards monitored wastewater for SARS-CoV-2 RNA. However, monitoring was primarily in high-income countries (65%) with less access to this valuable tool in low- and middle-income countries (35%). Data were not widely shared publicly or accessible to researchers to further inform public health actions, perform meta-analysis, better coordinate, and determine equitable distribution of monitoring sites. For WBE to be used to its full potential during COVID-19 and beyond, show us the data.

自宣布2019年全球冠状病毒病(COVID-19)大流行以来的一年中,已有超过1.1亿例病例和250万人死亡。从追踪脊髓灰质炎病毒等其他病毒社区传播的方法中学习,环境病毒学家和基于废水的流行病学(WBE)领域的研究人员迅速调整了他们现有的方法,以检测废水中的SARS-CoV-2 RNA。与COVID-19病例和死亡率数据不同,没有一个全球仪表板来跟踪全球范围内对SARS-CoV-2 RNA的废水监测。本研究对监测废水中SARS-CoV-2 RNA的大学、地点和国家的“covid - poops19”全球仪表板进行了为期一年的回顾。组装仪表板的方法结合了标准文献综述、Google表单提交和每日社交媒体关键字搜索。超过200所大学、1400个站点和55个国家的59个仪表板监测了废水中的SARS-CoV-2 RNA。然而,监测主要在高收入国家进行(65%),低收入和中等收入国家获得这一宝贵工具的机会较少(35%)。数据没有被广泛公开共享或供研究人员获取,以进一步为公共卫生行动提供信息、进行荟萃分析、更好地协调和确定监测点的公平分配。为使WBE在2019冠状病毒病疫情期间及以后充分发挥潜力,请向我们展示相关数据。
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引用次数: 110
Above- and below-ground microbiome in the annual developmental cycle of two olive tree varieties. 两个橄榄树品种年度发育周期中的地上和地下微生物组。
Pub Date : 2023-01-06 eCollection Date: 2023-01-01 DOI: 10.1093/femsmc/xtad001
Myrsini Kakagianni, Myrto Tsiknia, Maria Feka, Sotirios Vasileiadis, Kleopatra Leontidou, Nektarios Kavroulakis, Katerina Karamanoli, Dimitrios G Karpouzas, Constantinos Ehaliotis, Kalliope K Papadopoulou

The olive tree is a hallmark crop in the Mediterranean region. Its cultivation is characterized by an enormous variability in existing genotypes and geographical areas. As regards the associated microbial communities of the olive tree, despite progress, we still lack comprehensive knowledge in the description of these key determinants of plant health and productivity. Here, we determined the prokaryotic, fungal and arbuscular mycorrhizal fungal (AMF) microbiome in below- (rhizospheric soil, roots) and above-ground (phyllosphere and carposphere) plant compartments of two olive varieties 'Koroneiki' and 'Chondrolia Chalkidikis' grown in Southern and Northern Greece respectively, in five developmental stages along a full fruit-bearing season. Distinct microbial communities were supported in above- and below-ground plant parts; while the former tended to be similar between the two varieties/locations, the latter were location specific. In both varieties/locations, a seasonally stable root microbiome was observed over time; in contrast the plant microbiome in the other compartments were prone to changes over time, which may be related to seasonal environmental change and/or to plant developmental stage. We noted that olive roots exhibited an AMF-specific filtering effect (not observed for bacteria and general fungi) onto the rhizosphere AMF communities of the two olive varieties/locations/, leading to the assemblage of homogenous intraradical AMF communities. Finally, shared microbiome members between the two olive varieties/locations include bacterial and fungal taxa with putative functional attributes that may contribute to olive tree tolerance to abiotic and biotic stress.

橄榄树是地中海地区的标志性作物。其栽培特点是现有基因型和地理区域的巨大变异性。关于橄榄树的相关微生物群落,尽管取得了进展,但我们在描述这些植物健康和生产力的关键决定因素方面仍然缺乏全面的知识。在这里,我们确定了分别生长在希腊南部和北部的两个橄榄品种“Koroneiki”和“Chondrolia Chalkidikis”的地下(根层土壤、根)和地上(叶层和车皮层)植物区室中的原核、真菌和丛枝菌根真菌(AMF)微生物组,在整个结果季节的五个发育阶段。不同的微生物群落在地上和地下植物部分得到支持;前者在两个品种/地点之间往往相似,而后者是特定地点的。在这两个品种/地点,随着时间的推移,都观察到了季节性稳定的根系微生物组;相反,其他区室的植物微生物组容易随着时间的推移而变化,这可能与季节性环境变化和/或植物发育阶段有关。我们注意到,橄榄根对两个橄榄品种/地点/的根际AMF群落表现出AMF特异性过滤作用(在细菌和普通真菌中未观察到),导致同质的根际内AMF群落的聚集。最后,两个橄榄品种/地点之间共享的微生物组成员包括具有假定功能属性的细菌和真菌分类群,这些功能属性可能有助于橄榄树对非生物和生物胁迫的耐受性。
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引用次数: 1
An antimicrobial thiopeptide producing novel actinomycetes Streptomyces terrae sp. nov., isolated from subsurface soil of arable land. 从耕地地下土壤中分离出一种产生新型放线菌地链霉菌的抗菌硫肽。
Pub Date : 2023-01-01 DOI: 10.1093/femsmc/xtad014
Stanzin Choksket, Mahaldeep Kaur, Anil Kumar Pinnaka, Suresh Korpole

An antimicrobial producing Gram-positive, aerobic, nonmotile, and filamentous actinobacterial strain SKN60T was isolated from soil The isolate exhibited 99.3% and 99.0% identity with Streptomyces laurentii ATCC 31255T and S. roseicoloratus TRM 44457T, respectively, in 16S rRNA gene sequence analysis. However, the genome sequence displayed maximum ANI (88.45%) and AAI (85.61%) with S. roseicoloratus TRM 44457T. Similarly, the dDDH showed 33.7% identity with S. roseicoloratus TRM 44457T. It formed a cluster with S. roseicoloratus TRM 44457T and S. laurentii ATCC 31255T in phylogenomic tree. Cell wall analysis revealed the presence of diphosphatidylglycerol, phosphatidylethanolamine, and phosphatidylcholine as major polar lipids and diaminopimelic acid as diagnostic diamino acid. Major fatty acids were iso-C15:0, anteiso-C15:0, and iso-C16:0. The G+C content was found to be 72.3 mol%. Genome sequence analysis using antiSMASH database showed occurrence of a thiopeptide biosynthesis gene cluster with 94% similarity to berninamycin from S. bernensis UC5144. The mass of 1146 Da is identical with berninamycin. But subtle differences observed in leader peptide sequence of thiopeptide and berninamycin. Notably, S. bernensis is not validly reported and thus SKN60T is the only strain containing berninamycin BGC as no other phylogenetic relative had it. Additionally, strain SKN60T differed in phenotypic and genetic characteristics with all phylogenetic relatives of the genus Streptomyces. Therefore, it is proposed as a novel species with the name Streptomyces terrae sp. nov. strain SKN60T (=MTCC 13163T; = JCM 35768T).

从土壤中分离到一株革兰氏阳性、需氧、非运动和丝状放线菌SKN60T,经16S rRNA基因序列分析,该菌株与laurentii链霉菌ATCC 31255T和S. roseicoloratus TRM 44457T的同源性分别为99.3%和99.0%。结果表明,玫瑰色紫荆TRM 44457T的ANI和AAI最高,分别为88.45%和85.61%。与S. roseicoloratus TRM 44457T同源性为33.7%。在系统基因组树中与S. roseicoloratus TRM 44457T和S. laurentii ATCC 31255T形成集群。细胞壁分析显示,二磷脂酰甘油、磷脂酰乙醇胺和磷脂酰胆碱是主要的极性脂质,二氨基苯甲酸是诊断的二氨基酸。主要脂肪酸为iso-C15:0、前iso-C15:0和iso-C16:0。G+C含量为72.3 mol%。利用anti - smash数据库进行基因组序列分析,发现一个硫肽生物合成基因簇与伯纳霉素的相似性为94%。1146da的质量与贝尼霉素相同。但硫肽和贝尼霉素的前导肽序列存在细微差异。值得注意的是,没有有效的报道,因此SKN60T是唯一含有贝纳霉素BGC的菌株,因为没有其他系统发育亲缘关系。此外,菌株SKN60T在表型和遗传特征上与链霉菌属的所有亲缘系存在差异。因此,建议将其作为新种命名为Streptomyces terrae sp. nov.菌株SKN60T (=MTCC 13163T;= JCM 35768t)。
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引用次数: 0
The Acinetobacter baumannii website (Ab-web): a multidisciplinary knowledge hub, communication platform, and workspace. 鲍曼不动杆菌网站(Ab-web):一个多学科的知识中心、交流平台和工作空间。
Pub Date : 2023-01-01 DOI: 10.1093/femsmc/xtad009
Nabil Karah, Valeria Mateo-Estrada, Santiago Castillo-Ramírez, Paul G Higgins, Benjamin Havenga, Wesaal Khan, Sara Domingues, Gabriela Jorge Da Silva, Laurent Poirel, Patrice Nordmann, Cecilia Ambrosi, Chaoying Ma, Siobhán McClean, María Paula Quiroga, Verónica E Alvarez, Daniela Centron, Raffaele Zarrilli, Johanna J Kenyon, Thomas A Russo, Benjamin A Evans, Andres Opazo-Capurro, Rayane Rafei, Monzer Hamze, Ziad Daoud, Irfan Ahmad, Philip N Rather, Ruth M Hall, Gottfried Wilharm, Bernt Eric Uhlin

Acinetobacter baumannii is a Gram-negative bacterium increasingly implicated in hospital-acquired infections and outbreaks. Effective prevention and control of such infections are commonly challenged by the frequent emergence of multidrug-resistant strains. Here we introduce Ab-web (https://www.acinetobacterbaumannii.no), the first online platform for sharing expertise on A. baumannii. Ab-web is a species-centric knowledge hub, initially with 10 articles organized into two main sections, 'Overview' and 'Topics', and three themes, 'epidemiology', 'antibiotic resistance', and 'virulence'. The 'workspace' section provides a spot for colleagues to collaborate, build, and manage joint projects. Ab-web is a community-driven initiative amenable to constructive feedback and new ideas.

鲍曼不动杆菌是一种革兰氏阴性细菌,与医院获得性感染和疫情的关系日益密切。有效预防和控制这类感染通常面临多重耐药菌株频繁出现的挑战。在这里我们介绍Ab-web (https://www.acinetobacterbaumannii.no),这是第一个鲍曼不动杆菌专业知识分享的在线平台。Ab-web是一个以物种为中心的知识中心,最初有10篇文章,分为两个主要部分,“概述”和“主题”,以及三个主题,“流行病学”,“抗生素耐药性”和“毒性”。“工作空间”部分为同事提供了协作、构建和管理联合项目的场所。Ab-web是一个社区驱动的倡议,可以接受建设性的反馈和新想法。
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引用次数: 1
Organic and inorganic nutrients modulate taxonomic diversity and trophic strategies of small eukaryotes in oligotrophic oceans. 有机和无机营养物调节寡营养海洋中小型真核生物的分类多样性和营养策略。
Pub Date : 2023-01-01 DOI: 10.1093/femsmc/xtac029
Naomi Villiot, Amy E Maas, Alex J Poulton, Leocadio Blanco-Bercial

As the oligotrophic gyres expand due to global warming, exacerbating resource limitation impacts on primary producers, predicting changes to microbial assemblages and productivity requires knowledge of the community response to nutrient availability. This study examines how organic and inorganic nutrients influence the taxonomic and trophic composition (18S metabarcoding) of small eukaryotic plankton communities (< 200 µm) within the euphotic zone of the oligotrophic Sargasso Sea. The study was conducted by means of field sampling of natural microbial communities and laboratory incubation of these communities under different nutrient regimes. Dissimilarity in community composition increased along a depth gradient, with a homogeneous protist community within the mixed layer and distinct microbial assemblages at different depths below the deep chlorophyll maximum. A nutrient enrichment assay revealed the potential of natural microbial communities to rapidly shift in composition in response to nutrient addition. Results highlighted the importance of inorganic phosphorus availability, largely understudied compared to nitrogen, in constraining microbial diversity. Dissolved organic matter addition led to a loss of diversity, benefiting a limited number of phagotrophic and mixotrophic taxa. Nutrient history of the community sets the physiological responsiveness of the eukaryotic community to changing nutrient regimes and needs to be considered in future studies.

由于全球变暖导致的少营养环流扩大,加剧了资源限制对初级生产者的影响,预测微生物组合和生产力的变化需要了解群落对养分可用性的反应。本研究探讨了低营养马尾藻海富营养化带中有机和无机营养物如何影响小型真核浮游生物群落(< 200µm)的分类和营养组成(18S元条形码)。本研究通过野外取样和不同营养条件下的实验室微生物群落培养进行。群落组成差异沿深度梯度增大,混合层内原生生物群落均为均匀的,叶绿素最大值以下不同深度的微生物群落差异较大。一项营养富集试验揭示了天然微生物群落的潜力,以迅速改变组成,以响应营养添加。结果强调了无机磷有效性在限制微生物多样性方面的重要性,与氮相比,无机磷有效性在很大程度上尚未得到充分研究。溶解有机质的添加导致多样性的丧失,使有限数量的吞噬和混合营养类群受益。群落的营养史决定了真核生物群落对营养制度变化的生理反应,需要在未来的研究中加以考虑。
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引用次数: 1
Disparity in pseudohyphal morphogenic switching response to the quorum sensing molecule 2-phenylethanol in commercial brewing strains of Saccharomyces cerevisiae. 酿酒酵母商业酿酒菌株假菌丝形态发生切换对群体感应分子2-苯乙醇响应的差异。
Pub Date : 2023-01-01 DOI: 10.1093/femsmc/xtad002
Scott J Britton, Lisa J Rogers, Jane S White, Hedwig Neven, Dawn L Maskell

Saccharomyces cerevisiae can undergo filamentous growth in response to specific environmental stressors, particularly nitrogen-limitation, whereby cells undergo pseudohyphal differentiation, a process where cells transition from a singular ellipsoidal appearance to multicellular filamentous chains from the incomplete scission of the mother-daughter cells. Previously, it was demonstrated that filamentous growth in S. cerevisiae is co-regulated by multiple signaling networks, including the glucose-sensing RAS/cAMP-PKA and SNF pathways, the nutrient-sensing TOR pathway, the filamentous growth MAPK pathway, and the Rim101 pathway, and can be induced by quorum-sensing aromatic alcohols, such as 2-phenylethanol. However, the prevalent research on the yeast-pseudohyphal transition and its induction by aromatic alcohols in S. cerevisiae has been primarily limited to the strain Σ1278b. Due to the prospective influence of quorum sensing on commercial fermentation, the native variation of yeast-to-filamentous phenotypic transition and its induction by 2-phenylethanol in commercial brewing strains was investigated. Image analysis software was exploited to enumerate the magnitude of whole colony filamentation in 16 commercial strains cultured on nitrogen-limiting SLAD medium; some supplemented with exogenous 2-phenylethanol. The results demonstrate that phenotypic switching is a generalized, highly varied response occurring only in select brewing strains. Nevertheless, strains exhibiting switching behavior altered their filamentation response to exogenous concentrations of 2-phenylethanol.

酿酒酵母在特定的环境胁迫条件下,特别是氮限制条件下,可以进行丝状生长,因此细胞会进行假菌丝分化,这是一个细胞从单一椭球状外观转变为多细胞丝状链的过程。此前的研究表明,酿酒酵母的丝状生长受多个信号网络的共同调控,包括葡萄糖感应RAS/cAMP-PKA和SNF通路、营养感应TOR通路、丝状生长MAPK通路和Rim101通路,并可由群体感应芳香醇(如2-苯乙醇)诱导。然而,关于酿酒酵母-假菌丝转化和芳香醇诱导的流行研究主要局限于菌株Σ1278b。由于群体感应在商业发酵中的潜在影响,研究了商业酿酒菌株酵母向丝状表型转变的天然变异及其2-苯乙醇的诱导作用。利用图像分析软件对16株在限氮SLAD培养基上培养的商业菌株进行了全菌落成丝的计数;有些添加了外源2-苯乙醇。结果表明,表型转换是一个普遍的,高度变化的反应只发生在选择酿造菌株。然而,表现出切换行为的菌株改变了它们对外源浓度2-苯乙醇的成丝反应。
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引用次数: 2
Challenges in using transcriptome data to study the c-di-GMP signaling network in Pseudomonas aeruginosa clinical isolates. 利用转录组数据研究铜绿假单胞菌临床分离株c-di-GMP信号网络的挑战。
Pub Date : 2023-01-01 DOI: 10.1093/femsmc/xtad012
Melisa Gür, Jelena Erdmann, Anke Will, Ziwei Liang, Jens Bo Andersen, Tim Tolker-Nielsen, Susanne Häussler

In the Pseudomonas aeruginosa type strain PA14, 40 genes are known to encode for diguanylate cyclases (DGCs) and/or phosphodiesterases (PDEs), which modulate the intracellular pool of the nucleotide second messenger c-di-GMP. While in general, high levels of c-di-GMP drive the switch from highly motile phenotypes towards a sessile lifestyle, the different c-di-GMP modulating enzymes are responsible for smaller and in parts nonoverlapping phenotypes. In this study, we sought to utilize previously recorded P. aeruginosa gene expression datasets on 414 clinical isolates to uncover transcriptional changes as a result of a high expression of genes encoding DGCs. This approach might provide a unique opportunity to bypass the problem that for many c-di-GMP modulating enzymes it is not known under which conditions their expression is activated. However, while we demonstrate that the selection of subgroups of clinical isolates with high versus low expression of sigma factor encoding genes served the identification of their downstream regulons, we were unable to confirm the predicted DGC regulons, because the high c-di-GMP associated phenotypes were rapidly lost in the clinical isolates,. Further studies are needed to determine the specific mechanisms underlying the loss of cyclase activity upon prolonged cultivation of clinical P. aeruginosa isolates.

在铜绿假单胞菌型菌株PA14中,已知有40个基因编码二胍酸环化酶(DGCs)和/或磷酸二酯酶(PDEs),它们调节核苷酸第二信使c-di-GMP的细胞内池。虽然一般来说,高水平的c-二gmp驱动高运动表型向固定生活方式的转变,但不同的c-二gmp调节酶负责较小的和部分不重叠的表型。在这项研究中,我们试图利用先前记录的414个临床分离株的铜绿假单胞菌基因表达数据集来揭示编码DGCs的基因高表达导致的转录变化。这种方法可能提供了一个独特的机会来绕过许多c-二gmp调节酶的问题,即不知道在什么条件下它们的表达被激活。然而,虽然我们证明了临床分离株中sigma因子编码基因高表达和低表达亚群的选择有助于鉴定其下游调控,但我们无法确认预测的DGC调控,因为高c-di-GMP相关表型在临床分离株中迅速丢失。需要进一步的研究来确定长期培养临床铜绿假单胞菌分离株后环化酶活性丧失的具体机制。
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