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Pseudomonas Cyclic Lipopeptide Medpeptin:Biosynthesis and Modulation of Plant Immunity

Engineering 2023年第9期28卷 153-165页

作者： Yi-Lin Gu Jun-Zhou Li Yan Li Shen Cong Jing Wang Yi-Nan Ma Hai-Lei Wei State Key Laboratory of Efficient Utilization of Arid and Semi-arid Arable Land in Northern China Key Laboratory of Microbial Resources Collection and PreservationMinistry of Agriculture and Rural AffairsInstitute of Agricultural Resources and Regional PlanningChinese Academy of Agricultural SciencesBeijing 100081China Institute of Vegetables and Flowers Chinese Academy of Agricultural SciencesBeijing 100081China

The multifunctional secondary metabolites known as cyclic lipopeptides(CLPs),which are produced by a large variety of bacteria,have become a key category of plant immunity elicitors.Pseudomonas-CLPs(PsCLPs)are extremely diverse in structure and biological activity.However,an understanding of CLP-plant structure–function interactions currently remains elusive.Here,we identify medpeptin,a novel CLP from Pseudomonas mediterranea that consists of 22 amino acids.Medpeptin is synthesized by a non-ribosomal peptide synthase(NRPS)gene cluster and regulated by a quorum-sensing system.Further research indicates that medpeptin does not exhibit antimicrobial activity;instead,it induces plant cell death immunity and confers resistance to bacterial infection.Comparative transcriptome analysis and virus-induced gene silencing(VIGS)reveal a set of immune signaling candidates involved in medpeptin perception.Silencing of a cell-wall leucine-rich repeat extensin protein(NbLRX3)or a receptor-like protein kinase(NbRLK25)—but not BAK1 or SGT1—compromises medpeptin-triggered cell death and resistance to pathogen infection in Nicotiana benthamiana.Our findings point to a noncanonical mechanism of CLP sensing and suggest perspectives for the development of plant disease resistance.

关键词： Pseudomonas Cyclic lipopeptide Cell death Leucine-rich repeat extension(LRX) Medpeptin Receptor-like kinase(RLK)

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Enhanced fat degradation following the addition of a Pseudomonas species to a bioaugmentation product used in grease traps

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Journal of Environmental Sciences 2019年第3期31卷 174-188页

作者： Markella Tzirita Seraphim Papanikolaou Bríd Quilty School of Biotechnology Dublin City University (DCU) Department of Food Science and Human Nutrition Agricultural University of Athens National Institute for Cellular Biotechnology (NICB) DCU

The presence of fats, oils and greases(FOGs) in wastewater can lead to many problems including blockages. Investigation of a bioaugmentation product, consisting of Bacillus spp., to degrade butter(1%, W/V) and olive oil(1%, V/V) was performed in aerobic batch cultures for 13-day incubation. Gravimetric analysis of the remaining substrates showed slowly degradation of the oil after a 2-day lag, but no degradation of the butter. Addition of a Pseudomonas putida strain CP1 to the Bacillus spp. population promoted rapidly degradation of both fats after7 days of incubation. High fat accumulation revealed the potential use of the new bacterial mixture for production of added-value compounds. Lipase production only by the Bacillus spp.along with the analysis of the remaining lipids with thin layer chromatography and gas chromatography, suggested that the Bacillus spp. mainly only hydrolyzed the fat. The breakdown products were metabolized by the Pseudomonas putida CP1 performing preferential utilization of unsaturated fatty acids. Investigation of population dynamics using selective plating and a labeled Pseudomonas putida CP1::Tn7-gfp showed domination of the latter. The new mixture performed a successful cooperation with good potential for FOG treatment and an aggregative response desirable to fat degradation in grease traps.

关键词： Bacillus Bioaugmentation Biodegradation FOG Lipase Pseudomonas

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Phosphorylation of the Pseudomonas Effector AvrPtoB by Arabidopsis SnRK2.8 Is Required for Bacterial Virulence

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Molecular Plant 2020年第10期13卷 1513-1522页

作者： Lei Lei Danielle M.Stevens Gitta Coaker Department of Plant Pathology University of CaliforniaDavisDavisCAUSA

A critical component controlling bacterial virulence is the delivery of pathogen effectors into plant cells during infection.Effectors alter host metabolism and immunity for the benefit of pathogens.Multiple effectors are phosphorylated by host kinases,and this posttranslational modification is important for their activity.We sought to identify host kinases involved in effector phosphorylation.Multiple phosphorylated effector residues matched the proposed consensus motif for the plant calcium-dependent protein kinase(CDPK)and Snf1-related kinase(SnRK)superfamily.The conserved Pseudomonas effector AvrPtoB acts as an E3 ubiquitin ligase and promotes bacterial virulence.In this study,we identified a member of the Arabidopsis SnRK family,SnRK2.8,which interacts with AvrPtoB in yeast and in planta.We showed that SnRK2.8 was required for AvrPtoB virulence functions,including facilitating bacterial colonization,suppression of callose deposition,and targeting the plant defense regulator NPR1 and analyses receptor FLS2.Mass spectrometry analysis revealed that AvrPtoB phosphorylation occurs at multiple serine residues in planta,with S258 phosphorylation significantly reduced in the snrk2.8 knockout.AvrPtoB phospho-'null'mutants exhibited compromised virulence functions and were unable to suppress NPR1 accumulation,FLS2 accumulation,or inhibit FLS2-BAK1 complex formation upon flagellin perception.Taken together,these data identify a conserved plant kinase utilized by a pathogen effector to promote disease.

关键词： Arabidopsis Pseudomonas syringae effector AvrPtoB kinase SnRK2.8

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H-NS family protein MvaU downregulates phenazine-1-carboxylic acid(PCA)biosynthesis via binding to an AT-rich region within the promoter of the phz2 gene cluster in the rhizobacterium Pseudomonas strain PA1201

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Synthetic and Systems Biotechnology 2021年第4期6卷 262-271页

作者： Yun-Ling Fang Ying Cui Lian Zhou Chitti Thawai Tatheer Alam Naqvi Hong-Yan Zhang Ya-Wen He State Key Laboratory of Microbial Metabolism Joint International Research Laboratory of Metabolic and Developmental SciencesSJTU-Shanghai Nong Le Joint R&D Center on Biopesticides and BiofertilizersShanghai Jiao Tong University(SJTU)Shanghai200240China Department of Biology Faculty of ScienceKing Mongkut’s Institute of Technology LadkrabangBangkokThailand Department of Biotechnology COMSATS University IslamabadAbbottabad CampusAbbottabadPakistan Shanghai Nong Le Biological Products Company Limited Shanghai201419China

Histone-like nucleoid-structuring(H-NS)proteins are key regulators in gene expression silencing and in nucleoid compaction.The H-NS family member proteins MvaU in Pseudomonas aeruginosa are thought to bind the same AT-rich regions of chromosomes and function to coordinate the control of a common set of genes.Here,we explored the molecular mechanism by which MvaU controls PCA biosynthesis in P.aeruginosa PA1201.We present evidence suggesting that MvaU is self-regulated.Deletion of mvaU significantly increased PCA production,and PCA production sharply decreased when mvaU was over-expressed.MvaU transcriptionally repressed phz2 cluster expression and consequently reduced PCA biosynthesis.β-galactosidase assays confirmed that base pairing near the􀀀35 box is required when MvaU regulates PCA production in PA1201.Electrophoretic mobility shift assays(EMSA)and additional point mutation analysis demonstrated that MvaU directly bound to an AT-rich motif within the promoter of the phz2 cluster.Chromatin immunoprecipitation(ChIP)analysis also indicated that MvaU directly bound to the P5 region of the phz2 cluster promoter.MvaU repression of PCA biosynthesis was independent of QscR and OxyR in PA1201 and neither PCA or H2O2 were the environmental signals that induced mvaU expression.These findings detail a new MvaU-dependent regulatory pathway of PCA biosynthesis in PA1201 and provide a foundation to increase PCA fermentation titer by genetic engineering.

关键词： Pseudomonas Phenazine-1-carboxylic acid phz cluster MvaU Regulatory network

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Microbiologically influenced corrosion of titanium caused by aerobic marine bacterium Pseudomonas aeruginosa

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Journal of Materials Science & Technology 2019年第1期35卷 216-222页

作者： M.Saleem Khan Zhong Li Ke Yang Dake Xu Chunguang Yang Dan Liu Yassir Lekbach Enze Zhou Phuri Kalnaowakul Institute of Metal Research Chinese Academy of Sciences University of Chinese Academy of Sciences Corrosion and Protection Division Shenyang National Laboratory for Materials Science Northeastern University Laboratory of Microbial Biotechnology Faculty of Science and Technology Sidi Mohamed Ben Abdellah University Kasetsart University

Microbiologically influenced corrosion(MIC) is a big threat to the strength and safety of many metallic materials used in different environments throughout the world. The metabolites and bioactivity of the microorganisms cause severe deterioration on the metals. In this study, MIC of pure titanium(Ti) was studied in the presence of a highly corrosive aerobic marine bacterium Pseudomonas aeruginosa. The results obtained from electrochemical test showed that Ti was corrosion resistant in the abiotic culture medium after 14 d, while the increased corrosion current density(i_(corr)) obtained from polarization curves and the decreased charge transfer resistance(R_(ct)) from electrochemical impedance spectroscopy(EIS)indicated the accelerated corrosion of Ti caused by P. aeruginosa biofilm. For further confirmation of the above results, the surface of Ti was investigated using scanning electron microscopy(SEM), confocal laser scanning microscopy(CLSM) and X-ray photoelectron spectroscopy(XPS). According to the XPS results, TiO_2 was formed in both abiotic and biotic conditions, while unstable oxide Ti_2O_3 was detected in the presence of P. aeruginosa, leading to the defects in the passive film and localized corrosion. Pitting corrosion was investigated with the help of CLSM, and the largest pit depth found on Ti surface immersed in P. aeruginosa was 1.2 μm. Ti was not immune to MIC caused by P. aeruginosa.

关键词： Ti Microbiologically influenced corrosion Pseudomonas aeruginosa

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Pseudomonas sp.J1生物合成纳米硒机制研究

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广西大学学报（自然科学版） 2022年第1期47卷 219-228页

作者：张小萱任国栋刘畅陶文超张笑迎刘欢闵婷梅运军武汉轻工大学化学与环境工程学院湖北武汉430023 武汉轻工大学电气与电子工程学院湖北武汉430023 武汉轻工大学食品科学与工程学院湖北武汉430023

为了探究Pseudomonas sp.J1生物合成纳米硒的机制,采用电镜技术、X射线能谱分析法(EDS)及X射线光电子能谱分析法(XPS)对Pseudomonas sp.J1合成纳米硒过程及产物进行表征;通过添加谷胱甘肽抑制剂、改变培养基中NO_(3)^(-)、亚硒酸钠浓度... 详细信息

为了探究Pseudomonas sp.J1生物合成纳米硒的机制,采用电镜技术、X射线能谱分析法(EDS)及X射线光电子能谱分析法(XPS)对Pseudomonas sp.J1合成纳米硒过程及产物进行表征;通过添加谷胱甘肽抑制剂、改变培养基中NO_(3)^(-)、亚硒酸钠浓度以及培养温度等条件研究各因素对生物合成纳米硒的影响。结果表明:Pseudomonas sp.J1在胞质内合成纳米硒并向胞外转运;在生物合成纳米硒中谷胱甘肽起到了重要作用;亚硒酸钠浓度越高对细胞毒性越大;NO_(3)^(-)与SeO_(3)^(2-)形成竞争关系,不利于纳米硒的合成;当培养基中亚硒酸钠质量浓度为5 g/L且发酵温度为30℃时,Pseudomonas sp.J1转化效率最高,达到(47.73±2.15)% 。

关键词： Pseudomonas 纳米硒生物合成

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Selection of phages and conditions for the safe phage therapy against Pseudomonas aeruginosa infections

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Virologica Sinica 2015年第1期30卷 33-44页

作者： Victor Krylov Olga Shaburova Elena Pleteneva Sergey Krylov Alla Kaplan Maria Burkaltseva Olga Polygach Elena Chesnokova I.I. Mechnikov Research Institute for Vaccines & Sera RAMS Moscow 105064 Russian

The emergence of multidrug-resistant bacterial pathogens forced us to consider the phage therapy as one of the possible alternative approaches to treatment. The purpose of this paper is to consider the conditions for the safe, long-term use of phage therapy against various infections caused by Pseudomonas aeruginosa. We describe the selection of the most suitable phages, their most effective combinations and some approaches for the rapid recognition of phages unsuitable for use in therapy. The benefi ts and disadvantages of the various different approaches to the preparation of phage mixtures are considered, together with the specifi c conditions that are required for the safe application of phage therapy in general hospitals and the possibilities for the development of personalized phage therapy.

关键词： Pseudomonas aeruginosa bacteriophages phage therapy pseudolysogeny phage-bacteria interactions phage-phage interactions

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Production of rhamnolipids-producing enzymes of Pseudomonas in E. coli and structural characterization

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Frontiers of Chemical Science and Engineering 2017年第1期11卷 133-138页

作者： Kata Kiss Wei Ting Ng Qingxin Li Institute of Chemical & Engineering Sciences Agency for Science Technology and Research Jurong Island 627833 Singapore

Rhamnolipids are a class ofbiosurfactants that have a great potential to be used in industries. Five proteins/enzymes, namely RhlA, RhlB, RhlC, RhlG and RhlI, are critical for the production of rhamnolipids in Pseudomonas aeruginosa. Four of the 5 proteins except RhlC were successfully over-expressed in E. coli and three of them （RhlA, RhlB and RhlI） were purified and obtained in milligram quantities. The purified proteins were shown to be folded in solution. Homology models were built for RhlA, RhlB and RhlI. These results lay a basis for further structural and functional characterization of these proteins in vitro to favor the construction of super strains for rhamnolipids production.

关键词： rhamnolipids Pseudomonas RhlA RhlB,RhlI protein folding

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A rapid, simple, and sensitive immunoagglutination assay with silica nanoparticles for serotype identification of Pseudomonas aeruginosa

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Journal of Integrative Agriculture 2019年第8期18卷 1936-1943页

作者： DONG Xiu-mei TAO Jing LI Ting-ting ZHANG Ping ZHU Yan TANG Yu SU Rui-hong SHI Dong-fang College of Veterinary Medicine Northeast Agricultural University Harbin 150030 P.R.China Heilongjiang Key Laboratory for Animal Disease Control and Pharmaceutical Development College of Veterinary MedicineNortheast Agricultural University Harbin 150030 P.R.China

An agglutination test based on colored silica nanoparticles(colored SiNps) was established to detect serotypes of Pseudomonas aeruginosa. Monodisperse colored SiNps were used as agglutination test carriers. The colored SiNps were prepared through reverse microemulsion with reactive dyes, sensitized with 11 kinds of mono-specific antibodies against P. aeruginosa, and denoted as IgG-colored SiNps. Eleven kinds of IgG-colored SiNps were individually mixed with P. aeruginosa on a glass slide.Different serotypes of P. aeruginosa could be identified by agglutination test with evident agglutination. The P. aeruginosa could be detected in a range from 3.6×10^5 to 3.6×10^12 cfu mL^–1. This new agglutination test was confirmed to be a specific,sensitive, fast, easy-to-perform, and cost-efficient tool for the routine diagnosis of P. aeruginosa.

关键词： Pseudomonas aeruginosa colored silica nanoparticles agglutination test serotype detection

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十二烷基硫酸钠降解菌株Pseudomonas sp. SDS-2的降解能力及其低温降解活性的调控（英文）

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应用与环境生物学报 2019年第1期25卷 191-198页

作者：陈杨武张若木戴发志李旭东谭周亮中国科学院成都生物研究所中国科学院环境与应用微生物重点实验室成都610041 中国科学院成都生物研究所环境微生物四川省重点实验室成都610041 中国科学院大学北京100049

尽管生物法已广泛用于表面活性剂废水的处理,但低温对微生物的代谢活性产生明显不利影响,导致出水难以稳定达标.对筛选到的十二烷基硫酸钠(SDS)降解菌的降解能力进行考察,并对不同调控策略作用下该菌株的低温降解活性进行评估.对筛选到... 详细信息

尽管生物法已广泛用于表面活性剂废水的处理,但低温对微生物的代谢活性产生明显不利影响,导致出水难以稳定达标.对筛选到的十二烷基硫酸钠(SDS)降解菌的降解能力进行考察,并对不同调控策略作用下该菌株的低温降解活性进行评估.对筛选到的菌株进行16S rRNA基因序列测定与分析.该菌株在不同温度、pH、底物浓度、接种量下的降解能力以及不同调控策略(低温驯化、外源物质添加)下的低温降解活性均以化学需氧量(COD)的去除率间接表示.结果筛选到一株SDS降解菌,命名为SDS-2. 16S rRNA基因序列分析表明该菌株属于假单胞菌属(Pseudomonas sp.).该菌株的最佳生长条件为30℃、pH 9和120 mg/L氨浓度,而接种量对其降解活性无明显促进作用.当SDS初始浓度为2 500 mg/L时,该菌株对SDS的去除速率(以COD计)可达到355.3 mg L^(-1) h^(-1). 15℃下,长期驯化可使该菌株的降解活性达到30℃时的水平;10℃下,添加外源物质丁二酸钠和硝酸钾可使COD的去除率在48 h内分别提高25.3%和24.6%;外加蛋白胨和复合维生素可使COD的去除率在24 h内分别提高22.8%和11.7%.本研究筛选到的Pseudomonassp.SDS-2具有高的SDS降解活性,可为实际含SDS表面活性剂废水的处理提供微生物资源;同时,本研究中的调控策略亦可为SDS低温生物处理提供潜在处理方法.

关键词：十二烷基硫酸钠(SDS) Pseudomonas sp. SDS-2 低温驯化调控

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