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1.
Arch Microbiol ; 205(4): 123, 2023 Mar 20.
Article in English | MEDLINE | ID: mdl-36939906

ABSTRACT

An actinomycete, designated strain HSS6-12T, was isolated from hot spring sediment collected from Ranong province, Thailand. The strain showed taxonomic characteristics consistent with those of members of the genus Micromonospora. HSS6-12T produced a single spore directly on the substrate mycelium, and no aerial mycelium was detected. The isomer of diamino acid presented in cell wall peptidoglycan was meso-diaminopimelic acid. Arabinose, xylose, glucose, and ribose were detected in whole-cell hydrolysates. MK-10(H4), MK-9(H4), and MK-10(H6) were major menaquinones. Major cellular fatty acids were iso-C16:0, iso-C15:0, and iso-C17:0. Phospholipid profile was composed of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, and phosphatidylinositolmannosides. 16S rRNA gene analysis revealed that HSS6-12T shared the highest 16S rRNA gene sequence similarity with Micromonospora inositola DSM 43819T (99.3%). In contrast, the genome analysis showed that HSS6-12T formed a tight taxonomic position in a phylogenomic tree with Micromonospora endolithica DSM 44398T. Moreover, the average nucleotide identity-blast, the digital DNA-DNA hybridization, and the average amino acid identity values between HSS6-12T and M. inositola DSM 43819T and M. endolithica DSM 44398T were 83.1-84.0%, 27.5-28.7%, and 80.4-82.2%, respectively, indicating that HSS6-12T was different species with both closely related Micromonospora-type strains. In addition, HSS6-12T could be discriminated from its closely related type strains by many physiological and biochemical characteristics. Thus, HSS6-12T could be considered a novel species of the genus Micromonospora, and the name Micromonospora thermarum is proposed for the strain. The type strain is HSS6-12T (= BCC 41915T = JCM 17127T).


Subject(s)
Actinobacteria , Hot Springs , Micromonospora , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Phospholipids/analysis , Fatty Acids/analysis , Phylogeny , Vitamin K 2/chemistry , Actinobacteria/genetics , DNA , DNA, Bacterial/genetics , Bacterial Typing Techniques
2.
Arch Microbiol ; 205(6): 247, 2023 May 22.
Article in English | MEDLINE | ID: mdl-37212915

ABSTRACT

An actinomycete strain, AA8T, which produced a long straight chain of spores (verticillati type), was isolated from the rhizosphere soil of Mangifera indica in Bangkok, Thailand. A polyphasic taxonomic study was carried out to establish the taxonomic position of the strain. Strain AA8T formed a tight taxonomic position in the 16S rRNA gene tree with Streptomyces roseifaciens MBT76T. In contrast, the genome-based taxonomic analysis showed that strain AA8T shared low average nucleotide identity-BLAST (94.1%), the digital DNA-DNA hybridization (58.2%), and the average amino acid identity (93.6%) values with S. roseifaciens MBT76T. Moreover, a combination of physiological and biochemical properties indicated that strain AA8T was distinguished from all Streptomyces species with effectively published names. Strain AA8T, therefore, represents a novel species of Streptomyces, and the name Streptomyces telluris is proposed for the strain. The type strain is AA8T (= TBRC 8483T = NBRC 113461T). The chemical investigation led to the isolation of nine known compounds (compounds 1-9). Among these compounds, compound 7 (3,4-dihydroxybenzaldehyde) possesses strong antioxidant activity equal to ascorbic acid, a powerful antioxidative agent.


Subject(s)
Streptomyces , Fatty Acids/chemistry , Phospholipids/chemistry , Antioxidants , Sequence Analysis, DNA , RNA, Ribosomal, 16S/genetics , Diaminopimelic Acid/chemistry , Phylogeny , DNA, Bacterial/genetics , Bacterial Typing Techniques , Base Composition , Soil Microbiology , Thailand
3.
Article in English | MEDLINE | ID: mdl-37288648

ABSTRACT

An endophytic actinobacterium, designated strain PLAI 1-29T, was isolated from the root tissue of Zingiber montanum collected from Pathum Thani province, Thailand. Strain PLAI 1-29T was characterized using a polyphasic taxonomic approach. It typically exhibited morphological and chemotaxonomic properties of the genus Streptomyces. Strain PLAI 1-29T produced a spiral spore chain on aerial mycelium and grew at 15-40 °C, pH 6-10 on International Streptomyces Project 2 agar. The maximum NaCl concentration for growth was 9 % (w/v). Cells of strain PLAI 1-29T presented ll-diaminopimelic acid, arabinose, galactose and ribose. The detected phospholipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside. The major menaquinones were MK-9(H6) and MK-9(H8). The major cellular fatty acids were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The genome-based taxonomic details revealed the assignment of strain PLAI 1-29T to the genus Streptomyces and exhibited low threshold values for the delineation of a novel species by average nucleotide identity-blast (84.0%), average amino acid identity (80.0%) and digital DNA-DNA hybridization (27.6%) with its closest type strain, Streptomyces xinghaiensis S187T. Furthermore, several differential physiological and biochemical characteristics were detected between strain PLAI 1-29T and the closest type strain. Based on the combined phenotypic and genomic features, strain PLAI 1-29T (=TBRC 7645T=NBRC 113170T) is considered to represent a new Streptomyces species, for which we propose the name Streptomyces zingiberis sp. nov.


Subject(s)
Actinobacteria , Streptomyces , Fatty Acids/chemistry , Sequence Analysis, DNA , Phylogeny , Base Composition , RNA, Ribosomal, 16S/genetics , DNA, Bacterial/genetics , Bacterial Typing Techniques , Phospholipids/chemistry , Actinobacteria/genetics
4.
Article in English | MEDLINE | ID: mdl-36961870

ABSTRACT

An actinobacterium strain, SW21T, was isolated from seawater collected in the upper Gulf of Thailand. Cells were Gram-stain-positive, aerobic and rod-shaped. Growth was observed from 15 to 37 °C and at pH 6-8. Maximum NaCl for growth was 14 % (w/v). meso-Diaminopimelic acid, arabinose, galactose, glucose, rhamnose and ribose were detected in the whole-cell hydrolysate. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside were detected as the phospholipids in the cells. The major menaquinones were MK-9(H2) and MK-7(H2). The major cellular fatty acids were C16 : 0, C18 : 1 ω9c, C18 : 0 and C18 : 010-methyl (TBSA). The 16S rRNA gene sequence data supported the assignment of strain SW21T to the genus Gordonia and showed that Gordonia mangrovi KCTC 49383T (98.7 %) was the closest relative. Moreover, the average nucleotide identity-blast (85.5 %) and digital DNA-DNA hybridization (30.7 %) values between strain SW21T and its closest neighbour were below the threshold values for delineation of a novel species. The combination of genotypic and phenotypic data indicated that strain SW21T is representative of novel species of the genus Gordonia. The name Gordonia aquimaris sp. nov. is proposed for strain SW21T. The type strain is SW21T (=TBRC 15691T=NBRC 115558T).


Subject(s)
Actinobacteria , Gordonia Bacterium , Fatty Acids/chemistry , Thailand , RNA, Ribosomal, 16S/genetics , Phylogeny , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Sequence Analysis, DNA , Phospholipids , Seawater
5.
Article in English | MEDLINE | ID: mdl-37141114

ABSTRACT

An actinobacterium strain, PPF5-17T, was isolated from hot spring soil collected from Chiang Rai province, Thailand. The strain exhibited morphological and chemotaxonomic properties similar to those of members of the genus Micromonospora. Colonies of PPF5-17T were strong pinkish red and turned black after sporulation in ISP 2 agar medium. Cells formed single spores directly on the substrate mycelium. Growth was observed from 15 to 45 °C and at pH 5-8. Maximum NaCl concentration for growth was 3 % (w/v). PPF5-17T was found to have meso-diaminopimelic acid, xylose, mannose and glucose in the whole-cell hydrolysate. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositolmannosides were observed as the membrane phospholipids. MK-10(H6), MK-9(H6), MK-10(H4) and MK-9(H4) were the major menaquinones. The predominant cellular fatty acids were iso-C15 : 0, iso-C17 : 0, anteiso-C17 : 0 and iso-C16 : 0. PPF5-17T shared the highest 16S rRNA gene sequence similarity with Micromonospora fluminis LMG 30467T (99.3 %). A genome-based taxonomic study revealed that PPF5-17T was closely related to Micromonospora aurantinigra DSM 44815T in the phylogenomic tree with an average nucleotide identity by blast (ANIb) of 87.7 % and a digital DNA-DNA hybridization (dDDH) value of, 36.1 % which were below the threshold values for delineation of a novel species. Moreover, PPF5-17T could be distinguished from its closest neighbours, M. fluminis LMG 30467T and M. aurantinigra DSM 44815T, with respect to a broad range of phenotypic properties. Thus, PPF5-17T represents a novel species, for which the name Micromonospora solifontis sp. nov. is proposed. The type strain is PPF5-17T (= TBRC 8478T = NBRC 113441T).


Subject(s)
Actinobacteria , Hot Springs , Micromonospora , Fatty Acids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Phylogeny , Thailand , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Phospholipids/chemistry , Actinobacteria/genetics
6.
Article in English | MEDLINE | ID: mdl-35446756

ABSTRACT

A member of the genus Streptomyces, designated RCPT1-4T, was isolated from compost of Senna siamea (Lam.), collected from an agricultural area in Rayong province, Thailand. The spore morphology and the presence of ll-diaminopimelic acid in the peptidoglycan indicate that RCPT1-4T shows the typical properties of members of the genus Streptomyces. On the basis of the results of 16S rRNA gene sequence analysis, the strain should be classified as representing a member of the genus Streptomyces and was most closely related to Streptomyces fumigatiscleroticus NBRC 12999T with the highest 16S rRNA gene sequence similarity of 99.2 %, followed by Streptomyces spiralis NBRC 14215T (99.0 %). In addition, RCPT1-4T shared the highest average nucleotide identity by blast (ANIb) (86.0 %), and digital DNA-DNA hybridization (dDDH) (32.1 %) values with S. spiralis NBRC 14215T. Furthermore, several physiological and biochemical differences were observed between RCPT1-4T and the closely related type strains of species with validly published names. These taxonomic data indicated that RCPT1-4T could be considered to represent a novel species of the genus Streptomyces and the name Streptomyces sennicomposti sp. nov. is proposed for this strain. The type strain is RCPT1-4T (=TBRC 11260T=NBRC 114303T).


Subject(s)
Actinobacteria , Composting , Streptomyces , Actinobacteria/genetics , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Nucleic Acid Hybridization , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Thailand
7.
Int J Syst Evol Microbiol ; 72(12)2022 Dec.
Article in English | MEDLINE | ID: mdl-36748585

ABSTRACT

An actinobacterium strain PLK6-54T was isolated from Lankwai peat swamp forest soil collected from Yala province, Thailand. Strain PLK6-54T exhibited morphological and chemotaxonomic properties described for the genus Streptomyces. It formed a spiral spore chain directly on aerial mycelium. Growth was observed between 20 and 40 °C and at pH 5-8. The maximum NaCl for growth was 2 % (w/v). ll-Diaminopimelic acid, arabinose and ribose were detected in the whole-cell hydrolysate. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositolmannoside were detected as the phospholipids. The major menaquinones were MK-10(H2) and MK-9(H6). The major cellular fatty acids were iso-C16 : 0, anteiso-C15 : 0 and iso-C14 : 0. 16S rRNA gene sequence data supported the assignment of strain PLK6-54T to the genus Streptomyces and showed that Streptomyces rubidus NBRC 102073T (99.0 %) was the closest relative. Moreover, the average nucleotide identity-blast (85.5 %) and digital DNA-DNA hybridization (30.7 %) values reported between strain PLK6-54T and its closest neighbour were below the threshold values for delineation of a novel species. Strain PLK6-54T could be distinguished from related validly described Streptomyces species by several phenotypic properties. The combination of genotypic and phenotypic data indicated that strain PLK6-54T is representative of a novel species of the genus Streptomyces. The name Streptomyces acidipaludis sp. nov. is proposed for strain PLK6-54T. The type strain is PLK6-54T (=TBRC 11250T=NBRC 114297T).


Subject(s)
Fatty Acids , Streptomyces , Fatty Acids/chemistry , RNA, Ribosomal, 16S/genetics , Wetlands , Soil , Sequence Analysis, DNA , Phylogeny , Bacterial Typing Techniques , DNA, Bacterial/genetics , Base Composition , Phospholipids/chemistry , Forests
8.
J Appl Microbiol ; 132(2): 772-784, 2022 Feb.
Article in English | MEDLINE | ID: mdl-34260807

ABSTRACT

Extensively produced by members of the genus Streptomyces, piericidins are a large family of microbial metabolites, which consist of main skeleton of 4-pyridinol with methylated polyketide side chain. Nonetheless, these metabolites show differences in their bioactive potentials against micro-organisms, insects and tumour cells. Due to its close structural similarity with coenzyme Q, piericidins also possess an inhibitory activity against NADH dehydrogenase as well as Photosystem II. This review studied the latest research progress of piericidins, covering the chemical structure and physical properties of newly identified members, bioactivities, biosynthetic pathway with gene clusters and future prospect. With the increasing incidence of drug-resistant human pathogen strains and cancers, this review aimed to provide clues for the development of either new potential antibiotics or anti-tumour agents.


Subject(s)
Anti-Bacterial Agents , Antineoplastic Agents , Pyridines/pharmacology , Streptomyces , Anti-Bacterial Agents/pharmacology , Antineoplastic Agents/pharmacology , Multigene Family , Streptomyces/chemistry , Streptomyces/genetics
9.
Int J Mol Sci ; 23(13)2022 Jun 26.
Article in English | MEDLINE | ID: mdl-35806111

ABSTRACT

Bacteria and their eukaryotic hosts have co-evolved for millions of years, and the former can intercept eukaryotic signaling systems for the successful colonization of the host. The diffusible signal factor (DSF) family represents a type of quorum-sensing signals found in diverse Gram-negative bacterial pathogens. Recent evidence shows that the DSF is involved in interkingdom communications between the bacterial pathogen and the host plant. In this study, we explored the anti-inflammatory effect of the DSF and its underlying molecular mechanism in a zebrafish model. We found that the DSF treatment exhibited a strong protective effect on the inflammatory response of zebrafish induced by lipopolysaccharide (LPS). In the LPS-induced inflammation zebrafish model, the DSF could significantly ameliorate the intestinal pathological injury, reduce abnormal migration and the aggregation of inflammatory cells, inhibit the excessive production of inflammatory mediator reactive oxygen species (ROS) content, and prevent apoptosis. Through an RNA-Seq analysis, a total of 938 differentially expressed genes (DEGs) was screened between LPS and LPS + DSF treatment zebrafish embryos. A further bioinformatics analysis and validation revealed that the DSF might inhibit the LPS-induced zebrafish inflammatory response by preventing the activation of signaling in the Toll-like receptor pathway, attenuating the expression of pro-inflammatory cytokines and chemokines, and regulating the activation of the caspase cascade through restoring the expression of lysosomal cathepsins and apoptosis signaling. This study, for the first time, demonstrates the anti-inflammatory role and a potential pharmaceutical application of the bacterial signal DSF. These findings also suggest that the interkingdom communication between DSF-producing bacteria and zebrafish might occur in nature.


Subject(s)
Lipopolysaccharides , Zebrafish , Animals , Apoptosis , Bacteria , Inflammation/drug therapy , Lipopolysaccharides/toxicity , Lysosomes , Quorum Sensing/physiology , Toll-Like Receptors
10.
Int J Mol Sci ; 23(20)2022 Oct 18.
Article in English | MEDLINE | ID: mdl-36293339

ABSTRACT

The increasing prevalence of resistance in carbapenems is an escalating concern as carbapenems are reserved as last-line antibiotics. Although indiscriminate antibiotic usage is considered the primary cause for resistance development, increasing evidence revealed that inconsequential strains without any direct clinical relevance to carbapenem usage are harboring carbapenemase genes. This phenomenon indirectly implies that environmental microbial populations could be the 'hidden vectors' propelling carbapenem resistance. This work aims to explore the carbapenem-resistance profile of Vibrio species across diverse settings. This review then proceeds to identify the different factors contributing to the dissemination of the resistance traits and defines the transmission pathways of carbapenem resistance. Deciphering the mechanisms for carbapenem resistance acquisition could help design better prevention strategies to curb the progression of antimicrobial resistance development. To better understand this vast reservoir selecting for carbapenem resistance in non-clinical settings, Vibrio species is also prospected as one of the potential indicator strains for carbapenem resistance in the environment.


Subject(s)
Vibrio , beta-Lactamases , beta-Lactamases/metabolism , Carbapenems/pharmacology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Anti-Bacterial Agents/pharmacology , Vibrio/genetics , Vibrio/metabolism , Microbial Sensitivity Tests
11.
Mol Microbiol ; 114(5): 870-886, 2020 11.
Article in English | MEDLINE | ID: mdl-32757400

ABSTRACT

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot in crucifers. Our previous findings revealed that Xcc can degrade 4-hydroxybenzoic acid (4-HBA) via the ß-ketoadipate pathway. This present study expands on this knowledge in several ways. First, we show that infective Xcc cells induce in situ biosynthesis of 4-HBA in host plants, and Xcc can efficiently degrade 4-HBA via the pobA/pobR locus, which encodes a 4-hydroxybenzoate hydroxylase and an AraC-family transcription factor respectively. Next, the transcription of pobA is specifically induced by 4-HBA and is positively regulated by PobR, which is constitutively expressed in Xcc. 4-HBA directly binds to PobR dimers, resulting in activation of pobA expression. Point mutation and subsequent isothermal titration calorimetry and size exclusion chromatography analysis identified nine key conserved residues required for 4-HBA binding and/or dimerization of PobR. Furthermore, overlapping promoters harboring fully overlapping -35 elements were identified between the divergently transcribed pobA and pobR. The 4-HBA/PobR dimer complex specifically binds to a 25-bp site, which encompasses the -35 elements shared by the overlapping promoters. Finally, GUS histochemical staining and subsequent quantitative assay showed that both pobA and pobR genes are transcribed during Xcc infection of Chinese radish, and the strain ΔpobR exhibited compromised virulence in Chinese radish. These findings suggest that the ability of Xcc to survive the 4-HBA stress might be important for its successful colonization of host plants.


Subject(s)
Parabens/metabolism , Xanthomonas campestris/genetics , Xanthomonas campestris/metabolism , AraC Transcription Factor/genetics , Bacterial Proteins/metabolism , DNA, Bacterial/metabolism , Gene Expression Regulation, Bacterial/genetics , Parabens/chemistry , Trans-Activators/metabolism , Transcription Factors/metabolism , Virulence/genetics , Xanthomonas campestris/pathogenicity
12.
Mol Plant Microbe Interact ; 33(5): 705-714, 2020 May.
Article in English | MEDLINE | ID: mdl-32027580

ABSTRACT

Xanthomonadins are membrane-bound yellow pigments that are typically produced by phytopathogenic bacterial Xanthomonas spp., Xylella fastidiosa, and Pseudoxanthomonas spp. They are also produced by a diversity of environmental bacterial species. Considerable research has revealed that they are a unique group of halogenated, aryl-polyene, water-insoluble pigments. Xanthomonadins have been shown to play important roles in epiphytic survival and host-pathogen interactions in the phytopathogen Xanthomonas campestris pv. campestris, which is the causal agent of black rot in crucifers. Here, we review recent advances in the understanding of xanthomonadin chemical structures, physiological roles, biosynthetic pathways, regulatory mechanisms, and crosstalk with other signaling pathways. The aim of the present review is to provide clues for further in-depth research on xanthomonadins from Xanthomonas and other related bacterial species.


Subject(s)
Anisoles/chemistry , Xanthomonas campestris/chemistry , Biosynthetic Pathways , Signal Transduction
13.
Mol Plant Microbe Interact ; 33(3): 488-498, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31710580

ABSTRACT

Phenazine-1-carboxylic acid (PCA) is the primary active component in the newly registered, commercial biopesticide Shenqinmycin and is produced during fermentation by the engineered rhizobacterium strain Pseudomonas PA1201. Both phz1 and phz2 gene clusters contribute to PCA biosynthesis. In this study, we evaluated the role of OxyR in the regulation of PCA biosynthesis in PA1201. We first showed a functional link between oxyR expression and PCA biosynthesis. Deletion of oxyR and overexpression of oxyR both increase PCA biosynthesis. The molecular mechanisms underlying OxyR regulation of PCA production were investigated using several approaches. OxyR acts divergently in phz1 and phz2. Overexpression of oxyR activated the expression of phz1 and phz1-dependent PCA production. However, overexpression of oxyR had little effect on phz2-dependent PCA biosynthesis, while deletion of oxyR promoted phz2-dependent PCA production and exerted a negative effect on phz2 expression. Further, OxyR directly bound to the phz2 promoter region. In addition, the regulation of PCA biosynthesis by OxyR was associated with quorum sensing (QS) systems. Overexpression of OxyR positively regulated pqs QS system. Finally, transcriptomic analysis and subsequent genetic analysis revealed the small RNA phrS plays a key role in OxyR-dependent PCA accumulation. Specifically, OxyR directly binds to the phrS promoter region to positively regulate phrS expression wherein PhrS regulates the PCA positive regulator MvfR in order to control PCA biosynthesis.


Subject(s)
Pseudomonas aeruginosa/genetics , RNA/genetics , Trans-Activators/genetics , Bacterial Proteins/genetics , Gene Expression Regulation, Bacterial , Phenazines/metabolism , Promoter Regions, Genetic , Pseudomonas aeruginosa/metabolism , Quorum Sensing
14.
Appl Microbiol Biotechnol ; 104(3): 1013-1034, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31858191

ABSTRACT

The whole organisms can be packaged as biopesticides, but secondary metabolites secreted by microorganisms can also have a wide range of biological activities that either protect the plant against pests and pathogens or act as plant growth promotors which can be beneficial for the agricultural crops. In this review, we have compiled information about the most important secondary metabolites of three important bacterial genera currently used in agriculture pest and disease management.


Subject(s)
Bacteria/metabolism , Biological Control Agents , Secondary Metabolism , Agriculture/methods , Bacillus/metabolism , Crops, Agricultural , Pest Control, Biological , Pseudomonas/metabolism , Serratia/metabolism
15.
Phytopathology ; 110(2): 278-286, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31613175

ABSTRACT

A characteristic feature of phytopathogenic Xanthomonas bacteria is the production of yellow membrane-bound pigments called xanthomonadins. Previous studies showed that 3-hydroxybenzoic acid (3-HBA) was a xanthomonadin biosynthetic intermediate and also, that it had a signaling role. The question of whether the structural isomers 4-HBA and 2-HBA (salicylic acid) have any role in xanthomonadin biosynthesis remained unclear. In this study, we have selectively eliminated 3-HBA, 4-HBA, or the production of both by expression of the mhb, pobA, and pchAB gene clusters in the Xanthomonas campestris pv. campestris strain XC1. The resulting strains were different in pigmentation, virulence factor production, and virulence. These results suggest that both 3-HBA and 4-HBA are involved in xanthomonadin biosynthesis. When both 3-HBA and 4-HBA are present, X. campestris pv. campestris prefers 3-HBA for Xanthomonadin-A biosynthesis; the 3-HBA-derived Xanthomonadin-A was predominant over the 4-HBA-derived xanthomonadin in the wild-type strain XC1. If 3-HBA is not present, then 4-HBA is used for biosynthesis of a structurally uncharacterized Xanthomonadin-B. Salicylic acid had no effect on xanthomonadin biosynthesis. Interference with 3-HBA and 4-HBA biosynthesis also affected X. campestris pv. campestris virulence factor production and reduced virulence in cabbage and Chinese radish. These findings add to our understanding of xanthomonadin biosynthetic mechanisms and further help to elucidate the biological roles of xanthomonadins in X. campestris pv. campestris adaptation and virulence in host plants.


Subject(s)
Hydroxybenzoates , Parabens , Pigments, Biological , Xanthomonas campestris , Hydroxybenzoates/metabolism , Parabens/metabolism , Pigments, Biological/biosynthesis , Pigments, Biological/genetics , Plant Diseases/microbiology , Virulence Factors/genetics , Xanthomonas campestris/genetics , Xanthomonas campestris/metabolism , Xanthomonas campestris/pathogenicity
16.
Mol Plant Microbe Interact ; 32(2): 217-226, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30681910

ABSTRACT

Coenzyme Q (CoQ) is a lipid-soluble membrane component found in organisms ranging from bacteria to mammals. The biosynthesis of CoQ has been intensively studied in Escherichia coli, where 12 genes (ubiA, -B, -C, -D, -E, -F, -G, -H, -I, -J, -K, and -X) are involved. In this study, we first investigated the putative genes for CoQ8 biosynthesis in the phytopathogen Xanthomonas campestris pv. campestris using a combination of bioinformatic, genetic, and biochemical methods. We showed that Xc_0489 (coq7Xc) encodes a di-iron carboxylate monooxygenase filling the E. coli UbiF role for hydroxylation at C-6 of the aromatic ring. Xc_0233 (ubiJXc) encodes a novel protein with an E. coli UbiJ-like domain organization and is required for CoQ8 biosynthesis. The X. campestris pv. campestris decarboxylase gene remains unidentified. Further functional analysis showed that ubiB and ubiK homologs ubiBXc and ubiKXc are required for CoQ8 biosynthesis in X. campestris pv. campestris. Deletion of ubiJXc, ubiBXc, and ubiKXc led to the accumulation of an intermediate 3-octaprenyl-4-hydroxybenzoic acid. UbiKXc interacts with UbiJXc and UbiBXc to form a regulatory complex. Deletion analyses of these CoQ8 biosynthetic genes indicated that they are important for virulence in Chinese radish. These results suggest that the X. campestris pv. campestris CoQ8 biosynthetic reactions and regulatory mechanisms are divergent from those of E. coli. The variations provide an opportunity for the design of highly specific inhibitors for the prevention of infection by the phytopathogen X. campestris pv. campestris.


Subject(s)
Ubiquinone , Xanthomonas campestris , Escherichia coli/genetics , Saccharomyces cerevisiae/genetics , Ubiquinone/biosynthesis , Xanthomonas campestris/enzymology , Xanthomonas campestris/genetics
17.
Mol Plant Microbe Interact ; 32(2): 240-254, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30570452

ABSTRACT

Xanthomonas campestris pv. campestris uses the diffusible signal factor (DSF) family of quorum-sensing (QS) signals to coordinate virulence and adaptation. DSF family signals have been well-characterized using laboratory-based cell cultures. The in-planta QS signal used during X. campestris pv. campestris infection remains unclear. To achieve this goal, we first mimic in-planta X. campestris pv. campestris growth conditions by supplementing the previously developed XYS medium with cabbage hydrolysate and found that the dominant signal produced in these conditions was BDSF. Secondly, by using XYS medium supplemented with diverse plant-derived compounds, we examined the effects of diverse plant-derived compounds on the biosynthesis of DSF family signals. Several compounds were found to promote biosynthesis of BDSF. Finally, using an X. campestris pv. campestris ΔrpfB-Chinese cabbage infection model and an ultra-performance liquid chromatographic-time of flight-mass spectrometry-based assay, BDSF was found to comprise >70% of the DSF family signals present in infected cabbage tissue. BDSF at a concentration of 2.0 µM induced both protease activity and engXCA expression. This is the first report to directly show that BDSF is the predominant in-planta QS signal used during X. campestris pv. campestris infection. It provides a better understanding of the molecular interactions between X. campestris pv. campestris and its cruciferous hosts and also provides the logical target for designing strategies to counteract BDSF signaling and, thus, infection. Further studies are needed to get an exact idea about the DSF production dynamics of the wild-type strain inside the plant.


Subject(s)
Bacterial Proteins , Brassica , Signal Transduction , Xanthomonas campestris , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Brassica/microbiology , Signal Transduction/genetics , Virulence/genetics , Xanthomonas campestris/genetics , Xanthomonas campestris/physiology
18.
Mol Microbiol ; 110(1): 16-32, 2018 10.
Article in English | MEDLINE | ID: mdl-29995983

ABSTRACT

Xanthomonadins are yellow pigments that are produced by the phytopathogen Xanthomonas campestris pv. campestris (Xcc). A pig cluster is responsible for xanthomonadin biosynthesis. Previously, Xcc4014 of the cluster was characterized as a bifunctional chorismatase that produces 3-hydroxybenzoic acid (3-HBA) and 4-HBA. In this study, genetic analysis identified 11 genes within the pig cluster to be essential for xanthomonadin biosynthesis. Biochemical and bioinformatics analysis suggest that xanthomonadins are synthesized via an unusual type II polyketide synthase pathway. Heterologous expression of the pig cluster in non-xanthomonadin-producing Pseudomonas aeruginosa strain resulted in the synthesis of chlorinated xanthomonadin-like pigments. Further analysis showed that xanC encodes an acyl carrier protein (ACP) while xanA2 encodes a ATP-dependent 3-HBA:ACP ligase. Both of them act together to catalyse the formation of 3-HBA-S-ACP from 3-HBA to initiate xanthomonadin biosynthesis. Finally, we showed that xanH encodes a FabG-like enzyme and xanK encodes a novel glycosyltransferase. Both xanH and xanK are not only required for xanthomonadin biosynthesis, but also required for the balanced biosynthesis of extracellular polysaccharides and DSF-family quorum sensing signals. These findings provide us with a better understanding of xanthomonadin biosynthetic mechanisms and directly demonstrate the presence of extensive cross-talk among xanthomonadin biosynthetic pathways and other metabolic pathways.


Subject(s)
Anisoles/metabolism , Biosynthetic Pathways/genetics , DNA, Bacterial/genetics , Xanthomonas campestris/enzymology , Xanthomonas campestris/genetics , Acyl Carrier Protein/genetics , Acyl Carrier Protein/metabolism , Adenosine Triphosphate/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Glycosyltransferases/genetics , Glycosyltransferases/metabolism , Hydroxybenzoates/metabolism , Multigene Family/genetics , Polyketide Synthases/genetics , Polyketide Synthases/metabolism , Quorum Sensing
19.
Appl Microbiol Biotechnol ; 103(23-24): 9287-9303, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31707442

ABSTRACT

Synthetic chemical pesticides have been used for many years to increase the yield of agricultural crops. However, in the future, this approach is likely to be limited due to negative impacts on human health and the environment. Therefore, studies of the secondary metabolites produced by agriculturally important microorganisms have an important role in improving the quality of the crops entering the human food chain. In this review, we have compiled information about the most important secondary metabolites of fungal species currently used in agriculture pest and disease management.


Subject(s)
Anti-Infective Agents/metabolism , Biological Control Agents/metabolism , Crops, Agricultural/microbiology , Fungi/metabolism , Secondary Metabolism , Agriculture , Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Biological Control Agents/chemistry , Biological Control Agents/pharmacology , Crops, Agricultural/drug effects , Crops, Agricultural/growth & development , Fungi/classification , Pest Control, Biological , Pesticides/chemistry , Pesticides/metabolism , Pesticides/pharmacology
20.
J Ind Microbiol Biotechnol ; 46(7): 1025-1038, 2019 Jul.
Article in English | MEDLINE | ID: mdl-30989356

ABSTRACT

Coenzyme Q (ubiquinone) is a redox-active isoprenylated benzoquinone commonly found in living organisms. The biosynthetic pathway for this lipid has been extensively studied in Escherichia coli and Saccharomyces cerevisiae; however, little is known in Pseudomonas aeruginosa. In this study, we observed that CoQ9 is the predominant coenzyme Q synthesized by the Shenqinmycin-producing strain M18. BLASTP and domain organization analyses identified 15 putative genes for CoQ biosynthesis in M18. The roles of 5 of these genes were genetically and biochemically investigated. PAM18_4662 encodes a nonaprenyl diphosphate synthase (Nds) and determines the number of isoprenoid units of CoQ9 in M18. PAM18_0636 (coq7PA) and PAM18_5179 (ubiJPA) are essential for aerobic growth and CoQ9 biosynthesis. Deletion of ubiJPA, ubiBPA and ubiKPA led to reduced CoQ biosynthesis and an accumulation of the CoQ9 biosynthetic intermediate 3-nonaprenylphenol (NPP). Moreover, we also provide evidence that the truncated UbiJPA interacts with UbiBPA and UbiKPA to affect CoQ9 biosynthesis by forming a regulatory complex. The genetic diversity of coenzyme Q biosynthesis may provide targets for the future design of specific drugs to prevent P. aeruginosa-related infections.


Subject(s)
Biological Control Agents/metabolism , Pseudomonas aeruginosa/metabolism , Ubiquinone/biosynthesis , Biosynthetic Pathways/genetics , Pseudomonas aeruginosa/genetics
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