Journal of Microbiology

Enhancing Seed Germination of Cremastra appendiculata: Screening and Identification of Four New Symbiotic Fungi in the Psathyrellaceae Family: Zhangneng Pan, Jing Wang, Shanshan He, Haiyang Zhao, Xinyue Dong, Tao Feng, Yanyan Meng, Xiaojun Li; J. Microbiol. 2024;62(8):671-682. Published online June 28, 2024; DOI: https://doi.org/10.1007/s12275-024-00148-7

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Abstract: Several coprinoid fungi have been identified as promotors of Cremastra appendiculata seed germination, while others appear ineffective. This study aimed to discern which genera within the Psathyrellaceae family exhibit this capability and to identify the most effective coprinoid fungi for the cultivation of C. appendiculata. We collected 21 coprinoid fungi from diverse sources and symbiotically cultured them with C. appendiculata seeds. 9 fungi were found to induce seed germination and support seed development, specifically within the genera Coprinellus, Tulosesus, and Candolleomyces. In contrast, fungi that failed to promote germination predominantly belonged to the genera Coprinopsis and Parasola. Notably, four fungi-Coprinellus xanthothrix, Coprinellus pseudodisseminatus, Psathyrella singeri, and Psathyrella candolleana-were documented for the first time as capable of enhancing C. appendiculata seed germination. Strain 218LXJ-10, identified as Coprinellus radians, demonstrated the most significant effect and has been implemented in large-scale production, underscoring its considerable practical value. These findings contribute vital scientific insights for the conservation and sustainable use of C. appendiculata resources.

DNA vaccine dual-expressing viral hemorrhagic septicemia virus glycoprotein and C-C motif chemokine ligand 19 induces the expression of immune-related genes in zebrafish (Danio rerio): Jin-Young Kim , Hyoung Jun Kim , Jeong Su Park , Se Ryun Kwon; J. Microbiol. 2022;60(10):1032-1038. Published online August 1, 2022; DOI: https://doi.org/10.1007/s12275-022-2231-8

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Glycoprotein (G protein)-based DNA vaccines are effective in protecting aquaculture fish from rhabdoviruses but the degree of immune response they elicit depends on plasmid concentration and antigen cassette. Here, we developed a DNA vaccine using the viral hemorrhagic septicemia virus G (VG) gene and chemokine (C-C motif) ligand 19 (CCL19)a.2 regulated by the CMV promoter as the molecular adjuvant. After transfection of the prepared plasmid (pVG + CCL19) into epithelioma papulosum cyprini cells, mRNA expression was confirmed through quantitative real-time polymerase chain reaction. The vaccine was intramuscularly injected into zebrafish (Danio rerio), and 28 days after immunization, viral hemorrhagic septicemia virus (105 TCID50/10 μl/fish) was intraperitoneally injected. A survival rate of 68% was observed in the pVG + CCL19 group but this was not significantly different from the survival rate of fish treated with pVG alone, that is, without the adjuvant. However, the expression of interferonand cytokine-related genes in the spleen and kidney tissues of zebrafish was significantly increased (p < 0.05) on days 1, 3, 7, and 14 after immunization. Thus, CCL19a.2 induced an initial immune response as a molecular adjuvant, which may provide initial protection against virus infection before vaccination- induced antibody formation. This study provides insights on the functions of CCL19a.2 adjuvant in DNA vaccines.

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LncRNA activates immune response against Vibrio anguillarum in the intestine-liver axis of turbot (Scophthalmus maximus L.) by sponging miRNA in a ceRNA regulatory network
Xin Cai, Chengbin Gao, Alan J. Lymbery, Le Ma, Qiang Fu, Ranran Huang, Chao Li
Aquaculture.2023; 576: 739882. CrossRef
Determining transcriptomic response of kidneys of olive flounder to viral hemorrhagic septicemia virus infection using next-generation sequencing
Hyoung Jun Kim, Jeong Su Park, Se Ryun Kwon, Youngjin Park
Aquaculture.2023; 562: 738886. CrossRef
Integrative transcriptomic profiling reveals the key pathways in the regulation mechanism of fish intestine-spleen immunity during the bacterial challenges
Chengbin Gao, Xin Cai, Alan J. Lymbery, Le Ma, Min Cao, Chao Li
Aquaculture.2023; 568: 739320. CrossRef

Extracellular products-mediated interspecific interaction between Pseudomonas aeruginosa and Escherichia coli: Yang Yuan , Jing Li , Jiafu Lin , Wenjuan Pan , Yiwen Chu , Balakrishnan Prithiviraj , Yidong Guo , Xinrong Wang , Kelei Zhao; J. Microbiol. 2021;59(1):29-40. Published online December 23, 2020; DOI: https://doi.org/10.1007/s12275-021-0478-0

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Abstract

The Gram-negative pathogen Pseudomonas aeruginosa adopts several elaborate strategies to colonize a wide range of natural or clinical niches and to overcome the neighboring bacterial competitors in polymicrobial communities. However, the relationship and interaction mechanism of P. aeruginosa with other bacterial pathogens remains largely unexplored. Here we explore the interaction dynamics of P. aeruginosa and Escherichia coli, which frequently coinfect the lungs of immunocompromised hosts, by using a series of on-plate proximity assays and RNA-sequencing. We show that the extracellular products of P. aeruginosa can inhibit the growth of neighboring E. coli and induce a large-scale of transcriptional reprogramming of E. coli, especially in terms of cellular respiration- related primary metabolisms and membrane components. In contrast, the presence of E. coli has no significant effect on the growth of P. aeruginosa in short-term culture, but causes a dysregulated expression of genes positively controlled by the quorum-sensing (QS) system of P. aeruginosa during subsequent pairwise culture. We further demonstrate that the divergent QS-regulation of P. aeruginosa may be related to the function of the transcriptional regulator PqsR, which can be enhanced by E. coli culture supernatant to increase the pyocyanin production by P. aeruginosa in the absence of the central las-QS system. Moreover, the extracellular products of E. coli promote the proliferation and lethality of P. aeruginosa in infecting the Caenorhabditis elegans model. The current study provides a general characterization of the extracellular products-mediated interactions between P. aeruginosa and E. coli, and may facilitate the understanding of polymicrobial infections.

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Pigments from pathogenic bacteria: a comprehensive update on recent advances
Kusumita Acharya, Swarna Shaw, Sudipta Paul Bhattacharya, Shatarupa Biswas, Suman Bhandary, Arijit Bhattacharya
World Journal of Microbiology and Biotechnology.2024;[Epub] CrossRef
Selective detection of two bacterial species in a single collision system targeting metabolic products
Jun Lin, Qingwen Wang, Huike Tian, Qing Xin, Dong Zhang
Microchemical Journal.2024; 206: 111572. CrossRef
Effect of the Type VI Secretion System Secreted Protein Hcp on the Virulence of Aeromonas salmonicida
Hongyan Cai, Jiaying Yu, Ying Qiao, Ying Ma, Jiang Zheng, Mao Lin, Qingpi Yan, Lixing Huang
Microorganisms.2022; 10(12): 2307. CrossRef

Note] Antifungal Chitinase against Human Pathogenic Yeasts from Coprinellus congregatus: Yeeun Yoo Hyoung T. Choi; J. Microbiol. 2014;52(5):441-443. Published online February 17, 2014; DOI: https://doi.org/10.1007/s12275-014-3257-3

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Abstract

The inky cap, Coprinellus congregatus, produces mushrooms which become autolyzed rapidly to generate black liquid droplets, in which no cell wall is detected by microscopy. A chitinase (Chi2) which is synthesized during the autolytic phase of C. congregatus inhibits the growths of Candida al-bicans and Cryptococcus neoformans up to 10% at the con-centration of 10 μg/ml, about 50% at concentration of 20 μg/ml, and up to 95% at the concentration of 70 μg/ml. Upon treatment these yeast cells are observed to be severely de-formed, with the formation of large holes in the cell wall. The two yeast species show no growth inhibition at the concen-tration of 5 μg/ml, which means the minimum inhibitory concentrations for both yeast species are 10 μg/ml under these experimental conditions.

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Analysis of the Antifungal Potential of Macrocybe Titans Extract Against Candida Albicans
Fernanda CBN Pereira, Gabrielle C Peiter, Vivian EMS Justo, Gabrieli M Huff, Pollyanna CV Conrado, Mauro AP da Silva, Patrícia S Bonfim-Mendonça, Terezinha IE Svidzinski, Fabio R Rosado, Adriana Fiorini
Future Microbiology.2023; 18(6): 357. CrossRef
Disarming Fungal Pathogens: Bacillus safensis Inhibits Virulence Factor Production and Biofilm Formation by Cryptococcus neoformans and Candida albicans
François L. Mayer, James W. Kronstad, Yong-Sun Bahn, J. Andrew Alspaugh, Deborah Hogan
mBio.2017;[Epub] CrossRef

Allelic MHC Class I Chain Related B (MICB) Molecules Affect the Binding to the Human Cytomegalovirus (HCMV) Unique Long 16 (UL16) Protein: Implications for Immune Surveillance: Kanya Klumkrathok , Amonrat Jumnainsong , Chanvit Leelayuwat; J. Microbiol. 2013;51(2):241-246. Published online April 27, 2013; DOI: https://doi.org/10.1007/s12275-013-2514-1

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Abstract: Unique long 16 (UL16) is a viral glycoprotein produced in a host cell infected with human cytomegalovirus (HCMV). It down regulates surface expression of MICB, one of the NKG2D ligands, by forming stable intracellular complexes and retained in the endoplasmic reticulum. Down expression of MICB renders cells less susceptible to NK cell lysis via the NKG2D receptor. Diverse UL16 sequences were identified from different strains of HCMV. MICB is known to be polymorphic. It is not known whether these polymorphisms affect the interactions between these molecules leading to alteration of the immune surveillance of HCMV. The soluble Fc fusion variant UL16 proteins from four laboratory and clinical isolates (AD169, Toledo, PH, and TR) were produced. Four allelic MICB alleles (008, 003, 004, and 00502) were cloned and stable cell lines expressing these MICB alleles were produced. The binding activities of variant UL16 to allelic MICB proteins were determined by flow cytometry. The variants of UL16 proteins did not affect the binding activities to allelic MICB proteins. However, diverse MICB alleles differentially bound UL16. We found that MICB*008 which contains methionine and asparagine at the amino acid positions 98 and 113, respectively, in the alpha 2 domain showed decreased binding activities to UL16 when compared to MICB*003, 004, and MICB*00502 containing isoleucine and aspartic acid, respectively. This finding may imply that MICB*008 is a protective allele and involved in the immune surveillance of HCMV infected patients.

Secretion of Truncated Recombinant Rabies Virus Glycoprotein with Preserved Antigenic Properties Using a Co-Expression System in Hansenula polymorpha: Weidong Qian , Frank Aguilar , Ting Wang , Bingsheng Qiu; J. Microbiol. 2013;51(2):234-240. Published online April 27, 2013; DOI: https://doi.org/10.1007/s12275-013-2337-0

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Abstract: Rabies virus infection remains a serious public health threat in the developing world, where cost-concerns make widescale public health interventions impractical. The development of novel and inexpensive ELISA diagnostic antigens is critical in early detection and prevention of complications. The transmembrane glycoprotein (G) of rabies virus (RV) contains an external domain capable of inducing the synthesis of anti-rabies, virus-neutralizing antibodies, in infected or immunized hosts. In our study, the external G domain was synthesized and fused in-frame with a polyhistidine-tag coding sequence present in the expression plasmid. Soluble truncated recombinant G was secreted in Hansenula polymorpha (H. polymorpha) using H. polymorpha-derived calnexin (HpCNE1) overproduction and found to be correctly N-glycosylated. The truncated recombinant G was purified from cell culture supernatant by Ni-agarose affinity chromatography and when compared with the full-length glycoprotein, found to be similarly immunogenic in vaccinated rabbits. These results subsequently led us to explore the potential of truncated recombinant G as a diagnostic antigen in ELISA. Our results show that the truncated recombinant G can detect antibodies directed to both whole virion and native glycoprotein. More sophisticated applications of truncated recombinant G would profit from the correctly N-glycosylated and soluble monomer.

NOTE] Envelope Diversity, Characteristics of V3 Region and Predicted Co-Receptor Usage of Human Immunodeficiency Viruses Infecting North Indians: Raiees Andrabi , Rajesh Kumar , Manju Bala , Ambili Nair , Prakash SS , Vandana Kushwaha , Kalpana Luthra; J. Microbiol. 2012;50(5):869-873. Published online November 4, 2012; DOI: https://doi.org/10.1007/s12275-012-2136-z

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Abstract: Subtypes of human immunodeficiency virus type 1 circulating in 21 north Indian patients were characterized based on the partial sequence of the gp120 envelope protein. A majority of viruses (85.7%, 18/21) were subtype C, while 14.3% (3/21) were subtype A. Sequence analysis revealed that the V3 region was highly conserved compared with V4 and V5. The predicted use of co-receptors indicated exclusive usage of R5, except for two subtype A viruses (AIIMS279 and AIIMS281). Our results demonstrate conservation within the V3 loop of subtype C viruses, and suggest the emergence of non-clade C viruses in the north Indian population.

NOTE] The Activity of Phosphoinositide-Specific Phospholipase C Is Required for Vegetative Growth and Cell Wall Regeneration in Coprinopsis cinerea: Young Taek Oh , Chun-Seob Ahn , Kyung-Jin Lee , Jeong-Geun Kim , Hyeon-Su Ro , Jae Won Kim , Chang-Won Lee; J. Microbiol. 2012;50(4):689-692. Published online August 25, 2012; DOI: https://doi.org/10.1007/s12275-012-2004-x

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Abstract: Three isotypes of phosphoinositide-specific phospholipase C designated CcPLC1, CcPLC2, and CcPLC3 were identified in Coprinopsis cinerea, through a search of the genome sequence database. The functional role of the PI-PLCs were studied by using U73122, which specifically inhibits the activity of PI-PLC. The specificity of the inhibitor effect was confirmed by using an inactive structural analog U73433. The inhibition of PI-PLCs activity resulted in severely retarded germination of basidiospores and oidia, reduced hyphal growth, knobbly hyphal tips with many irregular side branches, and aberrant (branch-like structure) clamp cells. Furthermore, U73122 definitely inhibited cell wall formation. Here we report that PI-PLCs play important roles in various aspects of C. cinerea biology.

Effect of Glycosylation on the Biochemical Properties of beta-Xylosidases from Aspergillus versicolor: Alexandre Favarin Somera , Marita Gimenez Pereira , Luis Henrique Souza Guimaraes , Maria de Lourdes Teixeira de Moraes Polizeli , Hector Francisco Terenzi , Rosa Prazeres Melo Furriel , Joao Atilio Jorge; J. Microbiol. 2009;47(3):270-276. Published online June 26, 2009; DOI: https://doi.org/10.1007/s12275-008-0286-9

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Abstract: Aspergillus versicolor grown on xylan or xylose produces two beta-xylosidases with differences in biochemical properties and degree of glycosylation. We investigated the alterations in the biochemical properties of these beta-xylosidases after deglycosylation with Endo-H or PNGase F. After deglycosylation, both enzymes migrated faster in PAGE or SDS-PAGE exhibiting the same Rf. Temperature optimum of xylan-induced and xylose-induced beta-xylosidases was 45oC and 40oC, respectively, and 35oC after deglycosylation. The xylan- induced enzyme was more active at acidic pH. After deglycosylation, both enzymes had the same pH optimum of 6.0. Thermal resistance at 55oC showed half-life of 15 min and 9 min for xylose- and xylan-induced enzymes, respectively. After deglycosylation, both enzymes exhibited half-lives of 7.5 min. Native enzymes exhibited different responses to ions, while deglycosylated enzymes exhibited identical responses. Limited proteolysis yielded similar polypeptide profiles for the deglycosylated enzymes, suggesting a common polypeptide core with differential glycosylation apparently responsible for their biochemical and biophysical differences.

Molecular Analysis of the Copper-Responsive CopRSCD of a Pathogenic Pseudomonas fluorescens Strain: Yong-hua Hu , Hua-lei Wang , Min Zhang , Li Sun; J. Microbiol. 2009;47(3):277-286. Published online June 26, 2009; DOI: https://doi.org/10.1007/s12275-008-0278-9

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Abstract: CopRS/CopABCD is one of the known systems that control copper homeostasis in bacteria. Although CopRS/CopABCD homologues are found to exist in Pseudomonas fluorescens, the potential role of this system in P. fluorescens has not been investigated. In this study a genetic cluster, consisting of copR, S, C, and D but lacking copAB, was identified in a pathogenic P. fluorescens strain (TSS) isolated from diseased fish. The copRSCD cluster was demonstrated to be required for full copper resistance and regulated at the transcription level by Cu. Expression of copCD is regulated directly by the two-component response regulator CopR, which also regulates its own expression. Interruption of the regulated expression of copR affected bacterial growth, biofilm formation, and tissue dissemination and survival. A mutant CopR, which lacks the N-terminal signal receiver domain and is constitutively active, was found to have an attenuating effect on bacterial virulence when expressed in TSS. To our knowledge, this is the first report that suggests a link between CopR and bacterial pathogenicity in P. fluorescens.

Defining the N-Linked Glycosylation Site of Hantaan Virus Envelope Glycoproteins Essential for Cell Fusion: Feng Zheng , Lixian Ma , Lihua Shao , Gang Wang , Fengzhe Chen , Ying Zhang , Song Yang; J. Microbiol. 2007;45(1):41-47.; DOI: https://doi.org/2493 [pii]

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Abstract: The Hantaan virus (HTNV) is an enveloped virus that is capable of inducing low pH-dependent cell fusion. We molecularly cloned the viral glycoprotein (GP) and nucleocapsid (NP) cDNA of HTNV and expressed them in Vero E6 cells under the control of a CMV promoter. The viral gene expression was assessed using an indirect immunofluorescence assay and immunoprecipitation. The transfected Vero E6 cells expressing GPs, but not those expressing NP, fused and formed a syncytium following exposure to a low pH. Monoclonal antibodies (MAbs) against envelope GPs inhibited cell fusion, whereas MAbs against NP did not. We also investigated the N-linked glycosylation of HTNV GPs and its role in cell fusion. The envelope GPs of HTNV are modified by N-linked glycosylation at five sites: four sites on G1 (N134, N235, N347, and N399) and one site on G2 (N928). Site-directed mutagenesis was used to construct eight GP gene mutants, including five single N-glycosylation site mutants and three double-site mutants, which were then expressed in Vero E6 cells. The oligosaccharide chain on residue N928 of G2 was found to be crucial for cell fusion after exposure to a low pH. These results suggest that G2 is likely to be the fusion protein of HTNV.

Increase of Yeast Survival under Oxidative Stress by the Expression of the Laccase Gene from Coprinellus congregatus: Dongsik Kim , Eunju Kwak , Hyoung T. Choi; J. Microbiol. 2006;44(6):617-621.; DOI: https://doi.org/2466 [pii]

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Abstract: Coprinellus congregatus secreted a laccase isozyme when the culture was transferred to an acidic liquid medium (pH 4.1). The laccase cDNA gene (clac2) was used as a probe for cloning of the genomic laccase gene (lac2) including the promoter (Plac2). The open reading frame (ORF) of lac2 had 526 deduced amino acids and four conserved copper binding domains as other fungal laccases. Recombinant plasmid (pRSlac2p-cDNA) of lac2 cDNA with its own promoter was transformed in Saccharomyces cerevisiae. Expression of the transformed lac2 gene was induced by oxidative stress (H2O2) in yeast and the survival rate of the transformed yeast strain was greatly increased when compared with that of the control strain transformed with pRS316 yeast vector.

Genetic and Phenotypic Diversity of (R/S)-Mecoprop [2-(2-Methyl-4-Chlorophenoxy)Propionic Acid]-Degrading Bacteria Isolated fromSoils: Jong-Sung Lim , Mee-Kum Jung , Mi-Soon Kim , Jae-Hyung Ahn , Jong-Ok Ka; J. Microbiol. 2004;42(2):87-93.; DOI: https://doi.org/2040 [pii]

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Abstract: Twelve mecoprop-degrading bacteria were isolated from soil samples, and their genetic and phenotypic characteristics were investigated. Analysis of 16S rDNA sequences indicated that the isolates were related to members of the genus Sphingomonas. Ten different chromosomal DNA patterns were obtained by polymerase-chain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences from the 12 isolates. The isolates were found to be able to utilize the chiral herbicide mecoprop as a sole source of carbon and energy. While seven of the isolates were able to degrade both (R)- and (S)-mecoprop, four isolates exhibited enantioselective degradation of the (S)-type and one isolate could degrade only the (R)-enantiomer. All of the isolates were observed to possess plasmid DNAs. When certain plasmids were removed from isolates MP11, MP15, and MP23, those strains could no longer degrade mecoprop. This compelling result suggests that plasmid DNAs, in this case, conferred the ability to degrade the herbicide. The isolates MP13, MP15, and MP24 were identified as the same strain; however, they exhibited different plasmid profiles. This indicates that these isolates acquired different mecoprop-degradative plasmids in different soils through natural gene transfer.

Selection of Laccase Over-secreting Mutant in Coprinus congregature: Kim, Soon Ja , Choi, Hyoung Tae; J. Microbiol. 1995;33(2):146-148.

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Abstract: Coprinus congregatus has a membrane-associated laccase which is not secreted into culture media. A mutant monokaryon obtained, by U. V. irradiation followed by protoplast generation and regeneration method, was successfully isolated. When the mutant was grown on a agar plate or in a liquid medium, it secreted laccase while the wild type did not under the same growth conditions. The laccase of the mutant was compared with that of wild type did not under the same growth conditions. The laccase of the mutant was compared with that of wild type of native PAGE analysis, and showed identical mobility.

Production and Characterization of Monoclonal Antibodies to Human Immunodeficiency Virus Type 1 gp120 Envelope Glycoprotein: Choi, Eui Yul , Ryu, Ji Yoon , Lee, Yoon , Ha, Sung Gil , Chung, So Young , Park, Sang Yeol , Nham, Sang Uk , Lee, Young Ik , Park, Jin Seu; J. Microbiol. 1998;36(1):59-65.

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Abstract: Monoclonal antibodies to human immunodeficiency virus type 1 envelope glycoprotein gp 120(HIV-1 gp120) were produced and characterized. For immunogen recombinant gp120 polypeptide expressed in bacteria was prepared and injected into mice. From two fusion experiments, twenty hybridomas secreting monoclonal antibodies against the recombinant gp120 were initially screened by immunodot blot analysis. Among the antibodies, 15 of them showed strong reactivities with the recombinant protein expressed in bacteria in Western blot and thus it was tested if these could react with the recombinant protein expressed in insect cells. All of the 15 antibodies immunostained the protein band with varing degrees of reactivities. Next, we tested whether the antibodies recognize authentic gp120 protein expressed in mammalian cells. COS-1 cells were tranfected with the cDNA encoding gp120 protein, and the transiently ecpressed protein were analyzed with the mAbs by Western blot analysis and immunofluorescence microscopy. Six of the monoclonal antibodies reacted with the protein band of authentic gp120 expressed in mammalian cells in the Western blot, and five stained the cell periphery of the transfected COS-1 cells in immunofluorescence. The mAbs described in this study should prove to be useful tools for the biochemical, immunological and structural analysis of HIV-1 gp120 envelope glycoprotein.

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