Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About AEM
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Applied and Environmental Microbiology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About AEM
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Food Microbiology

Association of a d-Alanyl-d-Alanine Carboxypeptidase Gene with the Formation of Aberrantly Shaped Cells during the Induction of Viable but Nonculturable Vibrio parahaemolyticus

Wei-cheng Hung, Wann-Neng Jane, Hin-chung Wong
Wei-cheng Hung
aDepartment of Microbiology, Soochow University, Taipei, Taiwan, Republic of China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Wann-Neng Jane
bInstitute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan, Republic of China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hin-chung Wong
aDepartment of Microbiology, Soochow University, Taipei, Taiwan, Republic of China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/AEM.01723-13
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Additional Files
  • Fig 1
    • Open in new tab
    • Download powerpoint
    Fig 1

    Formation of aberrantly shaped cells in V. parahaemolyticus 1137 incubated at 4°C in MMS–0.5% NaCl for 12 h. (A) Cells observed under a light microscope. Bar, 5 μm. (B and C) Cells observed under a transmission electron microscope. Arrows indicate the bulging of aberrantly shaped cells. Bars, 0.2 μm.

  • Fig 2
    • Open in new tab
    • Download powerpoint
    Fig 2

    Effects of antibiotics on the formation of aberrantly shaped cells in V. parahaemolyticus 1137. Cultures were incubated at 4°C for 12 h in MMS with 0.5% NaCl, either without antibiotics (control) or with d-cycloserine (50 μg/ml) or cephalosporin C (10 μg/ml), and cell shapes were determined by microscopy. The cell shapes were classified into three groups: rod, coccoid, and other (nonrod, noncoccoid) aberrant shapes (15). Open bars, coccoid cells; hatched bars, aberrantly shaped cells; stippled bars, total of coccoid and aberrantly shaped cells. The values for the antibiotic-supplemented groups were compared with the corresponding values for the control group. Asterisks indicate significantly different values (P < 0.05). For each determination, a total of 200 bacteria from 20 randomly selected fields were examined.

  • Fig 3
    • Open in new tab
    • Download powerpoint
    Fig 3

    Effect of salinity on the formation of aberrantly shaped cells in V. parahaemolyticus 1137. Cultures were suspended in MMS with 0.5, 1.0, or 1.5% NaCl and were incubated at 4°C for 12 h. Coccoid cells (open bars) and aberrantly shaped cells (hatched bars) were counted under a light microscope. Their total was also determined (stippled bars). The values obtained at different salt concentrations were compared. Asterisks indicate significantly different values (P < 0.05). For each determination, a total of 200 bacteria from 20 randomly selected fields were examined.

  • Fig 4
    • Open in new tab
    • Download powerpoint
    Fig 4

    Expression of genes associated with cell wall synthesis in V. parahaemolyticus 1137. Strain 1137 was cultured in MMS–0.5% NaCl and was incubated at 4°C. The expression of target genes at 12 h relative to their expression at 0 h was determined by RT-qPCR.

  • Fig 5
    • Open in new tab
    • Download powerpoint
    Fig 5

    Formation of aberrantly shaped cells in the V. parahaemolyticus wild-type strain 1137, Δ2468 (dacB deletion mutant), Δ2468C (the complementary strain with dacB restored), and Δ2468V (dacB deletion mutant containing pSCB01). Cultures were suspended in MMS with 0.5% NaCl and were incubated at 4°C for 12 h, and aberrantly shaped cells were counted under a light microscope. The values for the mutant and complemented strains were compared to that for the wild type. Asterisks indicate significant differences (P < 0.05).

Tables

  • Figures
  • Additional Files
  • Table 1

    Bacterial strains and plasmids used in this study

    Strain or plasmidCharacteristics/sequenceSource or reference
    Strains
        V. parahaemolyticus
            1137Wild type, serotype O3:K6, KP+; clinical isolateThis study
            Δ24681137 ΔdacBThis study
            Δ2468C1137 ΔdacB containing pSCB01 dacBThis study
            Δ2468V1137 ΔdacB containing pSCB01This study
        E. coli
            XL1-BluerecA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacIqZΔM15 Tn10 (Tetr)]Stratagene
            SM10λpirthi thr leu tonA lacY supE recA::RP4-2-Tc::Mu λ pirR6K; Kmr46
    Plasmids
        pGEM-T EasyCloning vector; AprPromega
        pDS132R6K ori mobRP4 sacB Cmr47
        pBR328Vector derived from pBR325 and pMB1; Apr Cmr TcrRoche
        pSCB01Derived from pBR328 and pDS132; mobRP4 Apr Cmr TcrThis study
        pGEM-T dacBpGEM-T containing dacBThis study
        pDS132ΔdacBpDS132 with ΔdacBThis study
        pSCB01 dacBpSCB01 containing dacBThis study
  • Table 2

    Primers used in cloning experiments of this study

    PrimerSequence (5′ to 3′)Amplicon length (bp)
    Primer 1CTTCACTGACGGTGAAAAAG562
    Primer 2GGCCATTTTTATCTAAACCATAAGACAATAGCGGAAAAATAAGA
    Primer 3TCTTATTTTTCCGCTATTGTCTTATGGTTTAGATAAAAATGGCC579
    Primer 4AGTGTACATGCCAGTGATTG
    Primer CCAGTCTTTAGACTTGAATACTC2,025a
    Primer 1+CTATACGGTGTTGGTTTCTG3,338
    Primer 4+TGTGTCATCTATCGGCATAA
    Primer FTACGTTGGCCTCTACTCATT489
    Primer RCTTGGACACAGTTGTTGTTG
    Seq 1FTAACCCTAGAAAATCTCACT677
    Seq 1RTAATTCGTAGCGATTGTCTG
    Seq 2FGCATCAACCTATTCGTGTTA897
    Seq 2RATTTACTTTACTGTGGTGAA
    • ↵a Amplified by primers C and 4.

  • Table 3

    Selected genes associated with cell wall synthesis in V. parahaemolyticus

    LocusGeneProduct
    VP0454Penicillin-binding protein 3
    VP0535lspALipoprotein signal peptidase
    VP0457mraYPhospho-N-acetylmuramoyl-pentapeptide transferase
    VP0548tagEPutative ToxR-activated protein TagE, subfamily M23B unassigned peptidases (Vibrio harveyi)
    VP0564mepANon-penicillin-sensitive murein endopeptidase, d-alanyl-d-alanine endopeptidase; functions in hydrolyzing cell wall peptidoglycan
    VP0722rodAPenicillin-binding protein 2
    VP1385Hypothetical protein, cell wall endopeptidase, subfamily M23B
    VP1485nlpCLipoprotein NlpC, Spr peptidase (Vibrio parahaemolyticus), outer membrane protein
    VP2369mltAMurein transglycosylase A, catalyzes the cleavage of the β-1,4-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine residues
    VP2463ftsHCell division protein FtsH, family M41 unassigned peptidases (Vibrio parahaemolyticus)
    VP2468dacBd-Alanyl-d-alanine carboxypeptidase/endopeptidase, LMW PBP; removes terminal d-alanine from pentapeptide side chains
    VP2471Hypothetical protein, subfamily M23B nonpeptidase homologs (Vibrio sp. strain EJY3)
    VP2497mrcBPenicillin-binding protein 1B
    VP2658murAUDP-N-acetylglucosamine 1-carboxyvinyltransferase; adds enolpyruvyl to UDP-N-acetylglucosamine as a component of cell wall formation
    VP2751Penicillin-binding protein 1A
    VPA0517Hypothetical protein, zoocin A (Vibrio sp.), a PBPa
    VPA1649tagEPutative TagE protein, subfamily M23B unassigned peptidases (Vibrio alginolyticus); outer membrane protein related to metalloendopeptidases, cell envelope biogenesis
    • ↵a See reference 48.

Additional Files

  • Figures
  • Tables
  • Supplemental material

    Files in this Data Supplement:

    • Supplemental file 1 -

      Primers used in RT-qPCR (Table S1); preliminary data on the determination of antibiotic concentrations to be used in this study (Fig. S1).

      PDF, 126K

PreviousNext
Back to top
Download PDF
Citation Tools
Association of a d-Alanyl-d-Alanine Carboxypeptidase Gene with the Formation of Aberrantly Shaped Cells during the Induction of Viable but Nonculturable Vibrio parahaemolyticus
Wei-cheng Hung, Wann-Neng Jane, Hin-chung Wong
Applied and Environmental Microbiology Nov 2013, 79 (23) 7305-7312; DOI: 10.1128/AEM.01723-13

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Applied and Environmental Microbiology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Association of a d-Alanyl-d-Alanine Carboxypeptidase Gene with the Formation of Aberrantly Shaped Cells during the Induction of Viable but Nonculturable Vibrio parahaemolyticus
(Your Name) has forwarded a page to you from Applied and Environmental Microbiology
(Your Name) thought you would be interested in this article in Applied and Environmental Microbiology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Association of a d-Alanyl-d-Alanine Carboxypeptidase Gene with the Formation of Aberrantly Shaped Cells during the Induction of Viable but Nonculturable Vibrio parahaemolyticus
Wei-cheng Hung, Wann-Neng Jane, Hin-chung Wong
Applied and Environmental Microbiology Nov 2013, 79 (23) 7305-7312; DOI: 10.1128/AEM.01723-13
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • INTRODUCTION
    • MATERIALS AND METHODS
    • RESULTS AND DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About AEM
  • Editor in Chief
  • Editorial Board
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Ethics
  • Contact Us

Follow #AppEnvMicro

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

 

Print ISSN: 0099-2240; Online ISSN: 1098-5336