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
    • Latest Articles
    • Archive
    • Minireviews
  • Topics
    • Applied and Environmental Science
    • Clinical Science and Epidemiology
    • Ecological and Evolutionary Science
    • Host-Microbe Biology
    • Molecular Biology and Physiology
    • Therapeutics and Prevention
  • 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 mBio
    • Editor in Chief
    • Board of Editors
    • AAM Fellows
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • 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
mBio
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Latest Articles
    • Archive
    • Minireviews
  • Topics
    • Applied and Environmental Science
    • Clinical Science and Epidemiology
    • Ecological and Evolutionary Science
    • Host-Microbe Biology
    • Molecular Biology and Physiology
    • Therapeutics and Prevention
  • 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 mBio
    • Editor in Chief
    • Board of Editors
    • AAM Fellows
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
Research Article

Aspergillus fumigatus Photobiology Illuminates the Marked Heterogeneity between Isolates

Kevin K. Fuller, Robert A. Cramer, Michael E. Zegans, Jay C. Dunlap, Jennifer J. Loros
J. Andrew Alspaugh, Editor
Kevin K. Fuller
Department of Molecular and Systems Biology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Robert A. Cramer
Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michael E. Zegans
Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USADepartment of Surgery (Ophthalmology), Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jay C. Dunlap
Department of Molecular and Systems Biology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jennifer J. Loros
Department of Molecular and Systems Biology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USADepartment of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
J. Andrew Alspaugh
Duke University Medical Center
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
David Askew
University of Cincinnati
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
William Steinbach
Duke University Medical Center
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/mBio.01517-16
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Supplemental Material
  • Additional Files
  • FIG 1 
    • Open in new tab
    • Download powerpoint
    FIG 1 

    A. fumigatus isolates induce pigmentation or conidiation in response to light. Conidia of the indicated strain were point inoculated onto GMM (plate pictures) or RPMI 1640 (graphs) and incubated for 48 h either in constant darkness or under constant white light illumination. Conidiation values are normalized to the dark-grown sample of the indicated strain. Enumeration of conidia was performed in triplicate, and data were statistically analyzed by Student’s t test (*, P ≤ 0.05).

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

    The photoconidiation response is blue light and lreA dependent. (A) Conidia were point inoculated onto RPMI 1640 plates and incubated for 6 days in an alternating 12-h-light/12-h-dark photocycle (left column). Alternatively, plates were transferred to constant darkness following an initial 2-day incubation in the 12-h-light/12-h-dark environment (right column). (B) Conidia were spread across RPMI 1640 plates and incubated for 48 h either in the dark or under constant illumination under red or blue LEDs. Conidial counts are normalized to the respective dark-grown sample. Experiments were performed in triplicate, and total conidia were compared by Student’s t test (**, P ≤ 0.01; ***, P ≤ 0.001; ns, not significant). (C) Conidia were spread across RPMI 1640 plates and incubated for 48 h either in constant darkness or under constant white light illumination. Experiments were performed in triplicate, and error bars represent the mean conidial counts (±standard deviation). Groups were compared by Student’s t test (*, P ≤ 0.05). (D) Scheme of LreA-dependent regulation of asexual development in A. nidulans versus A. fumigatus.

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

    The photosensory control of germination and cell wall homeostasis differs between Af293 and CEA10. (A) Conidia were inoculated into GMM broth and incubated at 37°C for 8 h either in constant darkness or under constant illumination (blue plus red LEDs). A minimum of 100 conidia were scored for the presence or absence of a germ tube in each group, and light and dark samples were compared by the chi-square test (*, P ≤ 0.05). (B) Conidia of the indicated CEA10 genotype were spot inoculated onto GMM plus Congo red agar and incubated at 37°C for 48 h in constant darkness or under constant illumination (blue plus red LEDs).

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

    Virulence is variable across isolates and is independent of either the overt light response or lreA. (A) All animals were immunosuppressed with a single dose of triamcinolone acetonide (day −1) and inoculated intranasally with 1.0 × 106 conidia of the indicated isolate (CEA10, n = 8; Af293, n = 11; DCF-3 and DCF-6, n = 12). (B) Left, CEA10 background. The wild-type curve is from the same experiment as that depicted in panel A; the ΔlreA group was run concurrently with the WT and under the same protocol. Survival curves are not statistically significant (P = 0.16). Right, DCF-3 background. Groups of 10 CD-1 mice were immunosuppressed with two doses of Kenalog (days −1 and +3) and inoculated intranasally with the indicated genotype. Two independently isolated ΔlreA mutants are shown. WT versus ΔlreA-1, P = 0.14; WT versus ΔlreA-2, P = 0.03. All statistical analyses were performed with the log rank test.

Supplemental Material

  • Figures
  • Additional Files
  • Table S1 

    Strains used in this study Table S1, DOCX file, 0.1 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Figure S1 

    Photopigmentation and photoconidiation responses of additional A. fumigatus isolates. Conidia of the indicated strain were point inoculated onto GMM (plate pictures) or RPMI 1640 (graphs) and incubated for 48 h either in constant darkness or under constant white light illumination. Conidiation values are normalized to the dark-grown sample of the indicated strain. Enumeration of conidia was performed in triplicate, and data were statistically analyzed by Student’s t test (*, P ≤ 0.05). Download Figure S1, TIF file, 50.2 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Figure S2 

    (A) The photoconidiation response is more pronounced on RPMI 1640 medium (R) than on GMM (G). Experiments were performed in triplicate, and groups were compared by Student’s t test (*, P ≤ 0.05). (B) The overall conidiation levels are variable across the strains. Values are normalized to the Af293 DD sample. (C) The effect of light on growth rate is minimal for each of the tested isolates. Conidia were point inoculated on GMM and incubated for 3 days at 37°C in constant darkness or under constant white light illumination. Download Figure S2, TIF file, 66.9 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Table S2 

    Summary of photoresponsive behaviors in the analyzed isolates. Photopigmenting isolates are shaded. Table S2, DOCX file, 0.1 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Figure S3 

    (A) RT-PCR analysis of lreA expression. Strains were incubated in the dark for 48 h and then transferred to constant white light illumination for the indicated time points. (B) Schematic depiction of the split-marker deletion strategy of lreA is shown as well as an RT-PCR demonstrating loss of the lreA transcript in the putative knockouts. hph, hygromycin phosphotransferase gene. (C) Schematic depiction of the split-marker deletion strategy of fphA is shown as well as an RT-PCR demonstrating loss of the fphA transcript in either the CEA10 WT or ΔlreA background. (D) RT-PCR demonstrating the transcript of either lreA (top) or fphA (bottom) in the respective complemented strain. Download Figure S3, TIF file, 40.8 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Figure S4 

    Light decreases germination rates in most A. fumigatus isolates. Conidia were inoculated into GMM and incubated for 8 h either in constant darkness or under constant white light illumination. A minimum of 300 conidia were scored for the presence or absence of a germ tube in each group, and light and dark samples were compared by the chi-square test (*, P ≤ 0.05). Download Figure S4, TIF file, 9 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Figure S5 

    FphA protein sequence alignment based on Sanger sequencing from the indicated isolates. Download Figure S5, TIF file, 52.1 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Figure S6 

    Additional phenotypes of the CEA10 photoreceptor mutants. (A) Pigmentation. Conidia were point inoculated into GMM plates and incubated at 37°C for 48 h either in constant darkness or under constant illumination (blue plus red LEDs). Several independent isolates of the ΔfphA mutation are shown and are distinguished numerically (e.g., 5-1, 7-2). (B) Congo red sensitivity. Conidia were point inoculated onto GMM plates supplemented with the indicated concentration of Congo red. Plates were then incubated for 48 h at 37°C for 48 h either in constant darkness or under constant illumination (blue plus red LEDs). Download Figure S6, TIF file, 26.5 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Figure S7 

    Deletion of lreA in the Af293 background does not attenuate virulence. (A) Single-dose steroid model. Groups of 12 CD-1 mice were immunosuppressed with Kenalog-10 (day −1) and inoculated intranasally with 2.0 × 106 conidia of the indicated genotype. (B) Multidose steroid model. Groups of 16 CD-1 mice were immunosuppressed with two doses of Kenalog-10 (days −1 and +3) and inoculated intranasally with 1.5 × 106 conidia of the indicated genotypes. Survival curves are not statistically different (P = 0.22, log rank test). (C) Chemotherapy (neutropenia model). Groups of 17 CD-1 mice were immunosuppressed with cyclophosphamide (days −2 and +3) and Kenalog-10 (day −1) and inoculated intranasally with 2.0 × 106 conidia of the indicated genotypes. Survival curves are not statistically different (P = 0.4, log rank test). Download Figure S7, TIF file, 32.4 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • Figure S8 

    VeA protein alignment based on Sanger sequencing from the indicated isolates. Download Figure S8, TIF file, 63.3 MB.

    Copyright © 2016 Fuller et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

Additional Files

  • Figures
  • Supplemental Material
  • Supplementary Data

    Supplementary Data

    • Figure sf1, TIF - Figure sf1, TIF
    • Figure sf2, TIF - Figure sf2, TIF
    • Figure sf3, TIF - Figure sf3, TIF
    • Figure sf4, TIF - Figure sf4, TIF
    • Figure sf5, TIF - Figure sf5, TIF
    • Figure sf6, TIF - Figure sf6, TIF
    • Figure sf7, TIF - Figure sf7, TIF
    • Figure sf8, TIF - Figure sf8, TIF
    • Table st1, DOCX - Table st1, DOCX
    • Table st2, DOCX - Table st2, DOCX
PreviousNext
Back to top
Download PDF
Citation Tools
Aspergillus fumigatus Photobiology Illuminates the Marked Heterogeneity between Isolates
Kevin K. Fuller, Robert A. Cramer, Michael E. Zegans, Jay C. Dunlap, Jennifer J. Loros
mBio Sep 2016, 7 (5) e01517-16; DOI: 10.1128/mBio.01517-16

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 mBio 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.
Aspergillus fumigatus Photobiology Illuminates the Marked Heterogeneity between Isolates
(Your Name) has forwarded a page to you from mBio
(Your Name) thought you would be interested in this article in mBio.
Share
Aspergillus fumigatus Photobiology Illuminates the Marked Heterogeneity between Isolates
Kevin K. Fuller, Robert A. Cramer, Michael E. Zegans, Jay C. Dunlap, Jennifer J. Loros
mBio Sep 2016, 7 (5) e01517-16; DOI: 10.1128/mBio.01517-16
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • INTRODUCTION
    • RESULTS
    • DISCUSSION
    • MATERIALS AND METHODS
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About mBio
  • Editor in Chief
  • Board of Editors
  • AAM Fellows
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

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

Follow #mBio

@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

Copyright © 2019 American Society for Microbiology | Privacy Policy | Website feedback

Online ISSN: 2150-7511