hypovirulent strain  (ATCC)


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    ATCC hypovirulent strain
    Hypovirulent Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    c tropicalis atcc 66024 biofilm  (ATCC)


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    ATCC c tropicalis atcc 66024 biofilm
    C Tropicalis Atcc 66024 Biofilm, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    candida atcc 66024 cells  (ATCC)


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    ATCC candida atcc 66024 cells
    Candida Atcc 66024 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    c tropicalis atcc 66024  (ATCC)


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    ATCC c tropicalis atcc 66024
    C Tropicalis Atcc 66024, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    c tropicalis atcc 66024 cells  (ATCC)


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    ATCC c tropicalis atcc 66024 cells
    C Tropicalis Atcc 66024 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    c tropicalis atcc 66024 biofilms  (ATCC)


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    ATCC c tropicalis atcc 66024 biofilms
    C Tropicalis Atcc 66024 Biofilms, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    c tropicalis atcc 66024 cells  (ATCC)


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    ATCC c tropicalis atcc 66024 cells
    Effect of occidiofungin on Candida cell attachment. Relative viable cell number (bar graph) and metabolic activity (line graph) for cells exposed for 90 min to occidiofungin either immediately following treatment (left panel) or 48 h post-treatment (right panel). Data represent percent differences relative to untreated samples for (a) C. albicans ATCC 66027 and (b) C. tropicalis ATCC 66024. Averages reported for three biological replicates each containing technical triplicates. Error bars represent standard error of the mean. Significant differences, as determined using the post hoc Tukey HSD method, between occidiofungin exposed and untreated cells are indicated; * or § , P < 0.05; ** or §§ , P < 0.01.
    C Tropicalis Atcc 66024 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Occidiofungin inhibition of Candida biofilm formation on silicone elastomer surface"

    Article Title: Occidiofungin inhibition of Candida biofilm formation on silicone elastomer surface

    Journal: Microbiology Spectrum

    doi: 10.1128/spectrum.02460-23

    Effect of occidiofungin on Candida cell attachment. Relative viable cell number (bar graph) and metabolic activity (line graph) for cells exposed for 90 min to occidiofungin either immediately following treatment (left panel) or 48 h post-treatment (right panel). Data represent percent differences relative to untreated samples for (a) C. albicans ATCC 66027 and (b) C. tropicalis ATCC 66024. Averages reported for three biological replicates each containing technical triplicates. Error bars represent standard error of the mean. Significant differences, as determined using the post hoc Tukey HSD method, between occidiofungin exposed and untreated cells are indicated; * or § , P < 0.05; ** or §§ , P < 0.01.
    Figure Legend Snippet: Effect of occidiofungin on Candida cell attachment. Relative viable cell number (bar graph) and metabolic activity (line graph) for cells exposed for 90 min to occidiofungin either immediately following treatment (left panel) or 48 h post-treatment (right panel). Data represent percent differences relative to untreated samples for (a) C. albicans ATCC 66027 and (b) C. tropicalis ATCC 66024. Averages reported for three biological replicates each containing technical triplicates. Error bars represent standard error of the mean. Significant differences, as determined using the post hoc Tukey HSD method, between occidiofungin exposed and untreated cells are indicated; * or § , P < 0.05; ** or §§ , P < 0.01.

    Techniques Used: Cell Attachment Assay, Activity Assay

    Efficacy of occidiofungin on cells at different stages of biofilm development. The sensitivity of (a) C. albicans ATCC 66027 and (b) C. tropicalis ATCC 66024 cells to occidiofungin was reported as relatively viable cell number (bar graph) and metabolic activity (line graph). Occidiofungin was added to attached cells and biofilms monitored following a 48 h period (left panel) or added to cells in a 24 h biofilm and monitored following an additional 24 h growth period (right panel). Data from CFU and XTT assays are represented as percent differences relative to untreated biofilm cells with the average and standard error for three biological replicates each containing technical triplicates. Significant differences, as determined using the post hoc Tukey HSD method, between untreated and occidiofungin exposed biofilms are indicated; * or § , P < 0.05; ** or §§ , P < 0.01.
    Figure Legend Snippet: Efficacy of occidiofungin on cells at different stages of biofilm development. The sensitivity of (a) C. albicans ATCC 66027 and (b) C. tropicalis ATCC 66024 cells to occidiofungin was reported as relatively viable cell number (bar graph) and metabolic activity (line graph). Occidiofungin was added to attached cells and biofilms monitored following a 48 h period (left panel) or added to cells in a 24 h biofilm and monitored following an additional 24 h growth period (right panel). Data from CFU and XTT assays are represented as percent differences relative to untreated biofilm cells with the average and standard error for three biological replicates each containing technical triplicates. Significant differences, as determined using the post hoc Tukey HSD method, between untreated and occidiofungin exposed biofilms are indicated; * or § , P < 0.05; ** or §§ , P < 0.01.

    Techniques Used: Activity Assay

    Occidiofungin reduces cells released from a biofilm. (a) Average CFU/mL of dispersed cells from untreated and 0.5× MBIC 90 occidiofungin-treated C. albicans ATCC 66027 and C. tropicalis ATCC 66024 biofilms. (b) Dispersed cells from C. tropicalis biofilm treated with increasing concentration of occidiofungin presented as % viable cells. Data presented as average and standard error from three independent experiments. Significant differences, relative to untreated biofilm, were determined using the post hoc Tukey HSD method. * P < 0.05; ** P < 0.01.
    Figure Legend Snippet: Occidiofungin reduces cells released from a biofilm. (a) Average CFU/mL of dispersed cells from untreated and 0.5× MBIC 90 occidiofungin-treated C. albicans ATCC 66027 and C. tropicalis ATCC 66024 biofilms. (b) Dispersed cells from C. tropicalis biofilm treated with increasing concentration of occidiofungin presented as % viable cells. Data presented as average and standard error from three independent experiments. Significant differences, relative to untreated biofilm, were determined using the post hoc Tukey HSD method. * P < 0.05; ** P < 0.01.

    Techniques Used: Concentration Assay

    Occidiofungin killed cells are retained within the biofilm. Representative CLSM images of untreated and 3 h occidiofungin treated for (a) C. albicans ATCC 66027 and (b) C. tropicalis ATCC 66024 biofilm stained with Calcofluor White (left panel) and Live-or-Dye 640/662 TM viability dye (right panel) to visualize total and dead cells, respectively. Size bar: 50 microns.
    Figure Legend Snippet: Occidiofungin killed cells are retained within the biofilm. Representative CLSM images of untreated and 3 h occidiofungin treated for (a) C. albicans ATCC 66027 and (b) C. tropicalis ATCC 66024 biofilm stained with Calcofluor White (left panel) and Live-or-Dye 640/662 TM viability dye (right panel) to visualize total and dead cells, respectively. Size bar: 50 microns.

    Techniques Used: Staining

    Loss of actin cables in occidiofungin exposed biofilm cells. Preformed 24-h biofilm of Candida ATCC 66024 cells grown in RPMI media were left untreated (left) or treated with 0.5× MBIC 90 occidiofungin (right) for 3 h. (a) C. albicans ATCC 66027 (0.5× MBIC 90 ; 8 µg/mL) and (b) C. tropicalis ATCC 66024 (0.5× MBIC 90 ; 4 µg/mL) biofilm cells stained with Calcofluor White (top panel), Live-or-Dye viability stain (middle panel), and ActinGreen 488 (bottom panel) to visualize cell wall chitin, dead cells, and actin organization, respectively. Representative CLSM images of biofilm cells are shown. Size bar: 10 microns.
    Figure Legend Snippet: Loss of actin cables in occidiofungin exposed biofilm cells. Preformed 24-h biofilm of Candida ATCC 66024 cells grown in RPMI media were left untreated (left) or treated with 0.5× MBIC 90 occidiofungin (right) for 3 h. (a) C. albicans ATCC 66027 (0.5× MBIC 90 ; 8 µg/mL) and (b) C. tropicalis ATCC 66024 (0.5× MBIC 90 ; 4 µg/mL) biofilm cells stained with Calcofluor White (top panel), Live-or-Dye viability stain (middle panel), and ActinGreen 488 (bottom panel) to visualize cell wall chitin, dead cells, and actin organization, respectively. Representative CLSM images of biofilm cells are shown. Size bar: 10 microns.

    Techniques Used: Staining

    c tropicalis atcc 66024  (ATCC)


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    ATCC c tropicalis atcc 66024
    C Tropicalis Atcc 66024, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hypovirulent strain  (ATCC)


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    ATCC hypovirulent strain
    Hypovirulent Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ep721  (ATCC)


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    ATCC ep721
    Colony morphologies of wild-type Cryphonectria parasitica strain EP155 (left column) and mutant strain Δdcl2 (right column) following infection by hypoviruses CHV-1/EP713, CHV-1/Euro7, and <t>CHV-1/EP721.</t> The colony morphologies for the uninfected EP155 and Δdcl2 strains are shown at the top. Infections were initiated by transfection with the transcripts of the corresponding hypoviruses (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.
    Ep721, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Variations in Hypovirus Interactions with the Fungal-Host RNA-Silencing Antiviral-Defense Response"

    Article Title: Variations in Hypovirus Interactions with the Fungal-Host RNA-Silencing Antiviral-Defense Response

    Journal: Journal of Virology

    doi: 10.1128/JVI.00961-12

    Colony morphologies of wild-type Cryphonectria parasitica strain EP155 (left column) and mutant strain Δdcl2 (right column) following infection by hypoviruses CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721. The colony morphologies for the uninfected EP155 and Δdcl2 strains are shown at the top. Infections were initiated by transfection with the transcripts of the corresponding hypoviruses (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.
    Figure Legend Snippet: Colony morphologies of wild-type Cryphonectria parasitica strain EP155 (left column) and mutant strain Δdcl2 (right column) following infection by hypoviruses CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721. The colony morphologies for the uninfected EP155 and Δdcl2 strains are shown at the top. Infections were initiated by transfection with the transcripts of the corresponding hypoviruses (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.

    Techniques Used: Mutagenesis, Infection, Transfection

    Accumulation of dcl2 transcripts in response to different hypoviruses. The relative levels of dcl2 transcripts were measured by semiquantitative real-time RT-PCR for uninfected strain EP155 and strain EP155 infected by CHV-1/EP713, CHV-1/Euro7, CHV-1/EP721, their corresponding Δp29 mutants, and the chimeric CHV-1/EP713 and CHV-1EP721 viruses containing the swapped p29 coding domains 713/721p29 and 721/713p29, as indicated at the bottom. The values on the y axis were normalized to the dcl2 transcript level in uninfected strain EP155, set to a value of 1.
    Figure Legend Snippet: Accumulation of dcl2 transcripts in response to different hypoviruses. The relative levels of dcl2 transcripts were measured by semiquantitative real-time RT-PCR for uninfected strain EP155 and strain EP155 infected by CHV-1/EP713, CHV-1/Euro7, CHV-1/EP721, their corresponding Δp29 mutants, and the chimeric CHV-1/EP713 and CHV-1EP721 viruses containing the swapped p29 coding domains 713/721p29 and 721/713p29, as indicated at the bottom. The values on the y axis were normalized to the dcl2 transcript level in uninfected strain EP155, set to a value of 1.

    Techniques Used: Quantitative RT-PCR, Infection

    Hypovirus RNA accumulation in the C. parasitica Δdcl2 mutant strain. (A). Gel analysis of RNA isolated from hypovirus-infected wild-type C. parasitica strain EP155 and the Δdcl2 mutant strain. Approximately 10 μg of total nucleic acids isolated from each infected strain was loaded and subjected to electrophoresis in 1% agarose gels set at 15 V overnight. Lane 1 contains 0.5 μg of the in vitro-synthesized 12,734-nt transcript from infectious CHV-1/EP713 cDNA clone pRFL4 (see Materials and Methods). Lane M contains DNA size markers. Lanes 2 through 7 contain nucleic acids from strain EP155 infected with CHV-1/EP713, the Δdcl2 mutant infected with CHV-1/EP713, EP155 infected with CHV-1/Euro7, the Δdcl2 mutant infected with CHV-1/Euro7, EP155 infected with CHV-1/EP721, and the Δdcl2 mutant infected with CHV-1EP721, respectively. The migration positions of genomic DNA (gDNA) and the viral double-stranded replicative-form RNA (dsRNA) and positive-sense single-stranded viral RNA (+ssRNA) are indicated on the right. The bands migrating near the 8- to 10-kb markers in lane 2 (CHV-1/EP713-infected strain EP155) are defective viral RNAs commonly observed in infected wild-type C. parasitica strains but not observed in the RNA-silencing mutant Δdcl2 strain (38). Note the significant increase in accumulation of viral positive-sense ssRNA. (B). Relative accumulation of positive-sense ssRNA hypovirus RNA in infected wild-type C. parasitica strain EP155 and Δdcl2 mutant strain determined by real-time RT-PCR. The values on the y axis were normalized to the viral RNA levels in the infected wild-type strain EP155. The relative levels of positive-sense ssRNA for CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721 increased 33-, 32-, and 16-fold in the Δdcl2 strain relative to the levels in strain EP155, respectively.
    Figure Legend Snippet: Hypovirus RNA accumulation in the C. parasitica Δdcl2 mutant strain. (A). Gel analysis of RNA isolated from hypovirus-infected wild-type C. parasitica strain EP155 and the Δdcl2 mutant strain. Approximately 10 μg of total nucleic acids isolated from each infected strain was loaded and subjected to electrophoresis in 1% agarose gels set at 15 V overnight. Lane 1 contains 0.5 μg of the in vitro-synthesized 12,734-nt transcript from infectious CHV-1/EP713 cDNA clone pRFL4 (see Materials and Methods). Lane M contains DNA size markers. Lanes 2 through 7 contain nucleic acids from strain EP155 infected with CHV-1/EP713, the Δdcl2 mutant infected with CHV-1/EP713, EP155 infected with CHV-1/Euro7, the Δdcl2 mutant infected with CHV-1/Euro7, EP155 infected with CHV-1/EP721, and the Δdcl2 mutant infected with CHV-1EP721, respectively. The migration positions of genomic DNA (gDNA) and the viral double-stranded replicative-form RNA (dsRNA) and positive-sense single-stranded viral RNA (+ssRNA) are indicated on the right. The bands migrating near the 8- to 10-kb markers in lane 2 (CHV-1/EP713-infected strain EP155) are defective viral RNAs commonly observed in infected wild-type C. parasitica strains but not observed in the RNA-silencing mutant Δdcl2 strain (38). Note the significant increase in accumulation of viral positive-sense ssRNA. (B). Relative accumulation of positive-sense ssRNA hypovirus RNA in infected wild-type C. parasitica strain EP155 and Δdcl2 mutant strain determined by real-time RT-PCR. The values on the y axis were normalized to the viral RNA levels in the infected wild-type strain EP155. The relative levels of positive-sense ssRNA for CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721 increased 33-, 32-, and 16-fold in the Δdcl2 strain relative to the levels in strain EP155, respectively.

    Techniques Used: Mutagenesis, Isolation, Infection, Electrophoresis, In Vitro, Synthesized, Migration, Quantitative RT-PCR

    Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 and the Δdcl2 mutant strain. (A) Schematic diagrams of chimeric viruses. The genomes of CHV-1/EP713 and CHV-1/EP721 contain two open reading frames designated ORF A and ORF B, as indicated at the top. The coding regions of individual chimeric viruses derived from the CHV-1/EP713 genome are indicated as blue boxes, and the CHV-1/EP721 genome-derived coding regions are indicated as white boxes. The autocatalytic cleavage sites for p29 and p48 are indicated by asterisks. The 5′ and 3′ noncoding terminal regions are indicated as lines. (B) Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 (left column) and the Δdcl2 mutant strain (right column). Infections were initiated by transfection with transcripts of the corresponding chimeric virus (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.
    Figure Legend Snippet: Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 and the Δdcl2 mutant strain. (A) Schematic diagrams of chimeric viruses. The genomes of CHV-1/EP713 and CHV-1/EP721 contain two open reading frames designated ORF A and ORF B, as indicated at the top. The coding regions of individual chimeric viruses derived from the CHV-1/EP713 genome are indicated as blue boxes, and the CHV-1/EP721 genome-derived coding regions are indicated as white boxes. The autocatalytic cleavage sites for p29 and p48 are indicated by asterisks. The 5′ and 3′ noncoding terminal regions are indicated as lines. (B) Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 (left column) and the Δdcl2 mutant strain (right column). Infections were initiated by transfection with transcripts of the corresponding chimeric virus (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.

    Techniques Used: Mutagenesis, Derivative Assay, Transfection, Infection

    ep721  (ATCC)


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    ATCC ep721
    Colony morphologies of wild-type Cryphonectria parasitica strain EP155 (left column) and mutant strain Δdcl2 (right column) following infection by hypoviruses CHV-1/EP713, CHV-1/Euro7, and <t>CHV-1/EP721.</t> The colony morphologies for the uninfected EP155 and Δdcl2 strains are shown at the top. Infections were initiated by transfection with the transcripts of the corresponding hypoviruses (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.
    Ep721, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Variations in Hypovirus Interactions with the Fungal-Host RNA-Silencing Antiviral-Defense Response"

    Article Title: Variations in Hypovirus Interactions with the Fungal-Host RNA-Silencing Antiviral-Defense Response

    Journal: Journal of Virology

    doi: 10.1128/JVI.00961-12

    Colony morphologies of wild-type Cryphonectria parasitica strain EP155 (left column) and mutant strain Δdcl2 (right column) following infection by hypoviruses CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721. The colony morphologies for the uninfected EP155 and Δdcl2 strains are shown at the top. Infections were initiated by transfection with the transcripts of the corresponding hypoviruses (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.
    Figure Legend Snippet: Colony morphologies of wild-type Cryphonectria parasitica strain EP155 (left column) and mutant strain Δdcl2 (right column) following infection by hypoviruses CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721. The colony morphologies for the uninfected EP155 and Δdcl2 strains are shown at the top. Infections were initiated by transfection with the transcripts of the corresponding hypoviruses (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.

    Techniques Used: Mutagenesis, Infection, Transfection

    Accumulation of dcl2 transcripts in response to different hypoviruses. The relative levels of dcl2 transcripts were measured by semiquantitative real-time RT-PCR for uninfected strain EP155 and strain EP155 infected by CHV-1/EP713, CHV-1/Euro7, CHV-1/EP721, their corresponding Δp29 mutants, and the chimeric CHV-1/EP713 and CHV-1EP721 viruses containing the swapped p29 coding domains 713/721p29 and 721/713p29, as indicated at the bottom. The values on the y axis were normalized to the dcl2 transcript level in uninfected strain EP155, set to a value of 1.
    Figure Legend Snippet: Accumulation of dcl2 transcripts in response to different hypoviruses. The relative levels of dcl2 transcripts were measured by semiquantitative real-time RT-PCR for uninfected strain EP155 and strain EP155 infected by CHV-1/EP713, CHV-1/Euro7, CHV-1/EP721, their corresponding Δp29 mutants, and the chimeric CHV-1/EP713 and CHV-1EP721 viruses containing the swapped p29 coding domains 713/721p29 and 721/713p29, as indicated at the bottom. The values on the y axis were normalized to the dcl2 transcript level in uninfected strain EP155, set to a value of 1.

    Techniques Used: Quantitative RT-PCR, Infection

    Hypovirus RNA accumulation in the C. parasitica Δdcl2 mutant strain. (A). Gel analysis of RNA isolated from hypovirus-infected wild-type C. parasitica strain EP155 and the Δdcl2 mutant strain. Approximately 10 μg of total nucleic acids isolated from each infected strain was loaded and subjected to electrophoresis in 1% agarose gels set at 15 V overnight. Lane 1 contains 0.5 μg of the in vitro-synthesized 12,734-nt transcript from infectious CHV-1/EP713 cDNA clone pRFL4 (see Materials and Methods). Lane M contains DNA size markers. Lanes 2 through 7 contain nucleic acids from strain EP155 infected with CHV-1/EP713, the Δdcl2 mutant infected with CHV-1/EP713, EP155 infected with CHV-1/Euro7, the Δdcl2 mutant infected with CHV-1/Euro7, EP155 infected with CHV-1/EP721, and the Δdcl2 mutant infected with CHV-1EP721, respectively. The migration positions of genomic DNA (gDNA) and the viral double-stranded replicative-form RNA (dsRNA) and positive-sense single-stranded viral RNA (+ssRNA) are indicated on the right. The bands migrating near the 8- to 10-kb markers in lane 2 (CHV-1/EP713-infected strain EP155) are defective viral RNAs commonly observed in infected wild-type C. parasitica strains but not observed in the RNA-silencing mutant Δdcl2 strain (38). Note the significant increase in accumulation of viral positive-sense ssRNA. (B). Relative accumulation of positive-sense ssRNA hypovirus RNA in infected wild-type C. parasitica strain EP155 and Δdcl2 mutant strain determined by real-time RT-PCR. The values on the y axis were normalized to the viral RNA levels in the infected wild-type strain EP155. The relative levels of positive-sense ssRNA for CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721 increased 33-, 32-, and 16-fold in the Δdcl2 strain relative to the levels in strain EP155, respectively.
    Figure Legend Snippet: Hypovirus RNA accumulation in the C. parasitica Δdcl2 mutant strain. (A). Gel analysis of RNA isolated from hypovirus-infected wild-type C. parasitica strain EP155 and the Δdcl2 mutant strain. Approximately 10 μg of total nucleic acids isolated from each infected strain was loaded and subjected to electrophoresis in 1% agarose gels set at 15 V overnight. Lane 1 contains 0.5 μg of the in vitro-synthesized 12,734-nt transcript from infectious CHV-1/EP713 cDNA clone pRFL4 (see Materials and Methods). Lane M contains DNA size markers. Lanes 2 through 7 contain nucleic acids from strain EP155 infected with CHV-1/EP713, the Δdcl2 mutant infected with CHV-1/EP713, EP155 infected with CHV-1/Euro7, the Δdcl2 mutant infected with CHV-1/Euro7, EP155 infected with CHV-1/EP721, and the Δdcl2 mutant infected with CHV-1EP721, respectively. The migration positions of genomic DNA (gDNA) and the viral double-stranded replicative-form RNA (dsRNA) and positive-sense single-stranded viral RNA (+ssRNA) are indicated on the right. The bands migrating near the 8- to 10-kb markers in lane 2 (CHV-1/EP713-infected strain EP155) are defective viral RNAs commonly observed in infected wild-type C. parasitica strains but not observed in the RNA-silencing mutant Δdcl2 strain (38). Note the significant increase in accumulation of viral positive-sense ssRNA. (B). Relative accumulation of positive-sense ssRNA hypovirus RNA in infected wild-type C. parasitica strain EP155 and Δdcl2 mutant strain determined by real-time RT-PCR. The values on the y axis were normalized to the viral RNA levels in the infected wild-type strain EP155. The relative levels of positive-sense ssRNA for CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721 increased 33-, 32-, and 16-fold in the Δdcl2 strain relative to the levels in strain EP155, respectively.

    Techniques Used: Mutagenesis, Isolation, Infection, Electrophoresis, In Vitro, Synthesized, Migration, Quantitative RT-PCR

    Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 and the Δdcl2 mutant strain. (A) Schematic diagrams of chimeric viruses. The genomes of CHV-1/EP713 and CHV-1/EP721 contain two open reading frames designated ORF A and ORF B, as indicated at the top. The coding regions of individual chimeric viruses derived from the CHV-1/EP713 genome are indicated as blue boxes, and the CHV-1/EP721 genome-derived coding regions are indicated as white boxes. The autocatalytic cleavage sites for p29 and p48 are indicated by asterisks. The 5′ and 3′ noncoding terminal regions are indicated as lines. (B) Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 (left column) and the Δdcl2 mutant strain (right column). Infections were initiated by transfection with transcripts of the corresponding chimeric virus (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.
    Figure Legend Snippet: Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 and the Δdcl2 mutant strain. (A) Schematic diagrams of chimeric viruses. The genomes of CHV-1/EP713 and CHV-1/EP721 contain two open reading frames designated ORF A and ORF B, as indicated at the top. The coding regions of individual chimeric viruses derived from the CHV-1/EP713 genome are indicated as blue boxes, and the CHV-1/EP721 genome-derived coding regions are indicated as white boxes. The autocatalytic cleavage sites for p29 and p48 are indicated by asterisks. The 5′ and 3′ noncoding terminal regions are indicated as lines. (B) Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 (left column) and the Δdcl2 mutant strain (right column). Infections were initiated by transfection with transcripts of the corresponding chimeric virus (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.

    Techniques Used: Mutagenesis, Derivative Assay, Transfection, Infection

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    Colony morphologies of wild-type Cryphonectria parasitica strain EP155 (left column) and mutant strain Δdcl2 (right column) following infection by hypoviruses CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721. The colony morphologies for the uninfected EP155 and Δdcl2 strains are shown at the top. Infections were initiated by transfection with the transcripts of the corresponding hypoviruses (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.

    Journal: Journal of Virology

    Article Title: Variations in Hypovirus Interactions with the Fungal-Host RNA-Silencing Antiviral-Defense Response

    doi: 10.1128/JVI.00961-12

    Figure Lengend Snippet: Colony morphologies of wild-type Cryphonectria parasitica strain EP155 (left column) and mutant strain Δdcl2 (right column) following infection by hypoviruses CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721. The colony morphologies for the uninfected EP155 and Δdcl2 strains are shown at the top. Infections were initiated by transfection with the transcripts of the corresponding hypoviruses (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.

    Article Snippet: Hypovirus CHV-1/EP721 was identified in the Italian hypovirulent strain HI2 and then transferred by anastomosis to North American strain EP60 (ATCC 38765) isolated from Michigan (W. MacDonald, personal communication), resulting in a hypovirulent strain, EP721 (ATCC 66024).

    Techniques: Mutagenesis, Infection, Transfection

    Accumulation of dcl2 transcripts in response to different hypoviruses. The relative levels of dcl2 transcripts were measured by semiquantitative real-time RT-PCR for uninfected strain EP155 and strain EP155 infected by CHV-1/EP713, CHV-1/Euro7, CHV-1/EP721, their corresponding Δp29 mutants, and the chimeric CHV-1/EP713 and CHV-1EP721 viruses containing the swapped p29 coding domains 713/721p29 and 721/713p29, as indicated at the bottom. The values on the y axis were normalized to the dcl2 transcript level in uninfected strain EP155, set to a value of 1.

    Journal: Journal of Virology

    Article Title: Variations in Hypovirus Interactions with the Fungal-Host RNA-Silencing Antiviral-Defense Response

    doi: 10.1128/JVI.00961-12

    Figure Lengend Snippet: Accumulation of dcl2 transcripts in response to different hypoviruses. The relative levels of dcl2 transcripts were measured by semiquantitative real-time RT-PCR for uninfected strain EP155 and strain EP155 infected by CHV-1/EP713, CHV-1/Euro7, CHV-1/EP721, their corresponding Δp29 mutants, and the chimeric CHV-1/EP713 and CHV-1EP721 viruses containing the swapped p29 coding domains 713/721p29 and 721/713p29, as indicated at the bottom. The values on the y axis were normalized to the dcl2 transcript level in uninfected strain EP155, set to a value of 1.

    Article Snippet: Hypovirus CHV-1/EP721 was identified in the Italian hypovirulent strain HI2 and then transferred by anastomosis to North American strain EP60 (ATCC 38765) isolated from Michigan (W. MacDonald, personal communication), resulting in a hypovirulent strain, EP721 (ATCC 66024).

    Techniques: Quantitative RT-PCR, Infection

    Hypovirus RNA accumulation in the C. parasitica Δdcl2 mutant strain. (A). Gel analysis of RNA isolated from hypovirus-infected wild-type C. parasitica strain EP155 and the Δdcl2 mutant strain. Approximately 10 μg of total nucleic acids isolated from each infected strain was loaded and subjected to electrophoresis in 1% agarose gels set at 15 V overnight. Lane 1 contains 0.5 μg of the in vitro-synthesized 12,734-nt transcript from infectious CHV-1/EP713 cDNA clone pRFL4 (see Materials and Methods). Lane M contains DNA size markers. Lanes 2 through 7 contain nucleic acids from strain EP155 infected with CHV-1/EP713, the Δdcl2 mutant infected with CHV-1/EP713, EP155 infected with CHV-1/Euro7, the Δdcl2 mutant infected with CHV-1/Euro7, EP155 infected with CHV-1/EP721, and the Δdcl2 mutant infected with CHV-1EP721, respectively. The migration positions of genomic DNA (gDNA) and the viral double-stranded replicative-form RNA (dsRNA) and positive-sense single-stranded viral RNA (+ssRNA) are indicated on the right. The bands migrating near the 8- to 10-kb markers in lane 2 (CHV-1/EP713-infected strain EP155) are defective viral RNAs commonly observed in infected wild-type C. parasitica strains but not observed in the RNA-silencing mutant Δdcl2 strain (38). Note the significant increase in accumulation of viral positive-sense ssRNA. (B). Relative accumulation of positive-sense ssRNA hypovirus RNA in infected wild-type C. parasitica strain EP155 and Δdcl2 mutant strain determined by real-time RT-PCR. The values on the y axis were normalized to the viral RNA levels in the infected wild-type strain EP155. The relative levels of positive-sense ssRNA for CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721 increased 33-, 32-, and 16-fold in the Δdcl2 strain relative to the levels in strain EP155, respectively.

    Journal: Journal of Virology

    Article Title: Variations in Hypovirus Interactions with the Fungal-Host RNA-Silencing Antiviral-Defense Response

    doi: 10.1128/JVI.00961-12

    Figure Lengend Snippet: Hypovirus RNA accumulation in the C. parasitica Δdcl2 mutant strain. (A). Gel analysis of RNA isolated from hypovirus-infected wild-type C. parasitica strain EP155 and the Δdcl2 mutant strain. Approximately 10 μg of total nucleic acids isolated from each infected strain was loaded and subjected to electrophoresis in 1% agarose gels set at 15 V overnight. Lane 1 contains 0.5 μg of the in vitro-synthesized 12,734-nt transcript from infectious CHV-1/EP713 cDNA clone pRFL4 (see Materials and Methods). Lane M contains DNA size markers. Lanes 2 through 7 contain nucleic acids from strain EP155 infected with CHV-1/EP713, the Δdcl2 mutant infected with CHV-1/EP713, EP155 infected with CHV-1/Euro7, the Δdcl2 mutant infected with CHV-1/Euro7, EP155 infected with CHV-1/EP721, and the Δdcl2 mutant infected with CHV-1EP721, respectively. The migration positions of genomic DNA (gDNA) and the viral double-stranded replicative-form RNA (dsRNA) and positive-sense single-stranded viral RNA (+ssRNA) are indicated on the right. The bands migrating near the 8- to 10-kb markers in lane 2 (CHV-1/EP713-infected strain EP155) are defective viral RNAs commonly observed in infected wild-type C. parasitica strains but not observed in the RNA-silencing mutant Δdcl2 strain (38). Note the significant increase in accumulation of viral positive-sense ssRNA. (B). Relative accumulation of positive-sense ssRNA hypovirus RNA in infected wild-type C. parasitica strain EP155 and Δdcl2 mutant strain determined by real-time RT-PCR. The values on the y axis were normalized to the viral RNA levels in the infected wild-type strain EP155. The relative levels of positive-sense ssRNA for CHV-1/EP713, CHV-1/Euro7, and CHV-1/EP721 increased 33-, 32-, and 16-fold in the Δdcl2 strain relative to the levels in strain EP155, respectively.

    Article Snippet: Hypovirus CHV-1/EP721 was identified in the Italian hypovirulent strain HI2 and then transferred by anastomosis to North American strain EP60 (ATCC 38765) isolated from Michigan (W. MacDonald, personal communication), resulting in a hypovirulent strain, EP721 (ATCC 66024).

    Techniques: Mutagenesis, Isolation, Infection, Electrophoresis, In Vitro, Synthesized, Migration, Quantitative RT-PCR

    Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 and the Δdcl2 mutant strain. (A) Schematic diagrams of chimeric viruses. The genomes of CHV-1/EP713 and CHV-1/EP721 contain two open reading frames designated ORF A and ORF B, as indicated at the top. The coding regions of individual chimeric viruses derived from the CHV-1/EP713 genome are indicated as blue boxes, and the CHV-1/EP721 genome-derived coding regions are indicated as white boxes. The autocatalytic cleavage sites for p29 and p48 are indicated by asterisks. The 5′ and 3′ noncoding terminal regions are indicated as lines. (B) Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 (left column) and the Δdcl2 mutant strain (right column). Infections were initiated by transfection with transcripts of the corresponding chimeric virus (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.

    Journal: Journal of Virology

    Article Title: Variations in Hypovirus Interactions with the Fungal-Host RNA-Silencing Antiviral-Defense Response

    doi: 10.1128/JVI.00961-12

    Figure Lengend Snippet: Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 and the Δdcl2 mutant strain. (A) Schematic diagrams of chimeric viruses. The genomes of CHV-1/EP713 and CHV-1/EP721 contain two open reading frames designated ORF A and ORF B, as indicated at the top. The coding regions of individual chimeric viruses derived from the CHV-1/EP713 genome are indicated as blue boxes, and the CHV-1/EP721 genome-derived coding regions are indicated as white boxes. The autocatalytic cleavage sites for p29 and p48 are indicated by asterisks. The 5′ and 3′ noncoding terminal regions are indicated as lines. (B) Colony morphologies conferred by CHV-1/EP713–CHV-1/EP721 chimeric viruses in wild-type strain EP155 (left column) and the Δdcl2 mutant strain (right column). Infections were initiated by transfection with transcripts of the corresponding chimeric virus (3), and infected colonies were transferred to potato dextrose agar. The photographs were taken on day 7 of the experiment.

    Article Snippet: Hypovirus CHV-1/EP721 was identified in the Italian hypovirulent strain HI2 and then transferred by anastomosis to North American strain EP60 (ATCC 38765) isolated from Michigan (W. MacDonald, personal communication), resulting in a hypovirulent strain, EP721 (ATCC 66024).

    Techniques: Mutagenesis, Derivative Assay, Transfection, Infection