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Analysis of cytochrome b(5) reductase-mediated metabolism in the phytopathogenic fungus Zymoseptoria tritici reveals novel functionalities implicated in virulence.

Derbyshire MC, Michaelson L, Parker J, Kelly S, Thacker U, Powers SJ, Bailey A, Hammond-Kosack K, Courbot M, Rudd J - Fungal Genet. Biol. (2015)

Bottom Line: These defects coincided with alterations in fatty acid, sphingolipid and sterol biosynthesis observed through GC-MS and HPLC analyses.Our study reports the first functional characterisation of CBR gene family members in a plant pathogenic filamentous fungus.This also represents the first direct observation of CBR functional ablation impacting upon fungal sterol biosynthesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Biology and Crop Science, Rothamsted Research, West Common, Harpenden, Hertfordshire AL5 2JQ, UK. Electronic address: markcharder@gmail.com.

No MeSH data available.


Related in: MedlinePlus

ΔZtCBR1 mutants have altered sterol profiles. (A) Mean relative abundance of all ergosterol and all intermediates in the ergosterol biosynthetic pathway identified for WT and a ΔZtCBR1 strain. Bars represent standard error, all deviations from WT levels in the ΔZtCBR1 strain were significant (** p < 0.01, *** p < 0.001). (B) Diagram depicting the reactions catalysed by the enzymes CYP51 and Erg25 during sterol biosynthesis. Solid arrow represents usual direction of biosynthetic pathway. Perforated arrow represents an alternative route for the CYP51 substrate, eburicol, which may be followed more frequently if CYP51 activity is compromised. Solid boxes surround compounds that accumulated in ΔZtCBR1 relative to the WT. Red circles mark the sites of enzymatic alterations at each sterol biosynthesis step. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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f0040: ΔZtCBR1 mutants have altered sterol profiles. (A) Mean relative abundance of all ergosterol and all intermediates in the ergosterol biosynthetic pathway identified for WT and a ΔZtCBR1 strain. Bars represent standard error, all deviations from WT levels in the ΔZtCBR1 strain were significant (** p < 0.01, *** p < 0.001). (B) Diagram depicting the reactions catalysed by the enzymes CYP51 and Erg25 during sterol biosynthesis. Solid arrow represents usual direction of biosynthetic pathway. Perforated arrow represents an alternative route for the CYP51 substrate, eburicol, which may be followed more frequently if CYP51 activity is compromised. Solid boxes surround compounds that accumulated in ΔZtCBR1 relative to the WT. Red circles mark the sites of enzymatic alterations at each sterol biosynthesis step. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Mentions: In order to assess the effect of ZtCBR1 ablation on sterol biosynthesis, the sterol profile of the WT and ΔZtCBR1-1 strains were analysed using GC–MS. ΔZtCBR1-1 displayed a significant reduction in the relative abundance of the final product of the sterol pathway, ergosterol (ergosta-5,7,22-trienol), relative to the WT (p < 0.001). Several intermediate compounds in the sterol biosynthesis pathway including ergosta-5,8,22,24(28)-tetraenol, ergosta-5,8,22-trienol, ergosta-7,22-dienol and obtusifoliol (14α-dimethyl-5α-ergosta-8,24(28)-dienol) were significantly increased in relative abundance in the ΔZtCBR1-1 strain compared to the WT (p < 0.01, p < 0.01, p < 0.001 and p < 0.01 respectively). The substrate of the enzyme CYP51 (eburicol (4,4,14-trimethylergosta-8,24(28)-dienol)), which is a target of azole antifungals, accumulated in ΔZtCBR1-1 but was not detected in the WT (p < 0.001) (Fig. 8).


Analysis of cytochrome b(5) reductase-mediated metabolism in the phytopathogenic fungus Zymoseptoria tritici reveals novel functionalities implicated in virulence.

Derbyshire MC, Michaelson L, Parker J, Kelly S, Thacker U, Powers SJ, Bailey A, Hammond-Kosack K, Courbot M, Rudd J - Fungal Genet. Biol. (2015)

ΔZtCBR1 mutants have altered sterol profiles. (A) Mean relative abundance of all ergosterol and all intermediates in the ergosterol biosynthetic pathway identified for WT and a ΔZtCBR1 strain. Bars represent standard error, all deviations from WT levels in the ΔZtCBR1 strain were significant (** p < 0.01, *** p < 0.001). (B) Diagram depicting the reactions catalysed by the enzymes CYP51 and Erg25 during sterol biosynthesis. Solid arrow represents usual direction of biosynthetic pathway. Perforated arrow represents an alternative route for the CYP51 substrate, eburicol, which may be followed more frequently if CYP51 activity is compromised. Solid boxes surround compounds that accumulated in ΔZtCBR1 relative to the WT. Red circles mark the sites of enzymatic alterations at each sterol biosynthesis step. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4557397&req=5

f0040: ΔZtCBR1 mutants have altered sterol profiles. (A) Mean relative abundance of all ergosterol and all intermediates in the ergosterol biosynthetic pathway identified for WT and a ΔZtCBR1 strain. Bars represent standard error, all deviations from WT levels in the ΔZtCBR1 strain were significant (** p < 0.01, *** p < 0.001). (B) Diagram depicting the reactions catalysed by the enzymes CYP51 and Erg25 during sterol biosynthesis. Solid arrow represents usual direction of biosynthetic pathway. Perforated arrow represents an alternative route for the CYP51 substrate, eburicol, which may be followed more frequently if CYP51 activity is compromised. Solid boxes surround compounds that accumulated in ΔZtCBR1 relative to the WT. Red circles mark the sites of enzymatic alterations at each sterol biosynthesis step. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Mentions: In order to assess the effect of ZtCBR1 ablation on sterol biosynthesis, the sterol profile of the WT and ΔZtCBR1-1 strains were analysed using GC–MS. ΔZtCBR1-1 displayed a significant reduction in the relative abundance of the final product of the sterol pathway, ergosterol (ergosta-5,7,22-trienol), relative to the WT (p < 0.001). Several intermediate compounds in the sterol biosynthesis pathway including ergosta-5,8,22,24(28)-tetraenol, ergosta-5,8,22-trienol, ergosta-7,22-dienol and obtusifoliol (14α-dimethyl-5α-ergosta-8,24(28)-dienol) were significantly increased in relative abundance in the ΔZtCBR1-1 strain compared to the WT (p < 0.01, p < 0.01, p < 0.001 and p < 0.01 respectively). The substrate of the enzyme CYP51 (eburicol (4,4,14-trimethylergosta-8,24(28)-dienol)), which is a target of azole antifungals, accumulated in ΔZtCBR1-1 but was not detected in the WT (p < 0.001) (Fig. 8).

Bottom Line: These defects coincided with alterations in fatty acid, sphingolipid and sterol biosynthesis observed through GC-MS and HPLC analyses.Our study reports the first functional characterisation of CBR gene family members in a plant pathogenic filamentous fungus.This also represents the first direct observation of CBR functional ablation impacting upon fungal sterol biosynthesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Biology and Crop Science, Rothamsted Research, West Common, Harpenden, Hertfordshire AL5 2JQ, UK. Electronic address: markcharder@gmail.com.

No MeSH data available.


Related in: MedlinePlus