Azole sensitivity in Leptosphaeria pathogens of oilseed rape: the role of lanosterol 14α-demethylase

Thomas Sewell; Nichola J Hawkins; Henrik Stotz; Yongju Huang; Steven L Kelly; Diane E Kelly; Bart Fraaije; Bruce Fitt

doi:10.1038/s41598-017-15545-9

Azole sensitivity in Leptosphaeria pathogens of oilseed rape: the role of lanosterol 14α-demethylase

Thomas Sewell, Nichola J Hawkins, Henrik Stotz, Yongju Huang, Steven L Kelly, Diane E Kelly, Bart Fraaije, Bruce Fitt

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Abstract

Lanosterol 14-α demethy
lase is a key enzyme intermediating the biosynthesis of ergosterol in fungi,
and the target of azole fungicides. Studies have suggested that Leptosphaeria maculans and L.
biglobosa, the causal agents of phoma stem canker on oilseed rape, differ in their sensitivity to some
azoles, which could be driving pathogen frequency change in crops. Here we used CYP51 protein
modelling and heterologous expression to determine whether there are interspecific differences at the
target-site level. Moreover, we provide an example of intrinsic sensitivity differences exhibited by both
Leptosphaeria spp. in vitro and in planta. Comparison of homologous protein models identified highly
conserved residues, particularly at the azole binding site, and heterologous expression of LmCYP51B
and LbCYP51B, with fungicide sensitivity testing of the transformants, suggests that both proteins are
similarly sensitive to azole fungicides flusilazole, prothioconazole-desthio and tebuconazole. Fungicide
sensitivity testing on isolates shows that they sometimes have a minor difference in sensitivity in vitro
and in planta. These results suggest that azole fungicides remain a useful component of integrated
phoma stem canker control in the UK due to their effectiveness on both Leptosphaeria spp. Other
factors, such as varietal resistance or climate, may be driving observed frequency changes between
species.

Original language	English
Article number	15849
Journal	Scientific Reports
Volume	7
DOIs	https://doi.org/10.1038/s41598-017-15545-9
Publication status	Published - 20 Nov 2017

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10.1038/s41598-017-15545-9Licence: CC BY

Published_VersionFinal published version, 2.2 MBLicence: CC BY

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title = "Azole sensitivity in Leptosphaeria pathogens of oilseed rape: the role of lanosterol 14α-demethylase",

abstract = "Lanosterol 14-α demethylase is a key enzyme intermediating the biosynthesis of ergosterol in fungi,and the target of azole fungicides. Studies have suggested that Leptosphaeria maculans and L.biglobosa, the causal agents of phoma stem canker on oilseed rape, differ in their sensitivity to someazoles, which could be driving pathogen frequency change in crops. Here we used CYP51 proteinmodelling and heterologous expression to determine whether there are interspecific differences at thetarget-site level. Moreover, we provide an example of intrinsic sensitivity differences exhibited by bothLeptosphaeria spp. in vitro and in planta. Comparison of homologous protein models identified highlyconserved residues, particularly at the azole binding site, and heterologous expression of LmCYP51Band LbCYP51B, with fungicide sensitivity testing of the transformants, suggests that both proteins aresimilarly sensitive to azole fungicides flusilazole, prothioconazole-desthio and tebuconazole. Fungicidesensitivity testing on isolates shows that they sometimes have a minor difference in sensitivity in vitroand in planta. These results suggest that azole fungicides remain a useful component of integratedphoma stem canker control in the UK due to their effectiveness on both Leptosphaeria spp. Otherfactors, such as varietal resistance or climate, may be driving observed frequency changes betweenspecies.",

author = "Thomas Sewell and {J Hawkins}, Nichola and Henrik Stotz and Yongju Huang and {L Kelly}, Steven and {E Kelly}, Diane and Bart Fraaije and Bruce Fitt",

note = "Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Te images or other third party material in this article are included in the article{\textquoteright}s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article{\textquoteright}s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. {\textcopyright} The Author(s) 2017",

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AU - L Kelly, Steven

AU - E Kelly, Diane

AU - Fraaije, Bart

AU - Fitt, Bruce

N1 - Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Te images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2017

PY - 2017/11/20

Y1 - 2017/11/20

N2 - Lanosterol 14-α demethylase is a key enzyme intermediating the biosynthesis of ergosterol in fungi,and the target of azole fungicides. Studies have suggested that Leptosphaeria maculans and L.biglobosa, the causal agents of phoma stem canker on oilseed rape, differ in their sensitivity to someazoles, which could be driving pathogen frequency change in crops. Here we used CYP51 proteinmodelling and heterologous expression to determine whether there are interspecific differences at thetarget-site level. Moreover, we provide an example of intrinsic sensitivity differences exhibited by bothLeptosphaeria spp. in vitro and in planta. Comparison of homologous protein models identified highlyconserved residues, particularly at the azole binding site, and heterologous expression of LmCYP51Band LbCYP51B, with fungicide sensitivity testing of the transformants, suggests that both proteins aresimilarly sensitive to azole fungicides flusilazole, prothioconazole-desthio and tebuconazole. Fungicidesensitivity testing on isolates shows that they sometimes have a minor difference in sensitivity in vitroand in planta. These results suggest that azole fungicides remain a useful component of integratedphoma stem canker control in the UK due to their effectiveness on both Leptosphaeria spp. Otherfactors, such as varietal resistance or climate, may be driving observed frequency changes betweenspecies.

AB - Lanosterol 14-α demethylase is a key enzyme intermediating the biosynthesis of ergosterol in fungi,and the target of azole fungicides. Studies have suggested that Leptosphaeria maculans and L.biglobosa, the causal agents of phoma stem canker on oilseed rape, differ in their sensitivity to someazoles, which could be driving pathogen frequency change in crops. Here we used CYP51 proteinmodelling and heterologous expression to determine whether there are interspecific differences at thetarget-site level. Moreover, we provide an example of intrinsic sensitivity differences exhibited by bothLeptosphaeria spp. in vitro and in planta. Comparison of homologous protein models identified highlyconserved residues, particularly at the azole binding site, and heterologous expression of LmCYP51Band LbCYP51B, with fungicide sensitivity testing of the transformants, suggests that both proteins aresimilarly sensitive to azole fungicides flusilazole, prothioconazole-desthio and tebuconazole. Fungicidesensitivity testing on isolates shows that they sometimes have a minor difference in sensitivity in vitroand in planta. These results suggest that azole fungicides remain a useful component of integratedphoma stem canker control in the UK due to their effectiveness on both Leptosphaeria spp. Otherfactors, such as varietal resistance or climate, may be driving observed frequency changes betweenspecies.

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DO - 10.1038/s41598-017-15545-9

M3 - Article

SN - 2045-2322

VL - 7

JO - Scientific Reports

JF - Scientific Reports

M1 - 15849

ER -

Azole sensitivity in Leptosphaeria pathogens of oilseed rape: the role of lanosterol 14α-demethylase

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