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Conditional promoters for analysis of essential genes in Zymoseptoria tritici()

Development of new fungicides, needed for sustainable control of fungal plant pathogens, requires identification of novel anti-fungal targets. Essential fungal-specific proteins are good candidates, but due to their importance, gene deletion mutants are not viable. Consequently, their cellular role...

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Autores principales: Kilaru, S., Ma, W., Schuster, M., Courbot, M., Steinberg, G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academic Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4502454/
https://www.ncbi.nlm.nih.gov/pubmed/26092803
http://dx.doi.org/10.1016/j.fgb.2015.03.024
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author Kilaru, S.
Ma, W.
Schuster, M.
Courbot, M.
Steinberg, G.
author_facet Kilaru, S.
Ma, W.
Schuster, M.
Courbot, M.
Steinberg, G.
author_sort Kilaru, S.
collection PubMed
description Development of new fungicides, needed for sustainable control of fungal plant pathogens, requires identification of novel anti-fungal targets. Essential fungal-specific proteins are good candidates, but due to their importance, gene deletion mutants are not viable. Consequently, their cellular role often remains elusive. This hindrance can be overcome by the use of conditional mutants, where expression is controlled by an inducible/repressible promoter. Here, we introduce 5 inducible/repressible promoter systems to study essential genes in the wheat pathogen Zymoseptoria tritici. We fused the gene for enhanced green-fluorescent protein (egfp) to the promoter region of Z. tritici nitrate reductase (Pnar1; induced by nitrogen and repressed by ammonium), 1,4-β-endoxylanase A (Pex1A; induced by xylose and repressed by maltodextrin), l-arabinofuranosidase B (PlaraB; induced by arabinose and repressed by glucose), galactose-1-phosphate uridylyltransferase 7 (Pgal7; induced by galactose and repressed by glucose) and isocitrate lyase (Picl1; induced by sodium acetate and repressed by glucose). This was followed by quantitative analysis of cytoplasmic reporter fluorescence under induced and repressed conditions. We show that Pnar1, PlaraB and Pex1A drive very little or no egfp expression when repressed, but induce moderate protein production when induced. In contrast, Pgal7 and Picl1 show considerable egfp expression when repressed, and were strongly induced in the presence of their inducers. Normalising the expression levels of all promoters to that of the α-tubulin promoter Ptub2 revealed that PlaraB was the weakest promoter (∼20% of Ptub2), whereas Picl1 strongly expressed the reporter (∼250% of Ptub2). The use of these tools promises a better understanding of essential genes, which will help developing novel control strategies that protect wheat from Z. tritici.
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spelling pubmed-45024542015-07-21 Conditional promoters for analysis of essential genes in Zymoseptoria tritici() Kilaru, S. Ma, W. Schuster, M. Courbot, M. Steinberg, G. Fungal Genet Biol Article Development of new fungicides, needed for sustainable control of fungal plant pathogens, requires identification of novel anti-fungal targets. Essential fungal-specific proteins are good candidates, but due to their importance, gene deletion mutants are not viable. Consequently, their cellular role often remains elusive. This hindrance can be overcome by the use of conditional mutants, where expression is controlled by an inducible/repressible promoter. Here, we introduce 5 inducible/repressible promoter systems to study essential genes in the wheat pathogen Zymoseptoria tritici. We fused the gene for enhanced green-fluorescent protein (egfp) to the promoter region of Z. tritici nitrate reductase (Pnar1; induced by nitrogen and repressed by ammonium), 1,4-β-endoxylanase A (Pex1A; induced by xylose and repressed by maltodextrin), l-arabinofuranosidase B (PlaraB; induced by arabinose and repressed by glucose), galactose-1-phosphate uridylyltransferase 7 (Pgal7; induced by galactose and repressed by glucose) and isocitrate lyase (Picl1; induced by sodium acetate and repressed by glucose). This was followed by quantitative analysis of cytoplasmic reporter fluorescence under induced and repressed conditions. We show that Pnar1, PlaraB and Pex1A drive very little or no egfp expression when repressed, but induce moderate protein production when induced. In contrast, Pgal7 and Picl1 show considerable egfp expression when repressed, and were strongly induced in the presence of their inducers. Normalising the expression levels of all promoters to that of the α-tubulin promoter Ptub2 revealed that PlaraB was the weakest promoter (∼20% of Ptub2), whereas Picl1 strongly expressed the reporter (∼250% of Ptub2). The use of these tools promises a better understanding of essential genes, which will help developing novel control strategies that protect wheat from Z. tritici. Academic Press 2015-06 /pmc/articles/PMC4502454/ /pubmed/26092803 http://dx.doi.org/10.1016/j.fgb.2015.03.024 Text en © 2015 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kilaru, S.
Ma, W.
Schuster, M.
Courbot, M.
Steinberg, G.
Conditional promoters for analysis of essential genes in Zymoseptoria tritici()
title Conditional promoters for analysis of essential genes in Zymoseptoria tritici()
title_full Conditional promoters for analysis of essential genes in Zymoseptoria tritici()
title_fullStr Conditional promoters for analysis of essential genes in Zymoseptoria tritici()
title_full_unstemmed Conditional promoters for analysis of essential genes in Zymoseptoria tritici()
title_short Conditional promoters for analysis of essential genes in Zymoseptoria tritici()
title_sort conditional promoters for analysis of essential genes in zymoseptoria tritici()
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4502454/
https://www.ncbi.nlm.nih.gov/pubmed/26092803
http://dx.doi.org/10.1016/j.fgb.2015.03.024
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