The podosphaera fusca TUB2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research

The powdery mildew fungus Podosphaera fusca (synonym Podosphaera xanthii) is the main causal agent of cucurbit powdery mildew and one of the most important limiting factors for cucurbit production worldwide. Despite the fungus' economic importance, very little is known about the physiological a...

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Main Authors: Vela-Corcía, David, Bellón-Góme, Davinia, Lopez-Ruiz, Francisco, Tores, Juan, Perez-Garcia, Alejandro
Format: Journal Article
Published: Elsevier Ltd. 2014
Subjects:
Online Access:http://hdl.handle.net/20.500.11937/29198
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author Vela-Corcía, David
Bellón-Góme, Davinia
Lopez-Ruiz, Francisco
Tores, Juan
Perez-Garcia, Alejandro
author_facet Vela-Corcía, David
Bellón-Góme, Davinia
Lopez-Ruiz, Francisco
Tores, Juan
Perez-Garcia, Alejandro
author_sort Vela-Corcía, David
building Curtin Institutional Repository
collection Online Access
description The powdery mildew fungus Podosphaera fusca (synonym Podosphaera xanthii) is the main causal agent of cucurbit powdery mildew and one of the most important limiting factors for cucurbit production worldwide. Despite the fungus' economic importance, very little is known about the physiological and molecular processes involved in P. fusca biology and pathogenesis. In this study, we isolated and characterised the β-tubulin-encoding gene of P. fusca (PfTUB2) to develop molecular tools with different applications in powdery mildew research. PfTUB2 is predicted to encode a protein of 447 amino acid residues. The coding region is interrupted by six introns that occur at approximately the same positions as the introns present in other fungal TUB2-like genes. Once cloned, the PfTUB2 sequence information was used in different applications. Our results showed that the TUB2 gene is a good marker for molecular phylogenetics in powdery mildew fungi but it is unsuitable for the analysis of intraspecific diversity in P. fusca. The expression of PfTUB2 was proven to be stable in different temperature conditions, supporting its use as a reference gene in quantitative gene expression studies. Furthermore, an allele-specific PCR assay for the detection of resistance to methyl-2-benzimidazole carbamate (MBC) fungicides in P. fusca was developed based on the correlation between the single amino acid change E198A in β-tubulin and the MBC resistance phenotype. Lastly, PfTUB2 was used as a target gene in the development of a high-throughput method to quantify fungal growth in plant tissues.
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spelling curtin-20.500.11937-291982017-09-13T15:23:05Z The podosphaera fusca TUB2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research Vela-Corcía, David Bellón-Góme, Davinia Lopez-Ruiz, Francisco Tores, Juan Perez-Garcia, Alejandro High-throughput quantification Gene expression MBC resistance Allele-specific PCR Molecular phylogeny The powdery mildew fungus Podosphaera fusca (synonym Podosphaera xanthii) is the main causal agent of cucurbit powdery mildew and one of the most important limiting factors for cucurbit production worldwide. Despite the fungus' economic importance, very little is known about the physiological and molecular processes involved in P. fusca biology and pathogenesis. In this study, we isolated and characterised the β-tubulin-encoding gene of P. fusca (PfTUB2) to develop molecular tools with different applications in powdery mildew research. PfTUB2 is predicted to encode a protein of 447 amino acid residues. The coding region is interrupted by six introns that occur at approximately the same positions as the introns present in other fungal TUB2-like genes. Once cloned, the PfTUB2 sequence information was used in different applications. Our results showed that the TUB2 gene is a good marker for molecular phylogenetics in powdery mildew fungi but it is unsuitable for the analysis of intraspecific diversity in P. fusca. The expression of PfTUB2 was proven to be stable in different temperature conditions, supporting its use as a reference gene in quantitative gene expression studies. Furthermore, an allele-specific PCR assay for the detection of resistance to methyl-2-benzimidazole carbamate (MBC) fungicides in P. fusca was developed based on the correlation between the single amino acid change E198A in β-tubulin and the MBC resistance phenotype. Lastly, PfTUB2 was used as a target gene in the development of a high-throughput method to quantify fungal growth in plant tissues. 2014 Journal Article http://hdl.handle.net/20.500.11937/29198 10.1016/j.funbio.2013.12.001 Elsevier Ltd. restricted
spellingShingle High-throughput quantification
Gene expression
MBC resistance
Allele-specific PCR
Molecular phylogeny
Vela-Corcía, David
Bellón-Góme, Davinia
Lopez-Ruiz, Francisco
Tores, Juan
Perez-Garcia, Alejandro
The podosphaera fusca TUB2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research
title The podosphaera fusca TUB2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research
title_full The podosphaera fusca TUB2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research
title_fullStr The podosphaera fusca TUB2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research
title_full_unstemmed The podosphaera fusca TUB2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research
title_short The podosphaera fusca TUB2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research
title_sort podosphaera fusca tub2 gene, a molecular "swiss army knife" with multiple applications in powdery mildew research
topic High-throughput quantification
Gene expression
MBC resistance
Allele-specific PCR
Molecular phylogeny
url http://hdl.handle.net/20.500.11937/29198