Testing reference genes for transcript profiling in Uromyces appendiculatus during urediospore infection of common bean.
Identifieur interne : 000008 ( Main/Corpus ); précédent : 000007; suivant : 000009Testing reference genes for transcript profiling in Uromyces appendiculatus during urediospore infection of common bean.
Auteurs : Tobias I. LinkSource :
- PloS one [ 1932-6203 ] ; 2020.
English descriptors
- KwdEn :
- MESH :
- genetics : Basidiomycota.
- microbiology : Phaseolus, Plant Diseases.
- physiology : Basidiomycota.
- Gene Expression Profiling, Gene Expression Regulation, Fungal, Genes, Fungal, Host-Pathogen Interactions, Transcriptome.
Abstract
Uromyces appendiculatus is a major pathogen on common bean. Like other rust fungi, it uses effectors to influence its host plant. Effectors are assumed to possess characteristic expression profiles, reflecting their activity during the infection process. In order to determine expression profiles using RT-qPCR, stably expressed reference genes are necessary for normalization. These reference genes need to be tested. Using samples representing seven different developmental stages of the urediospore-based infection process we employed RT-qPCR to measure the expression of 14 candidate reference genes and determined the most suitable ones based on the range of Cq values and comparative calculations using the geNorm and NormFinder algorithms. Among the tested genes RPS14 had the smallest Cq range, followed by Elf1a and Elf3; geNorm rated Tub and UbcE2 best with CytB as a third and NormFinder found UbcE2, Tub and Elf3 as best reference genes. Combining these findings using equal weight for the rankings UbcE2, Elf3 and Tub can be considered the best reference genes. A combination of either two reference genes, UbcE2 and Tub or three reference genes, UbcE2, Tub, and Elf3 is recommended for normalization. However, differences between most genes were relatively small, so all tested genes can be considered suitable for normalization with the exception of RPS9, SDH, Ubc and PDK.
DOI: 10.1371/journal.pone.0237273
PubMed: 32760134
PubMed Central: PMC7410203
Links to Exploration step
pubmed:32760134Le document en format XML
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<term>Basidiomycota (physiology)</term>
<term>Gene Expression Profiling (MeSH)</term>
<term>Gene Expression Regulation, Fungal (MeSH)</term>
<term>Genes, Fungal (MeSH)</term>
<term>Host-Pathogen Interactions (MeSH)</term>
<term>Phaseolus (microbiology)</term>
<term>Plant Diseases (microbiology)</term>
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<front><div type="abstract" xml:lang="en">Uromyces appendiculatus is a major pathogen on common bean. Like other rust fungi, it uses effectors to influence its host plant. Effectors are assumed to possess characteristic expression profiles, reflecting their activity during the infection process. In order to determine expression profiles using RT-qPCR, stably expressed reference genes are necessary for normalization. These reference genes need to be tested. Using samples representing seven different developmental stages of the urediospore-based infection process we employed RT-qPCR to measure the expression of 14 candidate reference genes and determined the most suitable ones based on the range of Cq values and comparative calculations using the geNorm and NormFinder algorithms. Among the tested genes RPS14 had the smallest Cq range, followed by Elf1a and Elf3; geNorm rated Tub and UbcE2 best with CytB as a third and NormFinder found UbcE2, Tub and Elf3 as best reference genes. Combining these findings using equal weight for the rankings UbcE2, Elf3 and Tub can be considered the best reference genes. A combination of either two reference genes, UbcE2 and Tub or three reference genes, UbcE2, Tub, and Elf3 is recommended for normalization. However, differences between most genes were relatively small, so all tested genes can be considered suitable for normalization with the exception of RPS9, SDH, Ubc and PDK.</div>
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<Abstract><AbstractText>Uromyces appendiculatus is a major pathogen on common bean. Like other rust fungi, it uses effectors to influence its host plant. Effectors are assumed to possess characteristic expression profiles, reflecting their activity during the infection process. In order to determine expression profiles using RT-qPCR, stably expressed reference genes are necessary for normalization. These reference genes need to be tested. Using samples representing seven different developmental stages of the urediospore-based infection process we employed RT-qPCR to measure the expression of 14 candidate reference genes and determined the most suitable ones based on the range of Cq values and comparative calculations using the geNorm and NormFinder algorithms. Among the tested genes RPS14 had the smallest Cq range, followed by Elf1a and Elf3; geNorm rated Tub and UbcE2 best with CytB as a third and NormFinder found UbcE2, Tub and Elf3 as best reference genes. Combining these findings using equal weight for the rankings UbcE2, Elf3 and Tub can be considered the best reference genes. A combination of either two reference genes, UbcE2 and Tub or three reference genes, UbcE2, Tub, and Elf3 is recommended for normalization. However, differences between most genes were relatively small, so all tested genes can be considered suitable for normalization with the exception of RPS9, SDH, Ubc and PDK.</AbstractText>
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<CoiStatement>The author has declared that no competing interests exist.</CoiStatement>
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