Selection of reliable reference genes for gene expression studies in Botrytis cinerea
| Autor: | Wu Xiaoqing, Xie Xueying, Hetong Yang, Hongzi Zhou, He Ren, Yuping Lyu, Xinjian Zhang |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Hydrolases In silico Real-Time Polymerase Chain Reaction Microbiology 03 medical and health sciences Tubulin Gene Expression Regulation Fungal Reference genes Gene expression Molecular Biology Gene Glyceraldehyde 3-phosphate dehydrogenase Plant Diseases Botrytis cinerea Trichoderma Genetics Genes Essential biology Ubiquitin Oxalic Acid fungi Glyceraldehyde-3-Phosphate Dehydrogenases food and beverages biology.organism_classification Molecular biology Actins Housekeeping gene Reverse transcription polymerase chain reaction 030104 developmental biology Ubiquitin-Conjugating Enzymes biology.protein Botrytis NADP |
| Zdroj: | Journal of Microbiological Methods. 142:71-75 |
| ISSN: | 0167-7012 |
| DOI: | 10.1016/j.mimet.2017.09.006 |
| Popis: | Botrytis cinerea is an important plant pathogen causing grey mold disease in a wide range of plant species. The aim of this study was to identify reliable reference genes that can be used for the analysis of relative gene expression in B. cinerea with quantitative real-time reverse transcription PCR (qRT-PCR). Six commonly used housekeeping genes including actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin (UBQ), ubiquitin-conjugating enzyme (UCE), α-tubulin (α-TUB) and β-tubulin (β-TUB) were selected to test their expression stabilities in B. cinerea treated with different concentration of oxalic acid (1, 5 and 10mM) and confronted with antagonistic Trichoderma afroharzianum. Four in silico algorithms (geNorm, BestKeeper, NormFinder and Comparative ΔCt) were applied to evaluate the expression stabilities of these genes, and the UBQ gene was identified as the most stably expressed. It was used to normalize the expression levels of three genes related to oxalic acid production (NADPH, VEL1 and OAH) when B. cinerea was challenged by T. afroharzianum. The results of this study are useful for gene expression analysis in B. cinerea. |
| Databáze: | OpenAIRE |
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