Evaluation of reference genes for quantitative real-time PCR normalization in the Neopyropia (Pyropia)  oomycete pathogen Pythium porphyrae

Yang, Huichao; Yan, Yonggang; Weng, Peiwen; Sun, Congcong; Yu, Jiaxing; Tang, Lei; Li, Jie; Mo, Zhaolan

doi:10.1007/s10811-022-02865-1

Cited by 4 publications

(2 citation statements)

References 41 publications

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“…Previous studies also identified GAPDH as the recommended reference gene in different developmental stages of fungi Sparassis latifolia [28] and amphibian Andrias davidianus [29]. Additionally, GAPDH showed the best stability in Pythium porphyrae under conditions of salinity, pH and infection stages [30]. As far as we know, there are only a few reports selecting and validating the suitable reference genes to compare the gene expression between the mycelia and sclerotia of pathogens, such as S. sclerotiorum and Rhizoctonia solani.…”

Section: Discussionmentioning

confidence: 99%

“…In current study, EF1α was an ideal reference gene in S. rolfsii subjected to photoperiod and pH treatments. EF1α is also commonly used as a stably expressed reference gene in many species under various conditions, such as Mythimna separata under different photoperiod treatments [33], Pythium porphyrae under different pH treatments [30], Athetis dissimilis under different insecticide treatments [3] and Glycine max under different developmental stages and stress conditions [34]. Other than EF1α, PGK and β-TUB were also suitable reference genes in S. rolfsii across photoperiod and pH treatments, respectively.…”

Section: Discussionmentioning

confidence: 99%

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Selection and Validation of Reference Genes for Reverse-Transcription Quantitative PCR Analysis in Sclerotium rolfsii

Jiang,

Zhou,

Zhao

et al. 2023

IJMS

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Reference genes are important for the accuracy of gene expression profiles using reverse-transcription quantitative PCR (RT-qPCR). However, there are no available reference genes reported for Sclerotium rolfsii; it actually has a pretty diverse and wide host range. In this study, seven candidate reference genes (UBC, β-TUB, 28S, 18S, PGK, EF1α and GAPDH) were validated for their expression stability in S. rolfsii under conditions of different developmental stages, populations, fungicide treatments, photoperiods and pHs. Four algorithm programs (geNorm, Normfinder, Bestkeeper and ΔCt) were used to evaluate the gene expression stability, and RefFinder was used to integrate the ranking results of four programs. Two reference genes were recommended by RefFinder for RT-qPCR normalization in S. rolfsii. The suitable reference genes were GAPDH and UBC across developmental stages, PGK and UBC across populations, GAPDH and PGK across fungicide treatments, EF1α and PGK across photoperiods, β-TUB and EF1α across pHs and PGK and GAPDH across all samples. Four target genes (atrB, PacC, WC1 and CAT) were selected for the validation of the suitability of selected reference genes. However, using one or two reference genes in combination to normalize the expression of target genes showed no significant difference in S. rolfsii. In short, this study provided reliable reference genes for studying the expression and function of genes in S. rolfsii.

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Section: Discussionmentioning

confidence: 99%