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

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 ; it actually has a pretty diverse and wide host range. In this study, seven candidate reference genes (, , , , , and ) were validated for their expression stability in 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 . The suitable reference genes were and across developmental stages, and across populations, and across fungicide treatments, and across photoperiods, and across pHs and and across all samples. Four target genes (, , and ) 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 . In short, this study provided reliable reference genes for studying the expression and function of genes in .