Determination of Suitable RT-qPCR Reference Genes for Studies of Gene Functions in (Fallén).

The reverse transcription quantitative polymerase chain reaction (RT-qPCR) has been widely used to determine gene functions in (Fallén) (small brown planthopper). Selection of suitable reference gene(s) for normalizations of RT-qPCR data is critical for reliable results. To date, reports on identification of suitable reference genes are still very limited. is a destructive rice pest and it can transmit multiple viruses, including (RBSDV), (RSV), and (MRDV), to many important cereal crops worldwide. In this study, we examined the stablity of seven selected candidate reference genes in at different developmental stages, in different tissues, in RBSDV- or RSV-infected or in RBSDV-infected and ()-silenced . The RT-qPCR data representing individual candidate genes were analyzed using five different methods: the delta Ct method, geNorm, NormFinder, BestKeeper, and the RefFinder algorithm, respectively. The most stable reference gene for the specific condition was selected according to a comprehensive analysis using the RefFinder method. Ribosomal protein L5 () and are the most stably expressed genes in at different developmental stages. Alpha-1-tubulin () is the most stably expressed reference gene in different tissues of RBSDV viruliferous (RBSDV-V) or non-viruliferous (RBSDV-NV) . is the most stably expressed reference gene in RBSDV-V or RSV viruliferous (RSV-V) , while beta-tubulin () is the most stably expressed reference gene in RBSDV-V and -silenced . The selected reference genes were further investigated during analyses of RBSDV and gene expression in different tissues from RBSDV-V or RBSDV-NV . The stably expressed reference genes identified in this study will benefit future gene function studies using .