Selection of Appropriate Housekeeping Genes for Gene Expression Normalization in Hybrid Grouper (Epinephelus fuscoguttatus ♀ x E. lanceolatus ♂)

Since lots of researches using qPCR technique have addressed gene expression in groupers, the suitable housekeeping genes are needed to obtain reliable and precise results. In this study, by using different mathematical algorithms (Genorm, normFinder, BestKeeper and comparative Delta Ct method) and the comprehensive ranking software (RefFinder), the stability of seventeen housekeeping genes were assessed in eleven tissues of the hybrid grouper (Epinephelus fuscoguttatus female x E. lanceolatus male). Results showed that the stability of housekeeping genes were various in different tissues and the newly identified housekeeping genes were likely more stable than that of commonly used. In conclusion, the expression stability of housekeeping genes showed a tissue-specific manner, and the conserved oligomeric Golgi complex subunit 5 (cog5) and peroxisomal biogenesis factor 14 (pex14) could be universally used as appropriate housekeeping genes across different tissues in hybrid grouper. The present study provides the foundation for more accurate results of qPCR assay in hybrid grouper and other groupers.