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Background   L. decemlineata  is an exotic invasive insect pest, and invaded in Xinjiang Uygur autonomous region in China in the 1990s from Kazakhstan. It is a notorious defoliator of potato throughout most of the northern Xinjiang in current, and often causes extremely large yield losses of potato.    Results  The expression stability of nine  L. decemlineata  house-keeping genes ( Actin ,  ACT1  and  ACT2 ;  ADP-ribosylation factor ,  ARF1  and  ARF4 ;  TATA box binding protein ,  TBP1  and  TBP2 ;  ribosomal protein RP4  and  RP18 ;  translation elongation factor 1α EF1α ) was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) in seven developmental stages, three larval tissues and two insecticide treatments. The results were analyzed using three software programs: geNorm, NormFinder and BestKeeper. Although there was no consistent ranking observed among the house-keeping genes across the samples, the overall analysis revealed that  RP18, RP4, ARF1 , and  ARF4  were the four most stable house-keeping genes. In contrast,  ACT1  and  ACT2 , two of the most widely used reference genes, had the least stability. Our results suggest that the combined use of the four most stably expressed genes may produce optimal normalization for qRT-PCR.    Conclusions  The expression stability of the house-keeping genes varies among different developing stages, in different tissues and under different experimental conditions. Our results will enable a more accurate and reliable normalization of qRT-PCR data in  L. decemlineata.

Validation of reference genes for expression analysis by quantitative real-time PCR in Leptinotarsa decemlineata (Say)