Evaluation of Endogenous Reference Genes of Bactrocera (Tetradacus) minax by Gene Expression Profiling under Various Experimental Conditions

• Published in: The Florida entomologist 2014-06, Vol.97 (2), p.597-604
• Main Authors: Lue, Zhi-Chuang, Wang, Liu-Hao, Dai, Rui-Lin, Zhang, Gui-Fen, Guo, Jian-Ying, Wan, Fang-Hao
• Format: Article
• Language: English
• Subjects: Bactrocera
• ISSN: 0015-4040
• DOI: 10.1896/054.097.0235

Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is widely used for gene expression analysis in living organisms, and stably expressed endogenous reference genes are needed to obtain accurate results. Some commonly used reference genes varied among different experimental treatments. To obtain suitable reference gene for specific experimental conditions, the stability of 10 reference genes of Bactrocera (Tetradacus) minax (Enderlein) were evaluated in various development stages, and under temperatureand gamma -irradiation-stress conditions by semi-quantitative PCR. The present results indicated that the most stable candidate reference gene was RPL32 in eggs, 3rd instars, 1-, 90- and 160-day-pupae and newly emerged female and male adults (< 24 h), and under different stress conditions, i.e., 35 degree C temperature stress for 0 h, 1 h, 3 h and 5 h, and 4 degree C temperature stress for 0 h, 12 h, 36 h, 48 h and 60 h. GAPDH, G6PDH and RPL32 were ideal candidate endogenous genes under 35 degree C temperature stress for 0 h, 1 h, 3 h and 5 h, and under 4 degree C temperature stress for 0 h, 12 h, 36 h, 48 h and 60 h, and under a Gamma irradiation stress of 90 Gy. These results provide basic information for future studies of gene expression in B. minax, and should serve as a resource to screen reference genes for gene expression studies in other insect species.