Chinese Journal OF Rice Science >
Screening Reference Genes and Evaluating of Their Expression Stability for qRT-PCR Normalization in Chilo suppressalis (Lepidoptera: Pyralididae)
Received date: 2018-03-27
Revised date: 2018-05-15
Online published: 2019-01-10
【Objective】We aim to screen reference genes stably expressed in striped stem borers (SSB), Chilo suppressalis, under certain conditions, to lay a foundation for the study on the gene expression in SSB. 【Method】According to the results of SSB transcriptome data, 11 candidate genes from SSB were selected and their expression stability was detected under different treatment conditions (including development duration, tissues, feeding on different diets and rice varieties, dsRNA and mixed samples under temperature stress, insecticide treatment), by using real-time fluorescence quantitative PCR. The stability of the 11 candidate genes was evaluated by RefFinder, BestKeeper, GeNorm, NormFinder and ΔCt. 【Result】AK, RPL10 and EF1 were highly stable in different development durations; EF1, TUB and ACTB were highly stable in different tissues. TUB, RPL10 and EF1 were relatively stable under temperature stress. TF4 and ACTA were stable in insecticide treated samples. TUB, TF4, EF1 and RPL10 were stable in samples fed on different diets. TUB and EF1 were stable reference genes in samples fed on different rice varieties. TUB, AK, ACTB and EF1 were stable in the sample treated with dsRNA. EF1, TUB and ACTB were stable in mixed samples. 【Conclusion】 It provide a reference for selection of suitable reference gene under different test conditions, and also provide more reliable and accurate data for research in gene expression in rice SSB.
Key words： Chilo suppressalis; reference gene; qRT-PCR; gene expression
Hongxing XU, Guorong WANG, Yanhui LU, Yajun YANG, Xusong ZHENG, Junce TIAN, Zhongxian LÜ . Screening Reference Genes and Evaluating of Their Expression Stability for qRT-PCR Normalization in Chilo suppressalis (Lepidoptera: Pyralididae)[J]. Chinese Journal OF Rice Science, 2019 , 33(1) : 75 -84 . DOI: 10.16819/j.1001-7216.2019.8035
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