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Identification and testing of reference genes for Sesame gene expression analysis by quantitative real-time PCR

文献类型： 外文期刊

作者： Wei, Libin 1 ; Miao, Hongmei 1 ; Zhao, Ruihong 1 ; Han, Xiuhua 1 ; Zhang, Tide 1 ; Zhang, Haiyang 1 ;

作者机构： 1.Henan Acad Agr Sci, Henan Sesame Res Ctr, Zhengzhou 450002, Henan, Peoples R China

关键词： BestKeeper;GeNorm;NormFinder;Quantitative real-time PCR;Reference gene;Sesame (Sesamum indicum L.)

期刊名称：PLANTA （ 影响因子：4.116； 五年影响因子：4.316 ）

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收录情况： SCI

摘要： Sesame (Sesamum indicum L.) is an ancient and important oilseed crop. However, few sesame reference genes have been selected for quantitative real-time PCR until now. Screening and validating reference genes is a requisite for gene expression normalization in sesame functional genomics research. In this study, ten candidate reference genes, i. e., SiACT, SiUBQ6, SiTUB, Si18S rRNA, SiEF1α, SiCYP, SiHistone, SiDNAJ, SiAPT and SiGAPDH, were chosen and examined systematically in 32 sesame samples. Three qRT-PCR analysis methods, i. e., geNorm, NormFinder and BestKeeper, were evaluated systematically. Results indicated that all ten candidate reference genes could be used as reference genes in sesame. SiUBQ6 and SiAPT were the optimal reference genes for sesame plant development; SiTUB was suitable for sesame vegetative tissue development, SiDNAJ for pathogen treatment, SiHistone for abiotic stress, SiUBQ6 for bud development and SiACT for seed germination. As for hormone treatment and seed development, SiHistone, SiCYP, SiDNAJ or SiUBQ6, as well as SiACT, SiDNAJ, SiTUB or SiAPT, could be used as reference gene, respectively. To illustrate the suitability of these reference genes, we analyzed the expression variation of three functional sesame genes of SiSS, SiLEA and SiGH in different organs using the optimal qRT-PCR system for the first time. The stability levels of optimal and worst reference genes screened for seed development, anther sterility and plant development were validated in the qRT-PCR normalization. Our results provided a reference gene application guideline for sesame gene expression characterization using qRT-PCR system.

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