目的 从川续断Dipsacus asper中克隆并筛选出稳定的内参基因用于实时荧光PCR（qRT-PCR）的分析校正，为今后川续断功能基因的表达分析及调控机制研究提供基础。方法 从川续断转录组数据库中筛选并克隆肌动蛋白（Actin）、微管蛋白（Tubulin）、GAPDH基因家族内参基因的核心片段，以不同产地、不同组织部位和不同发育时期的川续断植株为材料，通过qRT-PCR获得各基因的表达信息，利用geNorm、NormFinder、BestKeeper、Delta CT、RefFinder共5个内参评价软件分析各基因的表达稳定性，综合评价并筛选出最佳的川续断内参基因。结果 克隆获得10个候选内参基因核心片段，分别属于Actin、Tubulin、GAPDH 3大基因家族，各家族基因序列间同源性较高；稳定性综合分析结果显示，DaACT103在不同产地及不同组织部位的表达稳定性较强且表达量相对较高，DaTUB5在不同发育时期的表达稳定性较强且表达量相对较低。结论 DaACT103和DaTUB5均可作为川续断的内参基因，其中，DaACT103适用于作为具有较高表达丰度基因的内参基因；DaTUB5适用于作为表达丰度较低基因的内参基因。

Objective To clone and screen the stable internal reference genes from Dipsacus asper for qRT-PCR analysis correction, so as to provide a preliminary basis for future research on expression analysis and regulation mechanism of D. asper functional genes. Methods The internal reference genes of Actin, Tubulin and GAPDH gene families were screened and cloned from D. asper transcriptome database. The D. asper plants from different origins, different tissues and different developmental stages were used to obtain expression information of each gene by qRT-PCR. The expression stability of each gene was analyzed by geNorm, NormFinder, BestKeeper, Delta CT and RefFinder, and the best genes were synthetically evaluated and screened. Results Ten core fragments for candidate internal reference genes were cloned, belonging to three gene families:Actin, Tubulin and GAPDH, with high homology among them. The results of stability analysis showed that the expression of DaACT103 was stable and relatively high in different regions and tissues, while the expression of DaTUB5 was stable and relatively low in different developmental stages. Conclusions DaACT103 and DaTUB5 are suitable as the internal reference genes for D. asper. DaACT103 is used as the internal reference gene with high abundances and DaACT105 is used as the internal reference gene with low abundances.

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国家自然科学基金资助项目（81860675）；贵州省高层次创新型人才（黔科合平台人才[2018]5638）；贵州省教育厅创新群体重大研究项目（黔教合KY字[2018]022）；现代农业产业技术体系建设专项（CARS-21）；贵州省国内一流建设学科项目（中药学）（GNYL[2017]008号）