目的 筛选出在北柴胡不同生长时期及不同器官中表达均稳定的内参基因并进行验证。方法 利用实时荧光定量PCR获得18个候选内参基因所有C_t值，通过3种不同算法（Bestkeeper、NormFinder、GeNorm）的软件对内参基因稳定性进行分析，采用皮尔森相关系数（Pearson correlation coefficient）分析3个软件给出的稳定性排名结果。结果 所有候选内参基因的C_t值相对宽泛，ADF1b、ADF5、ADF7、eIF2b和ACT2为最为合适的内参基因，而eIF6被认为稳定性最差的内参基因。3个软件计算结果均呈现显著性相关。结论 采用实时荧光定量PCR结合3种不同算法进行北柴胡内参基因的筛选及验证是可行的。在北柴胡柴胡分子遗传研究中，发掘到的内参基因对目的基因进行均一化处理有助于提高目的基因表达分析的精确性及可信度。

Objective To identify stable reference genes in different growth stages and different organs of Bupleurum chinese. Methods All C_t values of 18 candidate internal reference genes were obtained by real-time quantitative PCR. Three software (Bestkeeper, NormFinder, and GeNorm) based on different algorithms were used to analyze the stability of the internal reference gene. The Pearson correlation coefficient was also analyzed. Results The C_t values of all candidate genes were relatively broad. ADF1b, ADF5, ADF7, eIF2b, and ACT2 were the most stable reference genes, whereas the gene of eIF6 was the least stable of the reference gene. The results of three softwares showed significant correlation. Conclusion Real-time fluorescence quantitative PCR combined with three different algorithms for the screening and validation of the reference gene of B. chinese is feasible. The homogenization of the target gene by the reference genes of the present study is helpful to improve the accuracy and reliability of gene expression analysis in molecular genetic research of B. chinese.

国家自然科学基金资助项目（81603223）；四川省"十三五"育种攻关（2006NYZ0036-4-2）；四川省科技扶贫专项（2016NZYZF0071）