Li Jian, Han Jigang, Hu Yonghong, Yang Ji
Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, Fudan UniversityShanghai, China; Shanghai Key Laboratory of Plant Functional Genomics and Resources, Shanghai Chenshan Plant Science Research Center, the Chinese Academy of Science, Shanghai Chenshan Botanical GardenShanghai, China.
Shanghai Key Laboratory of Plant Functional Genomics and Resources, Shanghai Chenshan Plant Science Research Center, the Chinese Academy of Science, Shanghai Chenshan Botanical Garden Shanghai, China.
Front Plant Sci. 2016 Apr 21;7:516. doi: 10.3389/fpls.2016.00516. eCollection 2016.
Tree peony (Paeonia suffruticosa) is a perennial plant indigenous to China known for its elegant and vibrantly colorful flowers. A few genes involved in petal pigmentation have been cloned in tree peony. However, to date, there have been few studies on the comparison and selection of stable reference genes for gene expression analysis by quantitative reverse-transcription PCR (qRT-PCR) in this species. In this study, 10 candidate reference genes were evaluated for the normalization of qRT-PCR in three tree peony cultivars. GAPDH and UBC were identified as the top two most stable reference genes in 'Feng Dan' and 'Xi Shi,' and EF-1α/UBC was recommended to be the best combination for 'Que Hao.' The expression stability of various reference genes differed across cultivars, suggesting that selection and validation of reliable reference genes for quantitative gene expression analysis was necessary not only for different species but also for different cultivars. The results provided a list of reference genes for further study on gene expression in P. suffruticosa. However, in any case, a preliminary check on the accuracy of the best performing reference genes is requested for each qRT-PCR experiment.
牡丹（Paeonia suffruticosa）是一种原产于中国的多年生植物，以其优雅且色彩鲜艳的花朵而闻名。一些参与花瓣色素沉着的基因已在牡丹中被克隆。然而，迄今为止，关于该物种通过定量逆转录聚合酶链反应（qRT-PCR）进行基因表达分析时稳定内参基因的比较和选择的研究很少。在本研究中，对10个候选内参基因进行了评估，以用于三个牡丹品种qRT-PCR的标准化。甘油醛-3-磷酸脱氢酶（GAPDH）和泛素结合酶（UBC）被确定为‘凤丹’和‘西施’中最稳定的两个内参基因，而延伸因子-1α（EF-1α）/UBC被推荐为‘雀好’的最佳组合。不同品种中各种内参基因的表达稳定性不同，这表明不仅对于不同物种，而且对于不同品种，选择和验证用于定量基因表达分析的可靠内参基因都是必要的。这些结果提供了一份内参基因列表，用于进一步研究牡丹的基因表达。然而，无论如何，每个qRT-PCR实验都需要对表现最佳的内参基因的准确性进行初步检查。
