Fu Yanping, Niu Fei, Jia Hui, Wang Yanli, Guo Bin, Wei Yahui
Key Laboratory of Biotechnology of Shannxi Province, Key Laboratory of Resource Biology and Biotechnology in Western China (Ministry of Education), College of Life Science Northwest University Xi'an China.
Plant Direct. 2021 Nov 24;5(11):e362. doi: 10.1002/pld3.362. eCollection 2021 Nov.
() produces various types of effective lycopodium alkaloids, especially Huperzine A (HupA), which is a promising drug for the treatment of Alzheimer's disease. Numerous studies focused on the chemistry, bioactivities, toxicology, and clinical trials of HupA; however, the public genomic and transcriptomic resources are very limited for research, especially for the selection of optimum reference genes. Based on the full-length transcriptome datasets and previous studies, 10 traditional and three new candidate reference genes were selected in different tissue of . Then, two optimal reference genes and were confirmed by four analysis methods. In order to further verify the accuracy of the two reference genes, they were used to analyze the expression patterns of four HupA-biosynthetic genes (lysine decarboxylas, RS-norcoclaurine 6-O-methyltransferase, cytochrome P45072A1, and copper amine oxidase). The data suggested that the expression pattern of HupA-biosynthetic genes was consistent with them in transcriptome sequencing in different tissue of . This study identified that and provides the reliable normalization for analyzing the HupA biosynthetic gene expression in different tissues of on the transcriptional level.
（）能产生多种类型的有效石松生物碱，尤其是石杉碱甲（HupA），它是一种有前景的治疗阿尔茨海默病的药物。众多研究聚焦于石杉碱甲的化学、生物活性、毒理学及临床试验；然而，用于研究的公共基因组和转录组资源非常有限，尤其是用于选择最佳参考基因。基于全长转录组数据集和先前研究，在（）的不同组织中选择了10个传统的和3个新的候选参考基因。然后，通过四种分析方法确定了两个最佳参考基因（）和（）。为了进一步验证这两个参考基因的准确性，将它们用于分析四个石杉碱甲生物合成基因（赖氨酸脱羧酶、RS - 去甲乌药碱6 - O - 甲基转移酶、细胞色素P45072A1和铜胺氧化酶）的表达模式。数据表明，在（）不同组织的转录组测序中，石杉碱甲生物合成基因的表达模式与它们一致。本研究确定（）和（）为在转录水平分析（）不同组织中石杉碱甲生物合成基因表达提供了可靠的标准化方法。
