Evaluation of reference genes for qRT-PCR studies in the colchicine producing Gloriosa superba L.
Evaluation of reference genes for qRT-PCR studies in the colchicine producing Gloriosa superba L.
复制标题
DOI：
10.1007/s11816-023-00840-x
复制
发表时间：
2023-05-18
影响因子：
2.4
通讯作者：
中科院分区：
文献类型：
--
作者：
火百合是抗炎药物秋水仙素的两个主要来源之一。以前的研究表明，秋水仙素的产量在根茎中比在叶和根中更高。早期的前体取食和转录组分析已经为秋水仙素的生物合成提供了可能的途径和候选基因。利用实时定量逆转录聚合酶链式反应(qRT-PCR)技术比较分析不同组织中候选途径基因的表达水平，可以发现与其他组织相比，根茎中表达水平较高的基因，这可能表明基因产物在秋水仙素生物合成中的作用。归一化是qRT-PCR有效分析差异基因表达的重要步骤，具有广泛的应用前景。本研究从转录组数据集中选择候选参考基因，并对其进行分析，以确定秋水仙素生物合成相关基因正常化最稳定的基因。利用RefFinder软件，筛选出一个稳定的参考基因UBC22，用于调节大豆叶片、根和根茎中候选甲基转移酶(MT)基因的表达水平。以UBC22为参照基因，甲基转移酶GsOMT1、GsOMT3和GsOMT4在大豆根茎中的表达水平显著高于对照基因UBC22，而MT31794在根中的表达水平较高。综上所述，本研究提供了一个可行的参考基因表达分析系统，该系统可以帮助阐明秋水仙素的生物合成及其开发利用，从而提高秋水仙素的产量。网上版载有补充材料，可在10.1007/s11816023-00840-x查阅。
The flame lily, Gloriosa superba L., is one of the two primary sources of the anti-inflammatory drug, colchicine. Previous studies have shown that a higher level of colchicine production occurs in the rhizomes than in leaves and roots. Earlier precursor feeding and transcriptome analysis of G. superba have provided a putative pathway and candidate genes involved in colchicine biosynthesis. Comparative analysis of expression levels of candidate pathway genes in different tissues of G. superba using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) can reveal highly expressed genes in the rhizome compared to other tissues which could suggest roles of the gene products in colchicine biosynthesis. Normalization is an important step in effectively analyzing differential gene expression by qRT-PCR with broader applications. The current study selected candidate reference genes from the transcriptome datasets and analyzed them to determine the most stable genes for normalization of colchicine biosynthesis-related genes. Using RefFinder, one stable reference gene, UBC22, was selected to normalize gene expression levels of candidate methyltransferase (MT) genes in the leaves, roots, and rhizomes of G. superba. With UBC22 as reference gene, the methyltransferases, GsOMT1, GsOMT3, and GsOMT4 showed significantly higher expression levels in the rhizome of G. superba, while MT31794 was more highly expressed in the roots. In conclusion, the current results showed a viable reference gene expression analysis system that could help elucidate colchicine biosynthesis and its exploitation for increased production of the drug in G. superba. The online version contains supplementary material available at 10.1007/s11816-023-00840-x.
登录
查看更多内容
影响因子：
64.8
作者：
Nett RS;Lau W;Sattely ES
通讯作者：
Sattely ES
影响因子：
13.8
作者：
Larkin, Philip J.;Miller, James A. C.;Fist, Anthony J.
通讯作者：
Fist, Anthony J.
DOI：
10.3390/molecules21081078
发表时间：
2016-08-18
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Kishimoto S;Sato M;Tsunematsu Y;Watanabe K
通讯作者：
Watanabe K
影响因子：
64.8
作者：
Luo, Xiaozhou;Reiter, Michael A.;Keasling, Jay D.
通讯作者：
Keasling, Jay D.
DOI：
10.1126/science.aac9373
发表时间：
2015-09-04
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Galanie S;Thodey K;Trenchard IJ;Filsinger Interrante M;Smolke CD
通讯作者：
Smolke CD