Identification and Validation of Stable Reference Genes for Gene Expression Analysis in Sword-leaf Dogbane Using Quantitative Reverse Transcriptase Polymerase Chain Reaction

文献类型： 外文期刊

第一作者： Xu, Zongchang

作者： Xu, Zongchang;Zhang, Chengsheng;Li, Yiqiang;Wang, Meng;Zhou, Jinhui;Liu, Han

作者机构：

关键词： Apocynum venetum; flavonoid; luobuma; normalization; salt stress

期刊名称：JOURNAL OF THE AMERICAN SOCIETY FOR HORTICULTURAL SCIENCE （ 影响因子：1.144； 五年影响因子：1.617 ）

ISSN： 0003-1062

年卷期： 2018 年 143 卷 6 期

页码：

收录情况： SCI

摘要： Sword-leaf dogbane (Apocynum venetum) is a traditional Chinese herb with increasingly recognized potential to enhance health, but no study of stable reference genes in this herb has been reported. Based on a homologous cloning strategy, we have successfully cloned five candidate reference genes from sword-leaf dogbane: glyceraldehyde-3-phosphate dehydrogenase (AvGAPDH), beta tubulin (AvbTUB), polyubiquitin (AvUBQ), elongation factor 1-alpha (AvEF1 alpha), and actin (A vACTIN). Three distinct algorithms, geNorm, NormFinder, and BestKeeper, were used to estimate the expression stability of candidate reference primer pairs. We found that AvACTIN-2 and AvACTIN-3 presented the highest stability of expression in different tissue samples, and AvGAPDH-2 was most stable under salinity stress. In addition, we illustrated the application of these new reference genes by assaying the expression levels of two hyperoside biosynthesis terminal enzyme genes, flavonoid 3'-hydroxylase (F3'H) and flavonol synthase (FLS), under salinity stress. Our study is the first to report stable expression of internal reference genes in sword-leaf dogbane in multiple experimental sample sets.

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