Validation of reference genes for gene expression analysis of response to anthocyanin induction in cell cultures of Vitis davidii (Rom. Caill.) Foex
文献类型： 外文期刊
作者： Lai, Chengchun 1 ; Pan, Hong 1 ; Huang, Xiangui 1 ; Fan, Lihua 1 ; Duan, Changqing 2 ; Li, Shaozhen 3 ;
作者机构： 1.Fujian Acad Agr Sci, Inst Agr Engn & Technol, Fuzhou 350003, Fujian, Peoples R China
2.China Agr Univ, Coll Food Sci & Nutr Engn, Ctr Viticulture & Enol, Beijing 100083, Peoples R China
3.Beijing Huiyuan Food & Beverage Co Ltd, Beijing 101305, Peoples R China
关键词： Spine grape (Vitis davidii [Rom. Caill.] Foex.); Cell culture; Gene expression; Reference gene; Real-time quantitative PCR
期刊名称：IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-PLANT （ 影响因子：2.252； 五年影响因子：2.139 ）
ISSN： 1054-5476
年卷期： 2018 年 54 卷 6 期
页码：
收录情况： SCI
摘要： Real-time quantitative polymerase chain reaction (RT-qPCR) is an effective method for detecting changes of gene expression in plant cell metabolic regulation. A set of 15 reference gene candidates were selected for the present study of anthocyanin biosynthesis regulation, and stability. The suitability of their expression was evaluated in eight different experimental treatments in spine grape (Vitis davidii [Rom. Caill.] FoA << x.) cell cultures. The results indicated that SAND family protein (SAND) and V-type proton ATPase subunit G (VAG) were the most stable reference genes for culture duration, tubulin alpha-3/alpha-5 chain (alpha-tubulin) and tubulin beta-1 chain (beta-tubulin) for illumination conditions, ubiquitin-conjugating enzyme E2-17 kDa (UBQ) and VAG for UVB treatment, VAG and 60S ribosomal protein L18-2 (60SRP) for temperature treatment, AP47, clathrin adaptor complex subunit mu (AP-2) and 60SRP for cinnamic acid treatment, alpha-tubulin and UBQ for chitosan treatment, actin and alcohol dehydrogenase 2 (ADH2) for kinetin treatment, and beta-tubulin and elongation factor 1-alpha (EF1-alpha) for cell line. Finally, the reliability of the selected reference genes was confirmed by investigating the expression profiles of the target gene dihydroflavonol 4-reductase (DFR) in spine grape cell cultures. The results of the present study offer the most robust platform for the most precise and broad application of RT-qPCR to investigate gene expression associated with anthocyanin biosynthesis in spine grape cell cultures.
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