Assessment of reference gene stability in Rice stripe virus and Rice black streaked dwarf virus infection rice by quantitative Real-time PCR. Issue 1 (December 2015)

• Record Type: Journal Article
• Authors: Fang, Peng; Lu, Rongfei; Sun, Feng; Lan, Ying; Shen, Wenbiao; Du, Linlin; Zhou, Yijun; Zhou, Tong
• Is Part Of: Virology journal. Volume 12:Issue 1(2015)
• Page Start: 1
• Page End: 11
• Publication Date: 2015-12
• Subjects: Gene expression -- RT-qPCR -- Reference genes -- RSV- and RBSDV-infected rice; Virology -- Periodicals; 579.2
• DOI: 10.1186/s12985-015-0405-2
• Languages: English
• ISSNs: 1743-422X

Abstract

Abstract Background Stably expressed reference gene(s) normalization is important for the understanding of gene expression patterns by quantitative Real-time PCR (RT-qPCR), particularly forRice stripe virus (RSV) andRice black streaked dwarf virus (RBSDV) that caused seriously damage on rice plants in China and Southeast Asia. Methods The expression of fourteen common used reference genes of Oryza sativa L. were evaluated by RT-qPCR in RSV and RBSDV infected rice plants. Suitable normalization reference gene(s) were identified by geNorm and NormFinder algorithms. Results UBQ 10 + GAPDH andUBC + Actin1 were identified as suitable reference genes for RT-qPCR normalization under RSV and RBSDV infection, respectively. When using multiple reference genes, the expression patterns ofOsPRIb andOsWRKY, two virus resistance genes, were approximately similar with that reported previously. Comparatively, by using single reference gene (TIP41-Like ), a weaker inducible response was observed. Conclusions We proposed that the combination of two reference genes could obtain more accurate and reliable normalization of RT-qPCR results in RSV- and RBSDV-infected plants. This work therefore sheds light on establishing a standardized RT-qPCR procedure in RSV- and RBSDV-infected rice plants, and might serve as an important point for discovering complex regulatory networks and identifying genes relevant to biological processes or implicated in virus.