Selection of reference genes for quantitative real-time PCR expression studies in the apomictic and sexual grass Brachiaria brizantha

Abstract

Background: Brachiaria brizantha is an important forage grass. The occurrence of both apomictic and sexual reproduction within Brachiaria makes it an interesting system for understanding the molecular pathways involved in both modes of reproduction. Quantitative real time PCR (qRT-PCR) has emerged as an important technique to compare expression profile of target genes and, in order to obtain reliable results, it is important to have suitable reference genes. In this work, we evaluated eight potential reference genes for B. brizantha qRT-PCR experiments, isolated from cDNA ovary libraries. Vegetative and reproductive tissues of apomictic and sexual B. brizantha were tested to validate the reference genes, including the female gametophyte, where differences in the expression profile between sexual and apomictic plants must occur.

Reference gene selection for quantitative gene expression studies during biological invasions: A test on multiple genes and tissues in a model ascidian Ciona savignyi

“…Similar results were commonly obtained in many studies (e.g., Kuijk et al, 2007; Silveira et al, 2009;Guelke et al, 2015). Since the V value is dependent on both the diversity of samples and the stability of potential reference genes, dividing the data sets into more detailed sub-groups and applying as many reference genes as suggested by geNorm are the correspondingly feasible solutions (Sun et al, 2015).…”

Selection of reference genes for quantitative real-time PCR normalization in creeping bentgrass involved in four abiotic stresses

“…V3/4 values of PL and HL samples below 0.15 represented three reference genes necessary for qRT-PCR detection. However, the cut-off value of 0.15 was not sufficient, and higher values of V n / n?1 were confirmed in other studies (De Ketelaere et al 2006;Marum et al 2012;Silveira et al 2009). The data in current study showed slight variation between V3/4 (0.161) and V4/5 (0.152) in the total samples which revealed three reference genes may be necessary for efficient normalization.…”

Reference gene selection for qPCR in Ammopiptanthus mongolicus under abiotic stresses and expression analysis of seven ROS-scavenging enzyme genes

“…Normalization was performed using three different sets of reference genes: (1) two best reference genes selected by geNorm; (2) two best selected by NormFinder; (3) two best across all the treatments selected by both geNorm and NormFinder. Significant up-regulation with respect to control was shown by *P \ 0.05, Student's t test or **P \ 0.01, and significant down-regulation was shown by Many other tubulins were also thought to be variant, such as Arabidopsis Tub2 and Tub6 (Gutierrez et al 2008), potato Beta-tubulin (Nicot et al 2005) and B. brizantha Beta-tubulin (Silveira et al 2009). Elongation factor-1 alpha (EF1A) genes in barley and oat samples were ranked as the less reliable controls (Jarošová and Kundu 2010).…”