Recommended Reference Genes for Quantitative PCR Analysis in Soybean Have Variable Stabilities during Diverse Biotic Stresses

• DOI: 10.1371/journal.pone.0134890
• Publication Date: 2015-08-05T14:01:23Z
• Authors: Raman Bansal; Priyanka Mittapelly; Bryan J. Cassone; Praveen Mamidala; Margaret G. Redin...; Andy Michel
• Keywords: Reference Genes; Biotic stress

For real-time reverse transcription-PCR (qRT-PCR) in soybean, reference genes different tissues, developmental stages, various cultivars, and under stress conditions have been suggested but their usefulness for research on soybean biotic stresses occurring North-Central U.S. is not known. Here, we investigated the expression stabilities of ten previously recommended (ABCT, CYP, EF1A, FBOX, GPDH, RPL30, TUA4, TUB4, TUA5, UNK2) from Bean pod mottle virus (BPMV), powdery mildew (PMD), aphid (SBA), two‐spotted spider mite (TSSM). BPMV, PMD, SBA, TSSM are amongst most common pest problems other regions. Reference gene stability was determined using three software algorithms (geNorm, NormFinder, BestKeeper) a web-based tool (RefFinder). showed variability as well across stressors best were stress-dependent. ABCT FBOX found to be stable both BPMV SBA these had only minimal moderate during PMD stress. Expression TUA4 CYP stress; TUB4 Under analyzed, GPDH consistently unstable. all obtained pairwise variation (V2/3) values less than 0.15 which that combined use two would sufficient normalization. Further, demonstrated utility normalizing qRT-PCR data target validated current study. Following recommendations our study will enable an accurate reliable normalization