Screening of Reference Genes for qRT-PCR Analysis in Zingiber zerumet (L.) Smith Female Reproductive Organ after Pollination

Screening reference genes of pollinated Zingiber zerumet (L.) Smith female reproductive organ at development process is crucial, when we study and analyze the expression of key regulatory genes for Zingiber zerumet (L.) Smith abortion. According to the transcriptome database of Zingiber zerumet (L.) Smith and researches about traditional reference genes, we selected 10 relatively stable genes as candidate reference genes, including Actin-2 (ACT2), Actin-7 (ACT7), Beta tubulin-1 (TUB1), Beta tubulin-5 (TUB5), Alpha tubulin-3 (TUA3), Ubiquitin (UBQ), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Elongation factor 1-alpha (EF-1), Cyclophilin (CYP), Histone (H2A), and then analyzed their expression stability by qRT-PCR and three statistic programs: GeNorm, NormFinder and BestKeeper. The results showed that GAPDH and UBQ are the most stable genes during the development process of pollinated Zingiber zerumet (L.) Smith female reproductive organ. These two genes were suitable to be reference genes, and it was more accurate for qRT-PCR normalization analysis to utilize two reference genes at the same time. Consequently, GAPDH and UBQ were able to be reference genes for qRT-PCR normalization analysis in pollinated Zingiber zerumet (L.) Smith female reproductive organ at different development stages. This work is contributable to lay a foundation to study the molecular mechanism of abortive in Zingiber zerumet (L.) Smith, and also acts as a resource for other Zingiber to screen reference genes.