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Reverse transcription quantitative real-time PCR (RT-qPCR) is a common way to study gene regulation at the transcriptional level due to its sensibility and specificity, but it needs appropriate reference genes to normalize data . Ananas comosus var. bracteatus , with white-green chimeric leaves, is an important pantropical ornamental plant. Up to date, no reference genes have been evaluated in  Ananas comosus  var.  bracteatus . In this work, we used five common statistics tools (geNorm, NormFinder, BestKeeper, ΔCt method, RefFinder) to evaluate 10 candidate reference genes. The results showed that  Unigene.16454  and  Unigene.16459  were the optimal reference genes for different tissues,  Unigene.16454  and zinc finger ran-binding domain-containing protein 2 ( ZRANB2 ) for chimeric leaf at different developmental stages, isocitrate dehydrogenase [NADP] ( IDH ) and triacylglycerol lipase SDP1-like ( SDP ) for seedlings under different hormone treatments. The comprehensive results showed  IDH , pentatricopeptide repeat-containing protein ( PPRC ),  Unigene.16454,  and caffeoyl-CoA O methyltransferase 5-like ( CCOAOMT ) are the top-ranked stable genes across all the samples. The stability of glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) was the least during all experiments. Furthermore, the reliability of recommended reference gene was validated by the detection of porphobilinogen deaminase ( HEMC ) expression levels in chimeric leaves. Overall, this study provides appropriate reference genes under three specific experimental conditions and will be useful for future research on spatial and temporal regulation of gene expression and multiple hormone regulation pathways in  Ananas comosus  var.  bracteatus .

frontiers in genetics

Validation of Reference Genes for Quantitative Real-Time PCR Normalization in Ananas comosus var. bracteatus During Chimeric Leaf Development and Response to Hormone Stimuli