Abstract：Real-time fluorescence quantitative PCR (qPCR) is a common way to study gene regulation at the transcriptional level, but its accuracy depends on the normalization of data by appropriate reference genes. In order to obtain the stable expression of reference genes in quinoa (Chenopodium quinoa) under cold, drought stresses and abscisic acid (ABA), fluridone (FLU)(ABA inhibitor) treatments, five common statistics tools (geNorm, Normfinder, BestKeeper, ΔCt method, RefFinder) were used to evaluate 9 candidate reference genes. The results showed that the optimal reference genes for cold or drought stress were triphosphate guanosinase 3 (GTP638) and 26S proteasome (PRN483), PRN483 and dipeptidyl carboxypeptidase (DCP894) for ABA or fluridone treatments, ubiquitin-conjugating enzyme E2 (UBC822) and PRN483 were the optimal reference genes for cold, drought stresses and ABA, fluridone treatments. Moreover, the stability of the housekeeping gene β-actin (ACT878) was the weakest when subjected to cold, drought stresses, and ABA or fluridone treatments. Additionally, the reliability of the suggested reference gene was confirmed by the expression levels of 9-cis-epoxycarotenoid dioxygenase (NCED185) and oxidoreductase (OXI802). The results showed that when UBC822 or PRN483 were used as reference gene, the expression patterns of NCED185 and OXI802 were comparable, and they could respond to cold, drought stresses and ABA, fluridone treatments. However, with ACT878 as the reference gene, NCED185 and OXI802 did not exhibit a response to the diverse treatments. This research provides appropriate internal reference genes for gene expression analysis of quinoa, as well as technical support to explore related molecular mechanisms.
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