The present study was aimed at screening suitable reference genes for quantitative real-time polymerase chain reaction (qRT-PCR) in tiger puffer (Takifugu rubripes), an important aquaculture species in Asia and also a good model species for lipid research. Specifically, this reference gene screening was targeted at standardization of gene expression in different tissues (liver, muscle, brain, intestine, heart, eye, skin, and spleen) or under different nutritional conditions (starvation and different dietary lipid levels). Eight candidate reference genes (ribosomal protein L19 and L13 (RPL19 and RPL13), elongation factor-1 alpha (EF1α), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine guanine phosphoribosyl transferase1 (HPRT1), beta-2-Microglobulin (B2M), 18S ribosomal RNA (18SrRNA), and beta actin (ACTB)) were evaluated with four algorithms (geNorm, NormFinder, BestKeeper, and comparative ΔCt method). The results showed that different algorithms generated inconsistent results. Based on these findings, RPL19, EF1α, 18SrRNA, and RPL13 were relatively stable in different tissues of tiger puffer. During starvation conditions, ACTB/RPL19 was the best reference gene combination. Under different dietary lipid levels, ACTB/RPL13 was the most suitable reference gene combination. The present results will help researchers to obtain more accurate results in future qRT-PCR analysis in tiger puffer.

本研究旨在为亚洲重要的水产养殖物种和脂质研究的良好模式物种虎鲀 ( Takifugu rubripes )筛选合适的参考基因用于定量实时聚合酶链反应 (qRT-PCR) 。具体而言，该参考基因筛选针对不同组织（肝脏、肌肉、大脑、肠道、心脏、眼睛、皮肤和脾脏）或不同营养条​​件（饥饿和不同膳食脂质水平）下的基因表达标准化。8 个候选参考基因（核糖体蛋白 L19 和 L13（RPL19和RPL13）、延伸因子 1 α（EF1α）、甘油醛-3-磷酸脱氢酶（GAPDH ）)、次黄嘌呤鸟嘌呤磷酸核糖转移酶 1 ( HPRT1 )、β-2-微球蛋白 ( B2M )、18S 核糖体 RNA ( 18SrRNA ) 和 β 肌动蛋白 ( ACTB )) 使用四种算法（geNorm、NormFinder、BestKeeper 和比较 ΔCt 方法）进行评估. 结果表明，不同的算法产生的结果不一致。基于这些发现，RPL19、EF1α、18SrRNA和RPL13在河豚不同组织中相对稳定。在饥饿条件下，ACTB / RPL19是最佳的参考基因组合。不同膳食血脂水平下，ACTB /RPL13是最合适的参考基因组合。目前的结果将有助于研究人员在未来对河豚的 qRT-PCR 分析中获得更准确的结果。