Abstract：In order to screen out the reference genes suitable for different tissues of green sunfish Lepomis cyanellus, the expression levels of six candidate reference genes, ubiquitin folding modifier gene (ufm1), α-tubulin gene (TUBA), cytochrome b gene (CYTB), precursor protein transporter gene (SEC62), glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) and β-actin gene (ACTB), were detected in intestine, liver, muscle, brain, gill, gonad and other tissues in 9 samples of green sunfish by real-time fluorescence quantitative PCR (qRT-PCR). Stability of the candidate reference genes were evaluated by Norm Finder and BestKeeper software, and these algorithms were summarize by ReFinder to screen out the best reference genes. The results showed that the specific amplification products were found by quantitative PCR primers of the six candidate reference genes. The descending order of the expression levels of the six candidate reference genes in different tissues were ranked as CYTB>TUBA>ufm1>SEC62>ACTB>GAPDH, with the most stable reference gene of CYTB in the intestine, ufm1 in liver, muscle, gill and gonad, and TUBA in brain screened by ReFinder. The paired difference analysis by geNorm software revealed that at least two optimal candidate reference genes should be used. There was similar expression stability of ufm1 and TUBA in different tissues, with the overall best stability. The findings can be used for qRT-PCR analysis of gene expression differences in different tissues of green sunfish, and provide reference genes and theoretical basis for future gene expression analysis of green sunfish.
秦立鹏,曹露, 张惠满,刘青,刘少贞,宋晶. 绿太阳鱼不同组织qRT-PCR内参基因的筛选[J]. 水产科学, 2026, 45(2): 259-268.
QIN Lipeng, CAO Lu, ZHANG Huiman, LIU Qing, LIU Shaozhen, SONG Jing. Screening of qRT-PCR Reference Genes in Different Tissues of Green Sunfish Lepomis cyanellus. Fisheries Science, 2026, 45(2): 259-268.
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2026, Vol. 45 
