关键词: 茄子, 高温胁迫, 内参基因, 表达稳定性, 实时荧光定量PCR

Abstract:

The present study aimed at selecting the stable reference genes to ensure the reliability and accuracy in gene expression analysis of eggplant（Solanum melongena L.） under high temperature stress. By real-time fluorescence quantitative RT-PCR technology，we investigated the expression stability of eight candidate reference genes（SmEF1a，SmEF2，Sm40sRPS29，Sm60sRPL24，SmTRX，SmCKⅠ，SmDAHPSⅠ，SmUCP）from RNA-Seq database under high temperature stress and SmActin，and the stability of expression of nine reference genes were evaluated by GeNorm，NormFinder，BestKeeper and ReFinder，respectively. The results showed that，at different periods，different tissues and different lines of eggplant under high temperature，the expression abundance and stability of these nine reference genes are different. When analyzing the gene expression of thermo-sensitive line 05-1 and thermo-resistant line 05-4 under high temperature stress，SmEF1a and SmUCP were the best reference genes for different periods ，and SmEF1a and SmTRX could be used as best reference genes for different tissues. Meanwhile，SmTRX and SmEF2 could be used as best reference genes in lines or varieties with different thermo-tolerance. On the whole，SmEF1a and SmTRX exhibited the most stable expression among all of the tested samples，while SmActin and SmCKⅠexhibited the least stable expression under most of the experimental conditions. It was found to be feasible and efficient to identify stably expressed genes from transcriptome database under high temperature stress as candidate reference genes，which will be helpful for the study of expression of genes response to high temperature and the mechanisms of thermo-tolerance in eggplant.

Key words: Solanum melongena, high temperature stress, reference gene, expression stability, real-time fluorescence quantitative PCR