论文部分内容阅读
苹果果实着色期基因表达水平的变化对果实品质形成具有重要影响,选择适合的内参基因是提高实时荧光定量PCR分析准确性的首要条件。本试验以‘富士’苹果着色过程中不同取样时间的果皮为材料,通过q RT-PCR分析了常用候选持家基因β-actin、EF-1α、GAPDH和18S r RNA的表达变化,借助ge Norm和Norm Finder程序筛选出在果实着色期q RT-PCR分析的理想内参基因。结果表明,EF-1α的表达水平高且最稳定,其次是18S r RNA,而β-actin和GAPDH的相对表达水平较低;同时,用筛选的内参基因EF-1α和18S r RNA分析苹果花青苷合成途径的二氢黄酮醇-4-还原酶基因的表达水平,其表达规律比较一致,均呈现正态分布。因此,EF-1α和18S r RNA是研究苹果着色期表达的理想的内参基因。
The change of gene expression level in apple fruit during coloration has an important influence on the formation of fruit quality. Choosing suitable internal reference gene is the most important condition to improve the accuracy of real-time PCR. In this study, the expression changes of β-actin, EF-1α, GAPDH and 18S rRNA were analyzed by q RT-PCR using the pericarps of different sampling times during ’Fuji’application of apple. Using ge Norm and The Norm Finder program screened for the ideal reference gene for q RT-PCR analysis of fruit coloration. The results showed that the expression level of EF-1α was the highest and most stable, followed by 18S rRNA, while the relative expression levels of β-actin and GAPDH were lower. At the same time, the expression of EF-1α and 18S rRNA The expression level of dihydroflavonol-4-reductase gene in the pathway of cyanidin synthesis was consistent with the normal expression pattern. Therefore, EF-1α and 18S rRNA are the ideal internal control genes for studying the expression of apple during the coloration period.