菠萝蜜实时荧光定量PCR分析中内参基因的筛选  

Screening of Reference Genes for Real-time Fluorescence Quantitative PCR in Jackfruit

作　　者：汪永保[1] 余庆[1] 李映志[1] 李洪波[1] 叶春海[1] 

机构地区：[1]广东海洋大学农学院,广东湛江524088

出　　处：《热带作物学报》2014年第7期1374-1381,共8页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.30771484;No.31101506);广东省教育厅科技创新重点项目;广东省科技厅农业攻关项目(No.2009B20305006)

摘　　要：为筛选菠萝蜜实时荧光定量PCR研究用的内参基因,以菠萝蜜不同发育阶段的果实、叶和花序为材料,分析GAPDH、18S rRNA、UBQ、ɑ-tubulin和β-tubulin 5个内参基因的表达情况,并对其表达稳定性进行分析。结果表明,在各组织不同发育阶段中,5个内参基因的表达丰度变化存在差异;geNorm、NormFinder和BestKeeper 3种方法得出的结论有所不同,经RefFinder综合评价后,果实中最稳定的3个内参基因是UBQ、GAPDH、18S rRNA;叶片中最稳定的3个内参基因是UBQ、GAPDH、β-tubulin;花序中最稳定的3个内参基因是UBQ、GAPDH、ɑ-tubulin。To select appropriate reference genes for real time fluorescence quantitative experiment in jackfruit,samples of different developmental stages from the fruit,leaf and inflorescence were used to investigate the expression abundance and its expression stability of five reference genes,e.g.,GAPDH,18S rRNA,UBQ,atubulin and β-tubulin.Results showed that,at different developmental stages of jackfruit tissues,differences existed in the expression abundance of these five reference genes; Different conclusion in the stability of gene expression were obtained using geNorm,NormFinder and BestKeeper,and the comprehensive ranking by RefFinder indicated that,the three most stable reference genes in fruit were UBQ,GAPDH,18S rRNA,while the three most stable reference genes were UBQ,GAPDH and β-tubulin in leaf and UBQ,GAPDH and a-tubulin in inflorescence.

关 键 词：菠萝蜜 实时定量PCR 内参基因 表达稳定性 

分 类 号：S431.191[农业科学—农业昆虫与害虫防治]