浙江农业学报

• 园艺科学 • 上一篇    下一篇

猕猴桃实时荧光定量PCR分析中内参基因的筛选

  

  1. (浙江省农业科学院 园艺研究所,浙江 杭州 310021)
  • 出版日期:2015-04-25 发布日期:2015-04-30

Screening of reference genes for real-time quantitative PCR in kiwifruit

  1. (Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China)
  • Online:2015-04-25 Published:2015-04-30

摘要: 为筛选在猕猴桃中稳定表达的内参基因,以确保其基因表达分析结果的可靠性。应用实时荧光定量PCR技术分析了ACTB,TUB,18S rRNA和GAPDH四个常用植物内参基因在华特猕猴桃不同组织及果实不同发育时期的mRNA表达情况。经GeNorm软件分析发现,4种内参基因的表达稳定性各异,TUB和ACTB在6种不同组织中表达均稳定,TUB和ACTB组合适合作华特猕猴桃不同组织基因表达的内参基因;18S rRNA在果实不同发育时期的表达最稳定,18S rRNA和TUB组合最适合作为分析华特猕猴桃果实不同发育时期基因表达的内参基因。

关键词: 猕猴桃, 内参基因, qRT-PCR

Abstract: The present study aimed at selecting the stable reference genes to ensure the reliability and accuracy of gene expression analysis in kiwifruit for qRT-PCR. The expressions of four frequently-used reference genes (ACTB,TUB,18S rRNA,GAPDH) were analyzed in different tissues and fruits at different developmental stages, respectively. According to the analysis by GeNorm, the stabilities of the four candidate reference genes were different, TUB and ACTB were both stably expressed in six different tissues, which were recommended for genes expression normalization in different kiwifruit tissues. 18S rRNA was the most stable gene in different developmental stages of White kiwifruit, 18S rRNA and TUB were most suitable for normalization of mRNA expression levels in White kiwifruit.

Key words: kiwifruit, reference genes, qRT-PCR