组培条件下苹果实时定量PCR内参基因的筛选Selection of reference genes for Real-Time Quantitative PCR in apples(Malus domestica) in vitro
李君英;张鹤;韩永峰;邵建柱;孙建设;
摘要(Abstract):
【目的】筛选在组织培养条件下苹果苗中稳定表达的内参基因。【方法】应用实时荧光定量PCR技术,分析了18S r RNA,GAPDH,TUB,UBQ,ACT,EF-1α六个常用植物内参基因在苹果组培苗不同基因型、不同组织、不同继代时间的m RNA表达差异情况。供试的4个苹果基因型为‘长枝富士’‘短枝富士’‘红珍珠海棠’、砧木‘A8-11’;继代培养的不同组织为茎、叶、整株,生根培养的不同组织为根、茎、叶、整株;不同继代期间为10、20、30、40、50、60、70、80 d。【结果】经Ge Norm软件对6个内参基因的表达稳定性进行评价,ACT和EF-1α在苹果组培苗的不同组织和不同继代时间的基因表达分析中较稳定,UBQ和EF-1α在苹果组培苗不同品种中表达均稳定。【结论】组培条件下,苹果基因表达相关研究的理想内参基因为ACT、UBQ和EF-1α。
关键词(KeyWords): 苹果;组培苗;实时荧光定量PCR;内参基因
基金项目(Foundation): 国家苹果产业技术体系专项(CARS-28)
作者(Author): 李君英;张鹤;韩永峰;邵建柱;孙建设;
Email:
DOI: 10.13925/j.cnki.gsxb.20160097
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