摘要
葡萄果实发育后期基因表达水平的变化对果实品质形成具有重要影响,内参基因的选择是基因表达定量分析的关键影响因素。本试验以雷司令葡萄品种花后50、75和95 d的果实为材料,通过半定量RT-PCR,研究了常用持家基因Actin、Tubulin、GAPDH1、8 S rRNA的表达变化,探讨其作为葡萄果实发育后期基因表达半定量分析内参基因的可行性。结果表明:18 S rRNA的表达水平高,且最稳定,其次是Actin,而Tubulin和GADPH的相对表达水平较低;对VvTLP、GHF17 P和VvASR3个功能基因在不同果实材料中的表达水平进行半定量RT-PCR,VvTLP和VvASR的相对表达水平较高,GHF17 P的表达水平较低,整体上均呈现递增的趋势;以Actin和18 S rRNA为内参得到的归一化结果基本一致,在4个持家基因中18 S rRNA和Tubulin是优选的葡萄果实发育后期基因表达研究的内参基因。
The change of genes＇ expression level during berry ripening is crucial for the final quality of grapes.Internal reference gene selection is a key factor influencing the result of gene expression quantification.In this paper,berries of wine grape Riesling（Vitis vinifera L.） were sampled at 50,75 and 95 day after flowering（DAF）,with semi quantitative RT-PCR,the expression level of four commonly used housekeeping genes Actin,Tubulin,GAPDH and 18S rRNA were studied,and their feasibility as internal reference were evaluated.The results showed that 18S rRNA was with high and the most stable expression level,followed by Actin,whereas the expression level of Tubulin and GAPDH was relatively low.The expression level of three functional genes VvTLP,GHF17P and VvASR was investigated,among of them VvTLP and VvASR were with high expression level,and that of GHF17P was comparatively low.The quantification results with Actin and 18S rRNA as the internal control are similar,VvTLP,GHF17P and VvASR show up-regulated expression during ripening,18S rRNA and Tubulin are selected as internal references for gene expression level study of during grape berry development.
出处
《中国农业大学学报》
CAS
CSCD
北大核心
2010年第3期7-14,共8页
Journal of China Agricultural University
基金
国家现代葡萄产业技术体系项目(nycytx-30-04)
国家自然科学基金资助项目(30500347)
关键词
葡萄
发育后期
持家基因
半定量RT-PCR
grape berry
late development stage
housekeeping gene
semi quantitative RT-PCR
作者简介
通讯作者：赵晓，硕士研究生，E-mail：zhaoxiaocau@gmail．com
徐海英，研究员，主要从事葡萄育种研究，E—mail：haiyingxu63@sina．com
