果树学报2013，Vol.30Issue(1)：48-54,7.
‘琯溪蜜柚’荧光定量PCR内参基因的筛选
Validation of internal reference genes for qRT-PCR normalization in ‘ Guanxi Sweet Pummelo' (Citrus grandis)
摘要
Abstract
[Objective] The objective was to clone some housekeeping genes from 'Guanxi Sweet Pummelo' [Citrus grandis (L.) Osbeck], and select one or more suitable genes as reference genes. [Method]In this study, juice sacs of the fruits in different developmental stages and roots, stems and leaves of 'Guanxi Sweet Pummelo' were chosen as materials, then the expression stability of five candidate reference genes including actinl ,β-tubulin, 18S rRNA,EFl-α and Ubiquitin by real-time fluorescence quantitative reverse transcription PCR was analysed. Finally, four analytical softwares which are named geNorm, NormFinder, comparative Delta -CT and BestKeeper were used to assess the optimum reference genes. [Result]Our data showed that actin1 and β-tubulin are suitable reference genes for normalization during fruit developmental stages, while EFl-α and β-tubulin are proposed as good candidate genes across different tissues of 'Guanxi Sweet Pummelo'. [Conclusion]This work has validated optimal reference gene named β-tubulin, and lays the foundation for gene expression analysis of 'Guanxi Sweet Pummelo' in the future.关键词
‘琯溪蜜柚’/荧光定量PCR/内参基因Key words
'Guanxi Sweet Pummelo' [Citrus grandis (L.) Osbeck]/ Real-time fluorescence quantitative reverse transcription PCR/ Reference genes分类
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王梨嬛,潘永娟,杨莉,蔡盛华,黄新忠..‘琯溪蜜柚’荧光定量PCR内参基因的筛选[J].果树学报,2013,30(1):48-54,7.基金项目
国家自然科学基金青年科学基金(30900978) （30900978）
浙江师范大学博士科研启动基金(ZC304006122) （ZC304006122）
