花椰菜实时荧光定量PCR内参基因的筛选与评价  

Screening and Evaluation of Reference Genes for qRT-PCR in Cauliflower(Brassica oleracea L.var.botrytis L.)

作　　者：林珲[1] 裘波音 张前荣[1] 李大忠[1] 温庆放[1] Lin Hui;Qiu Boyin;Zhang Qianrong;Li Dazhong;Wen Qingfang(Fujian Engineering Research Center for Vegetables,Fujian Key Laboratory of Vegetable Genetics and Breeding,Crops Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou,350013)

机构地区：[1]福建省农业科学院作物研究所,福建省蔬菜遗传育种重点实验室,福建省蔬菜工程技术研究中心,福州350013

出　　处：《分子植物育种》2025年第1期105-112,共8页Molecular Plant Breeding

基　　金：国家现代农业产业技术体系基金项目(CARS-23-G-53);特色蔬菜育种攻关与产业化开发基金项目(zycxny-2021009);福建省农科院蔬菜遗传育种科技创新团队项目(CXTD2021038)共同资助。

摘　　要：为筛选适合花椰菜稳定表达的内参基因,利用实时荧光定量PCR技术,分析检测9个候选内参基因ACT1、ACT2、ACT7、TUB、EF-1α、EF-1β、GAPDH、UBQ和HIS在花椰菜不同组织、不同品种和花球不同发育时期处理下mRNA的表达稳定性。利用3个数据分析软件(geNorm,NormFinder和BestKeeper)综合评价9个候选内参基因的表达稳定性。结果表明,在花椰菜不同组织、不同品种和花球不同发育时期处理下,候选内参基因的表达稳定性存在差异。ACT2和EF-1α在不同组织处理下表达最为稳定;ACT7和TUB在不同品种处理下表达稳定性最好;ACT7和GAPDH在花球不同发育时期处理下被筛选为最合适的内参基因组合。结果表明,在特定条件下,获得合适的内参基因为花椰菜新基因表达分析的准确性提供了保障,并为十字花科芸薹属植物关键基因的挖掘提供了有益的参考。To select the most stable reference genes of cauliflower(Brassica oleracea L.var.botrytis L.)for quantitative real-time PCR,we assessed the mRNA expression in different tissues of nine candidate reference genes at different tissues,different cultivars,or various ballflower development stages.Genes assessed included ACT1,ACT2,ACT7,TUB,EF-1α,EF-1β,GAPDH,UBQ and HIS.The expression stabilites of these nine reference genes were evaluated by geNorm,NormFinder and BestKeeper.It was found that mRNA expression of the candidate reference genes differed significantly at different tissues,different cultivars or various ballflower development stages.In different tissues of cauliflower,ACT2 and EF-1αwere recommended as the reference genes for analyzing the mRNA expression;ACT7 and TUB were the most stable gene in different cultivars;ACT7 and GAPDH were stably expressed with various ballflower development stages.In conclusion,obtaining the appropriate internal reference genes under specific conditions provides a guarantee for the accuracy of new gene expression analysis in cauliflower and offers a useful reference for the mining of reference genes in Brassica plants of the Cruciferae family.

关 键 词：花椰菜 内参基因 实时荧光定量PCR 

分 类 号：S635.3[农业科学—蔬菜学]