西红花实时荧光定量PCR内参基因筛选  

Screening of Reference Genes Based on Real-Time Quantitative PCR Analysis in Saffron(Crocus sativus)

作　　者：周琳[1] 蔡友铭[1] 杨柳燕[1] 赵冰雪 李芳 张永春[1] ZhouLin;Cai Youming;Yang Liuyan;Zhao Bingxue;Li Fang;Zhang Yongchun(Shanghai Key Laborlory of Proected Horicuctburad Tchology,Farestry and Ponologyr Reseach Ietitwte,Shanghai Academy of Aprioulunal Sciences,Shanghai,201106;School of Ecological Techmalogy and Engineering,Shanghai Institute of Technology,Shanghai,201418;Agiculunal Colle of Anshun Uaiversity,Anshun,561000)

机构地区：[1]上海市农业科学院林木果树研究所,上海市设施园艺技术重点实验室,上海201106 [2]上海应用技术大学生态技术与工程学院,上海201418 [3]安顺学院农学院,安顺561000

出　　处：《分子植物育种》2020年第22期7467-7475,共9页Molecular Plant Breeding

基　　金：上海市现代农业产业技术体系建设[沪农科产字(2019)第8号];上海市农业科技成果转化项目[沪农科转字(2016)第2-2号]共同资助。

摘　　要：为筛选西红花不同组织间稳定表达的最佳内参基因,以西红花的根、球茎、主芽、花丝、叶片、侧芽共6个组织为试验材料,运用实时荧光定量PCR方法分析了18S核糖体RNA (18S rRNA)、肌动蛋白基因(ACT)、转录延伸因子-1α(EF1α)、3-磷酸-甘油醛脱氢酶基因(GAPDH)、微管蛋白基因(TUB)和多聚泛素基因(UBQ)共6个内参基因在不同组织中的表达差异情况。基于Ct值,利用geNorm、NormFinder和BestKeeper分析6个内参基因表达稳定性。结果表明,3个软件筛选出的最佳内参基因略有不同。综合分析结果显示,GAPDH和UBQ表达较为稳定,其次为18S rRNA和ACT,而EF1α和TUB不适合作为西红花不同组织的内参基因。本研究确定西红花q RT-PCR分析的合适内参基因为GAPDH和UBQ,可用于种球发育、花发育、次生代谢产物形成和积累等机理研究。In order to identify the suitable reference genes for the gene expression analysis in Crocus sativa, the expression of 6 housekeeping genes, including 18 S rRNA, ACT, EF1α, GAPDH, TUB and UBQ were assessed by using real time quantitative PCR in roots, corms, main buds, filaments, leaves and lateral buds of Crocus sativa.Based on the Ct value, the stability of 6 reference genes was analyzed by using geNorm, NormFinder and BestThe results showed that the suitable reference genes screened by the three softwares were slightly different. The results of comprehensive analysis showed that the expression of GAPDH and UBQ was stable, followed by 18 S rRNA and ACT, while EF1α and TUB were not suitable for different tissues of saffron. In this study, we determined that the suitable reference genes for saffron q RT-PCR analysis were GAPDH and UBQ, which can be used for the study of bulb development, flower development, secondary metabolite formation and accumulation.

关 键 词：西红花(Crocus sativus) 不同组织 内参基因 实时荧光定量PCR 

分 类 号：S567.239[农业科学—中草药栽培]