羽衣甘蓝不同组织及柱头发育实时荧光定量PCR内参基因的筛选  

Selection of Reference Genes for Real-time Fluorescence Quantitative PCR in Different Tissues and Stigma Development from Ornamental Kale

作　　者：李晗[1] 李治龙[1] 李晓屿 李玉花[1] 蓝兴国[1] 

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040

出　　处：《植物研究》2016年第4期565-572,共8页Bulletin of Botanical Research

基　　金：中央高校基本科研业务费专项基金项目(DL13CA13);国家自然科学基金项目(31070275)~~

摘　　要：以羽衣甘蓝(Brassica oleracea var.acephala)S_(13-b)S_(13-b)自交不亲和系为试材,利用实时荧光定量PCR(qRTPCR)技术检测Actin、Cpi6、EF-1β、GAPDH、Tub-α3、Tub-α6、Ubc7候选内参基因在不同组织和5个不同发育时期柱头的表达情况,并运用ge Norm和Best Keeper软件统计分析候选内参基因的表达稳定性。在不同组织中,ge Norm软件统计分析的结果表明Tub-α6和EF-1β的表达较稳定;Best Keeper软件统计分析的结果表明Ubc7和Tub-α6的表达较稳定。在不同发育柱头中,ge Norm软件统计分析的结果表明Actin和Ubc7的表达较稳定;Best Keeper软件统计分析的结果表明EF-1β和Tub-α6的表达较稳定。以筛选到的内参基因Tub-α6和Actin,分别分析羽衣甘蓝柱头S-位点糖蛋白基因(SLG)在不同组织和不同发育时期柱头的表达水平,结果显示出SLG主要在柱头中表达,而且SLG在柱头发育成熟时期表达量达到最高。We used real-time quantitative polymerase chain reaction to test the expression of Actin, eysteine proteinase inhibitor 6 （ Cpi6 ）, elongation factor 1-beta （ EF-1β）, glyceraldehyde-3-phosphate dehydrogenase （ GAPDH）, tubulin alpha-3 （ Tub-α3 ）, tubulin alpha-6 （ Tub-α6 ） and ubiquitin conjugating enzyme7 （ Ubc7 ）, and then identified the most suitable reference genes in a given set of tissues and different developmental stigmas of ornamental kale（Brassica oleracea var. acephala） S13.bS13_b homozygotes by using the geNorm and BestKeeper software programs. By a geNorm analysis, Tub-α6 and EF-lfl were the most suitable reference genes among the given set of tissues, and Actin and Ubc7 were the most stable genes during stigma development. By a BestKeeper analysis, Ubc7 and Tub-α6 were the most suitable reference genes among the given set of tissues, and EF-1β and Tub-α6 were the most stable genes during stigma development. Tub-α6 and Actin were the most suitable reference genes among the given set of tissues and during stigma development, respectively. Furthermore, the expression of SLG normalized with Tub-α6 or Actin showed that SLG was predominantly expressed in stigma among the given set of tissues and reached its highest level at approaching anthesis during stigma development.

关 键 词：羽衣甘蓝 实时荧光定量PCR 内参基因 柱头发育 SLG 

分 类 号：S635[农业科学—蔬菜学]