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.
出处
《植物研究》
CAS
CSCD
北大核心
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)~~
关键词
羽衣甘蓝
实时荧光定量PCR
内参基因
柱头发育
SLG
Brassica oleracea var. acephala
real-time quantitative PCR
reference genes
stigma development
SLG
