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
摘要： 以羽衣甘蓝(Brassica oleracea var. acephala)S13-bS13-b自交不亲和系为试材,利用实时荧光定量PCR(qRT-PCR)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geNorm和BestKeeper软件统计分析候选内参基因的表达稳定性。在不同组织中,geNorm软件统计分析的结果表明Tub-α6和EF-1β的表达较稳定;BestKeeper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,geNorm软件统计分析的结果表明Actin和Ubc7的表达较稳定;BestKeeper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。
关键词: 羽衣甘蓝, 实时荧光定量PCR, 内参基因, 柱头发育, SLG
Abstract: We used real-time quantitative polymerase chain reaction to test the expression of Actin, cysteine 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-1β 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.
Key words: Brassica oleracea var. acephala, real-time quantitative PCR, reference genes, stigma development, SLG
中图分类号:
S635
相关知识
百合体胚诱导、发育及不同组织实时定量PCR内参基因筛选
德国鸢尾花发育不同时期荧光定量内参基因的筛选及应用的制作方法
牡丹不同发育阶段种子和花瓣组织实时荧光定量PCR中内参基因的挖掘与筛选
蕙兰内参基因的筛选与A类花发育基因的表达特性研究
‘豆绿’牡丹的内参基因及其引物和应用
番茄细菌性斑点病菌实时荧光定量PCR检测方法的建立及应用
牡丹泛素延伸蛋白基因ubiquitin克隆及其作为内参基因的研究
桂花金属离子胁迫下lncRNA内参基因和应用
甲型H1N1流感病毒实时荧光定量PCR检测方法的建立
细菌16S rRNA基因实时荧光定量及分型方法的建立
网址: 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 https://m.huajiangbk.com/newsview1042939.html
| 上一篇: 研究发现：西兰花和羽衣甘蓝有助于 |
下一篇: 羽衣甘蓝的栽培与管理 |
