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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

来源：花匠小妙招时间：2024-12-11 20:44

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

原文链接: 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://www.huajiangbk.com/newsview1042939.html

分类：花卉