菜豆壳球孢侵染芝麻过程中内参基因的筛选

发布时间：2018-05-06 16:47

本文选题：芝麻茎点枯病菌 + qRT-PCR　；参考：《中国油料作物学报》2017年03期

【摘要】：为选择适合菜豆壳球孢(Macrophomina phaseolina)侵染芝麻过程中实时荧光定量PCR分析的内参基因,11个基因作为候选内参基因。经过PCR扩增效率筛选,β-肌动蛋白(ACTB)、泛素连接酶(UBC)、α-微管蛋白(TUBA)、γ-微管蛋白(TUBC)、3-磷酸甘油醛脱氢酶基因(GAPDH)、核糖体蛋白S5(RPS5-a,RPS5-b)和内部转录间隔区(ITS)8个基因符合要求,可用于稳定度筛选。利用实时荧光定量PCR技术,检测了这8个候选基因在菜豆壳球孢侵染芝麻8h、16h、24h和32h的表达情况。经Ge Norm软件分析,ACTB、GAPDH和RPS5-b等3个基因表达较稳定。经过最适内参基因数目分析,在菜豆壳球孢侵染芝麻过程中基因定量表达分析时,选择ACTB、GAPDH和RPS5-b的多基因组合作为内参,能够更准确地校正定量结果。
[Abstract]:In order to select the internal reference gene suitable for the real-time fluorescence quantitative PCR analysis of sesame seeds infected by Macrophomina phaseolina, 11 genes were used as candidate internal reference genes. After PCR amplification, eight genes, 尾 -actin ACTBN, ubiquitin ligase (UBCU), 伪 -tubulin (TUBAA), 纬 -tubulin (TUBC) 3-phosphoglyceraldehyde dehydrogenase (GAPDHN), ribosomal protein S5 (RPS5-aRPS5-b) and internal transcriptional spacer (ITS5), met the requirements and could be used for stability screening. The expression of these eight candidate genes was detected by real-time fluorescence quantitative PCR (PCR) in cultured sesame seeds infected with Phaseolus vulgaris for 24 h and 32 h, respectively. The expression of GAPDH and RPS5-b were stable by GE Norm software. According to the analysis of the optimum number of internal reference genes, the multi-genome cooperation of RPS5-b and ACTB GAPDH can be used to correct the quantitative results more accurately in the process of quantitative expression analysis of the genes in the process of infecting sesame seeds.
【作者单位】：河南省农业科学院植物保护研究所农业部华北南部农作物有害生物综合治理重点实验室河南省农作物病虫害防治重点实验室;
【基金】：国家自然科学基金(31301631) 农业部现代农业产业技术体系(CARS-15-1-05)
【分类号】：S435.653

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