无芒稗质体ACCase基因Real-timePCR定量方法的建立

Establishment of Real-time PCR Assay for Quantitative of Plastid ACCase in Beardless Barnyardgrass

为研究一种抗精喹禾灵无芒稗的抗药性机理,建立了一种以β-actin为内参基因利用实时荧光定量PCR技术测定抗性无芒稗质体ACCase基因相对表达水平的方法。分析结果表明,设计的实时定量引物特异性非常好,内参基因、目的基因的扩增效率分别为103.2%、104.3%,线性相关系数分别为0.993 0、0.991 2,完全满足Real-time PCR定量方法的要求。所建立的无芒稗质体型ACCase基因Real-time PCR定量方法将为研究质体型ACCase基因的表达和杂草对ACCase抑制剂的抗药性机制研究提供方法学参考。

In order to study the resistance mechanism of a quizalofop-ethyl-resistance beardless barnyardgrass, a relative quantitative analysis method was established based on real-time PCR technology and β-actin gene as the housekeeping gene. The results showed the specificity of the two sets of primers was very well. The amplification efficiencies of the primers for ^-actin and plastid ACCase gene was 103.2% and 104.3, while the correlation coefficients was 0.993 0 and 0.991 2, respectively. The results showed the real-time PCR quantitative method established in the article could he provided as a methodological reference for the study of the expression of plastid ACCase gene and the study of the mechanism of weed resistant to ACCase inhibitors.