荧光定量PCR技术检测花生种皮抗黄曲霉基因表达强度

依据已获得的基因芯片和cDNA研究结果,筛选上调表达并且表达差异系数在5.0以上的花生种皮基因序列设计引物,采用SYBR GreenⅠ实时荧光定量PCR法对上述基因进行相对定量分析,以验证基因芯片研究结果并为分离黄曲霉抗性相关基因奠定基础。结果表明:荧光定量PCR的结果与基因芯片的结果高度一致,目的基因在感病和抗病品种之间差异显著,其中BE87、AW03在抗病品种中相对表达量较高,可能在抵御黄曲霉侵染过程中发挥重要作用。

Detection of Expression Intensity of Aspergillus flavus Resistance Gene in Peanut Seed Capsule by Quantitative Real -Time PCR

According to the research results of obtained gene chip and cDNA library,the primers were designed based on the gene sequences of peanut seed capsule which up-regulated expression and the differential expression coefficient more than 5.0.The relative quantitative analysis of these genes was carried out by SYBR Green Ⅰ quantitative real-time PCR to identify the research results of gene chip and provide foundation for separating related genes resistant to Aspergillus flavus.The results showed that the quantitative real-time PCR results were consistent with the research results of gene chip on a high degree;there were significant differences between Aspergillus flavus resistant and susceptible varieties.Among which,BE87 and AW03 expressed more in Aspergillus flavus resistant varieties and maybe play an important role in defense from Aspergillus flavus infection.