应用实时荧光定量RT-PCR检测几种食用菌蛋白抗烟草花叶病毒的活性

Real-time fluorescent quantitative RT-PCR method to determine the antiviral activity of edible mushroom proteins against Tobacco mosaic virus(TMV)

根据烟草花叶病毒（Tobacco mosaic virus,TMV）外壳蛋白（CP）RNA的特异性序列设计TaqMan荧光探针及其引物,利用实时荧光定量RT-PCR检测食用菌蛋白抗TMV的活性。经食用菌蛋白处理后,TMV病毒汁液中TMV的RNA浓度下降了22.02%~87.93%,传统生物学测定的食用菌蛋白抗TMV的平均枯斑抑制率为10.88%～83.97%;相关性分析表明,实时荧光定量RT-PCR测定的病毒RNA浓度的下降与传统生物学方法测定的平均枯斑抑制率之间呈正相关（r=0.818 8）,具有较好的一致性。利用实时荧光定量RT-PCR检测蛋白抗烟草花叶病毒活性的方法,具有特异性好、快速、简便、重复性高的特点,适合于蛋白抗烟草花叶病毒活性的痕量快速高效检测。

The antiviral activity of edible mushroom proteins was determined by real-time fluorescent quantitative RT-PCR.TaqMan fluorescence probe and primers targeting the specific RNA sequence of TMV coat protein were designed and used to test the concentration of Tobacco mosaic virus RNA in samples.After the crude TMV extract was treated with proteins extracted from edible mushroom,the concentration of TMV RNA decreased 22.02%-87.93%.Traditional biology assessment of disease resistance revealed that inhibition rate of dry spots was 10.88%-83.97%.These two methods were well correlated with r=0.818 8.The research demonstrated that this method could be applied to determine the antiviral activity of edible mushroom proteins rapidly and effectively.