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白背飞虱Sogatella furcifera Horváth(WBPH)是南方水稻黑条矮缩病毒(southern rice black-streaked dwarf virus,SRBSDV)的主要传播介体。本实验以白背飞虱的三龄若虫、五龄若虫、雄虫和雌虫为材料,以18S为内参基因,SRBSDV的CP基因为目的基因,建立了基于MIQE指南的实时荧光定量PCR体系。使用该体系结合相对定量法,检测了不同虫态白背飞虱体内携带南方水稻黑条矮缩病毒的相对含量。结果表明,相同饲毒条件下,白背飞虱不同虫态体内SRBSDV的相对含量为雌虫>雄性>五龄若虫>三龄若虫。该法能准确定量白背飞虱体内南方水稻黑条矮缩病毒的相对含量。
Sogatella furcifera Horváth (WBPH) is a major transmission medium of southern rice black-streaked dwarf virus (SRBSDV). In this study, the third instar nymph, the fifth instar nymph, the male and the female of whitebacked planthopper were used as materials, and the 18S gene was used as internal reference gene. The CP gene of SRBSDV was used as the target gene, and a real-time fluorescence quantitative PCR system based on MIQE guidelines was established. Using this system combined with the relative quantification method, the relative content of southern black-streaked dwarf virus carrying in different forms of white-backed planthoppers was tested. The results showed that the relative content of SRBSDV in the different insect states of whitebacked planthopper was female> male> fifth instar nymph> third instar nymph under the same feeding conditions. The method can accurately quantify the relative content of southern rice black-streaked dwarf virus in the white-backed planthopper.