Phytopathology. 1998 Sep;88(9):885-9. doi: 10.1094/PHYTO.1998.88.9.885.
ABSTRACT An indirect immunofluorescent technique was developed to localize tobacco ringspot nepovirus (TRSV) in the vector nematode Xiphinema americanum sensu stricto. A population of this nematode that efficiently transmitted TRSV was given an acquisition access period of 10 days on TRSV-infected cucumber. Treatment of fragments of viruliferous nematodes with a polyclonal antiserum against TRSV followed by fluorescein isothiocyanate-conjugated goat anti-rabbit immunoglobulin G resulted in virus-specific bright fluorescence only in the lumen of the stylet extension and esophagus. Virus-specific fluorescent signals were observed in the virus-retention region of 44% of the nematode fragments examined. The percentage of nematodes labeled with virus-specific fluorescence increased as the acquisition access period increased from 0 to 22 days; the increase paralleled the increase in the transmission efficiency of the nematode population. Visualization of the entire virus-retention region of individual nematodes within a population of vector or nonvector nematodes provides a rapid and simple means of monitoring specific attachment of plant viruses.
摘要 本研究开发了一种间接免疫荧光技术，用于定位烟草环斑病毒（TRSV）在载体线虫 Xiphinema americanum s.s. 中的位置。将能够有效传播 TRSV 的线虫种群给予 10 天的 TRSV 感染黄瓜的获取访问期。用针对 TRSV 的多克隆抗血清处理携带病毒的线虫片段，然后用荧光素异硫氰酸酯标记的山羊抗兔免疫球蛋白 G 进行处理，结果仅在口针延伸和食道的内腔中观察到病毒特异性明亮荧光。在检查的线虫片段中，有 44%的线虫片段观察到具有病毒特异性的荧光信号。随着获取访问期从 0 天增加到 22 天，带有病毒特异性荧光的线虫百分比增加；这种增加与线虫种群的传播效率的增加相平行。在群体内的单个线虫或非载体线虫的整个病毒保留区域进行可视化，为监测植物病毒的特异性附着提供了一种快速而简单的方法。
