Lu Gang, Yao Min, Zhou Yijun, Tao Xiaorong
Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.
Institute of Plant Virology, Ningbo University, Ningbo, China.
Bio Protoc. 2020 Mar 20;10(6):e3565. doi: 10.21769/BioProtoc.3565.
Although many spherical and rod-shaped plant virus purification protocols are now available, only a few protocols on filamentous plant virus purification have been published. Here, we report a protocol for large-scale purification of Rice stripe virus (RSV) from RSV-infected rice tissues. RSV virions with high infectivity were first precipitated with polyethylene glycol (PEG) followed by pelleting through primary ultracentrifugation, ultracentrifugation in a glycerol cushion and ultracentrifugation in density gradient. The purified RSV virions can not only be viewed as filamentous particles under an electron microscope, but can also be acquired by insect vector through direct injection into insect body or through membrane feeding prior to transmission to rice plants.
尽管现在有许多关于球形和杆状植物病毒的纯化方案，但已发表的丝状植物病毒纯化方案却很少。在此，我们报告一种从感染水稻条纹病毒（RSV）的水稻组织中大规模纯化RSV的方案。首先用聚乙二醇（PEG）沉淀具有高感染性的RSV病毒粒子，然后通过一次超速离心、在甘油垫层中进行超速离心以及在密度梯度中进行超速离心来进行沉淀。纯化后的RSV病毒粒子不仅在电子显微镜下可被视为丝状颗粒，而且昆虫介体可以通过直接注射到昆虫体内或在传播到水稻植株之前通过膜饲法获取。
