Purification of Rice Stripe Virus

Gang Lu; Min Yao; Yijun Zhou; Xiaorong Tao

doi:10.21769/BioProtoc.3565

Purification of Rice Stripe Virus

Bio Protoc. 2020 Mar 20;10(6):e3565. doi: 10.21769/BioProtoc.3565.

Authors

Gang Lu^{1

2}, Min Yao^{1

3}, Yijun Zhou⁴, Xiaorong Tao¹

Affiliations

¹ Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.
² Institute of Plant Virology, Ningbo University, Ningbo, China.
³ Division of Antitumor Pharmacology, State Key Laboratory of Drug Research, Shanghai Institute of MateriaMedica, Chinese Academy of Sciences, Shanghai, China.
⁴ Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing, China.

Abstract

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.

Keywords: Density gradient centrifugation; Filamentous virion; Rice stripe virus; Ultracentrifugation; Virion purification.