Abstract
Northern cereal mosaic virus, grown in barley plants, was purified from grounding plant mate-rials suspended in 0.1M glycine solution at pH 6.5. Extracted juice was added to active carbon and DEAE cellulose powder as absorption materials for host cell components. The mixture was stirred for one to two minutes and filtered by suction through Celite (Hyflo-Supercel) in a Büchner funnel. The virus was then pelleted by centrifugation at 26, 000g. for one hour The resulting pellet was suspended in the glycine solution and centrifuged on a sucrose density at 41, 000g. for 45 minutes. The virus zone in sucrose density was withdrawn and further purified by density gradient electrophoresis, Viruses were almost completely separated from contaminating plant components by the electrophoresis.
When purified preparations of the virus were injected into its vector planthoppers (Laodelphax striatellus) with glass capillaries, a high percentage of survived planthoppers could transmit the virus to barley seedlings.
The virus particle was found to be bacilliform and 350 (330-380) nm×68nm in size. The virus was revealed to contain RNA. The lipid fraction extracted with chloroform-methanol (2:1) from a highly purified virus preparation was analyzed by thin layer chromatography. Sterol and phospholipids (phosphatidyl-ethanolamine and possibly phosphatidyl-serine) were detected. Antiserum to the purified virus (128-256 in titer) reacted positively with the semipurified preparations from four isolates of the same virus obtained from the northern and central parts of Japan. No serological differences were found among these four isolates.
