Comparative Purification of Two Luteoviruses

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Abstract
An Illinois vector-nonspecific isolate of barley yellow dwarf virus (BYDV) was purified from oats (Avena byzantina ''Coast Black'') and a California isolate of beet western yellows virus (BWYV) was purified from shepherd''s purse (Capsella bursa-pastoris). Root tissue yielded 4 times as much BYDV as shoots when ground to a very fine powder in lipid N2 with a mortar and pestle and homogenized in a blender prior to clarification. BYDV yields .**GRAPHIC**. = 8) from roots harvested in March-October averaged 0.7 mg/kg, while in November-February yields averaged 4.4 mg/kg. Average yields of 1.1 mg/kg .**GRAPHIC**. = 8) of BWYV were obtained when shoots were frozen in liquid N2, powdered in a blender and stirred for 24 h at room temperature in 0.1 M phosphate buffer (pH 6.0) with 0.5% sodium azide and 1.5% Rohament P, a macerating enzyme. The titer of BYDV peaked sharply in roots 10-14 days after inoculation; BWYV yields were consistently high from shoots harvested from 10 to > 20 days after inoculation. Purified BYDV and BWYV had A260/280 ratios of 1.89 and 1.65, respectively.