S U M M A R YThree biologically distinct strains of bean yellow mosaic virus (BYMVs, BYMVl and PV-2) were partially purified by centrifugation at relatively low g forces. Serologically these strains appeared to be distinct from each other but were related.Our recent isolation of the severe strain of bean yellow mosaic virus (BYMVs) from naturally infected sweet violet (Viola odorata L.) (Prowidenti & Granett, 1974) prompted us to compare serologically this isolate with two other known strains of the virus. Difficulties, however, were encountered in the purification of BYMVS with the method outlined by Damirdagh & Shepherd (1970) and successfully used for an authentic strain of BYMV by Uyemoto, Prowidenti & Schroeder (1972).This paper describes a reliable procedure for the partial purification of BYMVs, BYMVl (pea isolate I ; Hagedorn & Walker, 1950) and PV-2 (pea virus 2; Schroeder & Provvidenti, 1966) and reports their serological comparison in agar gel double diffusion tests.BYMVs and BYMVI were propagated in bean (Phaseolus vukaris L. cv. Red Kidney) and in pea (Pisum sativum L. cv. Ranger) while PV-2 was increased only in pea. Infected bean leaves were harvested 30-40 days after inoculation and pea leaves after 11-21 days. The three strains of BYMV were purified using a modification of the method recently proposed by Huttinga (1973).Each gram of chilled leaves was homogenized in a blender in a cold solution containing 7-5 ml of 0-1 M tris (hydroxy-methyl) aminomethane buffer, z ml of carbon tetrachloride and 2 ml of chloroform. The pH of the extractant was adjusted with thioglycollic acid to 6.5 for BYMVs and 8-5 for BYMVl and PV-2. The homogenate was centrifuged for 10 min at 6000 g (Sorvall RCzB, SS34 rotor, 7000 rev./min) to pellet most plant debris, after which the supernatant fluid was filtered through glass wool. The solution was then centrifuged at 20000g (Spinco Model L, 30 rotor, 15000 rev./min) for I h. The pelleted virus was resuspended in 0 -1 M tris buffer adjusted with HC1 to the same pH as the original extractant. Preparations were further clarified by centrifugation at 3000 g (SS34 rotor, 5000 rev./min) for 5 min. Higher centrifugal forces during either initial clarification or pelleting increased particle aggregation and breakage and reduced virus yield.
SUMMARY Plantago mottle virus (RMV), a member of the tymovirus group, was identified as the causal agent of a disease of pea (Pisum sativum) in New York State. The pea virus isolates were identical in host range and serology to the type strain from Plantago major. In susceptible pea genotypes symptoms were strongly influenced by ambient temperature; high temperature (35°C) reduced infectivity and suppressed symptoms, whereas low temperature (15 and 25°C) prolonged the incubation period but favoured the development of conspicuous leaf veinal chlorosis, mottle and necrosis. Resistance to P1MV was found in seventeen of twenty‐five domestic pea cultivars and in two of twelve foreign introductions. Many of the P1MV‐resistant lines were resistant also to bean yellow mosaic virus. The use of resistant cultivars and the apparent limited conditions for efficient transmission of this virus have minimized its importance to pea crops in New York State.
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