Purification and properties of lucerne Australian symptomless virus, a new virus infecting lucerne in Australia

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Summary The flexuous filamentous particles of wineberry latent virus (WLV) were found to measure 620. 12 nm and not 510. 12 nm as previously reported. Analysis of dsRNA from infected plants detected a major species of c. 5.7. 106 mol. wt and minor species of lower mol. wt. Purified virus particles formed a major and a minor buoyant density component in solutions of caesium salts with densities of 1.26 and 1.25 g cm‐3 in Cs2SO4 and 1.30 and 1.29 g cm‐3 in CsCl. The particles contained a single nucleic acid species, presumably single stranded RNA, and a single polypeptide of estimated mol. wt 2.78. 106 and 31 000 respectively. In indirect ELISA, purified particles of WLV and particles in plant sap failed to react specifically with antiserum to nine carlaviruses, 12 potexviruses, three capilloviruses or apple chlorotic leafspot closterovirus, nor was WLV found to react with several of these antisera in immunosorbent electron microscopy or immunoblots. In Marion and Olallie blackberry, WLV in mixture with raspberry bushy dwarf virus (RBDV), but not RBDV alone, induced veinal line‐pattern symptoms resembling those of calico disease reported from the USA.

Section: Methodsmentioning

confidence: 99%

Further properties of wineberry latent virus and evidence for its possible involvement in calico disease

Jones¹,

Mitchell²,

McGavin³

et al. 1990

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“…Analysis of virus particle protein was by SDS polyacrylamide gel electrophoresis followed by staining with Coomassie Brilliant Blue as described by Remah, Jones & Mitchell (1986). The molecular size markers used were the pre-stained low range protein standards from Bio-Rad.…”

Section: Ems Particle Purijkation and Virus Particlementioning

confidence: 99%

“…The molecular size markers used ranged from 0.36-9.5 kb (IBI). In some early experiments, ELV RNA was also analysed after denaturing in glyoxal as described by Remah et al (1986).…”

Section: Ems Particle Purijkation and Virus Particlementioning

confidence: 99%

Elderberry latent virus: its relationship to Pelargonium ringspot virus and its identification as a distinct member of the genus Carmovirus, family Tombusviridae

Jones¹,

McGavin²,

Brunt³

et al. 2000

The properties of Elderberry latent virus (ELV) and Pelargonium ringspot virus (PelRSV) were compared. The viruses were largely indistinguishable in herbaceous host range and symptomatology, particle morphology, sedimentation coefficient and RNA profiles and size. They were also very closely related serologically with SDI differences in agarose gel double-diffusion tests of 1 to 3. Purified virus particle preparations of each virus contained isometric particles c. 30 nm in diameter that sedimented as a major component with an so2,,, of 112-115s. Purified virus particle preparations contained a major and a minor ssRNA species that In polyacrylamide gel electrophoresis (PAGE) had estimated sizes of c. 3.8 kb and c. 1.6 kb respectively. Plants of Chenopodium quinoa infected with ELV or PelRSV each contained three dsRNA species of c. 3.8, 2.6 and 1.8 kbp, although the smallest of these species was not evident in all preparations. Protein from purified virus particle preparations contained a major polypeptide that, in SDS-PAGE, had an estimated Mr of 40 000 (40K). However, after storage of purified virus particles for 7-1 0 days, protein preparations from PelRSV particles also contained an additional major polypeptide of estimated A( of 37 000 that is probably derived by degradation of the 40K protein; this additional component was not observed in freshly prepared preparations of ELV. Neither virus was found to be related serologically to 16 other viruses with isometric particles and similar properties. These data, together with the recent finding by other researchers that the smallest RNA species is a sub-genomic RNA, suggests that both viruses are members of the genus Carmovirus, and that PelRSV is a minor variant of ELV. However, the taxonomic status of these two viruses is discussed in relation to recent brief reports comparing the nucleotide and amino acid sequences of these two viruses.

“…Protein and RNA were extracted from virus particle preparations purified by sucrose density gradient centrifugation and, in some instances, also by isopycnic centrifugation in caesium salts. Extraction of protein and RNA, and electrophoresis of protein in SDS in polyacrylamide gels and RNA in agarose gels was as described by Remah, Jones & Mitchell (1986).…”

Section: Polyacrylamide Gel Electrophoresis Of Virus Protein and Nuclmentioning

confidence: 99%

Properties of a host range and coat protein variant of broad bean true mosaic comovirus from Vicia faba

Jones¹,

Mitchell²

1994

Summary Unlike other described isolates of broad bean true mosaic comovirus (BBTMV), a variant, code name SB, infected some non‐leguminous plant species and, in N. benthamiana, induced systemic mottling and puckering of the leaves. However, like other described BBTMV isolates, purified SB particle preparations contained isometric particles c. 28 nm in diameter that sedimented as two nucleoprotein components with S20, w values of 90S and 109S; some preparations occasionally contained a component of c. 50S. Virus particles contained two ssRNA species which, when denatured in glyoxal, had estimated MT values of 2.1 × 106 and 1.3 × 106 and co‐electrophoresed with cowpea mosaic virus RNA‐1 and RNA‐2 respectively. Isolate SB was serologically indistinguishable from British and German isolates of BBTMV. However, SB virus particles contained a major polypeptide (L) of Mr between c. 31 000 and up to three minor ones (S) or Mr between c. 20 000 and 24 000. This contrasts with protein preparations from other BBTMV isolates that typically contain only two polypeptides of Mr c. 37 000 (L) and 21 000 (S). Following isopycnic centrifugation in CsCl, SB particles purified from pea separated into two major components with densities of 1.39 and 1.44 g cm‐3 and a minor component of estimated density 1.43 g cm‐3. In Cs2SO4, virus preparations separated into three major components with densities of 1.30, 1.32 and 1.36 g cm‐3 and a minor one of density 1.27 g cm‐3. In CsCl isopycnic gradients, SB particles purified from TV. benthamiana separated into two components with densities of 1.38 and 1.43 g cm‐3. During immuno‐electrophoresis in agarose gels, freshly prepared virus and preparations stored for up to 4 days at 4°C contained a single component that migrated rapidly to the anode, whereas similar preparations of an English isolate of BBTMV migrated as a single component that moved only slowly toward the anode but which, within 48 h, contained an additional component with a migration rate similar to that of isolate SB. Isolate SB is therefore a host range variant of BBTMV which, in comparison with previously described isolates of BBTMV, has an increased negative charge of its particles prior to any appreciable degradation of its S protein, and S protein that is degraded less rapidly. These features probably account for the anomalies observed in isopycnic centrifugation.