Biological and biochemical properties of an isolate of cherry rasp leaf virus from red raspberry

A mechanically transmissible virus obtained from symptomless plants of a red raspberry selection imported into Scotland from Quebec, Canada was indistinguishable serologically from a cherry isolate of cherry rasp leaf virus (CRLV). The raspberry isolate, CRLV-R, was graft transmitted to several virus indicator species and cultivars of Rubus without inducing noticeable symptoms. In Chenopodium quinoa sap, CRLV-R lost infectivity after dilution to 10^-5 or heating for 10 min at 60°C but was infective after 16 days (the longest period tested) at 18°, 4° or - 15°C. The virus particles are isometric, c. 28 nm in diameter, and were purified with difficulty from infected C. murale and C. quinoa plants. The particles comprise two nucleoprotein components with sedimentation coefficients of 89 and 115 S and are prone to aggregate during purification. When centrifuged to equilibrium in CS₂SO₄ solution, purified virus preparations formed two major components with p= 1·28 and 1·36 g/cm³. Virus particles contained two RNA species which, when denatured in glyoxal and electrophoresed in agarose gels, had estimated mol. wt of 2·56 × 10⁶ (RNA-1) and 1·26 × 10⁶ (RNA–2). Infectivity of CRLV-R RNA was abolished by treatment with proteinase K, suggesting that the RNA is linked to protein necessary for infectivity; RNA molecules contained polyadenylate. In reticulocyte lysates, CRLV-R RNA stimulated the incorporation of ³H-leucine, mainly into two polypeptides of estimated mol. wt 200 000 and 102 000. When electrophoresed in polyacrylamide gels, protein obtained from CRLV-R particles purified by centrifugation to equilibrium in Cs₂SO₄ separated into three bands with estimated mol. wt 26 000 , 23 000 and 21 000.     

Purification and particle properties of groundnut rosette assistor virus and production of a specific antiserum*

Purified preparations of the luteovirus, groundnut rosette assistor virus (GRAV), were made by treatment of groundnut leaf extracts with cellulase, followed by sucrose density gradient centrifugation. Yields of virus particles were about 0·5-1·0 mg/kg leaf material. The preparations contained isometric particles c. 28 nm in diameter with a sedimentation coefficient (s_{20, w}) of 115 S, a buoyant density in Cs₂SO₄ of 1·34 g/cm³, and A₂₆₀/A₂₈₀ of 1·86. The particles contained a single species of nucleic acid (presumably RNA), of mol. wt 2·09 × 10⁶and with no detectable polyadenylate sequence, and a single protein species, of mol. wt 24 × 10³. An antiserum produced in a rabbit had a titre of 1/256 in gel diffusion tests and detected GRAV in leaf extracts by ELISA. GRAV particles reacted in F(ab')₂-ELISA and immunosorbent electron microscopy (ISEM) tests with antisera to bean leaf roll, potato leafroll and tobacco necrotic dwarf luteoviruses, but did not react with antisera to carrot red leaf luteovirus.     

Purification and properties of ginger chlorotic fleck virus

A previously undescribed isometric virus, named ginger chlorotic fleck virus (GCFV), was detected in ginger (Zingiber officinale) imported into Australia from a number of countries. The geographical distribution of the virus is uncertain, but is thought to include India, Malaysia and Mauritius. The virus apparently does not occur in Australian commercial ginger plantings. The virus has isometric particles c. 30 nm in diameter, with a sedimentation coefficient of 111 S, and was readily purified from infected ginger with yields of 50–90 mg/kg leaf tissue. Purified preparations contained a major species of single-stranded RNA of mol. wt 1.50 × 10⁶ and a major coat protein species of mol. wt 29.0 × 10³. At pH 7, the particles formed a single zone in both caesium chloride and caesium sulphate gradients, with buoyant densities of 1.355 g cm^-3 (fixed virus) and 1. 297 g cm^-3 (unfixed virus), respectively. The virus particles migrated as two electrophoretic components and were labile when treated with 10 mM EDTA, 1 M NaCI, 10 mM tris pH 8.25 or when negatively stained with potassium phosphotungstate. GCFV was mechanically transmitted only to ginger, and was not transmitted by the aphids Myzus persicae. Pentalonia nigronervosa, Rhopalosiphum maidis or R. padi. Possible affinities of GCFV with the sobemo-virus group are discussed. The present cryptogram of GCFV is ^R/_l: ^1.5/₂₀: ^S/_S: ^S/_*.     

Olive latent ringspot virus, a newly recognised virus infecting olive in Italy

A manually transmissible virus, for which the name olive latent ringspot virus (OLRV) is proposed, was isolated from a symptomless olive tree. The virus was mechanically transmitted to test plants. Purified preparations of OLRV contained three classes of isometric particle, c. 28 nm in diameter, with sedimentation coefficients of 525 (T), 975 (M) and 1325 (B) and containing 0, 30 and 43% nucleic acid respectively. At equilibrium in CsCl gradients, the buoyant densities of T and M components were 1–29 and 1–43 g/cm³ respectively, whereas B component separated into two sub-components with buoyant densities of 1–51 g/cm³ (BJ and 1–52 g/cm³ (B₂). Particle preparations contained two species of single-stranded RNA with mol. wt 1–40 times 10⁶ and 2–65 times 10⁶, both necessary for infectivity. The coat protein of OLRV, dissociated under strong denaturing conditions, separated into four components in polyacrylamide gel electrophoresis. Over 75% of the protein was found in a band with mol. wt 57 600, but all four components were recognised as oligomers of a monomeric form with mol. wt 14 300. OLRV was serologically unrelated to 26 different isometric plant viruses including 17 recognised nepoviruses. Its properties strongly indicate that it is a hitherto undescribed member of the nepovirus group.     

Properties of broad bean yellow band virus, a possible new tobravirus

A virus was transmitted from broad bean plants in Apulia (Southern Italy) with leaves showing yellow rings, line patterns or yellow vein banding and malformations and necrosis of pods. Symptoms in some, but not all, test plants were similar to those induced by tobraviruses. Purified virus preparations contained two classes of rod-shaped particles containing c. 5% nucleic acid with sedimentation coefficients of 186S and 276S. After centrifugation to equilibrium in CsCl gradients, two components were resolved, with buoyant densities of 1·298 and 1·316 g/cm³. Unfractionated virus preparations contained two species of single-stranded RNA with mol. wts of c. 1·06 × 10⁶ and 2·48 × 10⁶ and one species of coat protein with mol. wt of c. 21 300. The modal lengths of the two classes of particles, both in plant sap and in purified preparations, were 77 nm (S particles) and 202 nm (L particles). L particles accumulated in infected cells in paracrystalline aggregates, whereas S particles were randomly distributed in the cytoplasm of cells. The virus was serologically unrelated to two isolates of tobacco rattle virus and two isolates of pea early-browning virus. The virus, named broad bean yellow band, is considered a distinct tobravirus.     

Some properties of pelargonium flower-break virus

A virus obtained from pelargonium cvs Irene and Paul Crampel appears to differ from any previously reported; although symptomless in most pelargonium cvs tested, it caused colour break in the flowers of two seedling clones. It seems uncommon in pelargoniums. The virus was readily transmitted by inoculation of sap, but not by Myzus persicae with short feeds, by dodder or through seed. It infected only fifteen of 100 species tested in six of thirty-five plant families. Pelargoniums were freed from the virus by heat-treatment. The virus remained infective after 10 min at 85 ^oC, 3 wk at 20 ^oC or 27 wk at 2 ^oC; it was infective at 1/500000 dilution of Nicotiana clevelandii or Chenopodium quinoa sap. Purified preparations were readily made by several methods, and contained isometric particles c. 30 nm diameter. Although a good antigen, the virus was serologically unrelated to any of forty-two isometric viruses. In immunoelectrophoresis, the virus moved as a single antigenic component towards the cathode. It gave a single, specific zone in density-gradient centrifugation, and one moving component (s⁰_{20 w}= 125 S) in analytical centrifugation. The virus contained one protein of mol. wt. c. 41000. The present cryptogram of the virus is (R)/*: */*:S/S:S/*, and the name pelargonium flower-break virus is proposed.     

Occurrence, purification and properties of narcissus tip necrosis virus

Narcissus tip necrosis virus (NTNV), a previously undescribed virus, was detected in the Netherlands and the United Kingdom in plants of twenty-one cultivars of trumpet, large-cupped, small-cupped, double, tazetta and poeticus narcissus. In some cultivars distinct leaf symptoms were sometimes associated with infection but in others infected plants remained symptomless and detection was dependent on serological tests. The virus was readily transmitted by manual inoculation to narcissus, but it failed to infect any of forty-six other plant species from fourteen families. Up to 50 mg of virus/kg of tissue were obtained by differential centrifugation of narcissus leaf extracts previously clarified with either diethyl ether, n-butanol or a mixture of n-butanol and chloroform. The virus particles are isometric, c. 30 nm in diameter, have a sedimentation coefficient (^s°₂₀ w) Of 123 S a buoyant density of 1·356 g/cm³, migrate as a single component in polyacrylamide gel electrophoresis, and contain a single RNA species of mol. wt 1·6×10⁶ and two major polypeptides of mol. wt 42000 and 39000. Although NTNV resembles tombusviruses it showed no serological relationship to the type member or six putative members of this group or to thirty-four other viruses with isometric particles. Its present cryptogram is R/*:1.6/(18):S/S:S/*.     

Purification and some particle properties of anthriscus yellows virus, a phloem-limited semi-persistent aphid-borne virus

Anthriscus yellows virus (AYV), a phloem-limited virus transmitted in the semi-persistent manner by the aphid Cavariella aegopodii, was purified by treatment of leaf extracts with cellulasc, followed by differential and sucrose density gradient centrifugation. ‘The preparations contained isometric particles c. 29 nm in diameter which were unstable unless stored in buffer at pH 8.0 containing 1 mM CaCl₂,. The particles sedimented as two components, ’full‘ nucleoprotein particles with A₂₆₀/A₂₈₀= 1.83 containing about 42% nucleic acid, and ’empty‘ protein shells with A₂₆₀,/A₂₈₀= 0.73; their buoyant densities in CsCl solutions were 1.52 and 1.27 g/cm³. Respectively. Yields of ihe nircleoprotein particles were c. 1.75 mg/kg leaf tissue. The particles contained a single species of RNA, of mol. wt 3.6 × 10 “(10 000 nucleotides). Particle protein preparations contained four electrophoretic species, of mol. wt (× 10³) 35.0, 28.3, 23.3 and 22.3.C. aegopodii did not transmit AYV from purified preparations. A rabbit injected with AYV preparations produced antibodies that coated AYV particles in electron microscope tests, but gave variable reactions in immunosorbent electron microscopy (ISEM), depending on the composition of the medium. No reactions were obtained in enzyme-linked inimunosorbent asjay (ELISA). No serological relationship was detected in ISEM between AYV and any of 10 viruses that resembled it in one or more properties.     

Purification and properties of elm mottle virus

A virus obtained commonly from Wych elm (Ulmus glabra) in Scotland showing ringspot and line-pattern leaf symptoms was serologically related to elm mottle virus (EMotV) from East Germany. The virus was seed-borne in elm and was transmitted by inoculation of sap to elm and twenty-one herbaceous species. No symptoms developed in infected elm seedlings kept in the glasshouse. In Chenopodium quinoa sap, EMotV lost infectivity after diluting to 10^-4, after 10 min at 60 ^oC, or 9 days at 18 ^oC. When purified from C. quinoa sap by clarification with n-butanol (8-5 %, v/v) and differential centrifugation, preparations contained quasi-spherical particles mostly 26–29 nm m diameter (mean = 28 nm) which sedimented as three nucleo-protein components with sedimentation coefficients (s^o₂o, w) of 83, 88 and 1 or S; most infectivity was associated with the 101 S component but infectivity was enhanced by adding the slower sedimenting components. When centrifuged to equilibrium in caesium chloride solution at 4 ^oC, purified virus preparations were largely degraded and contained many non-infective particles c. 15–22 nm in diameter, and intact infective particles which formed a band of density c. 1–34 g/cm³. Polyacrylamide gel electrophoresis indicated that EMotV contained a single major protein species of estimated mol. wt. 25000 and five RNA species of estimated mol. wt. 1–30, 1.15, 0–82, 0 39 and 0–30 times10⁶. Gel electrophoresis of RNA extracted from the separated components indicated that the 101 S component contained 1–30 x io⁶ mol. wt. RNA and the 83 S component 0–82 times 10⁶ mol. wt. RNA. In these and other properties, EMotV resembles the serologically unrelated tobacco streak virus.     

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

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 S_{20, 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 M_T 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 M_r between c. 31 000 and up to three minor ones (S) or M_r between c. 20 000 and 24 000. This contrasts with protein preparations from other BBTMV isolates that typically contain only two polypeptides of M_r 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.     

Purification and properties of Australian lucerne latent virus, a seed-borne virus having affinities with nepoviruses

An isolate of Australian lucerne latent virus (ALLV) from lucerne in New Zealand was mechanically transmitted to a few herbaceous hosts. It induced diagnostic symptoms in several species of the Chenopodiaceae, but was symptomless in most other hosts including lucerne and Trifolium subterraneum. It was seed transmitted in lucerne. When assayed to Chenopodium quinoa, infective C. quinoa sap lost infectivity after diluting to 10^-4, heating for 10 min at 55°C and storage for 4 days at 4°C. ALLV was purified from infected C. quinoa or pea plants by extracting sap in 0.1 m borate buffer (pH 7) containing 0.2% 2-mercaptoethanol and clarifying with 15% bentonite suspension, high and low speed centrifugation and sucrose density gradient centrifugation. Purified virus preparations contained isometric particles about 25 nm in diameter and sedimented as three virus components with sedimentation coefficients (s_20-w⁰) of 56 S, 128 S and 133 S. The 56 S component appeared to consist of nucleic acid-free protein shells. Polyacrylamide gel electrophoresis of virus preparations showed that ALLV contained a single protein species of mol. wt 55 000 and two RNA species of mol. wt 2.1 × 10⁶ and 2.4 × 10⁶. An antiserum to ALLV had an homologous titre of 1/256 to purified virus but failed to detect ALLV in infective sap of C. quinoa, pea or lucerne. Purified ALLV failed to react to antisera to 28 distinct isometric plant viruses including those to 10 nepoviruses.     

Pathology, soil transmission and characterization of cymbidium ringspot, a virus from cymbidium orchids and white clover (Trifolium repens)

Two strains of a virus, designated cymbidium ringspot virus (CyRSV), were isolated from cymbidium orchids and from Trifolium repens respectively in Britain. Experimentally infected cymbidiums developed slight chlorotic ring-mottle; T. repens developed flecks and mottling in the leaves, and slight stunting. Of 101 plant species tested, the cymbidium strain infected sixty-one (thirteen systemically) in twenty-three of thirty-five families; the clover strain infected sixty-four species (eighteen systemically) in twenty-two families. Both strains were propagated in Nicotiana clevelandii and assayed in Chenopodium quinoa. CyRSV was readily transmitted by inoculation of sap, and by foliage contact between plants, but not by the aphids Myzus persicae or Acyrtho-siphon pisum, nor through seed of T. incarnatum, Phaseolus vulgaris or N. clevelandii. Highly infective virus was released into soil from roots of infected N. clevelandii, and acquired by bait seedlings planted in such soil. Similar transmission occurred when purified virus was applied to the surface of sterilized soil containing bait plants; there was no evidence for any living soil vector. The virus was eliminated from 96 % of small cuttings taken from infected N. clevelandii plants grown at 35–37 °C for 9 wk. CyRSV was still infective in sap of N. clevelandii after dilution to 10?⁵-io–⁶ (only 2 × 10^_1 in cymbidium sap), or after 10min at 85–90 °C. It survived at least 10 months at c. 20 °C and more than 12 yr at 2 °C. Lyophilized sap was highly infective after over 13 yr at laboratory temperatures under high vacuum. Purified preparations made by clarification with n-butanol, followed by differential centrifugation and exclusion chromatography on controlled-pore glass beads, contained isometric particles c. 30 nm diam., with s°_20W= 137 S, and had a buoyant density in caesium chloride of 1–36 g/ml. The A 260/A 280 ratio was 1–55, and A max(26o)/A min(242) was 1–17. The virus contained c. 15 % of single-stranded RNA of mol. wt 1–7 × 10⁶; the nucleotide base ratios were: G27'8; A24/9; C2I-3; U26-I. There was one capsid polypeptide of mol. wt 43600. The virus was a good immunogen and a strongly reacting antigen in vitro; in Immunoelectrophoresis, each strain migrated as a single antigenic component towards the cathode. The cymbidium and clover strains were serologically closely related, although spurs were produced in immunodiffusion. No serological relationship was found to forty-three other isometric viruses, including eighteen tombusvirus isolates; CyRSV nevertheless shares many properties with tombusviruses, and we assign it provisionally to this group. The cryptogram is: R/r:1:7/15:S/S:S/O.     

Properties of Scottish isolates of cocksfoot mild mosaic virus and their comparison with others

Two isolates of cocksfoot mild mosaic virus obtained from cocksfoot (Dactylis glomerata) in Scotland differed in symptomatology, and apparently in host range, from isolates obtained in Germany and Wales. They were serologically more closely related to a Dutch isolate from cocksfoot, and to a Scottish isolate from timothy (Phleum pratense), than was the German isolate from cocksfoot. The Scottish isolate from timothy was somewhat more virulent than, but serologically closely related to a Welsh isolate from timothy. Particles of Scottish isolates from cocksfoot and timothy were best preserved for electron microscopy by fixation with osmium tetroxide. In 1.0 m KCl or 0.01 m ethylene diamine tetraacetate they were stable at pH 5.2–5.3 but unstable above pH 7; they were disrupted by 0.5% sodium dodecyl sulphate. The particles contained major and minor RNA components of mol. wt c. 1.5. 10⁶(RNA-1) and 0.5. 10⁶(RNA-2) respectively, together with polydisperse RNA of intermediate mol. wt and protein of mol. wt c. 27 000. In CsCl gradients, major and minor nucleoprotein components of density 1.39 and 1.38 g/ml respectively were distinguished. The less dense particles contained a larger proportion of intermediate-sized RNA molecules and of RNA- 2 , and a smaller proportion of RNA- 1 , than did the denser particles. Particles seem to contain either RNA-1 or various combinations of smaller RNA molecules. Despite the differences in antigenic constitution, symptomatology and particle stability between virus isolates obtained from cocksfoot and timothy in different countries, these isolates seem sufficiently similar to be considered one virus.     

Host range and properties of potato black ringspot virus

Potato black ringspot virus (PBRV), obtained from cultivated potato in Peru, was found to have a very wide host range resembling that of tobacco ringspot virus (TRSV-B), to which PBRV is distantly related serologically. However, PBRV caused the more severe symptoms in many species and, unlike TRSV B, infected Lycopersicon esculentum and Cyamopsis tetragonoloba. In Solanum tuberosum, PBRV caused necrotic spots and ringspots in systemically infected leaves in the year of infection and was readily transmitted through tubers to progeny plants, most of which developed no obvious symptoms although systemically infected. TRSV-B infected non-inoculated S. tuberosum leaves only sporadically, did not induce symptoms in them and was not transmitted through tubers to progeny plants. PBRV was cultured in Nicotiana clevelandii and infectivity was assayed in Cheno-podium amaranticolor or C. quinoa. Virus particles were purified from leaf extracts, after clarification using chloroform, by precipitation with 6% polyethylene glycol and differential centrifugation. Purified preparations contained 25 nm diameter isometric particles with somewhat angular outlines, sedimenting as three components (T, M and B) at 49, 84 and 117 S, and containing a single protein species of mol. wt 59 000. Preparations of PBRV nucleic acid contained two species, estimated by polyacrylamide gel electrophoresis in non-denaturing conditions to have mol. wt of about 25 10⁶ (RNA-1) and 15 10⁶ (RNA-2). Infectivity was associated with B particles, preparations of which contained RNA-1 and RNA-2, presumably in different particles. M particles contained RNA-2, were not infective and enhanced infectivity only slightly when added to B particles. Similar relative amounts of RNA-1 and RNA-2 were extracted from unfractionated virus using phenol or Pronase, but preparations obtained using phenol were much the more infective. PBRV has properties typical of nepoviruses; its present cryptogram is (R/1):2–5/41 + 15/28 or 2 1 5/46:S/S:S/*, nepovirus group.     

Properties of a carlavirus causing a latent infection of pepino (Solanum muricatum)

Pepino (Solanum muricatum) cuttings imported from Chile contained a latent virus which was transmitted by inoculation of sap to Chenopodium quinoa but not to 21 other species. The virus was transmitted by the aphid, Myzus persicae. In C. quinoa sap, the virus lost infectivity when diluted between 10^-3 and 10^-4, heated for 10 min between 65 and 70 °C, or stored at room temperature for 4 to 6 days. The virus particles were straight or slightly flexuous filaments 660 to 680 nm long. Up to 15 mg virus per 100 g C. quinoa leaves was obtained by clarification with a mixture of chloroform and carbon tetrachloride. Purified preparations had A_max/A_min= 1.11, A₂₆₀/A₂₈₀= 1–30, A^0.₂₆₀^1%= 2.8, and contained a single sedimenting component with a sedimentation coeficient of 149s and a buoyant density in CsCl of 1–318. The virus particles contained 5.5% of single-stranded RNA of mol. wt 2.4×10⁶ (estimated by gel electrophoresis of undenatured RNA) and sedimentation coefficient 38.5S, and a single polypeptide of mol. wt 33 000. The virus is distantly serologically related to potato S and carnation latent viruses and is considered a new member of the carlavirus group. The name pepino latent virus is proposed. The cryptogram for this virus is R/1: 2.4/5–5: E/E: S/Ve/Ap.     

Particle properties of parsnip yellow fleck virus

Particle preparations of parsnip yellow fleck virus (PYFV) isolates A-421 and P-121, representing the two major serotypes, were made by clarifying leal extracts with ether or butan-1-ol and concentrating the virus particles by precipitation with polyethylene glycol and differential centrifugation. The preparations contained c. 31 nm-diameter particles comprising two sedimenting components. Top component (T) consisted of stain-penetrable protein shells with A₂₆₀/A₂₈₀= 0.8–0.9, sedimentation coefficient (S₂₀) = 56 S (A-421) or 60 S (P-121), and buoyant density = 1.297 g/cm³. Bottom component (B) consisted of nucleoprotein particles, not penetrable by negative stain, with A₂₆₀/A₂₈₀= 1.9, sedimentation coefficient (S⁰_20.w) = 148 S (A-421) or 153 S (P-121), and buoyant density = 1.520 g/cm³ (A-421) or 1.490 g/cm³ (P-121). Yields of B component particles were up to c. 1 mg/100 g leaf tissue (both isolates); yields of T component particles were up to c. 0.6 mg (A-421) or 5.5 mg (P-121) per 100 g leaf tissue. PYFV particles were found to contain a single RNA species (mol. wt c. 3.4 × 10⁶, c. 9800 nucleotides), constituting 40% of the particle weight, and three polypeptide species, of mol. wt (× 10 ³) 30 , 26 and 24 (A-421) or 31 , 26 and 23 (P-121).     

Ullucus virus C, a newly recognised comovirus infecting Ullucus tuberosus (Basellaceae)

Ullucus virus C (UVC) is a comovirus prevalent in Ullucus tuberosus grown at high altitudes in the Bolivian and Peruvian Andes. It was transmitted mechanically to U. tuberosus (Basellaceae) and to five of 26 species from three of eight other families, infecting U. tuberosus symptomlessly but inducing conspicuous systemic infection in Chenopodium amaranticolor and C. quinoa. Sap from infected C. quinoa was usually infective after 10 min at 70 but not 75 °C, after dilution to 10^-7 but not 10^-8, and after 8 but not 16 wk at 20 °C. UVC was not transmitted by either of two aphid species (Aphis gossypii and Myzus persicae) or through seed of C. quinoa, but it was transmitted by leaf contact between infected and healthy plants. UVC has isometric particles which, in neutral phosphotungstate, are c. 28 nm in diameter. The particles sediment as three components (T, M and B) with sedimentation coefficients (s^?_{20, w}) of 51 S (T), 95 S (M) and 116 S (B). M component particles have a buoyant density (g cm^-3) in caesium chloride of 1.404, and B component particles separated into minor and major sub-components with densities of 1.409 and 1.463, respectively. T, M and B particles were serologically indistinguishable, and each contained similar relative amounts of two polypeptides of mol. wts 20 700 and 45 100. T particles contained only protein, but M particles also contained c. 30% ss-RNA of mol. wt 1–45 ×10⁶ and B particles c. 38% ss-RNA of mol. wt 2·2 × 10⁶. The virus is serologically distantly related to cowpea mosaic virus but, as it showed no relationship to any of 11 other similar viruses, it is probably a distinct member of the comovirus group.     

Properties of lucerne transient streak virus, and evidence of its affinity to southern bean mosaic virus

A virus obtained from naturally infected lucerne ( Medicago sativa ) in New Zealand reacted with antiserum to an Australian isolate of lucerne transient streak virus (LTSV). Some plants infected with New Zealand isolates showed yellow flecks along lateral veins of leaves; symptoms were transient in some lucerne plants but persistent in others. A New Zealand isolate (LTSV-NZ) infected 14 of 39 plant species tested by mechanical inoculation, but was not transmitted by five aphid species. In sap of Nicotiana clevelandii , LTSV-NZ was infective after storage for 4 wk at 20 ^oC, diluting to 10^-5, or heating for 10 min at 70 ^oC. Purified virus preparations contained a single electrophoretic component and a single sedimenting component (s_20w= 112 S ) which formed a single buoyant density component in CsCl (1.37 g cm^-3) but two density components in Cs₂SO₄ (1.26 and 1.32 g cm^-3). LTSV-NZ particles were stable in 10 ITIM EDTA at pH 5, but not at pH 8, being degraded into two sedimenting components of 105 S and 92 S. Particles contained c. 18% RNA in the form of one single-stranded RNA molecule of mol. wt 1–4 times 10⁶, and a polypeptide of mol. wt c. 32 400. LTSV-NZ was serologically unrelated to 24 other isometric plant viruses. However, its properties are similar to those of southern bean mosaic virus and allied viruses. The present cryptogram of LTSV is R/l: 1–4/(18):S/S:S/*.     

Crimson clover latent virus - a newly recognised seed-borne virus infecting crimson clover (Trigolium incarnatum)

Crimson clover latent virus (CCLV) was detected in five seed lots of crimson clover (Trifolium incarnatum) from Europe and in one from the United States of America. Ninety-seven per cent of all crimson clover plants examined were found to be infected but were without symptoms. Keeping crimson clover plants at 32–38°C for 34 days failed to free them from CCLV. The virus was not transmitted by Myzus persicae, but was transmitted by inoculation of sap to Chenopodium album, C. amaranticolor and C. quinoa. Twenty-four other plant species from seven families were not infected. CCLV was best propagated in C. quinoa in which it caused stunting and systemic chlorosis. Sap from infected C. quinoa was infective after dilution to 10^-2 but not 10^-3, after 10 min at 60°C but not 65°C, and after 20 days at 20°C. In neutral phosphotungstate, CCLV had isometric particles c. 26 nm in diameter with a hexagonal profile. About 20 to 80 A_1cm,260 units of purified virus were obtained from 1 kg of infected C. quinoa or C. amaranticolor leaves by extraction in 0.5 M phosphate buffer, pH 7.5, containing 0.01 M ethylene diamine tetra-acetate and 0.4% 2–mercaptoethanol and clarification with chloroform-butanol followed by two precipitations with polyethylene glycol (mol. wt 6000) and several cycles of differential centrifugation. Purified virus sedimented as three components with sedimentation coefficients (s°_{20, w}) of 52S, 101S and 122S. The 101S and 122S components had buoyant densities in CsCl of 1.438 and 1.495 g/cm³ respectively. From these values the nucleic acid content of the 101S and 122S components was estimated to be 32–35% and 40–41% respectively. The virus contained a single protein with an estimated mol. wt of 52 000 and two single-stranded RNA species of estimated mol. wt 1.6 × 10⁶ and 2.2 × 10⁶. CCLV was serologically unrelated to 31 other morphologically similar viruses. Although its vector is unknown, CCLV seems to have affinities with nepoviruses. The cryptogram of CCLV is R/1:2.2/40–41 + 1.6132–35:S/S:S/*.     

Some Properties of Cucumber Fruit Streak Virus

An isometric virus c. 30 nm in diameter with a single RNA species (mol.wt 1.45 × 10⁶) isolated from cucumber plants from the island of Crete (Greece) is described under the name of cucumber fruit streak virus (CFSV). The most evident symptom on naturally infected plants consisted of longitudinal chlorotic streak of the fruits. In glasshouse, the virus was soil-transmitted to C. sativus, and, mechanically, to a wide range of herbaceous hosts, most of which were infected only locally. Purified virus preparations sedimented as a single component with sedimentation coefficient of 132S. At equilibrium these preparations were homogeneous in CsCl gradients but formed two bands in Cs₂SO₄ gradients. Virus particles were stabilized by forces involving divalent cations, pH-dependent bonds and salt links between protein and RNA. Although some of the properties of CFSV are similar to those of other small spherical viruses with single RNA species there are differences which do not allow for the assignment of the virus to any of established taxonomic group of plant viruses.