Direct isolation of tobacco necrosis virus from soils

Summary In TNV-bearing soils, the virus occurred adsorbed to soil colloids in low levels. By direct assay, the TNV could be more readily isolated from the rhizosphere of naturally infected cluster bean plants. The level of reaction of the TNV isolated from the rhizosphere soil was the same as TNV-D (cb isolate) in precipitin ring tests with antisera against TNV-A and TNV-D. The phenomenon of release of TNV from the infected roots into the soil and adsorption of TNV particles to colloidal particles in the soil are discussed from the point of ecology and stability of TNV in soils.     

Partial purification and some properties of wineberry latent, a virus obtained from Rubus phoenicolasius

Wineberry latent virus (WLV) was obtained from a single symptomless plant of American wineberry (Rubus phoenicolasius) originally imported from the United States of America. On graft inoculation, WLV infected but induced no distinctive symptoms in several Rubus species including those used as indicators for known Rubus viruses. It was not seed-borne in wineberry. WLV was mechanically transmitted to several herbaceous species but induced local lesions in only a few; it was weakly systemic in some Chenopodium species. Infective C. quinoa sap lost infectivity after diluting to 10^-4, heating for 10 min at 70°C, and storage either for 8 days at 18°C or for 32 days at 4°C. Sap from infected plants contained flexuous filamentous particles c. 510°12 nm. WLV was partially purified by extracting infected C. quinoa leaves in 0·05 M tris-HCl buffer (pH ₇) containing 0·2% thio-glycerol and 10% (v/v) chloroform and concentrating virus by precipitation with 7% (w/v) polyethylene glycol (PEG, mol. wt 6000) and 0·1 NaCl. The virus was then pelleted through a 30% (w/v) sucrose pad containing 7% PEG+0·1 M NaCl and finally sedimented through a sucrose density-gradient. These preparations had A_260/280 ratios of 1·26, contained end to end aggregates of WLV particles and formed a partly polydispersed peak in the analytical ultracentrifuge. WLV did not react with antisera to four potex-viruses, or to apple chlorotic leaf spot or apple stem grooving viruses.     

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.     

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.     

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.     

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/*.     

Characterisation of a new strain of tobacco necrosis virus isolated from nutrient feeding solution*

A virus, isolated from the nutrient feeding solution of a bell pepper culture in a glasshouse, was characterised as a new serotype of tobacco necrosis necrovirus (TNV) and named TNV-nft. The virus was monopartite and contained a single RNA species and a single coat protein with molecular weights of 1.45± 10⁶ and 2.9 ± 10⁴ daltons, respectively. The particle had an apparent sedimentation coefficient of 110 S and a density of 1.37 g/cm³ in cesium chloride. In decoration tests antiserum prepared against TNV-nft reacted most strongly with the TNV strains Kassanis A and B, but at lower dilutions it also reacted with all other TNV-serotypes available. In Ouchterlony tests spur formation occurred with TNV strains Kassanis A and B, indicating that the TNV-nft strain was serologically distinct. The isolate was not associated with a satellite virus. Cloned cDNA prepared against viral RNA reacted exclusively with the viral RNA band in Northern blots.     

Studies on the occurrence of tomato bushy stunt virus in English rivers

Tomato bushy stunt virus (TBSV) of unknown source was isolated from water of the River Thames, near Oxford. The isolate designated TBSV-T was mechanically transmissible to several tomato (Lycopersicon esculentum) cvs and to other species including Petunia hybrida, pepper (Capsicum annuum). eggplant (Solanum melongena), Nicotiana clevelandii, Chenopodium amaranticolor and C. quinoa in which it caused systemic symptoms. It caused no infection of globe artichoke (Cynara scolymus) or Pelargonium domesticum. The virus was not adsorbed to soil and could be isolated from leachate of soil in which systemically-infected tomato or C. quinoa plants were grown. Tomato plants became infected when grown in soil watered with virus suspensions. TBSV-T was infective after 10 min at 80°C but not at 90°C and when diluted to 10^-5 but not to 10^-6. Purified virus preparations contained C. 30 nm isometric particles. In gel-diffusion serological tests, TBSV-T reacted with homologous anti-serum and with antiserum to petunia asteroid mosaic virus but not to pelargonium leaf curl virus. Seed-borne infection (50–65%) of TBSV was demonstrated in plants grown from seed of symptomlessly-infected tomato fruit. TBSV was isolated from symptomlessly-infected tomato fruit imported from Morocco during October-April 1981. One of the isolates (TBSV-M) was indistinguishable from TBSV-T in host range, symptomatology and serological reactions. TBSV was also found in tomato plants growing extraneously in primary settlement beds at sewage works; such plants having been derived from undigested seeds in sewage. Because of its ‘alimentary-resistance’ in man, it is possible that one ecological route whereby TBSV enters rivers is by man's consumption of TBSV-infected tomatoes and eventual sewage dispersal into rivers.     

The occurrence, hosts and properties of lilac chlorotic leafspot virus, a newly recognised virus from Syringa vulgaris

Lilac chlorotic leafspot virus (LCLV), a hitherto undescribed virus, was isolated from three of 65 lilacs (Syringa vulgaris) with chlorotic leafspotting symptoms growing in S.E. England. The virus was transmitted readily by sap-inoculation to 21 of 52 species from eight of 20 families, but it was not seed-borne in four hosts or transmitted in the semi-persistent manner by any of four aphid species. The virus was moderately stable in vitro; sap from Chenopodium quinoa was infective after 10 min at 60 but not 65 ^oC, after 8–16 days at 20 ^oC or 25–30 wk at 2 ^oC, and after dilution to 10^-3 but not 10^-4. Up to 180 mg of purified virus per kg leaf tissue were obtained from C. quinoa by clarification of buffered leaf extracts with 8% (v/v) n-butanol, followed by one cycle of differential centrifugation and molecular permeation chromatography on controlled pore glass beads (700 Å, 120–200 mesh). LCLV has fragile flexuous filamentous particles which, when intact, mostly measured c. 12-5 times 1500–1600 nm; the helical substructure (pitch c. 3–7 nm) was clearly visible on some particles mounted in uranyl acetate. The particles sedimented as a single component (sedimentation coefficient 96 S; buoyant density 1–302 g cm^-3) and contained c. 5% nucleic acid and a single polypeptide of mol. wt 27 times 10³. Although these properties place LCLV in the closterovirus group, the virus showed no serological relationship to any of six closteroviruses (beet yellows, beet yellow stunt, carnation necrotic fleck, apple chlorotic leafspot, apple stem grooving and potato virus T) and differed from other recognised or possible members of this group in host range and/or symptoms induced in indicator species. The infrequent occurrence of LCLV in lilac in S.E. England indicates that the virus could probably be eradicated by selecting only virus-free plant material for propagation.     

Properties of a new strain of tobacco streak virus from Clematis vitalba (Ranunculaceae)

A distinct strain of tobacco streak virus (TS V/Cle), isolated in Yugoslavia from wild Clematis vitalba showing chlorotic spots or yellow netting of the leaves and from many symptomless shrubs, is described. TSV/Cle was seed transmitted in C. vitalba (70%), and in the experimental hosts Chenopodium quinoa (80%), Nicotiana benthamiana and N. megalosiphon. It was also detected in the pollen of infected C. quinoa. Purified virus preparations mostly contained quasi-spherical particles measuring 24–26 × 28, 26–28 × 28–30 and 28–31 × 32–36 mn, and sedimented in sucrose density gradient and analytical centrifugation as three components with sedimentation coefficients of 76S, 87S and 98S. The virus contained a single polypeptide species of mol. wt of c. 25 000. Unfractionated TSV/Cle preparations contained four RNA species with mol. wts, estimated by gel electrophoresis in agarose, of 1.1 × 10⁶, 0.9 × 10⁶, 0.7 × 10⁶ and 0.3 × 10⁶. In comparative experiments, TSV/Cle differed from four reference strains of TSV (TSV/B, TSV/HF, TSV/RN, and TSV/Ro) in host range and in symptoms induced in some common hosts. In agar gel double diffusion tests it was more closely related to TSV/B and TSV/M (SDI = 5) than to TSV/HF (SDI = 7), TSV/RN (SDI = 7) or TSV/Ro (SDI = 5–8). Immunoelectrophoresis experiments clearly distinguished TSV/Cle from the reference strains. TSV/Cle strain was detected in C. vitalba plants from distant and climatically different regions in Yugoslavia.     

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.     

Further properties of black raspberry latent virus, and evidence for its relationship to tobacco streak virus

Purified preparations of an isolate of black raspberry latent virus (BRLV) contained quasispherical particles with a mean diameter of 28·5 nm; these particles were resolved into three sedimenting components (s_{20, w}= 82S, 95S and 104S), but when centrifuged to equilibrium in caesium chloride solution they formed a single infective band (σ= 1·35 g/cm³). During electrophoresis in polyacrylamide gels, virus particles separated into three classes, and virus RNA was resolved into three major (mol. wt 1·35, 1·10 and 0·85 × 10⁶) and one minor (mol. wt 0·4 × 10⁶) component. The protein from virus particles had an estimated mol. wt of 28000. Isolates of BRLV were found to be serologically related but not identical to some strains of tobacco streak virus. No symptoms developed in black raspberry seedlings infected with BRLV by mechanical inoculation, nor in eight red raspberry cultivars infected by graft inoculation. However, graft inoculation of BRLV to Rubus henryi, R. phoenicolasius and Himalaya blackberry induced symptoms typical of necrotic shock disease.     

Pathology and properties of tulip virus X, a new potexvirus

Tulip virus X (TVX), a previously undescribed mechanically transmissible virus, causes chlorotic and necrotic lesions in leaves and streaks of intensified pigmentation in tepals of tulip plants. The virus infected 22 of 42 other plant species in 10 of 14 families, but most host species were infected only erratically. TVX is best propagated in Chenopodium quinoa and assayed in C. amaranticolor. Spindleshaped inclusions were observed in epidermal cells of C. amaranticolor leaves. Leaf extracts from C. quinoa contained flexuous filamentous particles measuring c. 495 ×13 nm. The extracts were infective after dilution to 10^-9, after heating for 10 min at 60 °C but not at 65 °C, and after storage at c. 20 °C for 30 days or at -20 °C for 6 months. TVX particles were purified (500 μg/g C. quinoa leaf) from tissue extracts in 0.067 M phosphate buffer containing 10 mM EDTA at pH 7, by twice precipitating the virus with 8% polyethylene glycol in 0.2 M NaCl followed by differential centrifugation. The virus particles have a sedimentation coefficient (s_{20, w}) of 102 S. They contain a protein of mol. wt c. 22 500 and a nucleic acid that, when glyoxalated, migrates in agarose gel like single-stranded RNA of mol. wt 2.05 × 10⁶. TVX particles tend to aggregate, and evidence was obtained that a 118 S component which was consistently observed in purified preparations and in infective sap is an end-to-end dimer. A distant serological relationship was found between particles of TVX and those of viola mottle and hydrangea ringspot viruses, but no serological relationship was detected to nine other potexviruses. TVX is considered to be a distinct and definitive member of the potexvirus group.     

The relationship between the Andean strain of potato virus S and pepino latent virus

The titres obtained in microprecipitin tests with purified preparations of pepino latent virus (PepLV) and the Andean strain of potato virus S (PVS^A) using PepLV antiserum and two antisera to the ordinary strain of PVS (PVS°) indicated a close serological relationship between PepLV and PVS^A. Using antiserum to PVS°, both viruses were detected by ELISA when infective Chenopodium quinoa sap was diluted to 10^-5but not to 10^-6. Particles of both viruses were decorated equally well by antibodies to PVS^o, PVS^Aand PepLV in all virus-antiserum combinations. When PepLV was inoculated to C. quinoa, C. amaranticolor and potato plants, the symptoms induced closely resembled those of PVS^Ain these hosts. It is concluded that PepLV is an isolate of PVS^Afrom pepino.     

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.     

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/*.     

Peanut green mosaic virus — a member of the potato virus Y group infecting groundnut (Arachis hypogaea) in India

A virus, now named peanut green mosaic virus (PGMV), was isolated from groundnut (Arachis hypogaea) in India and identified as a member of the potato virus Y group by electron microscopy, aphid transmission, and its chemical properties. It was sap transmissible to 16 species of the Leguminosae, Solanaceae, Chenopodiaceae, Aizoaceae and Pedaliaceae; Phaseolus vulgaris was a good local lesion host. PGMV remained infective in buffered groundnut leaf sap at dilutions of 10^-3 after 3 to 4 days at 25 °C, or heating for 10 min to 55 °C but not 60 °C. PGMV was transmitted in the non-persistent manner by Aphis gossypii and Myzus persicae but was not seed-borne. Purified virus preparations contained flexuous filamentous particles c. 750 nm long which sedimented as a single component with a sedimentation coefficient (S°_20w) of 171S, and contained a single polypeptide (mol. wt 34 500 daltons) and one nucleic acid species (mol. wt 3.25 × 10⁶ daltons). PGMV is serologically unrelated to peanut mottle virus (PMV) and other viruses infecting leguminous crops. Infected leaves contained cylindrical, cytoplasmic inclusions.     

Effect of Fungicides on the Viability of Phylophthora cactorum Propagules m the Soil

Vein-clearing followed by downward rolling and necrosis of leaves and severe stunting of groundnut (Arachis hypogaea) plants were caused by cowpea mild mottle virus (CMMV). The virus was readily transmitted by mechanical sap inoculations to groundnut and to 10 plant species belonging to Leguminosae, Chenopodiaceae and Solanaceae. Chenopodium quinoa and Beta vulgaris were good diagnostic hosts. Diseased sap remained infective at 10^–3 but not 10^–4, when stored 8 to 9 days at 25 °C; for 10min at 75 °C but not 80°C. In limited tests, virus was not seed-transmitted m groundnut or soybean. Virus was transmitted by Bemisia tabaci but not by Aphis craccivora or Myzus persicae. An antiserum for CMMV was produced and virus was serologically related to CMMV reported on cowpea and groundnut crinkle virus (GCV) from West Africa. Employing carbon diffraction grating replica as a standard the modal length of virus particles to be 610 nm. Infected cells contained large number of virus particles associated with endoplasmic reticulum.     

Host range, properties and purification of raspberry bushy dwarf virus

Raspberry bushy dwarf virus (RBDV) was found in all plants of Lloyd George raspberry with bushy dwarf disease and occurred occasionally in plants of some other cultivars. It was transmitted by inoculation of sap to fifty-five other species in twelve families of flowering plants and infected most of them symptomlessly. It caused systemic symptoms in some species of Amaranthaceae, Chenopodiaceae and Cucurbitaceae, and necrotic local lesions in some Leguminosae. It did not induce bushy dwarf disease when returned to Lloyd George raspberry. Chenopodium quinoa was used for propagating the virus and Vigna cylindrica for local lesion assay. In C. quinoa sap, RBDV lost infectivity when diluted 10^-4, heated for 10 min at 65 °C or stored for 4 days at 22 °C. Preparations made by twice precipitating the virus at pH 4·8 and resuspending it at pH 7·0, followed by ultracentrifugation and exclusion chromatography in columns of 2 % agarose beads, contained isometric particles about 33 nm in diameter, which sedimented as two components, with sedimentation coefficients of 111 and 116S. Only a few particles, all of them disrupted, were seen in preparations mounted in phosphotungstate, but the particles were well preserved in uranyl formate provided that they were first dispersed in a saxlt such as MgCl₂ instead of distilled water. Many particles were oval in outline as though distorted during drying. No serological relationship was detected between RBDV and twenty-four other isometric viruses nor between RBDV and the filamentous virus apple chlorotic leafspot, to which it was previously thought to be related. An isolate of loganberry degeneration virus was serologically indistinguishable from RBDV.     

Particle purification and properties of cassava Ivorian bacilliform virus

A virus found in cassava from the north-west of the Ivory Coast was transmitted by inoculation with sap extracts to herbaceous species in six plant families. Chenopodium quinoa was used as a propagation host and C. murale was used for local lesion assays. The virus particles are bacilliform, c. 18 nm in diameter, with predominant lengths of 42,49 and 76 nm and a structure apparently similar to that found in alfalfa mosaic virus. Purified preparations of virus particles had A₂₆₀/A₂₈₀ of 1.7 ±0.05, contained one protein of M_rc. 22 000, and yielded three species of RNA with M_r (× 10^-6) of c. 0.7, 0.8 and 1.2. Although the virus particles were poorly immunogenic, an antiserum was produced and the virus was detected by enzyme-linked immunosorbent assay (DAS-ELISA) in leaf extracts at concentrations down to c. 6 ng/ml. Four other field isolates were also detected, including a strain which caused only mild systemic symptoms in C. quinoa instead of necrosis. The naturally infected cassava source plants were also infected with African cassava mosaic virus (ACMV) but when the new virus was cultured in Nicotiana benthamiana, either separately or together with ACMV, its concentration was the same. The new virus did not react with antisera to several plant viruses with small bacilliform or quasi-bacilliform particles, and alfalfa mosaic virus reacted only weakly and inconsistently with antiserum to the cassava virus. The new virus, for which the name cassava Ivorian bacilliform virus is proposed, is tentatively classified as the second member of the alfalfa mosaic virus group.