Host range, purification and some properties of rubus Chinese seed-borne virus

A previously undescribed virus, for which the name rubus Chinese seed-borne virus (RCSV) is proposed, was isolated from a single, symptomless plant of an unidentified Rubus species grown from seed collected in the wild in the People's Republic of China, Experimentally RCSV infected 23 out of 39 spp. in six out of eight families. The virus was seed-transmitted in Chenopodium quinoa (100%) and Nicotiana bigelowii (27%). RCSV was not transmitted by the nematodes Xiphinema diversicaudatum or X. index. The particles of RCSV were isometric, c. 30 nm in diameter with some penetrated by negative stains. In thin sections particles were found in double walled tubular structures with an outer membrane enclosing one or more tubules. In crude extracts some particles were found within single-walled tubules. Two virus-associated bands were seen in sucrose density gradients of purified preparations. The upper band was not infective and consisted of penetrated particles apparently devoid of nucleic acid. The lower, infective band was resolved into two components, of density 1.452 and 1.461 g/ml, in caesium chloride isopycnic gradients. There were two polypeptides (mol. wts c. 47 000 and 25 200 daltons) and two nucleic acid species (one of mol. wt c. 1.4 × 10⁶ daltons; the second was poorly defined by the methods used but was of higher molecular weight). RCSV was distantly related serologically (6–7 SDI) to the type isolate of strawberry latent ringspot virus (SLRV) and also reacted with antisera to serologicaly distinct grape and olive isolates of SLRV. It did not react with antisera to 10 other isometric 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.     

A distinct strain of lucerne Australian latent virus in white clover in New Zealand

A strain of lucerne Australian latent virus (LALV) was widespread in Trifolium repens (white clover) in Northland, New Zealand, occurring in 20–100% of plants tested on 23 of 26 farms. Infected plants showed chlorotic line patterns or were symptomless. In agar gel double diffusion serological tests, white clover isolates of LALV differed from lucerne isolates by two or three twofold dilution steps of antisera. In mechanical inoculation tests, white clover isolates did not infect lucerne and lucerne isolates did not systemically infect white clover. White clover and lucerne isolates also differed in their reactions in some other hosts. Six LALV isolates from white clover were indistinguishable in double diffusion serological tests and by enzyme-linked immunosorbent assay, although the isolates could be differentiated by some reactions in test plants. Purified LALV particles of lucerne and white clover isolates contained two nucleoprotein components and two RNA species of molecular weight 2.82 × 10⁶ (8150 nucleotides) and 2.46 × 10⁶ (7100 nucleotides) (estimated under denaturing conditions).     

Arracacha virus B, a second isometric virus infecting arracacha (Arracacia xanthorrhiza; Umbelliferae) in the Peruvian Andes

Arracacha virus B (AVB), a previously undescribed virus, was found together with arracacha virus A or with a 750 nm flexous filamentous virus in arracacha (Arracacia Xanthorrhiza; Umbelliferae) growing in the Huanuco region of the Peruvian Andes. AVB was transmitted by inoculation of sap to 30 species from eight families out of 45 species from 10 families tested. It was transmitted through seed of Chenopodium quinoa but not by Myzus persicae. AVB was best propagated in C. Quinoa or Tetragonia expansa and assayed in C. quinoa, C. murale or C. amaranticolor. Sap from infeted <C. Quinoa was occasionally infective after dilution to 10^-4 but not 10^-5, after 10 min at 65 d? C but not 70 d? C, and after 12 but not 14 days at 20 d? C. In neutral phosphotungstate, AVB has isometric partilces c. 26 nm in diameter with a hexagonal profile. About 50- 150 A1 cm₂₆₀ units of purified virus were obtained from 1 kg infected C. quinoa leaf by extraction in 0.5 M phosphate buffer at pH 7.5, containing 0.05 M ethylene-daiminetetra-acetate (EDTA) and 0.2% mercaptoethanol, and clarificatin with chloroform, followed by two precipitations with polyethylene glycol and three cylces of differential centrifugation. Purified virus coefficent (Sd?_{20 w},) of 126 S and A₂₆₀/A₂₈₀ ratio of 1.80, bnut formed two isopycnic bands in CsC1 of buoyant density 1.481 and 1.492 g/cm³ with estimated nucleic acid contents of 40 and 41% respectively. AVB particles contained two proteins of mol.wt 26 000 (major component) and 20 000. AVB was not serologically related to any of 20 other morphologically similar viruses. Its properties suggest that it does not fall into any recognised group of viruses. the cryptogram of AVB is */*:*/40–41:S/S:S/*     

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.     

Purification and some properties of strawberry mottle virus

Strawberry mottle virus (SMoV) (three isolates: HJ, 3E and N) were transmitted to Chenopodium quinoa plants by sap inoculation. All three isolates induced very similar symptoms consisting of chlorotic spots and ringspots in inoculated leaves, and vein chlorosis, mottling, and dwarfing of the upper leaves. SMoV isolate HJ was purified from infected C. quinoa by homogenisation with 10 mM phosphate buffer, pH 7.2 containing 5% Triton X-100, followed by differential, sucrose density-gradient and CsCl equilibrium density-gradient centrifugations. A fraction with a buoyant density of 1.42g^- cm^-3 after CsCl density-gradient centrifugation was highly infectious to C. quinoa and contained many isometric virus-like particles c. 37 nm in diameter. These virus-like particles were never found in fractions from uninfected preparations. Electrophoretic analysis of a fraction containing virus-like particles revealed that these particles might have a single coat protein subunit with the apparent molecular mass of 26 K daltons and one nucleic acid of 6.6 kilobases. Double-stranded RNA analysis of isolate HJ-infected or uninfected C. quinoa and Fragaria vesca var. semperflorens seedling line ‘Alpine’ plants showed that both infected plants had two infection-specific dsRNA bands of mol. wts 4.5 and 3.9 × 10⁶.     

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 elderberry latent virus, one of four sap-transmissible viruses obtained from American elder (Sambucus canadensis L.)

Cherry leaf roll, tomato black ring and two previously unrecorded viruses were transmitted to Chenopodium quinoa from Sambucus canadensis plants imported from the U.S.A. Of the two newly recognized viruses one, code-named elderberry virus A, has filamentous particles about 650 times 15 nm; the other, named elderberry latent virus (ELV), was transmitted to several herbaceous species but remained symptomless in elder and most other hosts. In C. quinoa sap ELV lost infectivity after dilution to 10^--⁵ to 10^-- ⁶ , 10 min at 85–90°C, and 7 days at 18°C. Infectivity of nucleic acid extracts was abolished by ribonuclease in 0.2 m sodium chloride. ELV was purified from C. quinoa leaf extracts that were clarified with chloroform, by precipitation at pH 5 and differential centrifugation. Purified preparations contained numerous isometric particles c. 30 nm in diameter and a few particles c. 17 nm in diameter. In 0.06 M phosphate buffer ELV sedimented as a major 112 S (calculated for infinite dilution) component and a 48 S minor component. ELV showed no serological relationship to twenty-seven other isometric plant viruses. Its present cryptogram is R/I: *I*:S/S:S/*.     

Effects of virus genotype and temperature on seed transmission of nepoviruses

Transmission of different nepoviruses through chickweed (Stellaria media) seed was differently affected by ambient temperature during seed production. Raspberry ringspot and tomato black ring (Scottish isolate) viruses were similarly and frequently transmitted at 14 , 18 and 22 ^oC, whereas arabis mosaic virus was transmitted most frequently at 14 ^oC, and strawberry latent ringspot and tomato black ring (German isolate) viruses at 22 ^oC. When infected by seed-borne nepoviruses, seedlings of S. media and other species were symptomless at 15–25 ^oC, and the viruses were therefore detected by inoculating sap to Chenopodium quinoa indicator plants. However, typical symptoms of arabis mosaic and tomato black ring viruses were induced by growing Nicotiana tabacum, N. clevelandii and C. quinoa seedlings infected with seed-borne virus at 33–37 ^oC during the third and fourth weeks after sowing, preceded and followed by periods at 15–25 ^oC. The proportion of N. tabacum seedlings developing symptoms was the same as that of untreated seedlings yielding sap-transmissible virus. Seed transmissibility of pseudo-recombinant isolates of raspberry ringspot and tomato black ring viruses, containing RNA-i from one virus strain and RNA-2 from another strain, depended greatly on the transmissibility of the strain contributing RNA-i. The source of RNA-2 had an additional but smaller influence. The satellite RNA (RNA-3) of tomato black ring virus was seed-transmitted in S. media and its occurrence in cultures did not affect the frequency of transmission of the virus. Results of testing the infectivity of extracts of seed from infected mother plants suggested that failure of seed transmission reflected failure to become established in the seed, not subsequent inactivation. Whereas seed transmissibility of raspberry ringspot virus is primarily dependent on information carried in RNA-i, transmissibility by nematode vectors, another property of major ecological importance, is determined by RNA-2. In the field, selection pressures presumably can act independently on the two parts of the genome but evidence was also obtained of selection for mutual compatibility of RNA-i and RNA-2.     

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.     

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.     

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.     

Host range, purification and properties of potato virus T

Potato virus T (PVT) infected nine species of tuber-bearing Solanum, most of them symptomlessly, and as a rule was transmitted through the tubers to progeny plants: two genotypes of S. tuberosum ssp. andigena were not infected. The virus was also transmitted by inoculation with sap to 37 other species in eight plant families. Chenopodium amaranticolor is useful as an indicator host, C quinoa as a source of virus for purification, and Phaseolus vulgaris as a local-lesion assay host; the systemic symptoms in Datura stramonium, Nicotiana debneyi and in these three species are useful for diagnosis. Attempts to transmit PVT by aphids failed, but the virus was transmitted through seed to progeny seedlings of four solanaceous species, and from pollen to seed of S. demissum. PVT was purified by clarifying sap with n-butanol or bentonite, followed by precipitation with polyethylene glycol, differential centrifugation and sedimentation in a sucrose density gradient. Purified preparations had an E260/E280 ratio of 1.18 and contained a single infective component with a sedimentation coefficient of 99 S. This component consisted of flexuous filamentous particles of about 640 times 12 nm that showed a characteristic substructure when stained with uranyl acetate. The virus particles contained a single species of infective single-stranded RNA, of molecular weight 2–2 times 106 daltons, and a single species of polypeptide of molecular weight about 27 000 daltons. PVT is serologically related to apple stem grooving virus but not to four other common potato viruses with flexuous filamentous particles. Apple stem grooving virus and PVT cause similar symptoms in several hosts, but also differ somewhat in host range and symptomatology. Apple stem grooving virus did not infect potato, caused additional symptoms in C. quinoa also infected with PVT, and its particles did not show the structural features specific to PVT. The two viruses are considered to be distinct. The cryptogram of PVT is R/1:2–2/(5): E/E: S/C.     

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.     

Parsnip mosaic virus, a new member of the potato virus Y group

Parsnip mosaic virus (PMV) occurs commonly in parsnip in Britain and is transmitted after acquisition access periods of 2–5 min by the aphids Cavariella aegopodii, C. theobaldi and Myzus persicae. It was transmitted by manual inoculation of sap, infecting parsnip, chervil, coriander and carrot plants systemically, and causing local lesions without subsequent systemic infection in eight Chenopodium spp., Spinacia oleracea, Gomphrena globosa, and Toreniafournieri. It lost infectivity in Chenopodium quinoa sap after dilution to 10^-3–10^-4, heating for 10 min at 55–58 °C, or storage at room temperature for 7–10 days. Preparations partially purified by n-butanol or chloroform clarification, followed by acid precipitation and/or chromatography on columns of 2% agarose beads, contained filamentous particles, many of which were aggregated or fragmented. Preparations made with chloroform and without acid precipitation contained unaggregated particles of 755 nm normal length, with a sedimentation coefficient of 149 S. PMV did not react with antisera to any of fourteen other viruses with filamentous particles. The present cryptogram for PMV is */*: */*:E/E:S/Ap.     

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/*.     

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 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.     

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.     

Properties of cocksfoot streak and cocksfoot cryptic, two viruses infecting cocksfoot (Dactylis glomerata) in Scotland

A Scottish isolate of cocksfoot streak virus (CSV-S) was found to have flexuous filamentous particles which, in sap of infected cocksfoot plants, had a modal length of 712 nm. It was transmitted from infected to healthy cocksfoot plants in a non-persistent manner by Myzus persicae and by mechanical inoculation of infective sap extracts containing an anti-oxidant. Apart from cocksfoot, mechanical inoculation of infective sap succeeded in infecting only four of 22 plant species tested. The infectivity of sap extracts containing 0.2% thioglycerol was lost after heating for 10 min at 55^oC but not 50^oC, storage at room temperature for 48 but not 24 hours, and after diluting 10^-2to 10^-3. Highly purified preparations of CSV-S particles sedimented as a single component with a sedimentation coefficient of 139S and had a buoyant density in rubidium bromide of 1.31 g/cm³. Virus particles were composed of one protein and one ssRNA species with estimated M_r of 31 000 and 3.2 times 10⁶respectively. In ELISA, an antiserum prepared to CSV-S detected the virus in all aerial parts of infected cocksfoot plants and, when present in the ratio of 1 infected leaf: 1000 healthy leaves. Both CSV-S-infected and -uninfected cocksfoot also contained a previously undescribed virus with isometric particles c. 30 nm in diameter. This virus, named cocksfoot cryptic virus (CCV), was seed-borne in two cvs of cocksfoot tested and its particles contained two dsRNA species of estimated M_r of 1.14 times 10⁶and 1.27 times 10⁶. Despite the fact that particles of CSV-S were largely free from CCV particles following exclusion chromatography on agarose beads prior to immunisation, immunoelectron microscopy (IEM) showed that the antiserum prepared to CSV-S also contained some antibodies to CCV. Evidence from IEM suggested a possible distant serological relationship of CCV to ryegrass and beet (BCV 1 or BCV 2, or both) cryptoviruses, all members of sub-group A of crypto viruses.