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.     

Melon rugose mosaic virus, the cause of a disease of watermelon and sweet melon

A mechanically transmissible virus with isometric particles c. 32 nm in diameter, was isolated from infected watermelons and sweet melons in the People's Democratic Republic of Yemen. Purified virus preparations contained two major sedimenting components with sedimentation coefficients of 61S and 117S. In isopycn ic centrifugation in CsCl the particles formed a single band of buoyant density 1.39 g cm^-3. Preparations of virus particles comprised of a single polypeptide of mol. wt c. 22 000 and ssRNA of mol. wt 2.1 × 10⁶. The virus was serologically related to three of six subgroups of tymoviruses tested. The name melon rugose mosaic virus is proposed for this newly described virus.     

Cowpea mild mottle, a newly recognized virus infecting cowpeas (Vigna unguiculata) in Ghana

Cowpea mild mottle virus (CMMV), a previously undescribed virus widespread in cowpeas (Vigna unguiculata) in the Eastern Region of Ghana, was seed-borne in V. unguiculata, Phaseolus vulgaris and Glycine max, but was not transmitted by twelve aphid species including Aphis craccivora, A. fabae, Acyrthosiphon pisum and Myzus persicae. CMMV was transmitted by inoculation of sap to eleven of seventeen members of the Papilionaceae causing very severe diseases in G. max and Arachis hypogaea, and to ten of fifty-one species within five of nineteen other families; it was best propagated in G. max and Nicotiana clevelandii, and assayed in Chenopodium quinoa. Sap from systemically infected G. max was infective after dilution to 10^-3 but not 10^-4, after 10 min at 65 °C but not at 70 °C, or after 4 days at 18 °C or 16 days at 2 °C. Lyophilized sap was infective after 3 years in vacuo. CMMV has straight to slightly flexuous, fragile filamentous particles, c. 13 × 650 nm which, in sap, are occasionally surrounded by a loose external spiral. About 5 mg of purified virus was obtained from 1 kg of leaf tissue of G. max or N. clevelandii by clarifying leaf extracts in 0.02 m borate buffer (pH 9.5) with chloroform, followed by two or three cycles of differential centrifugation, and density gradient centrifugation. Virus preparations had ultraviolet absorption spectra typical of a nucleoprotein containing c. 5 % nucleic acid, contained numerous particles without external spirals, which sedimented as a single component with a sedimentation coefficient (s°_{20, w}) of 165 × 4S, and contained a single polypeptide species with a molecular weight of 32000–33000. CMMV showed a distant serological relationship to carnation latent virus, but not to ten other morphologically similar viruses; it thus seems to be a distinct member of the carlavirus group, and has the cryptogram: */*:*/(5):E/E:S/*.     

The Natural Occurrence, Transmission, Properties and Possible Affinities of Cowpea Mild Mottle Virus

Cowpea mild mottle virus (CMMV), although thought to be of only local importance when first found in 1973 infecting cowpeas (Vigna unguiculata) in Ghana, has since been shown to have a very extensive geographical distribution and a wide natural host range. It occurs in Africa, Asia, South America and Oceania in tomato (Lycopersicon esculenturri) and a range of leguminous crops including cowpeas, groundnuts (Arachis hypogaea, soybean (Glycine max) and French beans (Phaseolns vulgaris). The virus has physico-chemical properties resembling those of aphid-borne carlaviruses; it has filamentous particles ca. 650 nm long which contain a single polypeptide of 31–33 KDa and single-stranded RNA of 2. 5 × 10⁶. Unlike carlaviruses, however, CMMV is transmitted in a non-persistent manner by whiteflies (Bemisia tabaci) and the particles occur in vivo in characteristic brush-like inclusions. It is also seed-borne in some, but not all, cultivars of cowpea and soybean, but seed-transmission is probably dependent upon the interaction of genotype, virus strain, time of infection and environmental factors. CMMV is serologically closely related to, and thus probably synonymous with, viruses previously designated groundnut crinkle, Psophocarpus necrotic mosaic, Voandzeia mosaic and tomato pale chlorosis. It is, however, serologically unrelated to 18 recognized carlaviruses; because it is also transmitted by whiteflies and induces the formation of unusual brush-like inclusions, it is probably best left unclassified or tentatively placed in a sub-group of the carlavirus group until the taxonomic significance of these features has been fully evaluated.     

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

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

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.     

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.     

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.     

Host range, purification and some properties of two carlaviruses from hop (Humulus lupulus): hop latent and American hop latent

Hop latent virus (HLV) occurs in virtually all commercial hop plants in England, without causing apparent symptoms. It was transmitted between hop plants in a non-persistent manner by the aphid Phorodon humuli, but was not seed-borne in hop. The virus infected six species in four families out of 40 in 13 families which were inoculated, but infection was systemic only in Dianthus deltoides and hop. Only Phaseolus vulgaris and Chenopodium murale developed symptoms. Purification of HLV from hop extracts was hampered by aggregation of virus particles but this was minimised either by resuspending pellets in phosphate-buffered saline containing Tween 20 or by avoiding ultra-centrifugation. Virus was purified from extracts treated with Triton X-100 by precipitation with polyethylene glycol (PEG) followed either by centrifugation through sucrose density gradients or by exclusion chromatography through columns of Sephadex G-25 and Sepharose 4B. Purified preparations contained filamentous particles c. 675 × 14 nm composed of c. 6% single stranded RNA of mol. wt c. 2.9 × 10⁶ and a single protein species of mol. wt c 33 000. Immunosorbent electron microscopy (IEM) decoration tests suggested that HLV was serologically related to carnation latent, Helenium virus S, lily symptomless and Nerine latent viruses. American hop latent virus (AHLV) was found in two introductions to England from Corvallis, USA in 1975 and 1976. It was transmitted between hop plants in the non-persistent manner by P. humuli. The virus infected 17 species in seven families out of 41 species in 13 families which were mechanically inoculated and was systemic in nine species. It did not cause symptoms in any of five English hop cultivars. C. quinoa was a convenient propagation host and countable local necrotic lesions and ringspots occurred in leaves of Datura stramonium. AHLV was purified by PEG precipitation and centrifugation in sucrose density gradients. Preparations contained filamentous particles c. 680 × 15 nm composed of c. 6% single-stranded RNA of mol. wt c. 3.0 × 10⁶ and a single protein species of mol. wt c. 33 000. In IEM decoration tests AHLV was serologically related to Nerine latent virus but did not react with antisera to 14 other carlaviruses.     

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/*.     

Studies on melon necrotic spot virus disease of cucumber and on the control of the fungus vector (Olpidium radicale)

Melon necrotic leaf spot virus (MNSV) caused a major outbreak of a leaf necrosis disease of hydroponically-grown cucumber plants at Humberside in 1983. The virus had c. 33 nm diam. particles which reacted serologically with MNSV antiserum of Dutch or American origin. Virus particles, which contained a single polypeptide (mol. wt 45 × 10³) and a presumed RNA species (mol. wt 1.5 × 10⁶), had a sedimentation coefficient (s_20.w) of 134 S and a buoyant density in caesium chloride of 1.35 g/cm³. The virus was mechanically transmissible, confined to species of Cucurbitaceae, transmitted by zoospores of Olpidium radicale and retained in the resting spores of the fungus. MNSV is thus both water-borne and soil-borne. O. radicale zoospores were killed in <5 min in suspensions containing 20 μg/ml of the surfactant Agral (alkyl phenol ethylene oxide). The disease did not reappear in 1984 when the cucumber crops were fed with nutrients containing 20μg/ml Agral.     

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/_*.     

Use of Clq enzyme-linked immunosorbent assay to detect plant viruses and their serologically different strains

Clq was prepared from bovine serum using a simple method involving repeated dialysis at low ionic strength in the presence of chelating agents (yield c. 3 mg/100 ml serum). It was viable when stored at -18°C for up to 2 months, and at 4°C for at least 10 wk in a storage buffer containing 10% sucrose. When used in Clq ELISA this test was as sensitive as the direct double antibody sandwich form of ELISA (direct ELISA) in detecting purified potato virus Y (PVY), with a limit of detection in both methods of c. 15 ng/ml, and slightly more sensitive in detecting purified cocksfoot mild mosaic virus (CMMV), with limits of detection of c. 15 ng/ml and c. 15–60 ng/ml respectively. Using an antiserum to one strain of each virus, Clq ELISA readily detected strains of PVY, CMMV, Andean potato latent virus (APLV) and barley yellow dwarf virus (BYDV). This included detection of APLV-Hu by APLV-Caj antibodies and CMMV(G) by PMV(S) antibodies, neither of which system gives detection in direct ELISA. Clq ELISA was therefore less specific than direct ELISA in detecting serologically different virus strains. Virus detection by Clq ELISA was inhibited when sap of tobacco, Nicotiana clevelandii and Setaria italica was used at low dilution. Inhibition by N. clevelandii sap was alleviated by using increased concentrations of virus specific antibody to detect APLV and plum pox virus. Also, extracting APLV infective N. clevelandii or CMMV infective S. italica saps in a minimum of buffer, centrifuging at low speed and diluting the supernatant before testing, partially overcame the inhibition. The inhibitory substance(s) in sap may act by preventing the binding of Clq to virus-antibody aggregates. Sap of wheat, oat and barley did not appear to have an inhibitory effect and BYDV was readily detected in naturally infected field grown plants of these species.     

Host range, purification and some properties of a new ilarvirus from Humulus japonicus

In 1986, during routine quarantine testing, an apparently undescribed virus was isolated from two of 21 plants of Humulus japonicus grown from seed imported into the UK from Beijing, Peoples Republic of China. The virus, for which we propose the name humulus japonicus virus (HJV), was transmitted mechanically to a wide range of herbaceous plants. No symptoms were seen in virus-infected H. japonicus plants. HJV infected, but did not become systemic, in the cultivated hop (Humulus lupulus) under our conditions although it has been detected serologically in both species of Humulus growing near Beijing. The virus was transmitted through seed of Chenopodium quinoa and was also associated externally with pollen of that species, but no pollen-transmission tests were conducted. HJV was easily purified. Virus particles comprised a single polypeptide (mol. wt c. 26 350) and four RNA molecules (mol. wts 1.31, 1.05, 0.75 and 0.39×10⁶). The three larger mol. wt RNAs were not infective in the absence of the smallest RNA. The particles were quasi-isometric and variable in size. Purified preparations of particles formed four bands in sucrose density gradients but (after fixing with formaldehyde) only a single band (with a density of 1.364 g/ml) in caesium chloride isopycnic gradients. These properties are similar to those of ilarviruses, and HJV was very distantly related serologically to prunus necrotic ringspot ilarvirus. We suggest, therefore, that HJV be regarded as a new member of the ilarvirus group. All known infected plants of H. japonicus at the site of introduction have been destroyed and the virus has probably been eliminated from there. Testing is continuing to confirm this.     

Some properties of an isolate of tobacco rattle virus from Northern Ireland

A virus obtained from soil in which potato plants had shown severe spraing symptoms induced symptoms on indicator plants typical of tobacco rattle virus (TRY). Purified virus preparations of a local-lesion isolate contained particles of two modal lengths, 192 nm and 94 nm containing RNA molecules of mol. wt 2.4 × 10⁶ and 1.23 × 10⁶. Virus coat protein had a mol. wt of c. 21 500. The virus was serologically distantly related to TRY (SYM) and pea early browning virus (PEBV) SP5, but did not react with TRY (CAM) or TRY (PRN) antisera. However, cDNA hybridisation indicated that the virus was more closely related to TRY (PRN) than either TRY (SYM) or PEBV (SP5). The virus isolate has been designated TRY (NI).     

Artichoke latent virus: characterisation, ultrastructure and geographical distribution

An isolate of artichoke latent virus (ALV-I) obtained from a symptomless artichoke plant in Southern Italy was characterised and compared with ALV isolates from other countries. ALV occurs in California and throughout the western part of the Mediterranean basin but of Mediterranean countries east of Italy, it was found only in Israel and Turkey. ALV-I was readily transmissible by inoculation of sap to a moderate range of hosts, was transmitted in a non-persistent manner by Aphis fabae, Brachicaudus cardui and Myzus persicae, but was not seed transmitted. The virus has flexuous rod-shaped particles measuring c. 12 nm × 746 nm with a sedimentation coefficient of 145 S and a buoyant density of 1·31 g/cm³. The particles contain single stranded RNA with a mol. wt of 3 × 10⁶ and protein composed of a single polypeptide species with a mol. wt of 33 000. Cylindrical cytoplasmic inclusions consisting of pinwheels and laminated aggregates were present in cells of naturally and artificially infected plants. ALV isolates from different geographical origin were indistinguishable from ALV-I biologically, morphologically, serologically and ultrastructurally. These properties place ALV in the Potyvirus group, but it was serologically unrelated to 12 other potyviruses 10 of which occur commonly in Italy.     

Garlic yellow streak virus, a potyvirus infecting garlic in New Zealand

In New Zealand, all garlic (Allium sativum) plants tested were infected by a virus with flexuous filamentous particles 700–800 nm long. This virus, called garlic yellow streak virus (GYSV), infected only two of 12 species tested and was transmitted to garlic by the aphid Myzus persicae in a non-persistent manner. In garlic sap, GYSV was infective at a dilution of 10^-4 but not 10^-3, after heating for 10 min at 60°C but not 65°C, and after 2 days but not 3 days at 25°C. The yield of virus, purified from naturally infected garlic, was 3–4 mg/kg fresh leaf. Preparations had A₂₆₀/A₂₈₀= 1.28 and A_man/A_min= 1.08. The virus particles had a sedimentation coefficient of 149S and a buoyant density in CsCl of 1.334 g/cm³. Mol. wt estimates for the virus nucleic acid were 2.95 × 10⁶ by electrophoresis in polyacrylamide gels and 3.46 × 10⁶ from the sedimentation coefficient (41.4S) in linear-log sucrose density gradients. Two polypeptides were detected in virus preparations; one (mol. wt 30 500) was possibly a breakdown product of the other (mol. wt 33 000). GYSV was serologically distantly related to onion yellow dwarf and leek yellow stripe viruses but was considered to be a separate virus because it differed from them in host range.     

Chenopodium necrosis: a distinctive strain of tobacco necrosis virus isolated from river water

A distinctive strain of tobacco necrosis virus (TNV) of unknown source was repeatedly isolated from water of the River Avon (Warwickshire) and two of its tributaries (R. Swift and R. Alne) using a technique developed for the concentration and isolation of water-borne bacteriophages. The same strain was isolated from the rivers Cam and Thames and from Lake Esthwaite (Cumbria) together with tomato bushy stunt virus. The TNV strain, designated Chenopodium necrosis (TNV-CN) was mechanically transmissible to C. amaranticolor and C. quinoa in both of which it caused local lesions and systemic infection. TNV-CN caused no infection when inoculated to tobacco (Nicotiana tabacum cv. White Burley) plants. The virus was not adsorbed to soil, could be isolated from leachate of soil in which systemically infected C. quinoa were grown and C. quinoa plants became infected when grown in soil watered with suspensions of the virus. The virus was not transmitted by Myzus persicae but was vectored by the zoospores of a lettuce isolate of Olpidium brassicae. TNV-CN was infective after 10 min at 85 °C., 3 wk at 20 °C and when diluted to 10^-8 but not 10^-9. Purified virus preparations contained c. 26 nm isometric virus particles. TNV-CN contained single-stranded RNA (mol. wt 1·5 × 10⁶) and one protein (mol. wt c. 26·4 × 10³) which co-electrophoresed in polyacrylamide gels with the protein of the D strain of TNV (TNV-D). Analytical centrifugation of TNV-CN indicated a single component virus with the same sedimentation coefficient (s₂₀, w= 115S) and buoyant density (1·385) in a CsCl gradient as those of TNV-D. TNV-CN and TNV-D were indistinguishable serologically.     

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.     

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.