Groundnut eyespot virus, a new member of the potyvirus group

A virus causing ‘eyespot’ leaf symptoms in groundnut plants was transmitted by sap-inoculation and by Aphis craccivora in the non-persistent manner. It infected 16 of 72 species from five of 12 families and was easily propagated in Arachis hypogaea and Physalis floridana. The virus has particles c. 13 × 755 nm and is serologically closely related to soybean mosaic and pepper veinal mottle viruses, and more distantly to four other potyviruses. The virus differs in host range, in vitro properties and serological properties from previously described strains of soybean mosaic and pepper veinal mottle viruses. It seems to be a distinct member of the potyvirus group and we propose the name groundnut eyespot virus.     

Viruses detected in Ullucus tuberosus (Basellaceae) from Peru and Bolivia

Ullucus tuberosus (Basellaceae) plants from 12 locations in the Andean highlands of Peru and Bolivia contained complexes of either three or four viruses. Specimens from six sites in Peru contained a potexvirus, a tobamovirus, a potyvirus and a comovirus, but those from another location lacked the potexvirus. All samples from five sites in Bolivia lacked the tobamovirus. The potexvirus (PMV/U) is a strain of papaya mosaic virus differing slightly from the type strain (PMV/T) in inducing milder symptoms in some common hosts and failing to infect a few other species. It symptomlessly infected U. tuberosus, and infected 15 of 29 species from seven of nine other families. PMV/U showed a close serological relationship to PMV/T and to boussingaultia mosaic virus and a distant relationship to commelina virus X, but it is apparently unrelated to any of ten other potexviruses. The tobamovirus (TMV/U) induced symptomless or inconspicuous infection in U. tuberosus, and infected 21 of 30 species from six of eight other families. It showed a very distant serological relationship to some strains of ribgrass mosaic, tobacco mosaic and tomato mosaic viruses, but failed to react with antisera to cucumber green mottle mosaic, frangipani mosaic, odontoglossum ringspot and sunn-hemp mosaic viruses. The potyvirus, tentatively designated ullucus mosaic virus (UMV), alone in U. tuberosus induced leaf symptoms indistinguishable from the chlorotic mottling and distortion found in naturally infected plants. UMV infected 12 of 20 species from four other families, and was transmitted in the non-persistent manner by Myzus persicae. It showed a distant serological relationship to only two (bidens mottle and alstroemeria mosaic) of 25 members or possible members of the potyvirus group tested. Some hosts and properties of the comovirus are described in an accompanying paper. None of the four viruses infected potato (Solanum tuberosum) and, with the possible exception of UMV, they differed from viruses reported previously to infect three other vegetatively propagated Andean crops (Oxalis tuberosa, Arracacia xanthorrhiza and Tropaeolum tuberosum).     

Glycine mosaic virus: a comovirus from Australian native Glycine species

Two strains of a virus designated Glycine mosaic virus (GMV) were found in Glycine clandestina and G. tabacina, legumes indigenous to Australia and the western Pacific region. When transmitted by sap inoculation, GMV infected mostly leguminous species, and caused mosaic and mottling symptoms. The virus was not found naturally in soybean G. max, but it infected all of the 21 cultivars tested. GMV has isometric particles of c. 28 nm diameter, and produces three components with sedimentation coefficients of 60 S (top), 103 S (middle), and 130 S (bottom). Both middle and bottom components are required for infectivity. The virions contain two major proteins with molecular weights of c. 21 500 and 42 000. GMV produces large aggregates of particles in the cytoplasm of the mesophyll cells of pea Pisum sativum, and also induces amorphous membrane-bound bodies and cytoplasmic vesicles. The type strain (from New South Wales) reacts with antisera to Echtes Ackerbohnenmosaik, broad bean stain, and a Californian isolate of squash mosaic virus. The GW strain (from Queensland) reacts with all of the latter antisera, as well as with antisera to cowpea mosaic virus (Sb and Ark strains), bean pod mottle, and red clover mottle viruses, and is serologically related to, but not identical with, the type strain. These properties clearly establish GMV as a new member of the comovirus group.     

Preliminary studies on sap-transmissible viruses of red clover (Trifolium pratense L.) in England and Wales

Red clover plants, collected from nine widely separated permanent pastures in England and Wales, were tested for sap-transmissible viruses. Viruses were identified by the symptoms they caused in test plants, by electron microscopy, and by serological tests. Of the 265 plants tested 14% were infected. Only pea mosaic virus was common and widespread; it was found in 8% of the plants, and in seven of the fields. Other viruses isolated were arabis mosaic, bean yellow mosaic, red clover mottle, and red clover vein mosaic; only red clover mottle virus produced diagnostic symptoms in red clover. No viruses were detected in seedlings grown from seed from eighty-nine commercial seed crops. Attempts to transmit red clover mottle virus by the Collembolan Sminthurus viridis L., which is common on red clover, failed.     

Glitoria yellow vein virus, a tymovirus from Kenya

Clitoria yellow vein virus (CYW) was found in Clitoria ternatea and Abrus precatorius in coastal districts of Kenya, but was not detected in food legume crops. When transmitted by inoculation of sap, CYW infected many species in the Papilionaceae, commonly causing yellowing of secondary and smaller leaf veins. All the economically important food legumes grown in the area of occurrence were very susceptible, so that CYW is potentially very important. The virus also infected okra (Hibiscus esculentus) and species in the Solanaceae, but none of many species of Cucurbitaceae. CYW is serologically closely related to cocoa yellow mosaic and kennedya yellow mottle viruses, and more distantly to okra mosaic and desmodium yellow mottle viruses. Other properties of CYW^ typical of the tymoviruses include particle morphology (particle diameter c. 28 nm; two components) with sedimentation coefficients of 50S (top) and 109S (bottom); molecular weight of protein sub-units c. 20000; thermal inactivation point c. 72 ^oC; and longevity in vitro c. 3 wk.     

Virus diseases of garden lupin in Great Britain

Two viruses occur widely in lupins in Britain. Alfalfa mosaic virus (AMV), of which two strains were isolated, was found mainly in named Russell varieties. Lupin mottle virus (LMV), a previously undescribed strain of the bean yellow mosaic virus (BYMV) common pea mosaic virus (CPMV) complex, was found more commonly in seedling lupins. Cucumber mosaic virus (CMV) was isolated once. The AMV strains were differentiated by their reaction in Phaseolus vulgaris; they were serologically closely related. Both AMV and LMV were aphid transmitted but not transmitted in lupin seed. LMV was distantly serologically related to both BYMV and CPMV. It cross-protected against BYMV but not against CPMV and it differed from both these viruses in some host reactions. The CMV isolate from lupins was similar to type CMV. It was transmitted both mechanically and by aphid, easily from cucumber to cucumber, but with difficulty from cucumber to lupin.     

Specificity of the effect of sap-transmissible viruses in increasing the accumulation of luteoviruses in co-infected plants

The concentration of potato leafroll luteovirus (PLRV) (c. 1300 ng/g leaf) in singly infected Nicotiana clevelandii plants was increased up to 10-fold in plants co-infected with each of several potyviruses, or with narcissus mosaic potexvirus, carrot mottle virus or each of three tobravirus isolates. With the tobraviruses, PLRV concentration was increased equally by co-infection with either NM-type isolates (coat protein-free cultures containing RNA-1) or M-type isolates (particle-producing cultures containing RNA-1 and RNA-2). In contrast, the accumulation of PLRV was not substantially affected by co-infection with either of two nepoviruses, cucumber mosaic cucumovirus, broad bean mottle bromovirus, alfalfa mosaic virus, pea enation mosaic virus or parsnip yellow fleck virus. The specificity of these interactions between PLRV and sap-transmissible viruses was retained in tests made in Nicotiana benthamiana and when beet western yellows luteovirus was used instead of PLRV.     

Broad-bean stain and true broad-bean mosaic viruses

Autumn-sown crops of broad beans (Vicia faba L.) in England often contain plants with some leaves characteristically distorted and with a chlorotic mosaic. From some of these plants true broad-bean mosaic virus was isolated in 1959 and 1960 but not in 1965 and 1966. From other plants a similar but distinct virus, which caused staining of the seeds and we call broad-bean stain virus, was isolated in 1960, 1965 and 1966. The two viruses were readily distinguished in serological tests, and in some test plants. Both were seed-borne, and spread in crops, but were not transmitted by several animal species tested as vectors. Both viruses have isometric particles about 25 mμ in diameter. Some of these particles contain about 35% ribonucleic acid, some about 26% and some of those of broad-bean stain virus contain none; these three types of particles had sedimentation coefficients of about 120–130 S, 100 S and 60 S respectively. The ribonucleic acid of each virus had molar base content of G 23%, A 26%, C 18% and U 32%. These two viruses are members of the cowpea mosaic group of plant viruses; broad-bean strain virus was serologically related to cowpea mosaic, F I, red-clover mottle, and squash mosaic viruses. The particles of all these viruses and of true broad-bean mosaic virus were similar in appearance, sedimentation behaviour, and nucleic acid content and composition. The nucleic acid of red-clover mottle virus had a molar base content of G 20%, A 29%, C 20%, U 30%.     

THE PROPERTIES OF TOMATO ASPERMY VIRUS AND ITS RELATIONSHIP WITH CUCUMBER MOSAIC VIRUS*

Chrysanthemum plants infected with tomato aspermy virus (TAV) produce severely distorted and discoloured flowers but show only slight leaf mottle.
TAV infected twenty-five of forty-five species (belonging to seventeen genera) tested and was transmitted by the aphid species Aulacorthum solarti, Macrosiphoniella sanborni and Myzus persicae .
Sap from infected tobacco leaves lost infectivity when diluted more than 1 in 10,000, when heated for 10 min. at above 65°C. and when stored for more than 42 hr. at 16–18°C.
Partial protection was obtained between TAV and two strains of cucumber mosaic virus. Evidence was obtained that this was true protection between related viruses and serological tests confirmed the view that TAV is a strain of cucumber mosaic virus. Evidence was obtained that this was true protection between related viruses and serological tests confirmed the view that TAV is a strain of cucumber mosaic virus.     

Peanut stripe virus - a new seed-borne potyvirus from China infecting groundnut (Arachis hypogaea)1

A new virus, peanut stripe (PStV), isolated from groundnut (Arachis hypogaea) in the USA, induced characteristic striping, discontinuous vein banding along the lateral veins, and oakleaf mosaic in groundnut. The virus was also isolated from germplasm lines introduced from the People's Republic of China. PStV was transmitted by inoculation of sap to nine species of the Chenopodiaceae, Leguminosae, and Solanaceae; Chenopodium amaranticolor was a good local lesion host. PStV was also transmitted by Aphis craccivora in a non-persistent manner and through seed of groundnut up to 37%. The virus remained infective in buffered plant extracts after diluting to 10^-3, storage for 3 days at 20°C, and heating for 10 min at 60°C but not 65°C. Purified virus preparations contained flexuous filamentous particles c. 752 nm long, which contained a major polypeptide of 33 500 daltons and one nucleic acid species of 3·1 × 10⁶ daltons. In ELISA, PStV was serologically related to blackeye cowpea mosaic, soybean mosaic, clover yellow vein, and pepper veinal mottle viruses but not to peanut mottle, potato Y, tobacco etch, and peanut green mosaic viruses. On the basis of these properties PStV is identified as a new potyvirus in groundnut.     

Isolation and Partial Characterization of Peanut Top Paralysis Virus (PTPV), a Destructive Virus Causing a New Disease in Peanut (Arachis hypogaea L.)

A destructive virus, causing top paralysis to peanut, was discovered in the wild germplasm collection growing in the USDA-ARS greenhouses, Stillwater, Oklahoma, USA. The symptoms observed on the wild plant were restricted to a few leaves as green batches in a light green to yellow background with some leaflets having lost most of the basal part of the laminae leaving the top portion rolling upwards forming a cone. The virus was mechanically transmitted to cultivated peanut ( Arachis hypogaea L,.) where it caused more severe and destructive symptoms including stunting, severe malformation of leaves and partial or complete disappearance of leaflet laminae. This virus differed in symptomology, host range, and/or serological reactivity from allpeanut viruses reported in the literature, particularly those causing leaf malformation and stunting. The virus induced necrotic local lesions on Phaseolus vulgaris L. cv. "Topcrop" and chlorotic local lesions with necrotic centres bordered withvery bright intense red color on Chenopodium amaranticolor. In both passive indirect enzyme-linked immunosorbent assay (PAS-ELISA) and Ouchterlony double immunodiffusion test, the virus did not react with antisera against brome mosaic, bean yellow mosaic, peanut stripe, potato Y, tobacco mosaic, watermelon mosaic 1, watermelon mosaic 2, wheat soilborne mosaic, wheat streak mosaic, and zucchini yellow mosaic viruses.
However, in reciprocal cross reactions the virus seemed to share a common antigenic determinant with a peanut mottle virus isolate from Oklahoma (PMV-OK). The virus had flexuous filamentous particles with a length of 750–850 nm, falling within the range reported for the potyvirus group. The virus was successfully purified and the molecular weight of its protein subunit was found to be 30000 d. A polyclonal antiserum was raised in rabbits against the virus and used for reciprocal serological tests.     

Nucleotide sequence of Chinese rape mosaic virus (oilseed rape mosaic virus), a crucifer tobamovirus infectious on Arabidopsis thaliana

The complete nucleotide sequence of Chinese rape mosaic virus has been determined. The virus is a member of the tobamovirus genus of plant virus and is able to infect Arabidopsis thaliana (L.) Heynh systemically. The analysis of the sequence shows a gene array that seems to be characteristic of crucifer tobamoviruses and which is slightly different from the one most frequently found in tobamoviruses. Based on gene organization and on comparisons of sequence homologies between members of the tobamoviruses, a clustering of crucifer tobamoviruses is proposed that groups the presently known crucifer tobamovirus into two viruses with two strains each. A name change of Chinese rape mosaic virus to oilseed rape mosaic virus is proposed.Abbreviations 2-ME 2-mercaptoethanol - EDTA ethylenediaminetetraacetic acid - SDS sodium dodecyl sulfate - UTR untranslated region - MP movement protein - CP capsid protein - CRMV Chinese rape mosaic virus - TVCV turnip vein clearing virus - PaMMV paprika mild mottle virus - PMMV-I pepper mild mottle virus (Italian isolate) - PMMV-S pepper mild mottle virus (Spanish isolate) - ToMV tomato mosaic virus - TMV tobacco mosaic virus - TMGMV tobacco mild green mosaic virus - ORSV odontoglossum ringspot virus - SHMV sunn hemp mosaic virus - CGMMV cucumber green mottle mosaic virus - ORMV oilseed rape mosaic virus     

Investigations of carnation viruses

Carnation vein mottle virus (CarVMV) is rare in glasshouse carnations in Britain, although locally common in Dianthus barbatus in private gardens. In Sim carnations free from other viruses, CarVMV caused slight diffuse chlorotic mottling in the younger leaves, decreased flower yield by c. 22%, and caused flower breaking in cvs William Sim and Dusty. In non-Sim cultivars Pink Shibiuya, Orchid Beauty and Vesta, leaf symptoms and flower breaking were more pronounced. In mixed infections with carnation mottle virus, symptoms were much more severe. CarVMV was not eliminated from carnation or D. barbatus plants grown for 4 wk at 37^oC, and only rarely from cuttings then taken from them, but it was readily eliminated by meristem-tip culture. Myzus persicae adults or nymphs acquired and transmitted the virus within a total time of 4 min, and remained infective for 30–60 min if feeding, or for 75 min if starved. The carnation aphid, M. persicae f. dianthi, transmitted the virus much less efficiently. The virus was not transmitted by dodder (Cuscuta campestris), or through seed of D. barbatus or Chenopodium quinoa. The maximum infective dilution in sap of D. barbatus, carnation and C. quinoa ranged from 10^-2 to 10^-5. The virus withstood 10 min at 60 but not 65^oC, up to 9 days at c. 18^oC or 3–4 wk at c. 2^oC. CarVMV infected twenty-two of 107 plant species in six of thirty-seven families; suscepts were confined to the Chenopodiaceae, Caryophyllaceae and closely allied families. C. quinoa was the best local lesion assay host. Seedling clones of D. barbatus, selected as resistant to carnation mottle virus, proved the best indicator and propagation species. Up to 50 mg virus/kg tissue were obtained by butanol clarification followed by differential and density gradient centrifugation. The preparations contained a single sedimenting component, s_20w= 144S, and had flexuous filamentous particles, c. 790 times 12 run; the particles contained a single polypeptide, mol. wt 34800, and 5% of a single-stranded ribonucleic acid (RNA) with nucleotide base ratios of G21: A25: C25: U29. Serologically CarVMV was related distantly to turnip mosaic (cabbage black ring strain), pea mosaic, watermelon mosaic (Strain 2) and bean yellow mosaic viruses, more closely to pepper veinal mottle virus, but unrelated to twelve other potyviruses. CarVMV is not at present a danger to carnation crops in Britain, but the recent trend of sending carnation plants to overwinter outdoors in warmer countries involves potential risks of more rapid spread by effective vector races of M. persicae.     

Discrimination by gel-diffusion serology of digitaria striate, maize sterile stunt and other rhabdoviruses of Poaceae

Eight rhabdoviruses from grass and cereal hosts and their antisera were used to examine virus relationships by gel-diffusion serology. Nucleocapsid (Nc) preparations from digitaria striate virus (DSV) and maize sterile stunt virus (MSSV) both contained a major protein of c. 52 OOO daltons, and antisera prepared to these readily discriminated related planthopper-transmitted rhabdoviruses. MSSV showed a moderately close relationship to barley yellow striate mosaic virus (BYSMV) when an antiserum prepared to whole virus was used, but the Nc antiserum showed clearer discrimination. Worthern cereal mosaic virus and DSV showed a distant relationship to BYSMV and MSSV. There was no serological relationship between any of these viruses and cereal chlorotic mottle virus, cynodon chlorotic streak virus, festuca leaf streak virus or maize mosaic virus.     

Structural fingerprinting: subgrouping of comoviruses by structural studies of red clover mottle virus to 2.4-A resolution and comparisons with other comoviruses

Red clover mottle virus (RCMV) is a member of the comoviruses, a group of picornavirus-like plant viruses. The X-ray structure of RCMV strain S has been determined and refined to 2.4 A. The overall structure of RCMV is similar to that of two other comoviruses, Cowpea mosaic virus (CPMV) and Bean pod mottle virus (BPMV). The sequence of the coat proteins of RCMV strain O were modeled into the capsid structure of strain S without causing any distortion, confirming the close resemblance between the two strains. By comparing the RCMV structure with that of other comoviruses, a structural fingerprint at the N terminus of the small subunit was identified which allowed subgrouping of comoviruses into CPMV-like and BPMV-like viruses.     

A Tobamovirus Causing Heavy Losses in Protected Pepper Crops in Spain

During a four-year (1982–1985) survey of plant viruses infecting pepper cultivars grown under plastic in the Southeastern region of Spain, a tobamovirus was found to be the major disease agent of this crop. The virus produces slight or no symptoms on the leaves, but causes chlorotic mottling, malformation and reduction in size with occasional necrosis on the fruits and was able to infect all commercial pepper cultivars tested, including those resistant to other tobamoviruses, causing a catastrophic disease. The biological and serological characterization of the virus showed that it is very similar to pepper mild mottle virus (PMMV) (Wetter et al. 1984) and therefore we have termed it as Spanish strain of PMMV (PMMV-S). The need of grouping all the so-called “pepper strains” of tobacco mosaic virus (TMV) as a new distinct member of the tobamovirus group with the name of PMMV is emphasized.     

Taxonomic patterns in the host ranges of viruses among grasses, and suggestions on generic sampling for host-range studies

Analyses of published host-range data for certain viruses reveal correlations with taxonomic groupings of grasses. Barley yellow dwarf virus (BYDV), cocksfoot mottle and phleum mottle viruses are found to have infected greater proportions of the festucoid grasses than of the non-festucoids to which they were inoculated. By contrast, all strains of sugarcane mosaic virus (SCMV) and of the closely related maize dwarf mosaic virus (MDMV) infected more non-festucoids than festucoids. In addition, infected plants from grass groups containing higher concentrations of genera susceptible to BYDV, SCMV and MDMV usually show clear symptoms, whereas infected plants from less susceptible groups are frequently symptomless. Some viruses, such as barley stripe mosaic, brome mosaic, cocksfoot streak and ryegrass mosaic, show no apparent preferences for particular grass groups. Samples of grasses employed in host-range studies are usually strongly biased towards festucoids. It is suggested that viruses ought to be adequately tested against genera from all the major groups, and a classified list of grass genera suitable for host-range studies is provided.     

Serological comparison of an Australian isolate of capsicum mosaic virus with capsicum tobamovirus isolates from Europe and America

A New South Wales isolate (Ca) of capsicum mosaic virus was tested against antisera to it and capsicum tobamovirus isolates from the Netherlands (P8, P11), USA (SL), Argentina (FO) and Sicily (PM). The comparison demonstrated that the four viruses Ca, P8, PM and SL are closely related to each other, forming a series of decreasing relationship to Ca in the above order. FO was related to these but insufficiently to be considered part of the group, and P11 was only slightly related to the others. The literature on serology of tobamoviruses in Capsicum spp. was collated and it is suggested that isolates from Sicily (pepper mild mottle), Australia (capsicum mosaic), The Netherlands (P8, P14) and USA (SL) be considered as strains of a virus distinct from both tobacco mosaic and tomato mosaic viruses and that these isolates all be referred to in future as strains of pepper mild mottle virus.     

Identification of Peanut Green Mosaic Virus Strains in India*

During field surveys, three peanut green mosaic virus isolates differing in symptomatology on groundnut and a few other hosts were collected. Ultrathin sections of infected groundnut leaflets showed cytoplasmic inclusions with pin wheels and scrolls. In enzyme-linked immunosorbent assay they reacted strongly with antisera to peanut green mosaic and soybean mosaic virus antisera, and moderately with adzuki bean mosaic and peanut stripe virus antisera. All isolates also reacted positively with antisera to peanut eye spot, blackeye cowpea mosaic, pea seed-borne mosaic, potato virus Y and tobacco etch viruses, and did not react with antisera to peanut mottle, bean yellow mosaic, bean common mosaic, clover yellow vein and sugarcane mosaic viruses. SDS-PAGE analysis of purified virus preparations of the three isolates showed a single polypeptide with mol. wt. of 34,500 daltons. Based on these results, the three isolates are identified as biologically distinct strains of peanut green mosaic virus.     

Immunodiagnosis of plant viruses by a virobacterial agglutination test

A new virobacterial agglutination (VBA) test for the immunodiagnosis of plant viruses is described. The test is based on the agglutination of Staphylococcus aureus cells by virus particles after treatment of the cells with homologous antiserum. The agglutination occurs within 1–5 min. The sensitivity of the test is 0·1-0·4 μg virus/ml and is not affected by the shape of the virus particle. The use of affinity purified antibodies for sensitisation of S. aureus cells increases the sensitivity of the reaction 50-fold and enables the detection of tobacco mosaic and cucumber green mottle mosaic viruses at a concentration of 2 ng/ml. The VBA test allows the estimation of potato viruses X, S, M and Y in the eyes and sprouts of infected tubers and in the leaves of infected plants. The diagnosis of carnation mottle virus in carnation plants and of mushroom viruses in mushroom (Agaricus bisporus) fruit-bodies and mycelium are also described.