The transmission of cocksfoot mottle and phleum mottle viruses by Oulema melanopa and O. lichenis.

Oulema melanopa and O. lichenis both transmit cocksfoot mottle and phleum mottle viruses with similar efficiencies. The viruses were serologically dissimilar and did not cross-protect against each other in barley. Both viruses were acquired after a few minutes feeding, but longer acquisition feeding periods increased both the efficiency of transmission and persistence in the vectors. Acquisition of either virus increased vector mortality whilst acquisition of both together did not. When both viruses were ingested, only one was transmitted. Each virus could be recovered from haemolymph and faeces, but regurgitation was not observed and could only be induced with the greatest difficulty. The results suggest possible circulative transmission of some beetle-borne viruses.     

Host-ranges of cocksfoot mottle and cynosurus mottle viruses

Cocksfoot mottle virus (CfMV) and cynosurus mottle virus (CyMV) had similar, but not identical, host-ranges within the Gramineae. Each infected fifteen out of forty-one species tested, considerably fewer than phleum mottle virus (PMV), but unlike PMV both infected Triticum aestivum (wheat), causing a lethal mottle. The species most useful for differentiating CfMV, CyMV and PMV are listed. Some species appear especially susceptible to virus infection.     

Epidemiology of cocksfoot mottle virus

Cocksfoot mottle virus (CFMV) was found to be widespread in cocksfoot seed crops in the East Midlands in the third harvest year and once present in the crop spread rapidly. Pot experiments with single cocksfoot plants showed that autumn infection with CFMV resulted in a significant reduction in the number of flowering tillers and weight and size of seed produced the following year. The spread of CFMV in experimental cocksfoot leys was studied; leys cut for conservation showed more infection than leys grazed by sheep, and increasing nitrogen application resulted in higher disease incidence. The disease increased rapidly in conserved leys in 1967 and its spread was associated with cutting by a forage harvester after the virus had been introduced into the crop in the autumn, possibly by the vector Lema melanopa. Animal grazing and cutting implements seemed to be more important agencies in disease spread than the vector.     

Some properties of cocksfoot mottle virus

Cocksfoot mottle virus (CFMV) was transmitted by manual inoculation of sap to cocksfoot (Dactylis glomerata L.), wheat, oats and barley, but not to nineteen other monocotyledonous and thirteen dicotyledonous plant species. The virus was also transmitted by cereal leaf beetles (Lema melanopa L.). Adult beetles infected plants more frequently than larvae, and remained infective for up to 2 weeks after they had fed on infected plants. Seed from infected cocksfoot and oat plants produced virus-free seedlings. The infectivity of sap was lost during 10 min. at 65° C., and 2 weeks at 20° C., but survived many months at — 15° C. Purified virus preparations, made by various methods, contained numerous nearly spherical particles, about 30 mμ in diameter. In electron micrographs some of the particles were penetrated by negative stain though most appeared intact. However, all the particles migrated together in a centrifugal (sedimentation coefficient = 118 S) or electrophoretic field. The ultraviolet absorption spectrum, and the phosphorus and nitrogen contents of the virus preparations, were typical of a nucleoprotein containing about 25 % nucleic acid. Serological tests failed to show any relationship between CFMV and eleven other viruses with particles of similar shape and size.     

The elimination of cocksfoot streak virus, cocksfoot mild mosaic virus and cocksfoot mottle virus from Dactylis glomerata by shoot tip and tiller bud culture

Shoot tips from 0.2 to 4.8 mm long and tiller buds 0.2 to 5.1 mm long were cultured from cocksfoot ( Dactylis glomerata ) plants known to be infected with combinations of cocksfoot streak, cocksfoot mild mosaic and cocksfoot mottle viruses. Regenerated plants were tested for the presence of viruses by electron microscopy, serology and the expression of symptoms. All viruses were eliminated by culturing shoot tips and tiller buds less than 1.2 mm long but the maximum explant size capable of regenerating healthy plants depended upon the infecting virus. Viruses were not detected in shoot tip and tiller bud samples of similar size to the explants that gave healthy plants. The method is of value in eliminating viruses from desirable stocks of D. glomerata that must be vegetatively propagated.     

Relationships, host-ranges and symptoms of some isolates of phleum mottle virus

Eleven isolates of phleum mottle virus, each from a different grass species, were grouped into five distinct strains: phleum mottle (PM), holcus transitory mottle (HTM), festuca mottle (FM), cocksfoot mild mosaic (CMM) and brome stem-leaf mottle (BSM). Their interrelationship was expressed three-dimensionally using serological differentiation indices. When 105 species of Gramineae were inoculated with each strain, eighty-one in forty genera became infected. These included festucoid, chloridoid, panicoid and andropogonoid species. Electron microscopy was more reliable than back-inoculation for detecting infection in plants that remained symptomless. Annual species were usually more severely affected than perennials. PM, HTM, FM, CMM and BSM did not appear to differ in host range, but sometimes differed in the symptoms induced in a given host. Each was symptomless in some hosts, caused a mild or severe mottle in others and mottling plus necrosis in a few. HTM was symptomless in most, and BSM in fewest species: BSM caused necrosis in most and HTM in fewest hosts.     

Association of cocksfoot mild mosaic and cocksfoot streak viruses in Dactylis glomerata and their simultaneous transmission by aphids

Macrosiphum (Sitobion) fragariae transmitted cocksfoot mild mosaic and cocksfoot streak viruses two to three times more frequently from plants in which they occurred together than from plants in which they occurred alone. In cells of doubly infected plants, particles of the two viruses commonly occurred in close association.     

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.     

Properties of cocksfoot streak and cocksfoot cryptic, two viruses infecting cocksfoot (Dactylis glomerate) 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 10-2 to 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 Mr of 31 000 and 3.2 times 106 respectively. 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 Mt of 1.14 times 106 and 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 cryptoviruses.     

Detection and characterization of defective interfering RNAs associated with the cocksfoot mottle sobemovirus

A new RNA of about 900 nt was found in the virions of cocksfoot mottle virus (CfMV) and in infected plants by RNA hybridization and RT-PCR. Structural features suggested that this RNA is a defective interfering RNA (diRNA). The CfMV diRNA was shown to consist of a 35-nt 5′-terminal genomic region, which formed a hairpin, and a 3′-terminal genomic region, which included the coat protein (CP) gene lacking the first 120 nt.In vitro translation of the diRNA started at the third Met codon to produce truncated CP. The CfMV diRNA was assumed totrans-activate synthesis of the CP subgenomic RNA (sgRNA).     

Infection of cereals and grasses by isolates of Polymyxa graminis (Plasmodiophorales)

The host range of isolates of Polymyxa was tested in mono-fungal sand cultures. Fourteen isolates of P. graminis, obtained from barley, wheat, oats or Poa annua and from several different countries, all infected barley and all but one infected wheat. Rye was also a good host, whereas oats (nine cultivars), Lolium multiflorum and Poa pratensis became only slightly infected. Wheat cultivars differed in susceptibility, with Galahad much more resistant than Avalon. Several common weed and pasture grasses were not infected by the two isolates tested. A range of wild Hordeum spp. were mostly susceptible to P. graminis and/or barley mild mosaic virus, which it transmits. An isolate of P. betae, used for comparison, caused slight infection on oats but not on other cereals. The variation within and between species of Polymyxa needs more detailed investigation.     

Infection and activation of monocytes by Marburg and Ebola viruses

In this study we investigated the effects of Marburg virus and Ebola virus (species Zaire and Reston) infections on freshly isolated suspended monocytes in comparison to adherent macrophages under culture conditions. Our data showed that monocytes are permissive for both filoviruses. As is the case in macrophages, infection resulted in the activation of monocytes which was largely independent of virus replication. The activation was triggered similarly by Marburg and Ebola viruses, species Zaire and Reston, as indicated by the release of the proinflammatory cytokines interleukin-1beta (IL-1beta), tumor necrosis factor alpha, and IL-6 as well as the chemokines IL-8 and gro-alpha. Our data suggest that infected monocytes may play an important role in the spread of filoviruses and in the pathogenesis of filoviral hemorrhagic disease.     

Determining the effectiveness of grazing and trampling by livestock in transmitting white clover mosaic and subterranean clover mottle viruses

Glasshouse and mini-sward experiments were done to determine the relative roles of grazing and trampling by livestock in transmitting white clover mosaic (WC1MV) and subterranean clover mottle (SCMoV) viruses between clover plants in pastures. Wounding due to grazing was simulated by repeatedly cutting plants with serrated scissors (glasshouse) or mowing (mini-swards), while wounding due to trampling was simulated by repeatedly bashing plants with the flat end of a wooden hammer handle (glasshouse) or rolling (mini-swards). In glasshouse experiments, cutting was more effective than bashing in transmitting WC1MV to white clover (Trifolium repens) plants but cutting and bashing transmitted it to subterranean clover (T. subterraneum) plants at similar rates. In an experiment with white clover mini-swards, mowing was more effective than rolling in transmitting WC1MV, and when both were combined, initially spread exceeded that obtained when the spread from mowing and rolling alone was added together. In glasshouse experiments, bashing was more effective than cutting in transmitting SCMoV to subterranean clover plants. In one experiment, neither mowing nor rolling spread SCMoV in mini-swards of subterranean clover. When transmission to subterranean clover cultivars which were ‘susceptible’ or ‘moderately susceptible’ to SCMoV was compared in glasshouse experiments, repeated bashing spread the virus more slowly to the ‘moderately susceptible’ cultivars. When mixed with ruminant saliva, infective sap containing WC1MV or SCMoV was still infective to clover plants after 4 wk storage at room temperature. When infective sap was allowed to dry naturally on a metal surface, SCMoV still infected clover plants when the dried sap was taken up in tap water after 4 but not 14 days, while WC1MV was infective after 24 h but not 4 days. These results suggest that grazing and mowing are more effective than trampling at transmitting WC1MV to white clover plants in pastures, while trampling is more effective at spreading SCMoV to subterranean clover. However, both transmitted WC1MV to subterranean clover at similar rates. Possible reasons for these differences are discussed in relation to differences in clover plant morphology and virus-specific factors.     

Influence of cocksfoot streak virus on the growth of cocksfoot swards

Small experimental swards of three types-healthy, 50% and 100% streak-virus-infected-were established from single tillers of randomly selected cocksfoot genotypes. In these swards 44% of infected plants died within 2 years compared with 21% of healthy plants. Mortality of infected plants was not increased by the presence of healthy plants in the same sward. In swards containing both healthy and infected plants, increased growth of healthy plants compensated for the low yield of infected ones only when the swards were frequently defoliated. Infrequent cutting apparently allowed infected plants to check the growth of healthy ones, and 50% infected swards yielded little more than 100% infected swards. This difference in response was attributed to differences in the growth habits of healthy and infected plants. Few initially healthy plants became infected during 2 years of the experiment.     

Infection of wild and cultivated Vaccinium spp. with blueberry leaf mottle nepovirus

A field survey was performed during the summers of 1991 and 1992 to determine whether blueberry leaf mottle virus (BLMV) infects wild blueberry bushes surrounding commercial blueberry fields and might act as a reservoir of the virus. Leaf samples from wild blueberries growing in wooded areas surrounding commercial plantings in Western central Lower Michigan were taken from bushes radiating out from the border of the commercial field into the wooded areas at 5, 50 and 100 m. BLMV was present in native bushes of Vaccinium corymbosum, V. myrtilloides and V. angustifolium, and two hybrids of V. corymbosum V. angustifolium. BLMV-infected pollen grains were detected in pollen traps located up to 2 miles from a source field.     

Relations of carrot red leaf and carrot mottle viruses with their aphid vector, Cavariella aegopodii

Studies with Scottish isolates of carrot red leaf (CRLV) and carrot mottle (CMotV) viruses confirmed the dependency of CMotV on CRLV for transmission by the aphid Cavariella aegopodii. CMotV was transmitted by aphids only when the two viruses were present in the same source plant, and its transmission was not assisted by anthriscus yellows virus, which acts as a helper for parsnip yellow fleck virus. Some test plants became infected with CRLV alone, and a few with CMotV alone. In winter, aphid transmission of CRLV and CMotV was greatly increased when the source plants received supplementary lighting whereas the CMotV infectivity of sap was not increased. C. aegopodii acquired CRLV and CMotV after minimum acquisition access times of 30 min and inoculated them after minimum inoculation access times of 2 min. There was a minimum latent period of 7–18 h. The viruses were retained by the aphid after moulting and are therefore circulative in the vector, but were not transmitted to progeny insects. Aphids allowed 24 h to acquire the viruses continued to transmit them for at least 12 days, but some aphids allowed 6 h or less for virus acquisition ceased to transmit after 3 or 4 days. CRLV is considered a tentative member of the luteovirus group.