Tolerance to ryegrass mosaic virus and its inheritance

Plant yield within and between four cultivars of perennial ryegrass infected with ryegrass mosaic virus (RMV) was closely related to symptom severity. Distribution of symptom severity was continuous in four perennial ryegrass and four Italian ryegrass cultivars infected with a severe RMV isolate, and also in another perennial ryegrass cultivar infected with a severe isolate of the virus, a mild one and one of intermediate severity. Symptom expression was polygenically inherited in both Italian (cv. RvP) and perennial (cv. S.24) ryegrass. Both additive and non-additive genetic variation was present in RvP, but the variation in S.24 was additive only. No significant maternal inheritance was present in either species.     

Transfer of polygenic resistance to ryegrass mosaic virus from perennial to Italian ryegrass by backcrossing

Polygenic resistance to ryegrass mosaic virus (RMV) was transferred from perennial to Italian ryegrass cv. Tribune by repeated cycles of backcrossing, polycrossing and selection. The resulting synthetic variety (Bb 2113) was highly resistant to infection by RMV whereas the Italian ryegrass cultivars Tribune, RvP, Bartolini, Augusta, Wilo and Dalita all were highly susceptible. Leaf width and length of Bb 2113 was similar to that of cv. Tribune and, like Tribune, Bb 2113 had awned florets. In a field trial, Bb 2113 had a similar persistency rating and number of inflorescences in the regrowth to cv. Tribune but produced 11% less dry matter. The results emphasise the need for a more efficient means of transferring small numbers of genes from one species of Lolium to another than backcrossing provides.     

The effect of ryegrass mosaic virus on the yield of perennial ryegrass swards

The influence of ryegrass mosaic virus (RMV) on yield of perennial ryegrass swards was examined under two harvesting frequencies and at two levels of nitrogen fertilizer, in a field trial planted with a clone of ryegrass, cv. S 321, using small plots established with proportions of healthy and infected plants. RMV reduced the height and the number of tillers of plants grown in pots and reduced the height also of plants in the field. The swards were planted in August and at a harvest 8 wk later RMV reduced the yield from 2–12 to 1–52 t dry matter (d.m.)/ha. In the first full harvest year, RMV caused only a small reduction in yield where no N fertilizer was applied and the application of a very small amount of fertilizer would have recouped the loss. On the other hand RMV severely restricted the increase in yield resulting from heavy dressings of the fertilizer. Thus where 400 kg N/ha was applied, RMV reduced the total annual yield from 18-6 to 13-8 t d.m./ha. The effect of the virus was especially great both in the Spring, the period of maximum herbage production, and when the level of fertilizer was high.     

Interaction between crown rust (Puccinia coronata) and two viruses of ryegrass

In Lemtal Italian and S.24 perennial ryegrass plants, two isolates of ryegrass mosaic virus (RMV) suppressed the amount of crown rust emerging on leaves inoculated with Puccinia coronata uredospores by up to 75% compared with the amount on virus-free plants. Severity of rust infection on barley yellow dwarf virus (BYDV) infected plants generally did not differ significantly from that on virus-free plants. When both RMV and BYDV were present, rust was restricted in Lemtal plants to a level intermediate between those occurring on plants infected by either virus alone, and in S.24 plants to a level below that obtained with either virus alone. The mean water soluble carbohydrate (WSC) content of Lemtal plants was reduced more than 20% by RMV, but was not significantly altered by BYDV. In S.24 plants the WSC content was increased by 10% by RMV and by 60% by BYDV. Rust reduced the WSC content of healthy and virus-infected plants, the reduction being positively correlated with the level of rust on the sampled leaves. In plants of Lemtal, but not of S.24, the degree of rust infection was positively correlated with the WSC content of leaves from rust-free control plants.     

Decreased severity of ryegrass mosaic virus after passage through resistant perennial ryegrass

After exposure to infection in the field, the proportion of plants showing distinct symptoms of ryegrass mosaic virus (RMV) was less in perennial than in Italian ryegrass. The perennial ryegrass cv. Mascot had a smaller proportion of plants with symptoms than cv. S.23. Far milder symptoms were induced in test plants by RMV from naturally infected perennial ryegrass plots than from Italian ryegrass plots. Within perennial ryegrass, RMV from cv. Mascot caused milder symptoms than that from cv. S.23. Severe RMV isolated from Italian ryegrass cv. Trident (RMVT) became milder after one passage through cv. Mascot, although not as mild as RMV obtained from field plots of cv. Mascot (RMVM). Families from two highly resistant perennial ryegrass clones and two randomly selected clones of cv. S.23 crossed in all possible combinations varied in symptom severity when inoculated with RMVT but not when inoculated with RMVM. Families inoculated with RMVT also yielded virus which varied in the severity of symptoms induced in test plants, families with severe symptoms yielding severer virus. Thus, much of the variation in the resistance of these clones could be attributed to infection with RMV of differing severity. Resistance was controlled by several genes which were mainly additive in their effect.     

Effect of cocksfoot mottle virus on resistant and susceptible cocksfoot cultivars grown alone or in combination with healthy or ryegrass mosaic virus-infected Italian ryegrass

Cocksfoot mottle virus (CfMV) was readily transmitted on cutting implements. However, after six harvests, only a quarter as many plants of the resistant cv. Cambria as of the susceptible cv. S.37 became infected. In addition to resistance to becoming infected, cv. Cambria also possessed some resistance once infected, and suffered less mortality than S.37. More CfMV-infected plants died when competing with healthy plants than when competing with other infected plants. Both CfMV and ryegrass mosaic virus (RMV) caused greater reductions in the yield of their respective hosts in mixed stands of cocksfoot and Italian ryegrass than in single species stands. CfMV reduced the proportion of cocksfoot in the mixture and RMV the proportion of ryegrass. However, CfMV-infected cv. Cambria competed better than CfMV-infected cv. S.37 with healthy or with RMV-infected ryegrass. It is concluded that grass cultivars selected for resistance to virus infection will slow down the rate of disease spread within the sward, and selecting for postinfection resistance will help maintain the aggressiveness and competitive ability of plants once they become infected.     

The occurrence and effects of red clover necrotic mosaic virus in red clover (Trffbliumpratense)

In 1976, red clover necrotic mosaic virus (RCNMV) was identified in red clover variety trials at the Scottish Colleges of Agriculture and at the trial centres of the National Institute of Agricultural Botany (NIAB) in Northumberland, Dyfed, Devon and Cambridge. In 1977, RCNMV was also found in two commercial crops of red clover in South Wales. The only previous finding of this virus in Britain was in 1971.
In red clover leaves RCNMV causes veinal chlorosis, often followed by severe necrosis and deformation; the plants become stunted. All cultivars tested were infected either in field or glasshouse experiments and three of the four most susceptible cultivars were tetraploids. Yield losses in cv. Hungaropoly averaged 57% over three cuts. RCNMV was transmitted manually but not through seed or by aphids {Acyrthosiphon pisum and Myzus persicae) or weevils (Apion spp. and Sitona lineatus). Seedlings became infected when grown in pots containing RCNMV-infected plants or soil from infected sites, and the roots of infected test seedlings contained an Olpidium sp. which may be the vector.
White clover mosaic virus (WCMV), also common in red clover at some sites, was less damaging than RCNMV and in a glasshouse experiment decreased yield by only 22%. An unidentified seed-borne virus with spherical particles c. 33 nm in diameter was the only virus detected in clover seedlings screened for RCNMV.     

Serological detection of ryegrass mosaic virus and ryegrass seed-borne virus

Ryegrass mosaic virus (RMV) was reliably detected in both perennial (S24) and Italian (S22) ryegrass, by enzyme-linked immunosorbent assay (ELISA) when plants had been infected for 8 wk. ELISA detected more infections in field-grown perennial ryegrass cv. Premo than either visual assessment or electron microscopy. However, with plants of Italian ryegrass cultivars only recently infected with RMV, positive reactions were more difficult to separate from the reactions of RMV-free plants, which varied considerably with cultivar, some giving high absorbance values. Immunosorbent electron microscopy showed that the RMV antiserum also contained antibodies to ryegrass seed-borne virus (RGSV), suggesting that these high values were caused by RGSV infection in the material tested.     

Effect of nitrogen on interactions between ryegrass mosaic virus (RMV) and Lolium species in mixed swards

The effect of rate of nitrogen applied to three Lolium perenne-based field swards on the development of RMV symptoms and on the impact of the virus on the growth and composition of the swards was examined over two growing seasons. Glasshouse experiments investigated the competition between Lolium spp. in relation to RMV infection and rate of nitrogen. In general, there was a higher incidence of RMV symptoms at the higher rates of nitrogen but the relationship was not consistent in all experiments. RMV infection only significantly reduced the cumulative total yield of the sward that contained the very susceptible Lolium multiflorum cv. Barverdi and was treated with the higher rate of nitrogen. However, yield losses were consistently greater at the higher rates of nitrogen. RMV altered the relative yields of sward components and the effect was greatest after an extended period of severe frost at the end of 1995. There is evidence that the frost severely restricted the yield of RMV-infected L. multiflorum tillers in 1996. Where RMV reduced the yield of susceptible species, there was significant compensatory growth by less susceptible companion species, i.e. L. perenne and Phleum pratense, but the compensation was generally incomplete, resulting in reduced yields for virus-infected swards. RMV had no significant effect on the overall quality of the three field swards but for cv. Barverdi grown in the glasshouse it significantly reduced the percentage organic matter and water soluble carbohydrate, and D value. The reductions were greater at the higher rates of nitrogen.     

The resistance of the perennial ryegrass cultivar Endura to ryegrass mosaic virus

The perennial ryegrass cv. Endura is particularly resistant to infection with two ryegrass mosaic virus isolates, RMV-Roth and RMV-Sax. A few plants appeared immune to RMV-Roth but were infected by RMV-Sax. Two plants that developed only mild symptoms after infection with RMV-Roth contained few virus particles, but passage through these resistant plants caused no detectable change in the virulence of RMV-Roth towards S 22 Italian ryegrass. In offsets derived from one of these resistant plants, RMV-Sax caused severe symptoms and attained a high virus concentration but it was unable to infect if the plant was already infected with RMV-Roth.     

Selection of perennial and Italian ryegrass plants resistant to ryegrass mosaic virus

Perennial ryegrass plants collected from fields and Italian ryegrass plants grown from seed were selected for resistance to infection by ryegrass mosaic virus (RMV) by repeated manual inoculation. Two of 108 perennial ryegrass plants and one of 150 Italian ryegrass plants were symptomless after seven and nine inoculations respectively. These three plants were propagated vegetatively. Plants of the two perennial ryegrass clones showed no symptoms after further manual inoculations with the initial isolate of RMV, or with an inoculum from infected plants collected from several fields, or after inoculation by viruliferous mites. Electron microscopy and back tests indicated that the plants were virus free. Some plants of the selected Italian ryegrass clone became infected after a further inoculation with mites or sap, but fewer than similarly inoculated unselected plants.     

Varietal differences in uptake of 32P labelled phosphate in clover plus ryegrass swards and monocultures

Lolium perenne cv. S.23, L. multiflorum cv. RvP, and Trifolium repens cvs S.184 and Olwen, were grown in mixed sward and monoculture during 1979. Whereas in mixtures grass roots absorbed more ³²P than clover roots, in monoculture clover generally absorbed more ³²P than grass roots. This showed that grass was a very strong competitor for uptake in mixed swards. Clover and grass monocultures absorbed most ³²P from 10 or 15 cm depth in the soil, while grass in mixtures absorbed most ³²P at 22.5 cm depth. Comparing varieties, in monocultures in June, Olwen was most active in absorbing ³²P at 15 cm. In August, Olwen absorbed more at 15 cm and 22.5 cm than S.184 or the grass varieties. Differences in absorption depth between varieties were less in mixtures than in monocultures. S.23 absorbed more ³²P in the late season than RvP, both in monoculture and in mixtures. Thus Olwen differed from S. 184 in depth and timing of uptake, whilst S.23 differed from RvP in time of uptake. Such varietal differences could be exploited by manipulation of depth and timing of fertiliser application to increase the precision of sward management.     

The elimination of ryegrass mosaic virus from Lolium multiflorum by meristem tip culture

Meristem tips were cultured from Lolium multiflorum plants infected with ryegrass mosaic virus (RMV). Meristem tips within the size range O'2-fi mm long were cultured on media with or without 2 ,4-D at 1 mg/1. The plants that regenerated in culture showed no symptoms over a 10 month period and no RMV particles were observed by electron microscopy. It was concluded that RMV had been eliminated. The method should prove useful in eliminating the virus from desirable genotypes used as parent material for seed production.     

Tolerance to ryegrass mosaic virus, its assessment and effect on yield

Symptom severity of eighteen populations of Italian ryegrass infected artificially in the glasshouse and naturally in the field with ryegrass mosaic virus (RMV) was strongly correlated. A smaller proportion of plants of the more tolerant populations showed symptoms in the field, but this was probably due to an association of tolerance with increasing incidence of symptomless infection rather than with resistance to infection. Under sward conditions, the yield of a sensitive genotype was reduced by 27% and that of two more tolerant ones by 15 and 13 %. The percentage dry matter yield loss of the most sensitive genotype was similar in all cuts, despite the appearance of extensive necrosis at the time of one cut. With the more tolerant genotype the yield loss varied from 7 to 25 % according to cut. Over the period of the experiment RMV infection did not increase plant mortality.     

A field study of the pattern of local spread of ryegrass mosaic virus in mown grassland

In a field experiment at Hurley, small areas of old ryegrass sward, known to be infected with ryegrass mosaic virus (RMV) were partially surrounded by newly-sown Italian ryegrass. The experiment was harvested by mowing on five occasions. Samples of Italian ryegrass taken when it was 11 months old showed a very marked increase in the levels of RMV close to the areas of old sward. Thus, 84% of the Italian ryegrass plants bordering the old sward to the north contained RMV, compared with 63% of plants only 2.7 m away. The corresponding values to the south were 68% and 39%. The much higher values encountered to the north are presumed to be caused by the movement of the mite vector of RMV (Abacarus hystrix) in relation to the prevailing wind. Two different mowers were used, a rotary mower and a reciprocating knife mower. We could demonstrate no increase in the level of RMV as a result of mowing from the infected sward and on to the new sowing and could demonstrate no difference in the infection levels as a result of using the different mowers. We conclude that in mown swards migration by the mite is responsible for all or nearly all of very local spread of RMV.     

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.     

Temporal dynamics of spread of four viruses within mixed species perennial pastures

Six mixed species, perennial pastures at two locations, A (four pastures) and B (two pastures), were sampled at regular intervals over periods of 10 to 22 months. The predominant plant species present were white clover (Trifolium repens), perennial ryegrass (Lolium perenne) and kikuyu grass (Pennisetum clandestinum). To determine the extent to which incidences of viruses transmitted in different ways change in the same pastures over time, samples of each plant species were taken at random on every visit and tested for virus presence. To help identify factors that might explain changes in virus incidence, records were also made of aphid presence, pasture management practices, grazing regimes, sward height and the relative proportions of different plant species within the swards. Samples of white clover were tested for presence of Alfalfa mosaic virus (AMV) and White clover mosaic virus (WCMV), ryegrass for Barley yellow dwarf virus (BYDV) and Ryegrass mosaic virus (RyMV), and kikuyu grass for BYDV and potyvirus infection. AMV and WCMV were detected in white clover, and BYDV and RyMV in ryegrass at both locations but often with wide incidence fluctuations for the individual viruses. AMV incidences in white clover ranged from 0% to 19% at A, and from 27% to 100% at B. WCMV incidences in white clover fluctuated between 9% and 46% at B, but never exceeded 1% at A. RyMV incidences in ryegrass fluctuated between 3% and 34% at A, and 19% and 73% at B. BYDV incidences in ryegrass ranged from 0% to 6% at A and 4% to 17% at B. In kikuyu grass, an unknown potyvirus and BYDV were detected twice (1% incidence) and once (4% incidence) respectively at B, and the unknown potyvirus only once (2% infection) at A. During repeated trapping of aphids in four pastures (two each at A and B), numbers of aphids caught varied widely between trapping dates and between individual pastures on the same trapping date. The species caught were Acyrthosiphon kondoi, A. pisum, Aphis craccivora, Rhopalosiphum padi and Therioaphis trifolii. Except in summer, when only T. trifolii was caught, A. craccivora was the most abundant. The trends in incidence for each virus within each pasture were compared with those from the other pastures sampled over identical periods to determine whether there was any commonality. For RyMV in ryegrass, overall incidence trends within the different pastures at both locations resembled each other during the same sampling periods. Within pastures at the same location there was commonality in incidence trends for RyMV and BYDV in ryegrass, but with AMV in white clover periods of similarity were rare even when pastures were adjacent and managed identically. Unravelling the individual effects of alterations in season, vector numbers, mowing, intermittent heavy grazing and pasture species composition on virus incidence proved difficult due to complex interactions between these and other factors influencing new spread or declining virus occurrence.     

Sitona weevils (Coleoptera: Curculionidae) as agents for rapid transfer of nitrogen from white clover (Trifolium repens L.) to perennial ryegrass (Lolium perenne L.)

The effects of root feeding by larvae of Sitona hispidulus (F.) (a common weevil pest of white clover) on the rate of transfer of nitrogen between plants of white clover (Trifolium repens L.) and perennial ryegrass (Lolium perenne L.) were investigated using a nutrient slant board technique. Clover plants, labelled with ¹⁵N were grown adjacent to ryegrass plants and were either infested with Sitona larvae or not infested. Ryegrass plants associated with the infested clover plants had a significantly higher dry matter yield and nitrogen content (75% and 74% respectively) than the uninvested plants, after 33 days exposure to insect herbivory. It was concluded that root feeding insects could play an important role in the cycling of nitrogen in grass/clover swards.     

Impact of the field slug Deroceras reticulatum on establishment of ryegrass and white clover in mixed swards

The relationship between numbers of Deroceras reticulatum and the establishment of ryegrass cv. Melle and white clover cv. Milkanova was studied on a mini-plot experiment. Assessments 12 and 33 days after sowing showed that Deroceras reticulatum had a proportionally greater impact on survival and growth of clover than on ryegrass. Slugs had no apparent effect on numbers or growth of three species of broad-leaved weeds (Senecio vulgaris, Cardamine hirsuta and Epilobium sp.) which occurred in the field experiment. In pot experiments where seedlings of ryegrass together with one of three clover cultivars, with or without chickweed (Stellaria media), were exposed to Deroceras reticulatum, seedlings of white clover cv. Aran were less affected by slug grazing than cvs S184 or Milkanova. The presence of different clover cultivars did not significantly influence the impact of slugs on growth of ryegrass or chickweed, nor did the presence or absence of chickweed influence the effect of slugs on ryegrass and clover seedlings. Slugs had less impact on the growth of chickweed than on ryegrass or clover. Thus slug grazing would make clover less able to compete with chickweed as well as the other weeds found in the mini-plot experiment.     

The effect of ryegrass mosaic virus on carbon assimilation and growth of ryegrasses

The growth of simulated swards of Italian and perennial ryegrass, artificially infected with ryegrass mosaic virus (RMV) or RMV-free, was investigated. During the build up of RMV infection, dry-matter yield was usually decreased in infected swards, and the effect was confirmed when single vegetative regrowths of swards were investigated in detail. The primary cause of lower yields in RMV-infected swards appears to be a decrease in net canopy photosynthesis (maximum decrease about 50%) and an associated increase in dark respiration (maximum increase about 50%). The decrease in net canopy photosynthesis is due primarily to a lowered rate of net photosynthesis of constituent leaves within the canopy. A secondary effect of RMV infection is to decrease tillering (maximum decrease about 30%) which results in a change in canopy structure and in particular a lower leaf area index. In the present experiments, lower light utilization by the swards is less important than the decrease in leaf photo-synthetic efficiency in lowering yield.