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.     

The effects of mixing Italian ryegrass (Lolium multiflorum) with perennial ryegrass (L. perenne) or red clover (Trifolium pratense) on the incidence of viruses

Mixing the ryegrass mosaic virus (RMV) resistant perennial ryegrass (Lolium perenne) cv. Endura with the susceptible Italian ryegrass (L. multiflorum) cv. RvP decreased infection of RvP wth RMV from 37% when grown alone to 22% when mixed. However, Endura yielded less than RvP and there was no yield benefit from mixing the two cultivars. Mixing red clover (Trifolium pratense) cv. Hungaropoly with RvP had no detectable effect on RMV incidence in RvP but did decrease the incidence of red clover necrotic mosaic virus in Hungaropoly from 9% to 1% and of white clover mosaic virus from 53-5% to 41%. The yield of the mixture was equal to that of RvP grown alone but given nitrogen fertiliser. The numbers of eriophyid mites, including Abacarus hystrix the vector of RMV, on ryegrass leaves were similar in pure and mixed swards. It is concluded that with herbage crops, the common practice of sowing mixtures of species may help control virus diseases.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

Effect of ryegrass mosaic virus on the quality of perennial ryegrass

In 1976 fields of perennial ryegrass located on two farms in the west of Scotland were found to be far more severely and extensively infected by ryegrass mosaic virus (RMV) than had been seen in earlier surveys of ryegrass fields in 1972 and 1973. The chemical analysis of samples of ryegrass from 10 of the fields examined in 1976 showed that RMV significantly reduced the organic matter content, organic matter digestibility, water soluble carbohydrate content and D value of the grass. Furthermore, the losses suffered by grass with the severe necrotic symptom of RMV were significantly greater than those incurred by the grass with the mild mottle only. A correlation analysis between water soluble carbohydrate level and D value showed that whilst the loss in water soluble carbohydrates in mottled grass accounted for much of the loss in digestibility this was not the case in the necrotic grass where loss of green tissue appeared to be a more important contributory factor.     

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.     

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.     

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.     

Hybrids between tetraploid Italian and perennial ryegrass

Summary Hybridization frequency was investigated between tetraploid perennial and Italian ryegrass (Lolium perenne X multiflorum) without emasculation by using genetic markers. The Italian phenotypes, fluorescentroots and awned florets, were dominant. About 82% of the plants in perennial X Italian and nearly 93 % of the plants in the reciprocal crosses were hybrids. The hybrids had a high multivalent frequency and involved homoeologous chromosome pairing. Aneuploids with 2n = 26, 27 and 29 chromosomes were present. The hybrids were highly fertile. The cytogenetic behaviour of these allopolyploids suggested that the genomes of the parental species have undergone little repatterning and have free genetic exchanges. The species maintained their self-incompatibility and cross-compatibility at the tetraploid level.     

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.     

Revisiting variation in leaf magnesium concentrations in forage grasses for improved animal health

Plant and Soil - An Italian ryegrass cultivar (Lolium multiflorum Lam. cv. Bb2067), selected and bred for increased leaf magnesium (Mg) concentration in the 1970s, reduced the incidence of...     

The transmission and effect on yield of ryegrass mosaic virus in a filtered air environment

The eriophyid mite vectors of ryegrass mosaic virus (RMV) were excluded from ryegrass by covering plots with a polyethylene film house ventilated by filtered air. Initially plots were manually inoculated to give different levels of RMV. During two seasons no mites were found within the house so that all transmission of virus was by sap. Rolling and frequent cutting of uninfected plots decreased yields, but surprisingly, increasing virus infection did not decrease yields significantly. Cutting transmitted virus within infected plots but not from infected to healthy plots, whereas combined rolling and cutting did transfer virus to healthy plots.     

Development of EST-derived CAPS and AFLP markers linked to a gene for resistance to ryegrass blast (Pyricularia sp.) in Italian ryegrass (Lolium multiflorum Lam.)

Ryegrass blast, also called gray leaf spot, is caused by the fungus Pyricularia sp. It is one of the most serious diseases of Italian ryegrass (Lolium multiflorum Lam.) in Japan. We analyzed segregation of resistance in an F₁ population from a cross between a resistant and a susceptible cultivar. The disease severity distribution in the F₁ population suggested that resistance was controlled by a major gene (LmPi1). Analysis of amplified fragment length polymorphisms with bulked segregant analysis identified several markers tightly linked to LmPi1. To identify other markers linked to LmPi1, we used expressed sequence tag-cleaved amplified polymorphic sequence (EST-CAPS) markers mapped in a reference population of Italian ryegrass. Of the 30 EST-CAPS markers screened, one marker, p56, flanking the LmPi1 locus was found. The restriction pattern of p56 amplification showed a unique fragment corresponding to the resistant allele at the LmPi1 locus. A linkage map constructed from the reference population showed that the LmPi1 locus was located in linkage group 5 of Italian ryegrass. Genotype results obtained from resistant and susceptible cultivars indicate that the p56 marker is useful for introduction of the LmPi1 gene into susceptible germplasm in order to develop ryegrass cultivars with enhanced resistance to ryegrass blast.     

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.