Studies on beet western yellows virus in oilseed rape (Brassica napus ssp. oleifera) and sugar beet (Beta vulgaris)

Oilseed rape (Brassica napus L. ssp. oleifera) was studied as a potential overwintering host for the sugar-beet yellowing viruses, beet yellows virus (BYV) and beet mild yellowing virus (BMYV), and their principal vector, Myzus persicae. In spring 1982, plants infected with a virus which reacted positively in enzyme-linked immunosorbent assay (ELISA) with BMYV antibody globulin were found in oilseed-rape crops; none of the plants contained virus which reacted with BYV antibody globulin. This virus was subsequently identified as beet western yellows virus (BWYV). No leaf symptoms could be consistently associated with infection of oilseed rape, but the virus was reliably detected by sampling any leaf on an infected oilseed-rape plant. Some isolates from oilseed rape did infect sugar beet in glasshouse tests, but the proportions of inoculated plants which became infected were low. Apparently there is therefore little danger of much direct transmission of BWYV by M. persicae from oilseed rape to sugar beet in spring. BWYV was introduced to and spread within oilseed-rape crops in autumn by M. persicae, and autumn-sown oilseed rape proved to be a potentially important overwintering host for M. persicae. In a survey of 80 autumn-sown crops of oilseed rape in East Anglia, northern England and Scotland in spring 1983, 78 were shown to be extensively infected with BWYV. Experimental plots of oilseed rape with 100% BWYV-infection yielded approximately 13.4% less oil than plots with 18% virus infection, the result of a decrease in both seed yield and oil content.     

The effects of yellowing viruses on yield of sugar beet in field trials, 1985 and 1987

The separate effects of beet yellows virus (BYV) and beet mild yellowing virus (BMYV) on yield of sugar-beet cultivars inoculated at different growth stages were assessed in field trials in 1985 and 1987. Early or mid-season inoculation decreased sugar yield by up to 47% for BYV, and up to 29% for BMYV. Infections after the end of July had no significant effect on yield. Both viruses caused significant increases in the juice impurities sodium, potassium and amino-nitrogen after infecting plants early in the season. Yield losses associated with infection were determined by the causative virus, the time of infection, and susceptibility of the sugar-beet cultivars.     

Efficient dsRNA-mediated transgenic resistance to Beet necrotic yellow vein virus in sugar beets is not affected by other soilborne and aphid-transmitted viruses

Rhizomania caused by Beet necrotic yellow vein virus (BNYVV) is one of the most devastating sugar beet diseases. Sugar beet plants engineered to express a 0.4 kb inverted repeat construct based on the BNYVV replicase gene accumulated the transgene mRNA to similar levels in leaves and roots, whereas accumulation of the transgene-homologous siRNA was more pronounced in roots. The roots expressed high levels of resistance to BNYVV transmitted by the vector, Polymyxa betae. Resistance to BNYVV was not decreased following co-infection of the plants with Beet soil borne virus and Beet virus Q that share the same vector with BNYVV. Similarly, co-infection with the aphid-transmitted Beet mild yellowing virus, Beet yellows virus (BYV), or with all of the aforementioned viruses did not affect the resistance to BNYVV, while they accumulated in roots. These viruses are common in most of the sugar beet growing areas in Europe and world wide. However, there was a competitive interaction between BYV and BMYV in sugar beet leaves, as infection with BYV decreased the titres of BMYV. Other interactions between the viruses studied were not observed. The results suggest that the engineered resistance to BNYVV expressed in the sugar beets of this study is efficient in roots and not readily compromised following infection of the plants with heterologous viruses.     

The control of Alternaria species on leaves of sugar beet infected with yellowing viruses

Leaves of virus-free sugar-beet plants rarely became infected with Alternaria spp. in two field experiments at Cambridge in 1965. Infection with beet yellows virus (BYV) increased susceptibility of plants to Alternaria only slightly but infection with beet mild yellowing virus (BMYV) increased it greatly. There was a close association between the severity of Alternaria symptoms, shown by different breeding lines and varieties of sugar beet, and the losses of sugar yield which they sustained after infection with BYV and BMYV. Many lines and varieties were resistant to Alternaria even when infected with BMYV and their resistance seemed to be inherited as a dominant character. Individual plants of any one line or variety differed greatly in resistance to Alternaria, suggesting that selection should improve the present level of resistance. Spraying the foliage of Alternaria-susceptible varieties with fungicides had little effect on the severity of Alternaria symptoms or on sugar yield. This was probably because the wet summer of 1965 was ideal for the spread of Alternaria and because rain washed the fungicide deposits from the sprayed leaves.     

The influence of primary infection date and establishment of vector populations on the spread of yellowing viruses in sugar beet

In three field experiments in 1985 and 1986, we studied the effect of the date of primary infection on the spread of beet yellows closterovirus (BYV) and beet mild yellowing luteovirus (BMW) from artificially inoculated sugar beet plants. Laboratory-reared vector aphids, Myzus persicae, were placed on these sources of virus. There was no substantial natural immigration of vectors or viruses. In two experiments, one with BMYV in 1985 and the other in BYV in 1986, populations of vector aphids remained low and there was little virus spread, i.e. c. 50 infected plants from one primarily infected source. The cause of this small amount of spread was the low number of vector aphids. In the third experiment, with BYV in 1986, large populations of M. persicae developed and there was substantial virus spread: c. 2000 infected plants in the plots which were inoculated before canopy closure. In later-inoculated plots in the same experiment, there was much less spread: c. 100 infected plants per virus source plant. Differences between fields in predator impact are implicated as the most probable factor causing differences in vector establishment and virus spread between these three experiments. Virus spread decreased with later inoculation in all three experiments. A mathematical model of virus spread incorporating results from our work has been used to calculate how the initial proportion of infected plants in a crop affects the final virus incidence. This model takes into account the effect of predation on the development of the aphid populations. The processes underlying the spread and its timing are discussed.     

Effects of the pyrethroid deltamethrin on the acquisition and inoculation of viruses by Myzus persicae

The pyrethroid, deltamethrin, alone or as an emulsifiable formulation, hindered infection of healthy plants with the persistent beet mild yellowing virus (BMYV) and both acquisition of, and infection with, the non-persistent potato virus Y (PVY) and the semi-persistent sugar beet yellows virus (BYV) by Myzus persicae in glasshouse tests.
Another pyrethroid, RU-15525, also protected against infection with PVY. Even sub-lethal amounts of deltamethrin decreased virus transmission by rapidly incapacitating the aphids, the effect being least with aphids most resistant to organophosphorous insecticides and to certain pyrethroids including deltamethrin. Demeton-S-methyl hindered infection only with BMYV. This work shows that deltamethrin restricts transmission of persistent, semi-persistent and perhaps more importantly of non-persistent viruses in the glasshouse, and has potential for doing the same in the field.     

The influence of size and duration of aphid infestation on host plant quality, and its effect on sugar beet yellowing virus epidemiology

This paper studies the influence of previous infestation on the host quality of sugar beet (Beta vulgaris L.) for aphids and the influence of previous infestation on sugar beet yellowing virus epidemiology. Sugar beet previously infested with Myzus persicae (Sulzer) or Aphis fabae Scopoli (Homoptera: Aphididae) had an improved host quality for subsequently infesting aphids of the same species. There was a significant negative relationship between the number of M. persicae infesting a plant and the proportion of those that died with a dark deposit in their stomachs, and a significant positive relationship between the number that settled on a plant and the number that infested it previously. Nymphs feeding on previously infested plants grew more rapidly than those on control plants. The beneficial effect of previous infestation persisted for at least 2 weeks and prolongation of the infestation beyond 2 weeks was of no further benefit to the aphids. Field grown sugar beet, previously colonised by M. persicae, was more susceptible to natural infestation by M. persicae up to 5 days after exposure. Previously infested plants were also more susceptible to infection with beet mild yellowing virus (BMYV) but not beet yellows virus (BYV), suggesting that the aphids on the previously infested sugar beet settled more readily and were more inclined to feed (and thus transmit BMYV) than aphids on the previously uninfested plants. The consequences for the control of sugar beet yellowing virus vectors are discussed.     

Breeding for resistance to infection with yellowing viruses in sugar beet

Differences in resistance to infection with beet yellows virus (BYV) and beet mild yellowing virus (BMYV) have been observed in virus-tolerant sugar-beet breeding material. The results of glasshouse virus-susceptibility tests usually agreed well with those of field experiments in which plants were exposed to artificial, or natural, infestation with viruliferous aphids. Breeding lines and varieties, which showed resistance to BYV when Myzus persicae Sulz, was used as vector, generally showed a similar resistance to this virus when Aphis fabae Scop. was used. Varieties which were resistant to infection with one virus were not necessarily resistant to the other, although some showed resistance to both BYV and BMYV. Preliminary results suggest that resistance to infection may be controlled by recessive genes which occur widely in sugar-beet cultivars. The mechanism of this form of resistance is not understood, but it does not appear to be closely associated with resistance to the aphid vectors of the viruses. The observed differences in resistance to infection demonstrate the possibility of breeding a sugar-beet variety in which two forms of resistance to virus yellows, tolerance and resistance to infection, are combined.     

The effects of Beet mild yellowing virus and Beet chlorosis virus on the yield of UK field-grown sugar beet in 1997,1999 and 2000

The separate effects of the aphid‐transmitted poleroviruses; Beet mild yellowing virus (BMYV) and Beet chlorosis virus (BChV), on the yield of field‐grown sugar beet were studied following different inoculation dates from May to July in 1997,1999 and 2000. Each sugar beet plant within the appropriate plots was infected with virus using at least 10 wingless viruliferous Myzus persicae per plant. In all 3 years, overall yield losses caused by BMYV were negatively correlated with time of infection with early season (May) inoculations causing 18–27% losses in sugar yield but late season losses only 4–15%. BChV decreased the sugar yield and sugar content of beet following early season inoculations, although the effects on sugar yield were more variable (range 8–24%) and the virus appeared to be less damaging compared to BMYV. However, inoculations with BChV in July of each year caused greater root and sugar losses than inoculations with BMYV at that time. Both poleroviruses increased the sodium content of the roots early in the season, although neither virus had an effect on potassium levels at any stage.     

Some effects of spraying with thiabendazole on the susceptibility of sugar beet to yellowing viruses and on their vector, Myzus persicae (Sulz.)

In a field experiment fewer sugar-beet plants became infected with aphid-transmitted yellowing viruses in plots that had been sprayed with solutions of thiabendazole lactate than in water-sprayed plots, after exposure to natural infestation with aphids. Subsequent glasshouse tests showed that foliar sprays of o·o1 % thiabendazole lactate in water significantly reduced the proportion of inoculated sugar-beet plants which became infected with beet yellows virus (BYV) or beet mild yellowing virus (BMYV) after inoculation with viruliferous Myzus persicae (Sulz.). This effect on virus transmission was not apparently due to a direct insecticidal action of thiabendazole, because adult aphids usually survived equally well on sprayed and unsprayed plants. Treatment of test plants with thiabendazole did not affect the transmission of beet mosaic virus to them by M. persicae. The fecundity of M. persicae was greatly reduced by transferring them to plants which had been sprayed with thiabendazole or by spraying them with thiabendazole before transfer to unsprayed plants. The fertility of adult Aphis fabae Scop, was also reduced by spraying with thiabendazole. The mechanisms whereby thiabendazole affected fecundity of aphids and transmission of viruses are not understood.     

Effects of sucrose sprays and darkness on aphid colonization of sugar beet and on aphid transmission of yellowing viruses

In the glasshouse, adult, apterous Myzus persicae (Sulz.) and Aphis fabae Scop, settled better and deposited more larvae on sucrose-sprayed sugar-beet plants than on water-sprayed plants. M. persicae settled badly and deposited few larvae on plants that were kept in the dark before or after infestation. The effects of darkness on aphids were reduced by spraying the host plants with 10% solutions of sucrose before infestation. Viruliferous M. persicae transmitted beet yellows virus (BYV) and beet mild yellowing virus (BMYV) less efficiently to dark-treated plants than to those grown in normal daylight. Spraying sugar beet with sucrose before inoculation with viruliferous M. persicae increased the proportion of successful BYV transmissions but only when the plants were dark-treated. The effects of sucrose and darkness on settling and larviposition of aphids and on virus transmission may be related to changes in the concentration of carbohydrates, particularly sugars, in the leaves.     

Detection of the luteoviruses, beet mild yellowing virus and beet western yellows virus, in aphids caught in sugar-beet and oilseed rape crops, 1990–1993

The incidence of beet mild yellowing luteovirus (BMYV) and non-beet-infecting strains of beet western yellows luteovirus (BWYV) in individual winged aphids, caught in yellow water-traps, in sugar beet during the spring and early summer, and in oilseed rape plots in the autumn, was monitored using monoclonal antibodies in ELISA tests from 1990 to 1993. Between 0% and 8% of the Myzus persicae trapped in sugar beet each year carried BMYV, whereas 0% to 4% caught in oilseed rape in the autumn contained this virus. In 1990, 6.5% of Macrosiphum euphorbiae trapped in sugar beet contained BMYV, but in subsequent years less than 1% were carrying virus. Much higher proportions (26–67%) of the M. persicae tested from sugar beet contained BWYV, and similar proportions tested from oilseed rape (24–45%) also carried this virus in the autumn. In contrast only 3–19% of the M. euphorbiae caught in sugar beet contained BWYV, and none in oilseed rape. In 1991 and 1992 large numbers of Breuicoryne brassicae were caught in the plot of oilseed rape, of which over 50% contained BWYV; none were carrying BMYV. In transmission studies between 1990 and 1992, 1% and 27% of M. persicae transmitted BMYV and BWYV respectively to indicator plants; subsequent ELISA tests on the same aphids showed that 3% and 33% respectively contained the two viruses. One percent of M. euphorbiae transmitted BMYV, but none were found to contain BMYV using ELISA; 15% transmitted BWYV whilst only 5% were found to carry the virus. In 1992 and 1993 the incidence of BMYV-infection in the sugar-beet fields in which aphids had been trapped ranged from 1.2%, in a field which had received granular pesticide (aldicarb) at drilling plus three foliar aphicidal sprays, to 39.5% in a field which had received only one foliar spray. In 1992 in a sugar-beet crop which had received no aphicidal treatments, and where 2.8% of immigrant M. persicae and 2.5% of M. euphorbiae contained BMYV, 11.6% of plants developed BMYV infection. Lowest levels of infection were associated with the use of granular pesticides at drilling. In 1990, 80% of oilseed rape plants in a field plot were infested with a mean of seven wingless M. persicae per plant by mid-December; 37% of these plants were infected with BWYV. The studies show that M. persicae is the principal vector of BWYV, and large proportions of winged M. persicae carry the virus, in contrast to BMYV, which is consistent with the common occurrence of BWYV in brassica crops such as oilseed rape.     

Localization of the beet mild yellowing virus inSinapis alba L.

Virus particles of isometric shape with a diameter of 26 nm were found in the sieve tubes and accompanying phloem cells in ultrathin sections prepared from the nerves of white mustard (Sinapis alba L.) leaves and roots infected with the beet mild yellowing virus (BMYV). BMYV particles were much more frequent in the roots ofSinapis alba plants. Isometric particles were not found in the leaves and roots of healthy mustard plants.     

Detection of BMYV and BWYV isolates using monoclonal antibodies and radioactive RNA probes, and relationships among luteoviruses

In order to discriminate between sugar beet infecting beet mild yellowing virus (BMYV) and other isolates of beet western yellows virus (BWYV), monoclonal antibodies (MAbs) and radioactive riboprobes were used. With MAbs prepared against BMYV or potato leafroll virus (PLRV) no distinction could be established between BMYV and BWYV. Seven probes were synthesised from a lettuce infecting BWYV isolate; their localisation in the genome is known and they cover almost its entire length. Probes from the '3 part of the genome hybridised with all BMYV and BWYV isolates whereas those from the '5 part did not recognise BMYV isolates, showing that a divergent '5 region exists in the genomes of BMYV and BWYV. Probes also readily detected the virus in single aphids. The relevance of this finding for epidemiological studies is discussed.
MAbs and riboprobes were also tested against other luteoviruses (PLRV; barley yellow dwarf virus (BYDV) MAV, PAV and RPV strains). The serological relationship between BMYV and PLRV was confirmed and an epitope common to PLRV and BYDV-RPV was found. Using probes, PLRV and BYDV-RPV were found to share domains of homology with BWYV. BYDV-PAV showed weak homology with BWYV, while BYDV-MAV showed none.     

Detection of beet yellows virus in sugar beet leaves and roots by enzyme-linked immunosorbent assay (ELISA)

Using the enzyme-linked immunosorbent assay (ELISA) beet yellows virus (BYV) could be detected reliably in the leaves of sugar beet andTetragonia expansa Pall. and in the roots of sugar beet. Specifio γ-globulin of BYV antiserum was coupled to horse radish peroxidase by periodate oxidation. Optimum dilutions of antigen (extract from infected leaves) were1: 50 to 1: 200 for BYV detection in sugar beet andT. expansa leaves and1: 2 to 1: 5 for detection in sugar beet roots. Extracts from beet roots are not to be purified by ultracentrifugation, however, by the described method virus can be demonstrated only in 80–90% of naturally infected sugar beet roots. The method is specific, no increase of extinction values was found in healthy or beet western yellows virus infected plants. Presence of virus can be demonstrated by visual as well as photometric evaluation. Results confirmed the suitability of peroxidase application for detection of plant viruses by ELISA.     

Identification of a second distinct strain of beet mild yellowing luteovirus using monoclonal antibodies and transmission studies

The variation among isolates of beet mild yellowing luteovirus (BMYV), collected from commercial crops of sugar beet during 1990, 1992 and 1993, was studied using monoclonal antibodies and transmissions to indicator species. The common strain of BMYV, which occurs throughout the sugar-beet root growing area, reacts with monoclonal antibodies MAFF 24, BWYV-BC-510H and BYDV-PAV-IL-1, and infects Capsella bursa-pastoris. A second strain, which failed to react with monoclonal antibody BYDV-PAV-IL-1 and which did not infect C. bursa-pastoris, was identified in 11% of sampled infected plants. The implications of the properties of this strain for the epidemiology of BMYV are discussed.     

SUGAR BEET YELLOWS: A PRELIMINARY STUDY OF THE DISTRIBUTION AND INTERRELATIONSHIPS OF VIRUSES AND VIRUS STRAINS FOUND IN EAST ANGLIA, 1955-57

Aphid transmissions to sugar beet seedlings from yellowed sugar beet leaves collected from commercial fields in East Anglia during the summers of 1955, 1956 and 1957, showed the occurrence of two yellowing viruses. One was sugar beet yellows virus (SBYV) and produced vein-etch and yellowing symptoms on beet seedlings in the glasshouse; the other produced yellowing but no etch. These two viruses were apparently unrelated, so that sugar beet tolerant to one of them would not necessarily be tolerant to the other. The second virus, called 'sugar beet mild yellowing virus' (SBMYV), decreased the root yield of sugar beet plants grown under glass, by as much as did the milder SBYV strains, but less than did the severe SBYV strains. The proportions of the two viruses in the samples differed from year to year and from place to place.     

Effect of virus yellows on yield of some monogerm cultivars of sugar beet

The yield of plants of monogerm cultivars of sugar beet artificially infected with both beet yellows and beet mild yellowing viruses was, on average, depressed 2–7% for every 100 ‘infected plant weeks’, equivalent to c. £25/ha at 1976 prices. The cv. Vytomo, previously recommended to growers as being tolerant of infection by virus yellows, had a high sugar content and abundant foliage but in field trials its actual yield of sugar was no greater when infected, and lower when virus-free, than that of some other monogerm cultivars.     

A controlled environment study of the effect of leaf physiological age on the movement of apterous Myzus persicae on sugar-beet plants

Apterous Myzus persicae were found to move frequently from leaf to leaf on sugar-beet plants in controlled environment conditions. It is suggested that aphid movement can be related to changes in the rate and content of translocate flow during leaf development. These changes make newly-emerged leaves nutritionally favourable to colonising aphids and make expanding leaves slowly wane in favourability during the process of ‘sink to source’ conversion leading to aphid dispersal from the leaf. Variation in temperature was not found to alter the rate of aphid movement or the period (measured in thermal time) that aphids spent on particular leaves. However, the lower temperature was found to increase the rate of aphid development, aphid size and fecundity; these effects could also be due to nutritional factors. This dispersal behaviour may be a tactic to maximise food intake by a polyphagous aphid and increase the probability that nymphs are deposited on nutritionally-favourable leaves. The implications of the interleaf dispersal of apterous M. persicae for within- and between-plant spread of beet yellows virus (BYV) and beet mild yellowing virus (BMYV) are discussed.     

Purification and Some Properties of an Isolate of Beet Yellows Virus from Ukraine

A purification procedure, which yielded up to 15–30 mg of beet yellows virus (BYV) per 100 g of infected Tetragonia expansa leaves, has been developed. The procedure included sap clarification with Triton X-100, and two cycles of ultracentrifugation through sucrose cushion, which contained PEG-6000 and NaCl. A specific antiserum was prepared, and BYV infection was successfully detected by the double-antibody sandwich (DAS) ELISA in infected sugar beet leaves and roots diluted up to 1 × 10⁵ and 1 × 10⁴, respectively. The virus concentration was demonstrated to decrease in infected sugar beet roots slowly during 7 months, thus allowing successful diagnosis of planting material in winter storage. BYV presence in Myzus persicae aphids was also reliably detectable using the DAS-ELISA. In a competitive DAS-ELISA test, the Ukraine and the British BYV isolates were found serologically indistinguishable.