The association of Xiphinema diversicaudatum (Micoletsky) with strawberry latent ringspot and arabis mosaic viruses in a raspberry plantation

An outbreak of strawberry latent ringspot virus (SLRV) in a plantation of Mailing Jewel raspberry coincided with the greatest abundance of the nematode vector, Xiphinema diversicaudatum. Arabis mosaic virus (AMV) was not detected in the crop but was, together with SLRV, in many weed species present. AMV was transmitted through the seed of Poa annua, Capsella bursa-pastoris and Senecio vulgaris and SLRV through the seed of Mentha arvensis. X. diversicaudatum were more numerous within the rows than between them and vertical sampling showed that most occurred between 4 and 12 in depth in both locations. Monthly sampling showed that egg laying occurred from April to July; populations increased to a peak in late autumn but declined during the winter, resulting in about a twofold annual increase in numbers. Females, males and juveniles transmitted AMV and SLRV to cucumber seedlings, and in the absence of plants the nematode retained AMV for 112 days and SLRV for 84 days.     

The transmission of strawberry latent ringspot virus by Xiphinema diversicaudatutn (Nematoda)

Strawberry latent ringspot virus (SLRV) was found in diseased rose bushes growing in a glasshouse where the soil contained Xiphinema diversicaudatum (Micol). Adult female, adult male and juvenile X. diversicaudatum all transmitted the virus to cucumber seedlings, and nematodes kept without plants for 32 days after acquiring SLRV later transmitted it. When transferred to fresh plants every 2–4 days for 3 weeks, single nematodes transmitted up to three times; one nematode did not transmit until 19 days after the transfers began. One adult X. diversicaudatum, out of 141 tested, transmitted both SLRV and arabis mosaic virus. In all respects X. diversicaudatum behaved as a vector of SLRV as it does as a vector of arabis mosaic virus.     

Transmission of the hop strain of arabis mosaic virus by Xiphinema diversicaudatum*

Hop plants became infected with the hop strain of arabis mosaic virus (AMV(H)) when grown in hopfield and woodland soil in which infected plants had been growing. Infection occurred in soil infested with the dagger nematode Xiphinema diversicaudatum, but neither in uninfested soil nor in soil previously heated to kill nematodes. X. diversicaudatum transferred direct from hop soils transmitted AMV(H) to young herbaceous plants and to hop seedlings; some of the hop seedlings developed nettlehead disease. A larger proportion of plants was infected using X. diversicaudatum obtained from a woodland soil and then given access to the roots of hop or herbaceous plants infected with AMV(H). AMV(H) was transmitted by adults and by larvae, in which the virus persisted for at least 36 and 29 wk, respectively. Difficulties were encountered in detecting AMV(H) in infected hop plants, due partly to the delay in virus movement from roots to shoots. Infection of hop shoots was seldom detected until the year after the roots were infested and sometimes nettlehead symptoms did not appear until the third year. Isolates of arabis mosiac virus from strawberry did not infect hop. The results are discussed in relation to the etiology and control of nettlehead and related diseases of hop.     

Epidemiology of three viruses infecting the rose in the United Kingdom

Studies on the epidemiology of arabis mosaic (AMV), prunus necrotic ringspot (PNRSV) and strawberry latent ringspot (SLRV) viruses were made in relation to commercial production of standard and bush roses. AMV or SLRV apparently induced either symptomless infection in rose cultivars and Rosa spp., or leaf symptoms ranging from small chlorotic flecks to severe chlorotic mosaic and, occasionally, plant death. Infection of R. canina ‘inermis’ or R. corymbifera by an isolate of SLRV from R. corymbifera also severely depressed flowering and hip formation. In addition, whereas this isolate could be graft-transmitted to all Rosa spp. tested, isolates from R. rugosa and R. multiflora failed to be graft-transmitted to R. canina ‘inermis’ or R. corymbifera. No difference was detected in graft-transmission tests of Rosa spp. with several isolates of AMV or PNRSV. In plantings of up to 7 yr none of the viruses was transmitted through pollen to healthy roses grown in nematode-free soil, and only SLRV was readily seed-transmitted, particularly in R. rugosa. Nevertheless, in soil containing viruliferous nematodes, AMV and/or SLRV were transmitted to c. 80% of healthy plants. AMV and particularly SLRV were each damaging to field-grown maiden rose bushes cv. Fragrant Cloud. SLRV delayed the onset of flowering, and reduced the number and size of blooms. Diseased bushes were less vigorous, and half or none of the AMV- or SLRV- infected bushes respectively, conformed to the British Standards Institution specifications for maiden bush roses. These results are discussed in relation to the commercial production of field-grown roses in the UK.     

Arabis mosaic and Prunus necrotic ringspot viruses in hop (Humulus lupulus L.)

Purified virus preparations made from nettlehead-diseased hop plants, or from Chenopodium quinoa, to which the virus was transmitted by inoculation of sap, contained polyhedral virus particles of 30 mμ diameter which were identified serologically as arabis mosaic virus (AMV). There were serological differences between AMV isolates from hop and from strawberry, and also differences in host range and in symptoms caused in C. quinoa and C. amaranticolor. AMV was always associated with nettlehead disease. The nematode Xiphinema diversicaudatum occurred in small numbers in most hop gardens, but was numerous where nettlehead disease was spreading rapidly. Preparations from nettlehead-affected hops also contained a second virus, serologically related to Prunus necrotic ringspot virus (NRSV), in mild and virulent forms which infected the same range of test plants but showed some serological differences. Mild isolates did not protect C. quinoa plants against infection by virulent isolates. Hop seedlings inoculated with virulent isolates of NRSV developed symptoms indistinguishable from those of split leaf blotch disease. Latent infection with NRSV was prevalent in symptomless hop plants. Nettlehead disease is apparently associated with dual infection of AMV and virulent isolates of NRSV. An unnamed virus with rod-shaped particles 650 mμ long was common in hop and was transmitted by inoculation of sap to herbaceous plants. Cucumber mosaic virus was obtained from a single plant of Humulus scandens Merr.     

Observations on the occurrence, pathogenicity and control of Pratylenchus vulnus, P. thornei and Xiphinema diversicaudatum associated with glasshouse roses

About half the nurseries and glasshouses in the Lea Valley of Hertfordshire were found to be infested with either or both Pratylenchus vulnus and Xiphinema diversicaudatum. The latter nematode probably occurred naturally in the soil on which the glasshouses were built but P. vulnus is thought to have been imported with rose rootstocks and is not known to occur outdoors in Britain. Both nematode species can cause decline of roses and even small numbers seem harmful. Numbers of P. vulnus can increase greatly, especially on rootstocks of Rosa canina, although numbers were often smaller when roses were severely damaged than on healthier crops. Numbers of X. diversicaudatum increased more slowly and R. canina was a better host for it than R. chinensis. X. diversicaudatum seemed to respond less quickly than P. vulnus to reduced host-plant vigour. The two rootstocks exhibited different host-status for the two species of nematode and cuttings of various rose cultivars showed different host-status to P. thornei, another species of lesion nematode which is not known to be pathogenic to roses. Two applications of dibromochloropropane liquid at the rate of 70 l/ha in a large volume of water maintained nematode densities at an acceptably low level, and growers who adopted this treatment as a supplement to pre-planting steam sterilisation and/or DD (dichloro-propane: dichloropropene) prolonged the productive life of their crops for several years.     

The isolation and identification of rhubarb viruses occurring in Britain

Virus-like symptoms were common in British crops of rhubarb. All plants tested of the three main varieties, ‘Timperley Early’, ‘Prince Albert’ and ‘Victoria’, were virus-infected. Turnip mosaic virus and a severe isolate of arabis mosaic virus (AMV) were obtained from ‘Timperley Early’; and ‘Prince Albert’ contained turnip mosaic virus, cherry leaf roll virus (CLRV), a mild isolate of AMV and, infrequently, cucumber mosaic virus (CMV). The main commercial variety ‘Victoria’ contained turnip mosaic virus, CLRV, a mild isolate of AMV and, infrequently, strawberry latent ringspot virus (SLRV). All the viruses were identified serologically. The rhubarb isolates did not differ markedly from other isolates of these viruses in herbaceous host reactions, properties in vitro or particle size and shape. A rhubarb isolate of CLRV was distinguished serologically from a cherry isolate of the virus. Turnip mosaic virus, CLRV and SLRV, were transmitted with difficulty, but AMV isolates were readily transmitted by mechanical inoculation. Turnip mosaic virus was also transmitted to rhubarb by Myzus persicae and Aphis fabae. CLRV was transmitted in 6–8% of the seed of infected ‘Prince Albert’ and ‘Victoria’ rhubarb and in 72% of the seed of infected Chenopodium amaranticolor. Mild isolates of AMV were also transmitted in 10–24% of the seed of infected ‘Prince Albert’ and ‘Victoria’ plants.     

The detection by serological methods of viruses infecting the rose

Homogenates of herbaceous test plants infected with arabis mosaic virus (AMV), prunus necrotic ringspot virus (PNRSV), or strawberry latent ringspot virus (SLRV), and purified virus preparations were used to assess the sensitivities of four serological methods (the enzyme-linked immunosorbent assay - ELISA, immunodiffusion in gels, the latex flocculation assay, and serologically specific electron microscopy -SSEM) for the detection of these viruses. The latex test was up to 250 times more sensitive than gel immunodiffusion, but SSEM and ELISA were respectively up to 1000 and 200 times more sensitive than the latex test. Gel immunodiffusion and latex tests failed to detect any of the viruses in infected roses. Although ELISA reliably detected PNRSV and SLRV when leaves from infected roses were homogenised in a leaf: buffer ratio of 1 g:10 ml, AMV was occasionally undetected. However, when a modified ELISA technique, which reduced non-specific reactions, was used some PNRSV-infected roses were also not detected. Detection by SSEM was c. twice as sensitive as ELISA for all three viruses in rose extracts. The relative advantages of ELISA and SSEM for the detection of plant viruses are discussed.     

Host range, purification and some properties of rubus Chinese seed-borne virus

A previously undescribed virus, for which the name rubus Chinese seed-borne virus (RCSV) is proposed, was isolated from a single, symptomless plant of an unidentified Rubus species grown from seed collected in the wild in the People's Republic of China, Experimentally RCSV infected 23 out of 39 spp. in six out of eight families. The virus was seed-transmitted in Chenopodium quinoa (100%) and Nicotiana bigelowii (27%). RCSV was not transmitted by the nematodes Xiphinema diversicaudatum or X. index. The particles of RCSV were isometric, c. 30 nm in diameter with some penetrated by negative stains. In thin sections particles were found in double walled tubular structures with an outer membrane enclosing one or more tubules. In crude extracts some particles were found within single-walled tubules. Two virus-associated bands were seen in sucrose density gradients of purified preparations. The upper band was not infective and consisted of penetrated particles apparently devoid of nucleic acid. The lower, infective band was resolved into two components, of density 1.452 and 1.461 g/ml, in caesium chloride isopycnic gradients. There were two polypeptides (mol. wts c. 47 000 and 25 200 daltons) and two nucleic acid species (one of mol. wt c. 1.4 × 10⁶ daltons; the second was poorly defined by the methods used but was of higher molecular weight). RCSV was distantly related serologically (6–7 SDI) to the type isolate of strawberry latent ringspot virus (SLRV) and also reacted with antisera to serologicaly distinct grape and olive isolates of SLRV. It did not react with antisera to 10 other isometric viruses.     

Effect of annual and perennial cropping regimes in microplots on the population density and vertical distribution of Xiphinema diversicaudatum and on soil porosity

Long-term trends in populations of Xiphinema diversicaudatum were measured in two microplot experiments in which annual and perennial crops of seven plant families were grown in monoculture. Numbers of X. diversicaudatum increased under perennial ryegrass, raspberry and strawberry but decreased under hop, spring barley, potato, sugar beet, cabbage, winter wheat and winter beans; these effects probably reflected differences in the inherent host status of the crops. There were fewer X. diversicaudatum at 40–60 cm below annual crops than below perennial crops, and there were differences between perennial crops in nematode vertical distribution. Soil pore space did not limit nematode population density; total pore space was uniformly large under all crops and there was no detectable instability to water slaking. Under ryegrass a greater proportion of soil volume was occupied by pores too small to accommodate X. diversicaudatum, but since ryegrass supported the second largest nematode population, it seems unlikely that this was seriously limiting.     

Some effects of oxamyl on the virus-vector nematodes Longidorus elongatus and Xiphinema diversicaudatutn

Oxamyl applied to field soil prevented Longidorus elongatus from acquiring and transmitting tomato black ring virus for at least 6 wk, although numbers of nematodes were not greatly decreased compared with the untreated control. Glasshouse and laboratory tests examined the effects of oxamyl on viruliferous L. elongatus and Xiphinema diversicaudatum. In these tests oxamyl (1.0 ppm) in the soil water largely prevented L. elongatus transmitting virus to bait plants over a period of 1 month. X. diversicaudatum was equally affected by smaller concentrations, 0.1 ppm being sufficient to inhibit virus transmission in one test. Inhibition of virus transmission was associated with a decrease in the number of root tip galls produced by nematode feeding, especially that of X. diversicaudatum. Few nematodes were seriously affected by oxamyl, except at the greatest concentration tested (100 ppm), when numbers of L. elongatus, but not of X. diversicaudatum, were decreased. In vitro-treated viruliferous nematodes protracted their stylets, but their subsequent ability to transmit virus was unimpaired.     

A damaging outbreak of arabis mosaic nepovirus in blackcurrant, the occurrence of other nepoviruses in Ribes species, and the demonstration that alfalfa mosaic virus is the cause of interveinal white mosaic in blackcurrant

In a crop of blackcurrant (Ribes nigrum), cv. Baldwin in Eire, chlorotic mottling and ringspot symptoms in leaves on plants and severe crop loss was associated with infection with arabis mosaic nepovirus (ArMV) and the presence in the soil of its nematode vector, Xiphinema diversicaudatum. This is only the second report of ArMV damaging a crop of blackcurrant. Tomato black ring (TBRV) and raspberry ringspot nepoviruses were detected in single plants of redcurrant (R. rubrum) in England and flowering currant (R. sanguineum) in Scotland respectively; each of these infected plants showed foliar chlorotic line-pattern symptoms. This is the first record of TBRV in redcurrant. A single blackcurrant plant in New Zealand showing symptoms typical of those described for interveinal white mosaic disease, contained alfalfa mosaic virus (AMV). When AMV particles were purified and concentrated from herbaceous test plants and mechanically inoculated to young blackcurrant plants, several became infected with AMV and most infected plants developed systemic symptoms typical of the original disease. This provides the strongest evidence to date that AMV is the causal agent of interveinal white mosaic disease.     

A comparison of the effectiveness of the four British virus vector species of Longidorus and Xiphinema

The frequency with which the four virus-vector species of longidoroid nematodes occurring in Britain transmitted their associated plant viruses were compared in a series of experiments using a standard procedure. In these tests Xiphinema diversicaudatum proved an effective vector of British isolates of arabis mosaic virus and strawberry latent ringspot virus and Longidorus attenuatus of an isolate of tomato black ring virus from England. In comparison, isolates of raspberry ringspot virus and tomato blackring virus from Scotland and of raspberry ringspot virus from England were transmitted much less readily by their respective vectors, L. elongatus and L. macrosoma. These differences in ability to transmit virus were not related to differences in feeding access on the virus source- or bait-plants, in the extent to which virus was retained within the nematode feeding apparatus or in the frequency with which virus was recovered from Longidorus in concurrent slash tests. Three Scottish isolates of raspberry ringspot and tomato black ring viruses were transmitted equally infrequently by two populations of L. elongatus and the frequency with which virus was transmitted was not greatly increased when the species of source or bait plants was changed.     

Infectibility of some new raspberry cultivars with arabismosaic and raspberry ringspot viruses and further evidence for variation in British isolates of these two nepoviruses

In field trials at sites of an outbreak of arabis mosaic nepovirus (AMV) in England and of raspberry ringspot nepovirus (RRV) in Scotland, the results of exposure of some new raspberry cultivars to natural infection with these viruses showed discrepancies from those obtained in graft inoculation tests using AMV-Lib and RRV-S, the Scottish type isolates. In particular, cv. Glen Prosen, which is immune to AMV-Lib and RRV-S, was infected with AMV and RRV in the field trials. Studies on these and other field isolates of AMV and RRV showed that they differed from the type isolates in Rubus host range and in symptomatology in herbaceous hosts. However, whereas four isolates of RRV found infecting Rubus were distinguishable by spur formation in gel double-diffusion serological tests, six AMV isolates were indistinguishable by this method. Immunoelectrophoresis of virus particles did not distinguish the six AMV isolates, but isolates RRV-MX and RRV-T were distinguishable from RRV-S and the English type isolate, RRV-E. Like the two RRV type isolates, RRV-MX contained a single electrophoretic component, but it migrated must faster whereas RRV-T contained two components, one with a migration rate similar to that of RRV-MX and the other similar to that of the type isolates. Polyacrylamide gel electrophoresis of protein preparations from highly purified virus particles of RRV isolates E, S and MX detected a single polypeptide of estimated mol. wt 54 × 10³, 54 × 10³ and 50 × 10³ respectively but that of isolate T contained two polypeptides of estimated mol. wt 54 × 10³ and 50 × 10³. These data suggest that RRV-T is a mixture of two isolates. In laboratory tests the nematode Xiphinema diversicaudatum transmitted four isolates of AMV efficiently whereas two populations of the nematode Longidorus elongatus were less efficient vectors of four RRV isolates. Neither vector species transmitted virus to any of nine raspberry cultivars. The results are discussed in relation to the control of nepoviruses in raspberry and to the biology of these viruses.     

Sites of virus retention in the alimentary tract of the nematode vectors, Xiphinema diversicaudatum (Micol.) and X. index (Thorne and Allen)

Electron microscopy of thin sections of Xiphinema diversicaudatum and X. index fed on plants infected respectively with arabis mosaic and grapevine fanleaf viruses showed that the viruses are retained as a monolayer of particles adsorbed on to the cuticle lining the lumina of the odontophore (stylet extension), anterior oesophagus and oesophageal bulb. During the moult of the nematode the cuticular lining is shed and together with the detached virus particles is ingested into the intestine through the oesophago-intestinal valve; this supports the limited experimental evidence that viruses transmitted by X. diversicaudatum and X. index are not retained through the moult.     

The long-term effects of four monocultural regimes on two field populations of the nematodes Xiphinema diversicaudatum and Longidorus spp.

Population density changes of Xiphinema diversicaudatum and several Longidorus spp. were observed under four cultural regimes in long-term field experiments. Numbers of X. diversicaudatum increased under strawberry and, to a lesser extent, ryegrass, but remained more or less unchanged in hop and fallow plots. A mixture of four Longidorus spp. increased in numbers under grass but stayed static, or declined slowly under the other cultural regimes. All changes in population density were slow to occur, taking 3 or 4 yr for differences to become statistically significant. Fluctuations in larval numbers were mainly responsible for overall population changes; numbers of adults showed little difference over 5 yr.     

An association of carbohydrates with particles of arabis mosaic virus retained within Xiphinema diversicaudatum

Sections through the odontophore of Xiphinema diversicaudatum showed two types of staining for carbohydrates using the periodic acid - thiosemicarbazide -silver proteinate (PA-TSC-SP) reaction. The first consisted of thin, localised, intensely-stained patches on the lining of the food canal of all the specimens examined. The second type, found only in nematodes exposed to AMV-infected plants, revealed cloud-like areas of carbohydrate - containing material associated with the stained patches on the lining of the food canal. By staining alternate sections with uranyl acetate/lead citrate, these carbohydrate clouds were shown to contain virus particles. Although the cloud material could have originated from either the plant or the nematode, sections through a pellet of partially purified virus particles prepared from plants did not stain for carbohydrate. The possible role of carbohydrate in virus retention and transmission is discussed.     

Tests for transmission of cherry leaf roll virus using Longidorus, Paralongidorus and Xiphinema nematodes

The ability of 10 nematode species to transmit three strains of cherry leaf roll virus (CLRV) was tested by three methods: (1) virus-infected source plants and virus-free bait plants were grown concurrently in nematode-infested soil, (2) as for (1) but virus source plants were removed before bait plants were planted, and (3) nematodes were extracted from soil after access to virus source plants, and were added to pots containing bait plants. The occurrence of galls on roots showed that nematodes fed both on source and on bait plants in all experiments and, in some experiments, CLRV was detected by direct assays (slash tests) of Longidorus elongatus, L. leptocephalus and Paralongidorus maximus. Although the nematodes readily transmitted control viruses, for which they are known to be vectors, CLRV was detected by root assays in only a few bait plants exposed to L. elongatus, L. macrosoma, Xiphinema diversicaudatum or L. leptocephalus + X. vuittenezi in tests by method 1. The recovery of CLRV in these tests is interpreted as being due to contamination. These results add to the increasing circumstantial evidence against the involvement of nematodes in the transmission of CLRV. Other possible mechanisms of spread are discussed.     

Do plant parasitic nematodes have differential effects on the productivity of a fast- and a slow-growing grass species?

We have examined the interaction between plant parasitic nematodes and plant species from different stages of grassland succession. In these grasslands, fertiliser application was stopped in order to restore the former nutrient-poor ecosystems. This management resulted in a reversed succession of high- to low-productivity. Nematodes isolated from a high-productive early-successional field and a low-productive late-successional field were inoculated to sterilised soil planted with seedlings of either Lolium perenne (a fast-growing early-successional species) or Festuca rubra (a slow-growing late-successional species). The experiment was performed at low and high supply rates of nutrients. We hypothesised that at a low nutrient supply rate the growth of L. perenne will be more reduced by nematode herbivory than the growth of F. rubra. Furthermore, we hypothesised that higher numbers of plant parasitic nematodes will develop under L. perenne. We found no support for our first hypothesis, because nematodes did not affect plant growth. Our results suggest that changes in the nutrient availability rather than plant parasitic nematodes affect plant succession in impoverished grasslands. On the other hand, plant species and nutrient supply rate significantly affected the density and composition of the plant parasitic nematode community. In line with our second hypothesis, plant parasitic nematodes reproduced better on the fast-growing L. perenne than on the slow-growing F. rubra. Our results, therefore, suggest that the succession of the plant parasitic nematode community is probably more affected by changes in the plant community than the other way round.