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.     

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.     

A satellite-like nucleic acid of arabis mosaic virus associated with hop nettlehead disease

An isolate of arabis mosaic virus (AMV) from a hop plant with symptoms of nettlehead disease induced unusually severe symptoms when transmitted to Chenopodium quinoa. This isolate, called AMV-Ta, yielded particle preparations in which up to 80% of the nucleic acids consisted of a species of low molecular weight (SNA), estimated to be about 75 000 daltons by polyacrylamide gel (PAG) electrophoresis. An isolate free of detectable S-NA (AMV-To) was derived from AMV-Ta by inoculating plants with the two high molecular weight genomic RNA species of AMV (2–8 times 10⁶ and 1–3 times 10⁶ daltons; Murant, 1981) separated from S-NA by PAG electrophoresis. This isolate induced much milder symptoms in C. quinoa. Hop seedlings inoculated with AMV-Ta, either mechanically or by nematodes, developed characteristic nettlehead symptoms. Hop seedlings similarly inoculated with AMV-To remained free of nettlehead symptoms. Two species of S-NA associated with hop nettlehead isolates of AMV were detected at two sites in Kent, and two West Midlands sites. At both sites in Kent and at one of the West Midlands sites, the occurrence of the S-NA species was closely correlated with the incidence of nettlehead symptoms. At the other site in the West Midlands, the occurrence of nettlehead symptoms was too erratic to allow positive correlation of SNA with symptom development. Our results show that S-NA plays an active part in symptom production in experimentally inoculated plants of both hops and C. quinoa. In addition, the close correlation between the occurrence of AMV with additional nucleic acid species, and the incidence of nettlehead symptoms in commercially grown hops, suggests a role for S-NA in the aetiology of this disease.     

Production and use of cDNA clones from arabis mosaic virus

Arabis mosaic virus (AMV) genomic RNAs were converted to dsDNA and cloned into bacterial plasmids. Insert sizes of cDNA clones ranged from 0·2 to 3·2 kbp. Restriction enzyme mapping identified clones representing at least 90% of the RNA-2 genome. A 0·9 kbp clone specific to RNA-1 was also identified. Northern blot hybridisations of AMV RNAs with clones from either RNA-1 or RNA-2 showed no cross reactions. The sensitivity of virus detection in dot hybridisation was 15 pg of purified genomic RNA and 40 pg of purified virus particles. The possibility of using cDNA clones for the detection of AMV in strawberry sap was demonstrated. Two AMV dsRNAs corresponding to genomic RNAs in size were isolated from infected cucumber plants and reacted in hybridisation experiments.     

Ultrastructural studies on the nematode Xiphinema diversicaudatum: Egg shell formation

The ultrastructure of the formation of the egg shell in the longidorid nematode Xiphinema diversicaudatum is described. Upon fertilization a vitelline membrane, which constitutes the vitelline layer of the egg shell, is formed. The chitinous layer is secreted in the perivitelline space, between the vitelline layer and the egg cell membrane. On completion of the chitinous layer, the material of the lipid layer is extruded from the egg cytoplasm to the outer surface, through finger-like projections. Both chitinous and lipid layers are secreted by granules in the egg cytoplasm that disappear as the layers are completed. Chitinous and lipid layers are formed during the passage of the egg through the oviduct. The vitelline layer is enriched with secretions produced by the oviduct cells and then by phospholipids secreted by the cells of the pars dilatata oviductus. The inner uterine layer is also formed by deposition of secretory products apposed on the egg shell in the distal uterine region and Z-differentiation. In the proximal part of the uterus, the egg has a discontinuous electron-dense layer, the external uterine layer. Tangential sections between chitinous and uterine layers revealed the presence of holes, possibly egg pores, delimited by the two uterine layers.     

Ultrastructural studies on the nematode Xiphinema diversicaudatum: Oogenesis and fertilization

Oogenesis and fertilization in longidorid nematodes has been examined for the first time at electron microscope level in Xiphinema diversicaudatum. Oogonia in the germinative zone of the ovary are irregularly shaped and lie adjacent to each other or separated by processes of the epithelial cells of the ovary. Developing oocytes pass in single file up to the growth zone and fibrogranular formation occurs around their nucleus. The perinuclear deposits remain until the oocyte is fully grown. Oocytes increase rapidly in volume because of the production of secretory granules. Three types of granules are recognizable. Type 1 granules are spherical, amorphous in structure and delimited by a lighter area, probably consisting of lipoprotein. Type 2 granules, electron lucent, arranged in groups, are lipid inclusions. Type 3 are dense spheres and may represent yolk bodies. The two last are then utilized by the developing embryo. Mature oocytes assume a smooth, cylindrical configuration as they traverse the oviduct. A cone of fertilization seems to be formed at the distal pole of the oocyte, where the sperm penetrates. The sperm totally penetrates the oocyte, through an invagination formed at the oocyte surface. The oocyte continues to undergo two unequal cytoplasmic divisions, resulting in the formation of a female pronucleus and two polar bodies. Under the stimulus of fertilization, a new egg cell membrane is produced, the first one becoming the vitelline envelope.     

Cross-protection between arabis mosaic virus and grapevine fan leaf virus isolates in Chenopodium quinoa

Cross-protection experiments were performed in Chenopodium quinoa using arabis mosaic virus (ArMV) and grapevine fanleaf virus (GFLV) isolates. Two factors were specially studied, namely the time interval and the distance between the two inoculations, respectively, with the hypovirulent isolate and with the hyper virulent challenge isolale. ArMV-S clearly protected C. quinoa from a super infection with GFLV-F13 as shown by a diminution, or even suppression, of the synthesis of the coat protein and the nucleic acids of the GFLV-F13 isolate. In the homologous interaction between GFLV isolates (GH and F13), protection was also observed. In the interaction between GFLV-GH and ArMV-862, by contrast, symptoms were typical of the hyper virulent ArMV-862 and the amount of coat protein of ArMV-862 was normal.     

Transmission of carrot mosaic virus by aphids

The transmission of the carrot mosaic virus (CMV) by the aphidsAcyrtJiosiphon pisum HARRÍS,Cavariela aegopodii SCOP, andMyzus persicae SULZ was proved experimentally. It was observed simultaneously that CMV has a non-persistent character. CMV can be transmitted already 2 min after acquisition feeding by the aphidsMyzus persicae SDLZ andCavariella aego-podii Scop. When the time of acquisition feeding is prolonged to 4 min, CMV is transmitted also by aphidAcyrthosiphon pisum HAREÍs. The host range of the investigated virus wasalso determined and its transmission to 8 plant species, belonging to 4 families, was achieved. On the basis of studies of the vector virus relationship and of the host range, further proof was given for the different character of the Australian Carrot motley dwarf virus, theApivm virus 1 Roland and CMV. The experiments showed that preliminary starving of the aphids for 1 h increases their ability to transmit the virus by 3–3%.     

Apple mosaic virus isolated from hop plants in Japan

A strain of apple mosaic virus was isolated from hop plants in Japan. The virus was purified from young hop plants and back-inoculated to virus-free hop plants obtained by meristem tip culture. Inoculated plants developed chlorotic spots, ringspots and a band pattern accompanied by necrosis in the inoculated and systemically infected leaves. Shoot tips of infected plants sometimes became necrotic and these symptoms resembled those of a ring- and band-pattern mosaiclike disease prevalent in hop gardens in Japan. Since apple mosaic virus was recovered from infected plants, it is likely that the virus was the causal agent of this disease. Agar gel double diffusion tests and ELISA showed the hop virus to be serologically closely related to apple mosaic virus (ApMV), and distantly related to prunus necrotic ringspot virus (PNRSV). The virus had a narrow host range, and infected only cucumber of 18 species of Cucurbitaceae, Chenopodiaceae, Leguminosae or Solanaceae inoculated. It produced chlorotic spots on the inoculated cotyledons of cucumber, but no systemic infection. By contrast, ApMV from apple and PNRSV from peach had wide host ranges and infected cucumber plants systemically.     

Nucleotide sequence predicts circularity and self-cleavage of 300-ribonucleotide satellite of arabis mosaic virus

The nucleotide sequence of the satellite of arabis mosaic virus was determined using the satellite RNA encapsidated in virions. The 300-nucleotide long sequence showed extensive homology (50%) with that of the 359-nucleotide satellite RNA of tobacco ringspot virus, which occurs both in a linear and a circular form. This homology also revealed the presence of conceived sequences believed to mediate self-cleavage of the latter as well as other viral satellite RNAs. A circular form of the arabis mosaic virus satellite can be isolated from infected tissues and partially converts to the linear form upon elution from denaturing gels.     

Transmission of Nepoviruses by Xiphinema americanum-group Nematodes

The transmission of North American nepoviruses by putative species belonging to the Xiphinema americanum-group is reviewed. Xiphinema americanum sensu stricto, X. californicum, and X. rivesi each transmit cherry rasp leaf (CRLV), tobacco ringspot (TobRSV), and tomato ringspot nepovirus (TomRSV), and X. bricolensis is a vector of TomRSV. The apparent lack of specificity in the transmission of North American nepoviruses by X. americanum-group species markedly contrasts with the specific associations between European nepoviruses and their vector nematode species. Two complementary projects are described examining the taxonomic identity of putative species in the X. americanum-group, their morphological and genetic relationships, their ontogeny, and their ability to transmit viruses.     

Transmission of Pepino mosaic virus by the Fungal Vector Olpidium virulentus

Transmission of Pepino mosaic virus (PepMV) by the fungal vector Olpidium virulentus was studied in two experiments. Two characterized cultures of the fungus were used as stock cultures for the assay: culture A was from lettuce roots collected in Castellón (Spain), and culture B was from tomato roots collected in Murcia (Spain). These fungal cultures were maintained in their original host and irrigated with sterile water. The drainage water collected from irrigating these stock cultures was used for watering PepMV‐infected and non‐infected tomato plants to constitute the acquisition–source plants of the assay, which were divided into six different plots: plants containing fungal culture A (non‐infected and PepMV‐infected); plants containing fungal culture B (non‐infected and PepMV‐infected); PepMV‐infected plants without the fungus; and plants non‐infected either with PepMV and the fungus. Thirty‐six healthy plants grouped into six plots, which constituted the virus acquisition–transmission plants of the assay, were irrigated with different drainage waters obtained by watering the different plots of the acquisition–source plants. PepMV was only transmitted to plants irrigated with the drainage water collected from PepMV‐infected plants whose roots contained the fungal culture B from tomato with a transmission rate of 8%. No infection was detected in plants irrigated with the drainage water collected from plots with only a fungus or virus infection. Both the virus and fungus were detected in water samples collected from the drainage water of the acquisition–source plants of the assay. These transmission assays demonstrated the possibility of PepMV transmission by O. virulentus collected from tomato crops.     

A satellite RNA of arabis mosaic nepovirus and its pathological impact

Three RNA species were encapsidated in arabis mosaic nepovirus from lilac (ArMV-L): the two genomic RNAs, RNA-1 and -2, and an RNA-3, Mr. 0.4 × 10⁶. The genomic RNAs from ArMV-L separated by gel electrophoresis from RNA-3 were infectious and, during ten passages of this isolate (ArMV-SF) RNA-3 was not produced; only RNA-1 and RNA-2. When inoculated with the genomic species of ArMV-L, or with those of other ArMV isolates, RNA-3 replicated but did not do so in the absence of RNA-1 and RNA-2. The RNA-3 did not share extensive regions of nucleotide sequences with the genomic RNAs but, like them, was polyadenylated and was linked to a protein (VPg) which did not seem essential for replication. When the pathogenicity of ArMV-L and ArMV-SF was compared in 42 plant species/cultivars representing 36 genera and 14 families, the following differences were observed: in three species of legume, disease progressed more rapidly when RNA-3 was present but in species from five families, the presence of RNA-3 was correlated with symptom amelioration. When RNA-3 was present, the amounts of ELISA detected antigen were not consistently different in tip leaves of Nicotiana megalosiphon, Chenopodium amaranticolor, C. quinoa or Pisum sativum from those in leaves where RNA-3 was absent.     

Mechanical transmission of Apple mosaic virus in Australian hop (Humulus lupulus) gardens

The transmission of Apple mosaic virus (ApMV; hop, H and intermediate, I serotypes) in Australian hop cultivars was assessed in glasshouse and field trials. Under field conditions, the rate of ApMV transmission was halved when contact between neighboring plants was prevented by early season applications of paraquat to restrict basal shoot growth. However, in a separate field trial the presence of root grafts between hop plants, which may contribute to virus transmission, was also suggested. In glasshouse trials, ApMV was transmitted successfully to hop by the mechanical inoculation of infective sap, simulated pruning, foliar contact, and root grafting, but not by root contact. The rate of mechanical transmission of ApMV to the hop cultivar ‘Victoria’ was greater than to other hop cultivars commonly grown in Australia. However, success of mechanical transmission of ApMV also appeared to be influenced by the cultivar from which inoculum was obtained. ApMV was detected throughout the year in all tissues, in chronically infected field grown plants of cultivar ‘Victoria’, suggesting a uniform virus distribution. The reliability of ApMV detection by serology did not decline in ‘Victoria’ plants later in the growing season as occurred in other cultivars.     

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.     

Host status of some plants for Xiphinema diversicaudatum (Micol.) and their susceptibility to viruses transmitted by this species

Forty plant species were grown in pots containing viruliferous Xiphinema diversicaudatum (Micol.) for 15 wk to assess the host range of the nematode in relation to infection with arabis mosaic (AMV) and strawberry latent ring-spot (SLRV) viruses. Host status for the nematode was determined mainly from changes in total populations, but the presence of eggs in the uteri of females and changes in the numbers of adults provided additional criteria. The nematode multiplied on relatively more woody perennials than on herbaceous crop plants or weeds. Chrysanthemum coronarium was the only plant on which numbers declined significantly below those on the controls. Most plant species became infected with either AMV or SLRV. Neither virus was detected in eight out of thirteen species of trees and shrubs although four were good hosts for the nematode. Galling or distortion of the terminal region of fine feeder roots, associated with X. diversicaudatum feeding, was seen on many of the experimental plants.