THE PROPERTIES OF TOMATO ASPERMY VIRUS AND ITS RELATIONSHIP WITH CUCUMBER MOSAIC VIRUS*

Chrysanthemum plants infected with tomato aspermy virus (TAV) produce severely distorted and discoloured flowers but show only slight leaf mottle.
TAV infected twenty-five of forty-five species (belonging to seventeen genera) tested and was transmitted by the aphid species Aulacorthum solarti, Macrosiphoniella sanborni and Myzus persicae .
Sap from infected tobacco leaves lost infectivity when diluted more than 1 in 10,000, when heated for 10 min. at above 65°C. and when stored for more than 42 hr. at 16–18°C.
Partial protection was obtained between TAV and two strains of cucumber mosaic virus. Evidence was obtained that this was true protection between related viruses and serological tests confirmed the view that TAV is a strain of cucumber mosaic virus. Evidence was obtained that this was true protection between related viruses and serological tests confirmed the view that TAV is a strain of cucumber mosaic virus.     

SOME EFFECTS OF HIGH TEMPERATURE ON THE SUSCEPTIBILITY OF PLANTS TO INFECTION WITH VIRUSES

When plants were kept at 36°C. for some time before inoculation, their susceptibility to infection by five mechanically transmissible viruses was greatly increased. When kept at 36° after inoculation, fewer local lesions were produced than at lower temperatures, but the effects of the post-inoculation treatment differed with different viruses. Tomato spotted wilt and tobacco mosaic viruses multiply in plants at 36°, and the post-inoculation treatment reduced the local lesions they caused to numbers that varied between 10 and 90% of the control; these two viruses also have large thermal coefficients of heat inactivation. By contrast, tobacco necrosis, tomato bushy stunt and cucumber mosaic viruses, were much affected by post-inoculation treatment, lesion formation being completely prevented by exposure to 36° for a day or more. These three viruses appear not to multiply in plants at 36°, and although they have high thermal inactivation points, they have small temperature coefficients of thermal inactivation.
The extent to which lesion formation was affected by pre- or post-inoculation exposure of plants to 36° depended not only on the length of the treatment, but also on the physiological condition of the plants.
The symptoms of infected plants changed considerably if kept at 36°. At 36° Nicotiana glutinosa , inoculated with tobacco mosaic virus, gave chlorotic local lesions instead of necrotic ones, and became systemically infected. When systemically infected plants were brought to ordinary glasshouse temperature, the infected tissues all collapsed and died in a day.     

THE EFFECT OF TEMPERATURE ON INFECTION WITH STRAINS OF CUCUMBER MOSAIC VIRUS

Cucumber mosaic virus strains differed in their ability to multiply in plants at 37° C. Some strains multiplied in inoculated leaves and produced systemic symptoms in plants at this temperature; plants systemically infected with one such strain remained infected after prolonged treatment at 37° C. Other strains did not appear to multiply in inoculated leaves at 37° C. and heat treatment was successful in freeing plants from infection with these. Tests with one strain of each type showed both to be rapidly inactivated in expressed sap at 37° C.
Strains of cucumber mosaic virus forming small necrotic local lesions in leaves of french bean var. Canadian Wonder, produced many fewer lesions in plants kept after inoculation at 25° C. for 24 hr. and then at 15° C. than in plants kept continuously at the lower temperature.     

RASPBERRY YELLOW DWARF, A SOIL-BORNE VIRUS

An apparently undescribed virus, provisionally named raspberry yellow dwarf virus (RYDV), was isolated from naturally infected raspberry, strawberry, blackberry and several weed species by mechanical inoculation of sap to Chenopodium amaranticolor. The severe disease it caused in Malling Exploit raspberry usually occurred patchily in otherwise normal plantations: these patches increased in size from year to year. RYDV was differentiated from raspberry ringspot and tomato black ring viruses by the symptoms produced in C. amaranticolor , tobacco and Petunia hybrida. RYDV lost infectivity when sap was heated for 10 min. at 61° C., diluted 10^-5or kept for 15 days at 18° C. RYDV was precipitated without inactivation by acetone and by ammonium sulphate.
Isolates of RYDV from different plants and localities, and of different virulence, were identified by plant-protection and serological tests. Such tests gave no evidence that RYDV was related to raspberry ringspot, tobacco ringspot, tomato black ring or cucumber mosaic viruses.
Raspberry and sugar-beet plants became systemically infected with RYDV when grown under glass in soil from a field where the disease had occurred in raspberry plants, and where the virus persisted in the soil for 3 years after the raspberry plants were removed. RYDV seems to be widely disseminated in England but recently introduced and rare in eastern Scotland.
Like raspberry ringspot and tomato black ring viruses, RYDV causes symptoms of the ringspot type in tobacco, has a wide natural and experimental host range, is soil-borne and of local importance. Such features seem characteristic of ringspot viruses as a group.     

SAP-TRANSMISSIBLE MOSAIC DISEASES OF SOLANACEOUS CROPS IN TRINIDAD

Four sap-transmissible viruses were isolated from cultivated Solanaceae in Trinidad: (1) tobacco mosaic virus, from tobacco, tomato and sweet pepper; (2) cucumber mosaic virus, from tobacco and petunia; (3) 'pepper vein-banding virus', probably related to pepper mosaic viruses in Puerto Rico and Brazil, from peppers and tobacco; (4) 'egg-plant mosaic virus', possibly related to the tobacco ring-spot virus, from egg-plant and tomato. Pepper vein-banding virus causes leaf-crinkling and vein-banding in Physalis floridana , petunia, various Nicotiana spp. and most peppers; the Large Bell Hot pepper is killed; tomato and egg-plant are immune. Egg-plant mosaic virus produces mosaic, ring-spotting, or both, on different solanaceous species. It also gives local and systemic ring-spotting on Chenopodium hybridum and necrotic local lesions on the primary leaves of cowpea (var. Black-eye); cucumber is a symptomless carrier. Only cucumber mosaic virus was found naturally infecting non-solanaceous hosts, cucumber and certain common wild plants.
The thermal inactivation point of pepper vein-banding virus is 62° C, its dilution end-point 2×10^-5 and its longevity in vitro 6 day s at 23–30° C.; corresponding values for egg-plant mosaic virus are 78° C., 10^-6 and over 3 weeks. Aphisgossypii transmits cucumber mosaic and pepper vein-banding, but not egg-plant mosaic, of which Epitrix sp. is an occasional vector. Tobacco mosaic, as elsewhere, probably has no regular insect vectors in Trinidad.     

Disease intensity of some tomato viroses in Serbia

In this paper we present the data on the disease intensity of the tomato plants grown in glass and plastic-houses, and in the open field. The infection was caused by the following viruses: Tomato mosaic virus (ToMV), Tobacco mosaic virus (TMV), Tomato spotted wilt virus (TSWV), Alfalfa mosaic virus (AMV), Potato virus X (PVX), Potato virus Y (PVY), Tomato black ring virus (TBRV), Tomato ringspot virus (ToRSV), Tomato aspermy virus (TAV), and Cucumber mosaic virus (CMV). These viruses represented most frequent tomato pathogens in Serbia. According to the obtained results, it could be concluded that 92.94% of the tested tomato plants grown in glass and plastic-houses, and 89.82% grown in the open field were infected by one of the above viruses. Most of the plant samples were infected by two or more viruses. The most frequent viruses — tomato pathogens in Serbia were ToMV, PVY and TMV.     

ANEMONE MOSAIC–A VIRUS DISEASE

The name anemone mosaic is proposed for a previously unrecorded virus disease of Anemone coronaria L.; infected plants have mottled leaves, and broken and distorted flowers. This virus can cause winter browning, and can contribute to crinkle in anemones.
The virus infected forty-seven out of ninety plant species tested; it was transmitted by mechanical inoculation, and by four of the six aphid species tested. Most aphids ceased to be infective within 30 min. when continuing to feed after leaving an infected plant.
Properties in vitro varied according to conditions of the tests; the thermal inactivation point was always below 62°C., the dilution end-point did not exceed 1/2500, and the virus inactivated at 18°C., the fewer than 72 hr.
Intracellular inclusion bodies were produced in all hosts examined.
Anemone mosaic virus is very similar to viruses placed in the turnip virus 1 group of Hoggan & Johnson, and is serologically related to cabbage black ringspot virus, although AMV infection did not protect plants against infection with cabbage black ring-spot virus.
Weeds naturally infected with AMV were found in anemone plantations, and this virus was detected, together with cucumber mosaic and tobacco necrosis viruses, in corms imported into this country.     

Biological properties of pseudorecombinant and recombinant strains created with cucumber mosaic virus and tomato aspermy virus.

Cucumber mosaic virus (CMV) and tomato aspermy virus (TAV) are closely related cucumoviruses. We have made pseudorecombinant viruses in which the RNAs 3 of these two viruses have been exchanged and recombinant viruses containing chimeric RNA 3 molecules, in which the coat proteins and the 3'-end regions of CMV and TAV have been exchanged, giving rise to recombinants designated RT3 and TR3. The replication properties and the cell-to-cell and long-distance movement patterns of these pseudorecombinant and recombinant viruses were examined in different hosts. All the viruses were able to replicate and accumulate RNA 4 in protoplasts. The pseudorecombinants and the R1R2RT3 recombinant infected tobacco systemically, but the R1R2TR3 recombinant was not detectable, even in the inoculated leaves. Comparison of the abilities of the viruses to replicate in protoplasts and intact cucumber plants suggests that cell-to-cell movement factors are also encoded by RNAs 1 and/or 2. Major determinants of symptom severity in Nicotiana glutinosa are localized on the 3' part of RNA 3, and in Nicotiana benthamiana, more severe symptoms were observed with the T1T2R3 strain than with the others tested.     

SOME EFFECTS OF CHANGING TEMPERATURE AND OF VIRUS INHIBITORS ON INFECTION BY CUCUMBER MOSAIC VIRUS

Whereas the spinach strain of cucumber mosaic virus fails to multiply and cause symptoms in tobacco plants kept above 30° C., the yellow strain infects at 36° C. and causes more severe symptoms than at 20° C. Increasing the temperature up to 28° C. increases the initial rate at which the spinach strain multiplies, but the virus later reaches much higher concentrations in leaves at lower temperatures, presumably because it is rapidly inactivated at 28° C. Exposing inoculated plants to 36° C. for 6 hr. decreases the number of infections by the spinach strain when the exposure starts within 6 hr. of inoculation, but not afterwards.
Pancreatic ribonuclease inhibits infections by strains of cucumber mosaic virus; inhibition is greatest when the enzyme is present in the inoculum, and when applied to inoculated leaves its effect decreases rapidly with increasing time after inoculation.
Infection by and the multiplication of strains of cucumber mosaic virus in tobacco are only slightly affected by thiouracil and greatly by azaguanine, whereas strains of tobacco mosaic virus are inhibited much more by thiouracil than by azaguanine. Like tobacco mosaic virus, cucumber mosaic virus multiplies more when inoculated leaves are floated in nutrient solutions than in water, but unlike tobacco mosaic virus, its multiplication is not inhibited by thiouracil more in nutrient solutions than in water.     

VIRUSES OF COWPEA, VIGNA UNGUICULATA L. (WALP.), IN NIGERIA

The cowpea strain of tobacco mosaic virus was isolated from a range of leguminous hosts at Ibadan, but was rare in cultivated crops. Systemic symptoms in species infected experimentally are described.
A new virus of cowpea was also found in Nigeria. The physical properties (thermal inactivation point 56° C., dilution end-point 1/50,000 and longevity in vitro 4 days at 25° C.) differ from those for cowpea viruses reported elsewhere and the name cowpea yellow mosaic virus is proposed. This virus produces local lesions in French bean ( Phaseolus vulgaris L.) and local and systemic lesions in Bengal bean ( Mucuna aterrima Holland), but does not infect other leguminous hosts. The virus was purified and an antiserum prepared against it.
Both viruses are transmitted by a beetle ( Ootheca mutabilis Sahlb.) which loses infectivity within 48 hr. of leaving plants infected with either or both viruses.     

Comparative Analysis of Tissue Tropism of Bipartite Geminiviruses

Abutilon mosaic virus (AbMV), a bipartite geminivirus of the genus Begomovirus, has been vegetatively propagated for many years in Abutilon sellovianum in which it is strictly phloem-restricted. Using in situ hybridization and immunological analyses, the tissue tropism of AbMV in the laboratory host Nicotiana benthamiana was compared with that of two other bipartite begomoviruses, African cassava mosaic virus (ACMV) and tomato golden mosaic virus (TGMV). Analysis of the first systemically infected leaves and longitudinal sections of axillary and flower buds revealed that all three viruses are initially confined to the vascular traces, although both ACMV and TGMV are later detectable in nearly all tissue types. In contrast, AbMV remained strictly phloem-limited in this host throughout the course of infection. The ability of ACMV and TGMV to move out of N. benthamiana phloem tissues is correlated with the development of severe symptoms in comparison with the mild symptoms associated with AbMV infection. It was also demonstrated that Sida micrantha mosaic virus, a virus that is closely related to AbMV, is phloem-limited in Malva parviflora even though it induces severe leaf curl, stunting and necrosis in this host. The present data demonstrate that bipartite begomoviruses can exhibit strikingly different patterns of tissue tropism.     

The appearance and spread of mosaic infection in the tomato crop and the relation to seed transmission of the virus

Appearance and spread of infection with mosaic-inducing viruses were studied for three seasons in tomato crops under glass. Comparison was made between the reactions of plants raised from virus-free seed and those of plants raised from virus-infected seed, on plots distributed at random in a house in which no precautions against entry and spread of virus were taken. Freedom from mosaic infection was maintained longest in plants raised from virus-free seed. An experiment was carried out after steam sterilization of the soil and under exceptionally favourable weather conditions. Appearance of mosaic symptoms occurred later in the life of the plants in this season and plants raised from virus-free seed did not react differently from other plants.
The location of plants first showing mosaic symptoms was related to the depth and texture of soil beneath those plants.
Tests were made of the apparent virus content of infected tomato seed during germination and differences were found in the persistence of virus during germination in seeds of differing origin.
Apparent, 'delayed' seed transmission of mosaic-inducing viruses occurs in the tomato crop, but as yet, this condition can only be interpreted in terms of differences in the resistance of plants raised from seed of differing origin to the multiplication and systemic spread of those viruses. The use of virus-free seed taken from well-nourished vigorous plants is essential to the production of a virus-free tomato crop under commercial conditions.     

Salicylic acid-dependent expression of host genes in compatible Arabidopsis-virus interactions

Plant viruses elicit the expression of common sets of genes in susceptible hosts. Studies in Arabidopsis (Arabidopsis thaliana) and tomato (Lycopersicon esculentum) indicate that at least one-third of the genes induced in common by viruses have been previously associated with plant defense and stress responses. The genetic and molecular requirements for the induction of these stress and defense-related genes during compatible host-virus interactions were investigated with a panel of Arabidopsis mutant and transgenic plants defective in one or more defense signaling pathways. pad4, eds5, NahG, npr1, jar1, ein2, sid2, eds1, and wild-type Columbia-0 and Wassilewskija-2 plants were infected with two different viruses, cucumber mosaic virus and oilseed rape mosaic virus. Gene expression was assayed by a high-throughput fiber-optic bead array consisting of 388 genes and by RNA gel blots. These analyses demonstrated that, in compatible host-virus interactions, the expression of the majority of defense-related genes is induced by a salicylic acid-dependent, NPR1-independent signaling pathway with a few notable exceptions that did require NPR1. Interestingly, none of the mutant or transgenic plants showed enhanced susceptibility to either cucumber mosaic virus or oilseed rape mosaic virus based on both symptoms and virus accumulation. This observation is in contrast to the enhanced disease susceptibility phenotypes that these mutations or transgenes confer to some bacterial and fungal pathogens. These experimental results suggest that expression of many defense-related genes in compatible host plants might share components of signaling pathways involved in incompatible host-pathogen interactions, but their increased expression has no negative effect on viral infection.     

Selection for virus resistance in tomato exposed to tissue culture procedures

Protocols elaborated with the objective of achieving valuable material for selection procedure of variants with virusresistance traits in tomato genotypes are presented. Preliminary results are demonstrated in the domain of testing for variability in somaclones obtained through indirect adventitous organogenesis initiated on leaf explants of cultivated tomato (Lycopersicon esculentum Mill.). Somaclones were grown in greenhouse conditions and variation of their symptoms upon infection with tomato mosaic (ToMV) or cucumber mosaic (CMV) respectively was observed. Tests for resistance to the local isolates of the above cited viruses were performed using enzyme linked immunosorbent assay and back inoculation onto diagnostic plants. Screening data are presented. Desirable variants were selected from cultivars ‘Moneymaker’, ‘Potentat’ and ‘Rutgers’. Some of the ‘Moneymaker’ somaclones exhibited increased tolerance to cucumber mosaic virus, a few seemed to be even fully resistant though most were susceptible as donor plants. The most favourable somaclonal lines are actually further tested and monitored for changes in horticultural characteristics. The described procedure of searching for resistance trait in specific pathogen-free (SPF) plants regenerated from infected tissue looks promising and thus can serve as aid in attaining appropriate objectives of breeding programme. Additionaly experiments were initiated to obtain somaclones from cultivars ‘Beta’, ‘Krakus’ and Stevens Rodade hybrid via regeneration of isolated protoplasts. To this end the callus stage was obtained from all donors.     

Replication of cucumber mosaic virus satellite RNA in vitro by an RNA-dependent RNA polymerase from virus-infected tobacco.

An RNA-dependent RNA polymerase purified from tobacco infected with cucumber mosaic virus catalyzes the synthesis of (-) and (+) strands of the viral satellite RNA, CARNA 5, but fails to replicate the satellite RNA of peanut stunt virus (PSV). The enzyme replicates the genomic RNAs of the three principal cucumoviruses CMV, PSV and tomato aspermy virus (TAV) with varying efficiencies. The specificity with which CMV RdRp replicates different sequence-unrelated RNA templates suggests that the site of their recognition requires secondary or higher level structural organization.     

Viruses infecting winter oilseed rape (Brassica napus ssp. oleifera)

Turnip mosaic virus (TuMV) and cauliflower mosaic virus (CaMV) have been found infecting field crops of winter oilseed rape (Brassica napus ssp. oleifera) in South Warwickshire. Other viruses found include broccoli necrotic yellows virus (BNYV) and a member of the beet western yellows virus group. Systemic leaf symptoms caused by TuMV varied within and between cultivars; the three predominant reaction types were classified as necrotic, mosaic and immune. Some recently introduced cultivars of oilseed rape were more severely affected by TuMV infection than older cultivars. Reactions to CaMV were less varied and immunity was not found. The seed yield from TuMV and CaMV-infected plants was less than that of healthy control plants. This effect was due to infected plants producing either fewer seeds, smaller seeds or both. Germination of seeds from infected plants was unaffected if sown soon after harvest. After storage for one year the germination of seed from a virus infected plant was significantly less than that of seed from a virus-free plant. All commercial cultivars tested were experimentally susceptible to turnip yellow mosaic virus (TYMV) and some American strains of cucumber mosaic virus (CMV).     

THE TRANSMISSION OF PLANT VIRUSES BY BITING INSECTS, WITH PARTICULAR REFERENCE TO COWPEA MOSAIC

Experiments on the virus-vector relationship of the Trinidad cowpea mosaic virus, transmitted by Ceratoma ruficornis , gave the following results: ability to infect decreased with increasing time after ceasing to feed on infected plants, but vectors remained infective for 14 days (much longer than the longevity in vitro of the virus at glasshouse shade temperatures of 23–31°C.); the beetles transmitted more consistently after longer feeding on infected plants, though feeds of under 5 min. made them efficient vectors; the proportion of plants infected increased with the amount of feeding damage on them; fasting the vectors before feeding on infected plants increased voracity but had no effect on their ability to transmit; beetles became infective immediately after feeding on infected plants. Cowpeas were infected by inoculation with macerated infective vectors or with juice regurgitated by vectors. There is no evidence that aphids or other sucking insects can transmit the virus. It seems similar to squash mosaic and turnip yellow mosaic, for vectors of all three viruses probably transmit by regurgitating infective juice during feeding.     

PROPERTIES AND HOST RANGE OF TURNIP CRINKLE, ROSETTE AND YELLOW MOSAIC VIRUSES

Three isolates of turnip yellow mosaic virus and two other flea-beetle transmitted viruses, turnip crinkle and turnip rosette, have many similar properties: thermal inactivation end-point between 80 and 90° C.; dilution end-point greater than 10^-4; longevity in vitro at about 20° C. at least 30 days. All were transmitted by mechanical inoculation to a wide range of cruciferous host plants, including many weeds. Turnip yellow mosaic virus infected only Reseda odorata outside the Cruciferae , whereas rosette virus infected a few and crinkle virus many non-cruciferous hosts.