Studies on the occurrence of tomato bushy stunt virus in English rivers

Tomato bushy stunt virus (TBSV) of unknown source was isolated from water of the River Thames, near Oxford. The isolate designated TBSV-T was mechanically transmissible to several tomato (Lycopersicon esculentum) cvs and to other species including Petunia hybrida, pepper (Capsicum annuum). eggplant (Solanum melongena), Nicotiana clevelandii, Chenopodium amaranticolor and C. quinoa in which it caused systemic symptoms. It caused no infection of globe artichoke (Cynara scolymus) or Pelargonium domesticum. The virus was not adsorbed to soil and could be isolated from leachate of soil in which systemically-infected tomato or C. quinoa plants were grown. Tomato plants became infected when grown in soil watered with virus suspensions. TBSV-T was infective after 10 min at 80°C but not at 90°C and when diluted to 10^-5 but not to 10^-6. Purified virus preparations contained C. 30 nm isometric particles. In gel-diffusion serological tests, TBSV-T reacted with homologous anti-serum and with antiserum to petunia asteroid mosaic virus but not to pelargonium leaf curl virus. Seed-borne infection (50–65%) of TBSV was demonstrated in plants grown from seed of symptomlessly-infected tomato fruit. TBSV was isolated from symptomlessly-infected tomato fruit imported from Morocco during October-April 1981. One of the isolates (TBSV-M) was indistinguishable from TBSV-T in host range, symptomatology and serological reactions. TBSV was also found in tomato plants growing extraneously in primary settlement beds at sewage works; such plants having been derived from undigested seeds in sewage. Because of its ‘alimentary-resistance’ in man, it is possible that one ecological route whereby TBSV enters rivers is by man's consumption of TBSV-infected tomatoes and eventual sewage dispersal into rivers.     

Serological Studies on Mung Bean Yellow Mosaic Virus

Particles of mung bean yellow mosaic virus (MYMV) were purified by a method that yields up to 3 mg per kg of systemically infected Phaseolus vulgaris“Top Crop” and used to prepare antiserum. MYMV antiserum prepared gave a single precipitin line and had a titre of 1/512 with homologous virus in gel double-diffusion tests. MYMV was shown to be serologically related to other whitefly-transmitted viruses, bean golden mosaic virus, tobacco leaf curl virus and cassava latent virus.     

Natural Infection of Tomato and Pelargonium in Germany by a Tombusvirus Originally Described from Pepper in Morocco

Two serologically apparently identical tombusviruses were isolated from glasshousegrown tomato plants showing mottle and malformation of the upper leaves and stunting of the shoot tips and, at a low rate, from a pelargonium plant, respectively. Both plant species were received from growers in southern Germany. In agar gel double diffusion tests and immuno-electrophoresis, the two virus isolates could not be distinguished from a tombusvirus (MPV) isolated from pepper in Morocco by F ischer and L ockhart (1977), although there were some minor differences in the reported host responses. In the tomato planting, strong symptoms were produced only temporarily. The majority of the 104 tomato plants of 15 different cultivars which had become infected by MPV following mechanical inoculation of the leaves showed only mild symptoms in contrast to a similar group of plants which developed severe stunting and foliar deformation when infected with the type strain of tomato bushy stunt virus. MPV was translocated only occasionally to the upper parts of tomato seedlings which had been grown in virus-containing soil, although root infections were more frequent. MPV is apparently not a major threat to tomato cultivation in Germany.     

Detection of Petunia Asteroid Mosaic, Carnation Ringspot and Tobacco Necrosis Viruses in Ditches and Drainage Canals in a Grapevine-Growing Area in West Germany

A number of virus isolates were obtained from ditches and drainage canals in the Palatinate grapevine-growing area (‘Rheinpfalz’) in West Germany. By means of the immunoelectron microscopical decoration and the agar gel double diffusion tests these isolates were identified as petunia asteroid mosaic virus, carnation ringspot virus and tobacco necrosis virus strain D, respectively.     

Purification,electron microscopy and serology of virus isolated fromEuonymus europaea

“Euonymus mosaic virus”, purified from cucumber cotyledons by the differential and density-gradient centrifugation, shows typical nucleoprotein absorption spectrum. Electron microscopy reveals isometric virus particles of about 37 nm diameter. No reaction of purified “Euonymus mosaic virus” was observed with antisera against a raspberry ringspot virus, tobacco ringspot virus, cherry leaf roll virus, strawberry latent ringspot virus, tomato ringspot virus, elm mosaic virus, arabis mosaic virus, tomato bushy stunt virus and watermelon mosaic virus.     

Discrimination by gel-diffusion serology of digitaria striate, maize sterile stunt and other rhabdoviruses of Poaceae

Eight rhabdoviruses from grass and cereal hosts and their antisera were used to examine virus relationships by gel-diffusion serology. Nucleocapsid (Nc) preparations from digitaria striate virus (DSV) and maize sterile stunt virus (MSSV) both contained a major protein of c. 52 OOO daltons, and antisera prepared to these readily discriminated related planthopper-transmitted rhabdoviruses. MSSV showed a moderately close relationship to barley yellow striate mosaic virus (BYSMV) when an antiserum prepared to whole virus was used, but the Nc antiserum showed clearer discrimination. Worthern cereal mosaic virus and DSV showed a distant relationship to BYSMV and MSSV. There was no serological relationship between any of these viruses and cereal chlorotic mottle virus, cynodon chlorotic streak virus, festuca leaf streak virus or maize mosaic virus.     

Identification of tomato bushy stunt virus in cherry and plum trees showing fruit pitting symptoms

The fruit pitting symptoms on cherries, plums and prunes were investigated from the standpoint of their etiology. Tomato bushy stunt virus (TBSV) was isolated from pitted fruits of these plants and from their leaves and identified by means of biological and serological methods. Both isolates reacted with antisera againstPetunia and artichoke strain of this virus. In addition, the etiology of pseudopox disease of plum and that of cherry detrimental canker is discussed.     

Purification and properties of arabis mosaic and tomato bushy stunt viruses isolated from lilac (Syringa vulgaris L.)

The paper gives more detailed characteristics of Arabis mosaic virus (AMV) and tomato bushy stunt virus (TBSV) isolated from lilac, the latter being identified in lilac (from plants suffering from yellow ring disease) for the first time. The isolate of TBSV from lilac, from which an antiserum with a titre of 1024 was prepared, is closely related to the artichoke strain. Information is given about two types of ringspot disease and about chlorotic ringspot of lilac. Whereas in the leaves of lilac suffering from ringspot disease (of ring mosaic type) the presence of AMV was demonstrated, the sap transmission from the leaves diseased with ringspot of linepattern (and wave-like mosaic) type failed; from the leaves affected by chlorotic ringspot a mixture of AMV and cherry leaf roll virus was identified. In addition, the polyetiological nature of “spring” mosaic and necrotic mosaic of lilac, in which bacteriumPseudomonas syringae van Hall, was found is dealt with. The TBSV was also identified in the isolate of necrotic mosaic.Additional index words: Lilac ringspot, chlorotic ringspot, yellow ring, “spring” mosaic, necrotic mosaic, cherry leaf roll virus,Pseudomonas syringae van Hall.     

Purification and serology of the W strain of cucumber mosaic virus

During studies on the purification of cucumber mosaic virus (strain W) it was found that preparations were most infective and stable when made from tobacco leaves (10–12 days after inoculation) homogenized in phosphate buffer containing EDTA and thioglycollic acid and clarified with diethyl ether. The preparations were further purified by centrifugation in sucrose density gradients containing EDTA at pH 9.0 and were then stable at 2 °C for > 100 days. When mounted in neutralized ammonium molybdate they were shown to consist of predominantly intact particles. In tube and ring precipitin tests and in agar gel-diffusion tests, specific precipitation with homologous antiserum occurred only in media containing alkaline adjusted solutions (ammonium molybdate and dipotassium hydrogen phosphate).     

A structural comparison of concanavalin a and tomato bushy stunt virus protein

Summary Significant structural equivalence has been found among the polypeptide folds of the two tomato bushy stunt virus (TBSV) subunit domains and concanavalin A. This suggests gene duplication in the TBSV coat protein and leads to speculation on common functional properties of concanavalin A and viral coat proteins.Non-standard abbreviations TBSV tomato bushy stunt virus - SBMV southern bean mosaic virus     

Natural infection of sugar beet with tomato bushy stunt virus

A virus transmissible toChenopodium quinoa was isolated from leaves of sugar beet showing large chlorotic ring spots and line pattern. The virus was serologically unrelated to tobacco necrosis virus and tomato black ring virus or to its beet ringspot strain either. A positive result was obtained with antiserum against tomato bushy stunt virus. Reactions of herbaceous indicators and properties of the virus in crude sap were in accordance with the serological diagnosis. A survey of natural hosts of tomato bushy stunt virus demonstrated recently by the authors is given.     

Natural Infection of Cucumbers by Zucchini Yellow Mosaic Virus in Lebanon

An elongated virus was isolated in the Sin-El-Fil Area east of Beirut from cucumber plants showing severe mottling. The particles were 799 nm long after fixation in glutaraldehyde, but were degraded to shorter pieces in unfixed preparations. Infected cells contained cylindrical inclusions with pinwheel and scroll elements accompanied by proliferated endoplasmic reticulum. The virus was purified and an antiserum was prepared. Different serological tests (slide precipitin test, immunoelectronmicroscopical decoration test, immunosorbent electron microscopy) indicated that it was closely related or identical with zucchini yellow mosaic virus (ZYMV) and more distantly related to watermelon mosaic virus 2 and bean yellow mosaic virus. ZYMV isolates from Italy, France and the Lebanon differ in some host reactions.     

THE INFLUENCE OF LIGHT INTENSITY ON THE SUSCEPTIBILITY OF PLANTS TO CERTAIN VIRUSES

Reducing the light intensity under which plants were grown in summer to one-third increased their susceptibility to infection with tobacco necrosis, tomato bushy stunt, tobacco mosaic and tomato aucuba mosaic viruses. With the first two viruses shading increased the average number of local lesions per leaf by more than ten times and by more than five times with the second two.
Reducing the light intensity increased the virus content of sap from leaves inoculated with Rothamsted tobacco necrosis virus by as much as twenty times. As it also reduced the total solid content of sap by about one-half, purification was greatly facilitated; crystalline preparations of the virus were readily made from shaded plants but not from unshaded controls.
Reducing the light intensity also increased the virus content of systemically infected leaves; the greatest effect was with tomato bushy stunt virus with which increases of up to ten times were obtained, but with tobacco mosaic and aucuba mosaic viruses there were also significant increases.
The importance of controlled illumination in raising plants for virus work and the possible mechanisms responsible for the variations in susceptibility are discussed.     

The relationship of potato black ringspot virus to tobacco ringspot and allied viruses

The relationship between potato black ringspot virus (PBRV), isolates of tobacco ringspot virus from blueberry (TRSV-B), cherry (TRSV-C) and calico-diseased potato (TRSV-P), and eucharis mottle virus (EuMV) was examined in tests of three types. In gel-diffusion precipitin tests, the reaction end-points of antisera, and spur formation, indicated that PBRV and TRSV-P are very closely related but not identical antigenically, as are TRSV-B and TRSV-C, and that these two pairs are more distantly related to each other and to EuMV. In plant-protection tests in Nicotiana angustifolia, PBRV, TRSV-B and EuMV conferred protection against the homologous virus but not against one another. PBRV, but not TRSV-B, conferred protection against TRSV-P. In tests with the two RNA species of PBRV, infectivity increased greatly when preparations of RNA-1 and RNA-2 were mixed, and both species are probably needed for infection. Infectivity did not increase when RNA-1 or RNA-2 of PBRV was mixed with RNA-2 or RNA-1, respectively, of TRSV-B; the two viruses seem too distantly related to form pseudo-recombinants. It is concluded that PBRV and tobacco ringspot virus should be considered separate viruses, and that TRSV-P should be considered a strain of PBRV. EuMV should perhaps be recognised as a virus distinct from, but related to, PBRV and tobacco ringspot virus.     

Purification and some properties of oat golden stripe virus

Oat golden stripe virus (OGSV) was maintained in oats by mechanical inoculation and purified by extraction of leaves in borate buffer, two cycles of centrifugation through sucrose cushions and isopycnic centrifugation in CsCl. An antiserum with a titre of 1/1024 in precipitin tests was prepared. Particle length distribution was bimodal with median values, respectively, of 150 and 300 nm from dip preparations. Measurements from immunosorbent electron microscopy (ISEM) and purified preparations showed that the particles had partially degraded during these procedures. The virus sedimented as two components of 168 S and 218 S and had a buoyant density of 1321 g cm^-3. Four isolates of OGSV reacted with the antiserum. Antiserum to members and possible members of the furovirus group were tested in ISEM decoration tests and in ELISA. OGSV was related to soil-borne wheat mosaic virus but not to beet necrotic yellow vein virus, hypochoeris mosaic virus or potato mop-top virus.     

Serological Comparison of Isolates of Cherry Leaf Roll Virus from Diseased Beech and Birch Trees in a Forest Decline Area in Germany with Other Isolates of the Virus

Abstract Two isolates of cherry leaf roll virus (CLRV), one from diseased beech and one from diseased birch trees in an area with forest decline near Bonn in West Germany, were found to be serologically closely related, but not indentical as assessed by spurformation of precipitin lines in agarose gel double diffusion tests. Such tests also distinguished these German CLRV isolates from ten other distinct CLRV isolates obtained from different natural hosts and from various countries. The German beech isolate was most similar serologically to isolates from walnut and from birch in England and the German birch isolate to an English cherry isolate and an isolate from Sambucus racemosa in Finland. These results provide further evidence of the antigenicdiversity of CLRV.     

Comparative analysis of the capacity of tombusvirus P22 and P19 proteins to function as avirulence determinants in Nicotiana species

We have used an agroinfiltration assay for a comparative study of the roles of tombusvirus P22 and P19 proteins in elicitation of hypersensitive response (HR)-like necrosis and the role of P19 in silencing suppression in Nicotiana species. The advantage of agroinfiltration rather than expression in plant virus vectors is that putative viral avirulence proteins can be evaluated in isolation, eliminating the possibility of synergistic effects with other viral proteins. We found that tombusvirus P22 and P19 proteins elicited HR-like necrosis in certain Nicotiana species but, also, that Nicotiana species could recognize subtle differences in sequence between these proteins. Furthermore, Nicotiana species that responded with systemic necrosis to virion inoculations responded to agroinfiltration of tombusvirus P19 with a very weak and delayed necrosis, indicating that the rapid HR-like necrosis was associated with putative resistance genes and a plant defense response that limited the spread of the virus. Tombusvirus P19 proteins also appeared to differ in their effectiveness as silencing suppressors; in our assay, the P19 proteins of Cymbidium ringspot virus and Tomato bushy stunt virus were stronger silencing suppressors than Cucumber necrosis virus P20. Finally, we show that agroinfiltration can be used to track the presence of putative plant resistance genes in Nicotiana species that target either tombusvirus P19 or P22.     

The AU-rich RNA recombination hot spot sequence of Brome mosaic virus is functional in tombusviruses: implications for the mechanism of RNA recombination

RNA recombination can be facilitated by recombination signals present in viral RNAs. Among such signals are short sequences with high AU contents that constitute recombination hot spots in Brome mosaic virus (BMV) and retroviruses. In this paper, we demonstrate that a defective interfering (DI) RNA, a model template associated with Tomato bushy stunt virus (TBSV), a tombusvirus, undergoes frequent recombination in plants and protoplast cells when it carries the AU-rich hot spot sequence from BMV. Similar to the situation with BMV, most of the recombination junction sites in the DI RNA recombinants were found within the AU-rich region. However, unlike BMV or retroviruses, where recombination usually occurred with precision between duplicated AU-rich sequences, the majority of TBSV DI RNA recombinants were imprecise. In addition, only one copy of the AU-rich sequence was essential to promote recombination in the DI RNA. The selection of junction sites was also influenced by a putative cis-acting element present in the DI RNA. We found that this RNA sequence bound to the TBSV replicase proteins more efficiently than did control nonviral sequences, suggesting that it might be involved in replicase "landing" during the template switching events. In summary, evidence is presented that a tombusvirus can use the recombination signal of BMV. This supports the idea that common AU-rich recombination signals might promote interviral recombination between unrelated viruses.     

Bell Pepper Mottle Virus, a Distinct Tobamovirus Infecting Pepper

Bell pepper mottle virus (BPeMV) can be distinguished by symptomatology and host range from other tobamoviruses but a reliable identification needs serological tests. The relationships of BPeMV to tobacco mosaic virus (TMV), Odontoglossum ringspot virus (ORSV), tobacco mild green mosaic virus (TMGMV), and pepper mild mottle virus (PMMV) were investigated using precipitin drop tests on slides, immunodiffusion gel tests, double antibody sandwich enzyme-linked immunosorbent assay (ELISA), and indirectELISA using enzyme-linked goat anti-rabbit globulins for the determination of antiserum titers and serological differentiation indices (SDI). Comparisons of SDIs and amino acid composition data demonstrated that BPeMV is a new species of the tobamovirusgroup. BPeMV, ORSV, PMMV, and TMGMV form a cluster within the genus (group) and could be considered as a subgenus of tobamoviruses.     

Top 10 plant viruses in molecular plant pathology

Many scientists, if not all, feel that their particular plant virus should appear in any list of the most important plant viruses. However, to our knowledge, no such list exists. The aim of this review was to survey all plant virologists with an association with Molecular Plant Pathology and ask them to nominate which plant viruses they would place in a 'Top 10' based on scientific/economic importance. The survey generated more than 250 votes from the international community, and allowed the generation of a Top 10 plant virus list for Molecular Plant Pathology. The Top 10 list includes, in rank order, (1) Tobacco mosaic virus, (2) Tomato spotted wilt virus, (3) Tomato yellow leaf curl virus, (4) Cucumber mosaic virus, (5) Potato virus Y, (6) Cauliflower mosaic virus, (7) African cassava mosaic virus, (8) Plum pox virus, (9) Brome mosaic virus and (10) Potato virus X, with honourable mentions for viruses just missing out on the Top 10, including Citrus tristeza virus, Barley yellow dwarf virus, Potato leafroll virus and Tomato bushy stunt virus. This review article presents a short review on each virus of the Top 10 list and its importance, with the intent of initiating discussion and debate amongst the plant virology community, as well as laying down a benchmark, as it will be interesting to see in future years how perceptions change and which viruses enter and leave the Top 10.