Properties of a resistance-breaking strain of potato virus X

During indexing of a potato germplasm collection from Bolivia, a strain of potato virus X (PVX), X_HB, which failed to cause local lesions in inoculated leaves of Gomphrena globosa was found in 7% of the clones. X_HB was transmitted by inoculation of sap to 56 species from 11 families out of 64 species from 12 families tested. It was best propagated in Nicotiana glutinosa or N. debneyi; Montia perfolia and Petunia hybrida were useful as local lesion hosts. Inoculated leaves of G. globosa plants kept at 10°, 14°, 18°, 22°, or 26 °C after inoculation were always infected symptomlessly. X_HB caused a mild mosaic, systemic chlorotic blotching or symptomless infection in 16 wild potato species and eight Andean potato cultivars, systemic necrotic symptoms in clone A6 and cultivar Mi Peru, and bright yellow leaf markings in cultivar Renacimiento. It caused necrotic local lesions in inoculated leaves of British potato cultivars with the PVX hypersensitivity gene Nb but then invaded the plants systemically without causing further necrosis; with gene Nx systemic invasion occurred but no necrotic symptoms developed. These reactions resemble those of PVX strain group four. X_HB differed from other known strains of PVX in readily infecting PVX-immune clones 44/1016/10, G. 4298.69 and USDA 41956, cultivars Saphir and Saco, and Solanum acaule PI 230554. X_HB had slightly flexuous filamentous particles with a normal length of 516 nm. It was transmitted readily by plant contact and it partially protected G. globosa leaves from infection with X_CP, a group two strain of PVX. Sap from infected N. glutinosa was infective after dilution to 10^--⁶ but not 10^--⁷ after 10 min at 75° but not 80 °C and after 1 yr at 20 °C. X_HB was readily purified from infected N. debneyi leaves by precipitation with polyethylene glycol followed by differential centrifugation. Microprecipitin tests showed that X_HB and X_CP are closely related serologically.     

Host range and some properties of potato mop-top virus

Potato mop-top virus (PMTV) was transmitted by inoculation of sap to twenty-six species in the Solanaceae or Chenopodiaceae and to Tetragonia expansa; species in eleven other plant families were not infected. The virus was cultured in inoculated leaves of Nicotiana tabacum cv. Xanthi-nc or in N. debneyi. Diagnostic local lesions were produced in Chenopodium amaranticolor. In winter, ten solanaceous species were slowly invaded systemically but the first leaves infected were those immediately above inoculated leaves. When transmitted to Arran Pilot potato by the vector Spongospora subterranea, PMTV induced all the main types of shoot and tuber symptoms found in naturally infected plants. Isolates of PMTV from different sources differed considerably in virulence. PMTV-containing tobacco sap lost infectivity when heated for 10 min at 80 °C, diluted to 10^-4, or stored at 20 °C for 14 weeks. Infectivity was partially stabilized by 0·02% sodium azide. When sap was centrifuged for 10 min at 8000 g, infectivity was mainly in the sediment. Infective sap contained straight rod-shaped particles about 20 nm wide, with lengths up to 900 nm and crossbands at intervals of 2·5 nm. Many of the particles were aggregated side-to-side, and the ends of most seemed damaged. The slight infectivity of phenol-treated leaf extracts was abolished by pancreatic ribonuclease. The present cryptogram of PMTV is R/*:*/*:E/E:S/Fu.     

East African strains of cowpea aphid-borne mosaic virus

Cowpea aphid-borne mosaic virus (CAMV) was isolated for the first time in East Africa where three distinct strains, type, veinbanding and mild, were differentiated by host range and serology. The three strains infected 17/38, 18/37 and 10/35 legume species, and 11/21, 7/21 and 3/19 non-legume species, respectively. The viruses were propagated in cowpea and assayed in Chenopodium amaranticolor. Isolates of all three strains had similar in vitro properties: dilution end point between 10^-3 and 10^-4; thermal inactivation point between 56 and 58 °C; longevity in vitro between 2 and 3 days. Infectivity of sap from frozen leaves was high after 4 wk but much less after 7 wk; infectivity was largely precipitated by 50% acetone but inactivated by 50% ethanol. High yields of virus were consistently obtained from cowpea by extracting systemically infected leaves in 0.5 m sodium citrate containing 1% mercaptoethanol (pH 8.1), and clarifying with 8.5 ml n-butanol/100 ml sap. Virus preparations contained numerous unaggregated and aggregated virus particles c. 750 nm long and contained components with sedimentation coefficients (s°_{20, w}) of 150S and 175S (presumably unaggregated and aggregated particles, respectively). CAMV is serologically distantly related to bean common mosaic virus, but not to bean yellow mosaic or eight other morphologically similar viruses. It is a typical but distinct member of the potato virus Y group.     

Leaf mottling of Spartina species caused by a newly recognised virus, spartina mottle virus

The cause of a previously undocumented leaf mottling of Spartina species was investigated. Negatively stained preparations of sap from mottled leaves revealed flexuous particles 725 × 12 nm. Pinwheels with associated laminar inclusion bodies were observed in thin sections of affected mesophyll cells. The virus was purified from infected Spartina anglica plants and had a sedimentation coefficient in 0·015 m borate of 150S. The virus was transmitted by inoculation of sap to healthy Spartina anglica, but not to a range of other graminaceous or dicotyledonous species tested. It was distantly serologically related to agropyron mosaic virus, but not to other viruses with similar morphology; the name spartina mottle virus is proposed.     

The tomato calcium-permeable channel 4.1 (SlOSCA4.1) is a susceptibility factor for pepino mosaic virus

The hyperosmolality-gated calcium permeable channel 4.1 (OSCA4.1) belongs to an evolutionarily conserved small family of mechano-sensitive channels. OSCA members may represent key players in plant resistance to drought and to pathogen infection but are scarcely studied. After screening for resistance to pepino mosaic virus (PepMV) a collection of 1000 mutagenized tomato families, we identified a mutant showing no symptoms and reduced virus accumulation. Resistance was mapped to chromosome 2 between positions 46 309 531 to 47 044 163, where a missense mutation caused the putative truncation of the OSCA4.1 protein. A CRISPR/Cas9 slosca4.1 mutant was resistant to PepMV, but not to tobacco mosaic virus or potato virus X. Inoculation of mutant and wild type tomato protoplasts showed that resistance was expressed in single cells, suggesting a role for SlOSCA4.1 in early viral function(s); congruently, SlOSCA4.1 re-localized to structures reminiscent of viral replication complexes. We propose that SlOSCA4.1 contributes to the correct regulation of the Ca²⁺ homeostasis necessary for optimal PepMV infection. PepMV is a pandemic virus that causes significant losses in tomato crops worldwide. In spite of its importance, no tomato-resistant varieties have been deployed yet; the mutant identified here has great potential to breed tomato varieties resistant to PepMV.     

Properties of some defective strains of tobacco mosaic virus and their behaviour as affected by inhibitors during storage in sap

From the type strain of tobacco mosaic virus, defective strains were isolated that produced chlorotic or ringspot type symptoms in tobacco and were difficult to transmit without carborundum in the inoculum. Their concentration was less than 0–1 μg/ml of sap instead of the usual 2 mg/ml with the type strain. Phenol extracts of infected leaves were a little more infective than extracts in buffer, whereas phenol extracts of leaves infected with type strain were very much less infective than extracts in buffer. Electron microscopy of infective sap rarely showed any virus particles, but preparations concentrated by ultracentrifugation contained virus particles, many of which were broken or seemed inadequately assembled. Changing the ambient temperature at which infected plants were kept from 20 to 35°C did not increase the amount or improve the appearance of the virus. Some of the strains were inactivated during heating for 10 min between 70 and 80 °C. Undiluted sap lost its infectivity in 3 days at 20 °C, as did the type strain when diluted to 0–1 μg/ml in sap from healthy leaves. This is because substances that inhibit infection were produced by microbes in the sap. The ability of sap from healthy leaves to inhibit infection increased by more than twenty-five times when left 3 days at 20 °C. Infectivity of appropriate mixtures of type strain and aged sap was restored by diluting them in buffer. Sodium azide at 0·02% in sap prevented formation of the inhibitor. The infectivity of the defective strains increased when inoculated together with the type strain.     

Some hosts, properties and possible affinities of a labile virus from Hypochoeris radicata (Compositae)

Hypochoeris mosaic virus (HMV) is common in Hypochoeris radicata (‘cat's ear’) in western Canada. It infected 10 of 53 mechanically inoculated species in five of twelve families, but was not transmitted by aphids or through seed or soil. Sap from infected Nicotiana clevelandii was sometimes infective after dilution to 10^-1 and occasionally 10², after 10 min at 45 but not 50°C, and after 1 but not 2 days at 20°C. Infectivity of crude nucleic acid extracts from infected leaves was rapidly abolished by RNase but not by DNase. Host sap contained very few rod-shaped particles or particle fragments mostly 21.0–22.5 nm in diameter, and up to 420 nm long but with predominant lengths of 120–140 and 240–260 nm. Many rods in purified virus preparations were less than 240 nm long, and the majority were c. 140 nm or shorter. The particles had a helical substructure with a pitch of 2.58 nm and contained a single type of protein of estimated mol. wt 24.5 × 10³. HMV showed no serological relationship to eight morphologically similar viruses (beet necrotic yellow vein, broad bean necrosis, barley stripe mosaic, peanut clump, potato mop-top, Nicotiana velutina mosaic, wheat soil-borne mosaic and defective strains of tobacco mosaic). It is probably a hitherto undescribed tobamovirus.     

Eggplant mosaic virus, and its relationship to Andean potato latent virus

Eggplant mosaic virus (EMV), obtained from Solanum melongena L. from Trinidad, is readily transmitted by inoculation of sap to several solanaceous and a few non-solanaceous plant species. Purified preparations of EMV contain isometric particles 30 nm in diameter, and with sedimentation co efficients of either 111 or 53 S. The particles have thirty-two major morphological subunits. EMV is closely serologically related to Andean potato latent virus and has a similar host range, but is more virulent. Also, whereas EMV accumulates fastest in Nicotiana clevelandii leaves at 20–24 °C, Andean potato latent virus accumulates fastest at 15 °C, and fails to attain a serologically detectable concentration at 24 °C. A few symptomatologically or serologically distinguishable strains of EMV were obtained. EMV has properties typical of viruses of the Andean potato latent subgroup of the turnip yellow mosaic group of viruses, and its present cryptogram is */*:*/*:S/S:S/Cl.     

Narcissus mosaic virus

Narcissus mosaic virus (NMV) is widespread in British crops of trumpet, large-cupped and double daffodils, but was not found in Narcissus jonquilla or N. tazzeta. Many commercial daffodil cultivars seem totally infected, and roguing or selection is therefore impracticable. Strict precautions by breeders and raisers to prevent infection of new cultivars is recommended. Healthy daffodil seedlings were readily infected with NMV by mechanical inoculation, but the virus was not detected in them until 17 months after inoculation, when a mild mosaic appeared. NMV infected twenty-eight of fifty-three inoculated plant species; only five (Nicotiana clevelandii, Gomphrena globosa, Medicago sativa, Trifolium campestre and T. incarnatum) were infected systemically, and NMV was cultured in these and assayed in Chenopodium amaranticolor and Tetragonia expansa. The virus was not transmitted to and from G. globosa or N. clevelandii by three aphid species, or through the seeds of Narcissus, G. globosa and N. clevelandii but was transmitted by handling. G. globosa sap was infective at a dilution of 10 ^-5 but not at 10^-6, when heated for 10 min. at 70° C. but not at 75° C, and after 12 weeks at 18° C, or 36 weeks at 0–4° C. NMV withstood freezing in infected leaves and sap, and purified preparations and freeze-dried sap remained infective for over 2 years. NMV was precipitated without inactivation by ammonium sulphate (313 g./l.) but was better purified by differential centrifugation of phosphate-buffer extracts treated with n-butanol. Such virus preparations from G. globosa, N. clevelandii, C. amaranticolor and T. expansa were highly infective, serologically active, produced a specific light-scattering zone when centrifuged in density-gradients and contained numerous unaggregated particles with a commonest length of 548–568 mμ. Antisera prepared in rabbits had precipitin tube titres of 1/4096. NMV was detected in three experimental hosts but not in narcissus sap. Unlike some viruses with elongated particles, NMV precipitates with antiserum in agar-gel. Purified preparations reacted with antiserum to a Dutch isolate of NMV but not with antisera to seven other viruses having similar particles and in vitro properties, or to narcissus yellow stripe virus.     

Peanut stripe virus - a new seed-borne potyvirus from China infecting groundnut (Arachis hypogaea)1

A new virus, peanut stripe (PStV), isolated from groundnut (Arachis hypogaea) in the USA, induced characteristic striping, discontinuous vein banding along the lateral veins, and oakleaf mosaic in groundnut. The virus was also isolated from germplasm lines introduced from the People's Republic of China. PStV was transmitted by inoculation of sap to nine species of the Chenopodiaceae, Leguminosae, and Solanaceae; Chenopodium amaranticolor was a good local lesion host. PStV was also transmitted by Aphis craccivora in a non-persistent manner and through seed of groundnut up to 37%. The virus remained infective in buffered plant extracts after diluting to 10^-3, storage for 3 days at 20°C, and heating for 10 min at 60°C but not 65°C. Purified virus preparations contained flexuous filamentous particles c. 752 nm long, which contained a major polypeptide of 33 500 daltons and one nucleic acid species of 3·1 × 10⁶ daltons. In ELISA, PStV was serologically related to blackeye cowpea mosaic, soybean mosaic, clover yellow vein, and pepper veinal mottle viruses but not to peanut mottle, potato Y, tobacco etch, and peanut green mosaic viruses. On the basis of these properties PStV is identified as a new potyvirus in groundnut.     

Partial purification and some properties of wineberry latent, a virus obtained from Rubus phoenicolasius

Wineberry latent virus (WLV) was obtained from a single symptomless plant of American wineberry (Rubus phoenicolasius) originally imported from the United States of America. On graft inoculation, WLV infected but induced no distinctive symptoms in several Rubus species including those used as indicators for known Rubus viruses. It was not seed-borne in wineberry. WLV was mechanically transmitted to several herbaceous species but induced local lesions in only a few; it was weakly systemic in some Chenopodium species. Infective C. quinoa sap lost infectivity after diluting to 10^-4, heating for 10 min at 70°C, and storage either for 8 days at 18°C or for 32 days at 4°C. Sap from infected plants contained flexuous filamentous particles c. 510°12 nm. WLV was partially purified by extracting infected C. quinoa leaves in 0·05 M tris-HCl buffer (pH ₇) containing 0·2% thio-glycerol and 10% (v/v) chloroform and concentrating virus by precipitation with 7% (w/v) polyethylene glycol (PEG, mol. wt 6000) and 0·1 NaCl. The virus was then pelleted through a 30% (w/v) sucrose pad containing 7% PEG+0·1 M NaCl and finally sedimented through a sucrose density-gradient. These preparations had A_260/280 ratios of 1·26, contained end to end aggregates of WLV particles and formed a partly polydispersed peak in the analytical ultracentrifuge. WLV did not react with antisera to four potex-viruses, or to apple chlorotic leaf spot or apple stem grooving viruses.     

Peanut green mosaic virus — a member of the potato virus Y group infecting groundnut (Arachis hypogaea) in India

A virus, now named peanut green mosaic virus (PGMV), was isolated from groundnut (Arachis hypogaea) in India and identified as a member of the potato virus Y group by electron microscopy, aphid transmission, and its chemical properties. It was sap transmissible to 16 species of the Leguminosae, Solanaceae, Chenopodiaceae, Aizoaceae and Pedaliaceae; Phaseolus vulgaris was a good local lesion host. PGMV remained infective in buffered groundnut leaf sap at dilutions of 10^-3 after 3 to 4 days at 25 °C, or heating for 10 min to 55 °C but not 60 °C. PGMV was transmitted in the non-persistent manner by Aphis gossypii and Myzus persicae but was not seed-borne. Purified virus preparations contained flexuous filamentous particles c. 750 nm long which sedimented as a single component with a sedimentation coefficient (S°_20w) of 171S, and contained a single polypeptide (mol. wt 34 500 daltons) and one nucleic acid species (mol. wt 3.25 × 10⁶ daltons). PGMV is serologically unrelated to peanut mottle virus (PMV) and other viruses infecting leguminous crops. Infected leaves contained cylindrical, cytoplasmic inclusions.     

Studies on a virus causing stem grooving and graft-union abnormalities in Virginia Crab apple*

A virus was transmitted from apple trees to Nicotiana glutinosa and Chenopodium spp. and back to a range of woody indicators in which it affected only Virginia Crab; symptoms were grooves in the xylem, and swelling and necrosis of the scion immediately above the union with the stock. The virus was distinct from that causing stem pitting in Virginia Crab, because although easily detectable in several apple varieties, it was not found in many trees infected with stem pitting virus. The stem grooving virus has flexous particles 600–700 m/μ long, a heat inactivation point of 67 °C, a dilution end-point of 10^-3 in N. glutinosa sap and remains infective for at least 2 days at 20 °C.     

Purification and Partial Characterization of Isometric Virus-like Particles in Kalanchoe Species

Isometric virus-like particles (IVLP) were detected in crude sap from Kalanchoe pinnata, K. daigremontiana and K. tubiflora plants showing a mild mosaic on the leaves. These particles of 35 nm in diameter were transmitted mechanically to several test plants but not to healthy Kalanchoe. Air temperatures above 30 °C hindered the infection process. The IVLP were purified from systematically infected Nicotiana benthamiana using Triton X-100 as clarifying agent followed byP, EG precipitation. IVLP were degraded by organic solvents and formed aggregates in the presence of 2 mmol/1 CaCl₂. The particles occurred in relatively low concentration in plant sap and lost infectivity in leaves frozen at -70 °C for one week and in purified preparations kept at 4 °C. In buffer crude sap of N. benthamiana IVLP had a thermal inactivation point between 45 and 50 °C on a longevity in vitro of 20 h at 25 °C. Particles contained one nucleoprotein component witha molecular weight of 46,000 daltons and a ssRNA species which, when denatured, had a molecular weight of 1.2 × 10⁶. IVLP purified preparations exhibited a typical nucleoprotein absorption spectrum with a maximum at 254–260 nm and a minimum at 240,–243 nm and a A 260/280 ratio of 1.56. The buoyant density of the IVLP was 1.32 g/ml calculated by isopycnic centrifugation on CsCl. Ultrastructural studies in infected leaves of K. pinnata indicated that IVLP caused an increase in chloroplast volume, distortion of the grana and reduced the number of thylakiods per grana. IVLP infection also impared the diurnal pattern of synthesis and hydrolysis of starch, characteristic of CAM plants. The non-serological reaction of the IVLP with antisera specific to members of 7 different groups of spherical viruses as well as the combination of physicochemical properties and host range, exhibited by these particles impeded their taxonomic location. In nature, young Kalanchoe plantlets acquire the IVLP through their physical connections with the infected mature leaves.     

The relationship between the Andean strain of potato virus S and pepino latent virus

The titres obtained in microprecipitin tests with purified preparations of pepino latent virus (PepLV) and the Andean strain of potato virus S (PVS^A) using PepLV antiserum and two antisera to the ordinary strain of PVS (PVS°) indicated a close serological relationship between PepLV and PVS^A. Using antiserum to PVS°, both viruses were detected by ELISA when infective Chenopodium quinoa sap was diluted to 10^-5but not to 10^-6. Particles of both viruses were decorated equally well by antibodies to PVS^o, PVS^Aand PepLV in all virus-antiserum combinations. When PepLV was inoculated to C. quinoa, C. amaranticolor and potato plants, the symptoms induced closely resembled those of PVS^Ain these hosts. It is concluded that PepLV is an isolate of PVS^Afrom pepino.     

The distribution of poplar mosaic virus in hybrid poplars and virus detection by ELISA

Purified poplar mosaic virus (PMV) at a concentration of 8 ng/ml was readily detected by enzyme-linked immunosorbent assay (ELISA). Bioassay in Nicotiana megalosiphon was more sensitive (detecting 1–4 ng/ml) and latex flocculation less sensitive (c. 25 ng/ml) than ELISA assays. While the foliar sap of fresh, naturally-infected poplars (e.g. Populus. euramericana cv. Robusta) was not infective at dilutions greater than 2 . 10–², ELISA easily detected PMV antigen when sap was diluted 4 . 10–³ in buffer or when one part of infected tissue was triturated with 99 parts healthy leaf. Furthermore, although sap from poplar leaves stored at -20 °C for 6 months was not infective, PMV was still detectable in ELISA tests. PMV antigen in poplar leaves was not all pelleted after centrifugation for 2.5 h at 130 000 g yet parallel tests using unbuffered sap from systemically infected Nicotiana megalosiphon foliage showed that infectivity was restricted to the pellet. In poplar foliage, the concentration of PMV antigen was generally greatest where symptoms were most obvious; least antigen was detected in the overwintering leaves located at the bases of long shoots. In winter, when root and inner bark tissue in the trunk was an erratic source of PMV, the virus was readily detected in buds, the concentration being greatest in the bases, including the meristem, of terminal buds. Propagation from single node cuttings of P. euramericana cv. Regenerata allowed the selection of clones that consistently showed either ‘severe’ or ‘mild’ foliar symptoms. The associated virus isolates also infected another poplar clone causing symptoms characteristic of their source. ELISA consistently detected less PMV antigen in field-grown cv. Regenerata than in cv. Robusta foliage, but this was reversed when the associated virus isolates were propagated in Nicotiana glutinosa at 24 °C. During 6 yr, 21 out of 127 poplars at a site in Western England, became infected with PMV. By contrast, in Eastern England, none of 46 were infected. The aphids Pterocomma populea and Myzus persicae did not transmit PMV.     

Purification and properties of sweet potato mild mottle, a white-fly borne virus from sweet potato(Ipomoea batatas) in East Africa

A virus obtained from sweet potatoes in Kenya, Uganda and Tanzania was transmitted by inoculation of sap and by whiteflies (Bemisia tabaci). It infected forty-five of 119 plant species in fourteen of thirty-six plant families. It was propagated in Nicotiana glutinosa and N. tabacum, in which diagnostic symptoms of vein clearing, leaf curling and distortion developed. Cheno-podium quinoa was a good local lesion host. Different seedling lines of sweet potato differed greatly in their susceptibility to infection and in symptoms produced; some developed leaf mottling and were stunted, some were symptomless, and some appeared immune. The virus was transmitted by dodder (Cuscuta campestris) but not by aphids, or through seed of Ipomoea nil or N. clevelandii. Sweet potato sap contained strong inhibitors of infection, and a low concentration of virus. Virus-free cuttings of sweet potato were obtained by thermotherapy (4–5 wk at 35 °C), or by meristem-tip culture. The virus remained infective in sap of N. tabacum after dilution to 10-³, or after 10 min at 55 °C (but not 60 °C), 3 but not 7 days at 18 °C, or 42 but not 49 days at 2 °C. Infectivity was abolished by sonication or u.v. irradiation, by 2% formaldehyde or 2% tri-sodium orthophosphate, and was greatly decreased by 20 % CHC1₃ or 20 % ether. Purified virus preparations were obtained from N. tabacum by clarifying phosphate buffer extracts with n-butanol, virus precipitation with polyethylene glycol, and differential centrifugation. The virus sedimented as one band in density gradients, and produced a single sedimenting boundary in analytical centrifugation (s°20, w = 1555)- It contained one polypeptide species of mol wt 37700, and preliminary digestion experiments suggested a single-stranded RNA. Antisera prepared against the virus reacted specifically in precipitin tube tests with titres of 1/16384, but no serological relationships could be found between the virus and fourteen viruses of the potato virus Y group. Electron micrographs showed straight, filamentous particles c. 950 nm long when mounted in MgCl_a, but 800–900 nra long in EDTA. The present cryptogram is: (R/i):*/*:E/E:S/Al. This virus is probably the same as Sheffield's virus B.     

Some hosts and properties of dahlia mosaic virus

Dahlia mosaic virus (DMV) infected twenty-five of the eighty-five plant species from four of eighteen families inoculated, but only dahlias were found naturally infected. DMV infected fourteen members of the Solanaceae, Amaranthaceae and Chenopodiaceae, and eleven of twenty-nine Compositae. Verbesina encelioides was the best plant for diagnosis, assay and source of virus. Systemically infected hosts contained ovoid intracellular inclusions 2–5–10 μm in diameter which were shown by electron microscopy to consist of a finely granular, vacuolated matrix containing numerous virus particles. V. encelioides sap was sometimes infective after dilution to 1/2000 but not 1/3000, after heating for 10 min to 75 °C but not 80 °C, and after 4 days at 18 °C or 32 days at 2 °C. Sap from infected dahlia, Zinnia elegans or Ageratum houstonianum rapidly became non-infective, but extracts made with 0·05 M sodium thioglycollate or 0·03 M sodium diethyldithiocarbamate remained infective for 24–48 h at 18 °C. Some purified preparations remained infective for up to 3 years at 2 °C. DMV was best purified from V. encelioides by one or more cycles of differential centrifugation, followed by density-gradient centrifugation and further concentration. Composition, molarity, and pH of the extracting buffer had little effect on yield of virus. Best yields were obtained from extracts stored with 8-5% (v/v) n-butanol at 2 °C for 10–14 days. Purified preparations were infective at dilutions up to 1/5000, had ultraviolet absorption spectra typical of a nucleoprotein (Å 260/280 = 1·47), probably contained DNA, and had a single sedimenting component having isometric particles c. 50 nm in diameter with a sedimentation coefficient of 254 S. The cryptogram of DMV is (D)/*:*/(16):S/S:S/Ap. DMV is serologically closely related to cauliflower mosaic virus, but the viruses are distinct pathogens. The two viruses have similar properties, size, shape and other characteristics, and together with at least three others form a small but apparently homogeneous group of aphid-borne viruses.     

Properties of a tomato isolate of Pelargonium zonate spot virus

A manually transmissible virus isolated from tomato plants with stunting, unfruitfulness, malformation and yellow rings and line patterns of the leaves was indistinguishable from Pelargonium zonate spot virus (PZSV) in biological, physico-chemical and serological properties. The tomato isolate (PZSV-T) of PZSV was seed transmitted in Nicotiana glutinosa and was detected in the pollen of this host. In sap of N. glutinosa PZSV-T lost infectivity after diluting 10^-1 to 10^-2, heating for 10 min at 35 to 40 °C or storage at 25 °C for 7 h. Virus particles were quasi-spherical with a diameter ranging between 25 and 35 nm with a modal value of 29 nm. Particles sedimented as three components (TV, MV and BV) with sedimentation coefficients of 80S (TV), 90S (MV) and 118S (BV); component BV is probably an aggregate of TV. Particles were unstable in CsCl and CS₂SO₄ but formaldehyde-stabilised particles banded at a common density of 1–268 g/cm³ in Cs₂SO₄. Particles contained a single protein species with mol. wt of c. 23000 and c. 18% single stranded RNA present as two species with mol. wts of c. 1.25 × 10⁶ (RNA-1) and 0.95 × 10⁶ (RNA-2). Mixtures of RNA-1 + RNA-2 were infectious and this infectivity was not enhanced by the addition of coat protein. Virus particles had a Tf (mid point of extinction when heated) of 63 °C and were readily dissociated by 0.1% SDS. PZSV-T was serologically unrelated to alfalfa mosaic and to 32 isometric viruses including five ilarviruses. Some properties of PZSV resemble those of ilarviruses but others are sufficiently different to suggest that it may not be a member of this virus group.     

Immunodiagnosis of plant viruses by a virobacterial agglutination test

A new virobacterial agglutination (VBA) test for the immunodiagnosis of plant viruses is described. The test is based on the agglutination of Staphylococcus aureus cells by virus particles after treatment of the cells with homologous antiserum. The agglutination occurs within 1–5 min. The sensitivity of the test is 0·1-0·4 μg virus/ml and is not affected by the shape of the virus particle. The use of affinity purified antibodies for sensitisation of S. aureus cells increases the sensitivity of the reaction 50-fold and enables the detection of tobacco mosaic and cucumber green mottle mosaic viruses at a concentration of 2 ng/ml. The VBA test allows the estimation of potato viruses X, S, M and Y in the eyes and sprouts of infected tubers and in the leaves of infected plants. The diagnosis of carnation mottle virus in carnation plants and of mushroom viruses in mushroom (Agaricus bisporus) fruit-bodies and mycelium are also described.