Reproduction of sugar beet mosaic and tobacco mosaic viruses in anthocyanized beet plants

During our studies on the interaction of anthocyanins and plant virus diseases, reproduction of sugar beet mosaic (SBMV) and tobacco mosaic viruses (TMV) was investigated. Experiments were carried out in leaves of sugar beet,Beta vulgaris cv. Dobrovicka N and its spontaneous anthocyanized mutant. SBMV induces a systemic infection while TMV is responsible for primary local symptoms in sugar beet leaves only. Our quantitative analyses onAmaranthus caudatus L. andChenopodium quinoa Wilid. showed a significant decrease in concentration of SBMV in juice extracted from anthocyanized beet plants as compared with extracts from normal green infected plants. Significant differences were also obtained when SBMV — containing juice was tested in mixtures with healthy extracts from anthocyanized and normal green plants. Also the intensity of TMV symptoms in beet leaves was considerably decreased in leaves of antho-eyanized plants.     

Detection of Infectious Tobamoviruses in Forest Soils

Our objectives were to evaluate elution and bait plant methods to detect infectious tobamoviruses in forest soils in New York State. Soils were collected from two forest sites: Whiteface Mountain (WF) and Heiberg Forest (HF). The effectiveness of four buffers to elute tomato mosaic tobamovirus (ToMV) from organic and mineral fractions of WF soil amended with ToMV was tested, and virus content was assessed by enzyme-linked immunosorbent assay (ELISA). The effectiveness of Chenopodium quinoa (Willd.) bait plants to detect the virus also was tested. Both methods then were utilized to detect tobamoviruses in 11 WF and 2 HF soil samples. A phosphate buffer (100 mM, pH 7.0) eluted more ToMV from soil than the other buffers tested. Mineral soil bound more virus than organic soil. Virus recoveries from virus-amended organic and mineral soils were 3 and 10%, respectively, and the detection sensitivity was 10 to 20 ng/g of soil. Roots of bait plants grown in all virus-amended soils tested positive by ELISA, and virus concentrations averaged 10 ng/g. Both ToMV and tobacco mosaic tobamovirus (TMV) were transmitted to C. quinoa by elution from one of two HF soil samples but not from the WF soil samples. A tobamovirus was detected by bait planting in 12 of 73 (16%) root extracts representing 5 of 13 soil samples (38%). Tobamovirus-like particles were seen by transmission electron microscopy in 6 of 12 infected root extracts. Tobamoviruses occur in forest soils in New York State. Abiotic soil transmission to trees may permit localized spread and persistence of these viruses in forest ecosystems.     

Serological studies of Bratislava mosaic virus of grape vine

Bratislava mosaic virus was transmitted mechanically toChenopodium quinoa. The virus was isolated by means of gradient ultracentrifugation and the, proteins were distributed on an automatic recording photometer. Single fractions were taken with a fraction collector and used as antigen. At the same time, antigen from leaves of the variety Sylván zelený infected with the Bratislava mosaic virus was prepared by means of the gradient ultracentrifugation. The antisera were obtained from rabbits immunized with individual antigens. The antisera were tested with the saps ofC. quinoa infected with the Bratislava mosaic virus, by the method of double diffusion into agar according to Ouchterlony, and with adjusted saps from suckers infected with the Bratislava mosaic virus, by the method of double diffusion into agar according to Oudin. A specific reaction can be obtained in both cases only under the assumption that all conditions mentioned in the study are strictly kept.     

The Systemic Infection by Tobacco Mosaic Virus of Tobacco Plants Coutaining the N Gene at Temperatures Below 28°C

Tobacco plants containing the N-gene are occasionally systemically infected with tobacco mosaic virus (TMV) at tempreratures below 28°C, but contain low concentrations of virus: they often fail to set seed, and can outlive healthy control plants. Infection is thus similar to that induced when N-gene tobacco plants are grafted onto systemically infected tobacco lacking the N-gene. Shoots from systemically infected N-gene plants can induce systemic infection in other graft-inoculated N-gene plants. Stem sections of N-gene tobacco plants act as good conduits for TMV between plants lacking the N-gene, and girdling experiments suggest that virus movement probably occurs in the phloern.     

Transmission of sowbane mosaic virus by Thrips tabaci in the presence and absence of virus-carrying pollen

When pollen of sowbane mosaic sobemovirus (SoMV)-infected Chenopodium amaranticolor was dusted onto C. amaranticolor and C. quinoa test seedlings which were then infested with 5–10 adult Thrips tabaci, SoMV was transmitted to 25%C. amaranticolor and 88% of C. quinoa plants. Five hours access of T. tabaci to pollen-contaminated C. quinoa seedlings was sufficient for SoMV transmission, but 1 h was not. SoMV was also transmitted when T. tabaci was mixed with pollen of C. amaranticolor and then placed on C. quinoa test seedlings. Further, plant-to-plant transmission of SoMV occurred in the absence of virus-carrying pollen when T. tabaci was caged on infected non-flowering C. amaranticolor or C. quinoa for 1–8 days and then adults transferred to C. quinoa test seedlings. This is the first time that thrips have been shown to be a vector of SoMV, and that a virus outside the ilarvirus group is reported to be transmitted using pollen and thrips.     

Kristalloide Einschlüsse im Zytoplasma pflanzlicher Zellen

Summary In healthy as well as dahlia mosaic sick plants ofVerbesina encelioides, Sanvitalia procumbens, Zinnia elegans, Calendula spec. andDahlia hybrids, leaf cell vacuoles are found in the marginal cytoplasm which contain protein crystals. They are single membrane-limited products of the endoplasmatic reticulum. They may be found mainly in the older leaves and especially in those of virus infected plants. The crystalline structures consist of tetragonally arranged tubules of 105 Å in diameter, separated by an interspace about 35 Å wide. There are similar structures in virus infected plants ofFragaria vesca, but not inChenopodium quinoa, where the vacuoles contain no bodies. This cell organelle is compared with crystalline inclusions already described. Its significance and relations to the virus disease are discussed.     

Mechanical transmission of beet yellows virus toChenopodium quinoa Willd. andChenopodium foliosum (Moench) Asch.

Beet yellows virus (BYV) was mechanically transmitted by sap from sugar-beet plants, infected with BYV, to the plants ofChenopodium quinoa Willd. and ofChenopodium foliosum (Moench) Asch. Mechanical transmission of BYV to the plants ofTetragonia expansa Murr. failed. Infectious material was homogenized in phosphate buffer with veronal and EDTA, pH 7–8. Experimental plants were darkened three days before infection and kept at a temperature of 5°C. Plants ofC. quinoa Willd. were decapitated. Back transmissions fromC. quinoa Willd. andC. foliosum (Moench) Asch. infected with BYV, to sugar-beet plants were carried out by the aveidMyzus persicae Sulz. These transmissions were positive. Filamentous particles of BYV, of an average length 1275 nm, were found in plants ofC. quinoa Willd. andC. Foliosum (Moench) Asch., infected with BYV.     

Interaction of cucumber mosaic virus and potato virus y with tobacco mosaic virus

A study was performed on the interaction of cucumber mosaic virus (CMV) of potato virus Y (PVY) with tobacco mosaic virus (TMV). Interference was evaluated using tobacco plantsNicotiana tabacum cv. Java responding to CMV and PVY with a systemic infection and to TMV with local necrotic lesions. The decrease in TMV — induced lesion number gave evidence of a decrease in susceptibility caused by the previous infection with CMV or PVY, the decrease of lesion enlargement demonstrated a decreased TMV reproduction in the plants previously infected with CMV or PVY. The interference concerned was incomplete, as evaluated from reproduction of the challenging TMV and from the decrease in susceptibility of the host to TMV brought about by the first infection with CMV or PVY.     

Dark and light green tissues of tobacco leaves systemically infected with tobacco mosaic virus

There are significant changes in the structure of the upper tobacco (Nicotiana tabacum L.) leaves systemically infected with tobacco mosaic virus (TMV) especially in the light green tissue (LGT). Dark green areas (DGI) had intermediate status between healthy tissue and LGT. DGI contained significantly less infectious TMV and viral antigen than the LGT. The DGI, LGT and healthy tissues did not differ in the permeability of cell membranes and in the set of acidic pathogenesis-related (PR) proteins but the total content of PR-proteins in the healthy plants was higher than in the infected ones with the DGI being intermediate between healthy tissue and LGT. The crude leaf extracts from DGI and LGT showed less total ribonuclease activity and ribonuclease isozymes in comparison with control.     

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

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

Reduced Photosystem II Activity and Accumulation of Viral Coat Protein in Chloroplasts of Leaves Infected with Tobacco Mosaic Virus

We previously reported (A Reinero, RN Beachy 1986 Plant Mol Biol 6:291-301) that coat protein (CP) of tobacco mosaic virus (TMV) accumulates in chloroplasts of systemically infected leaves. To determine the significance of such interaction we examined electron transport rates in chloroplasts containing different levels of TMV-CP. Tobacco (Nicotiana tabacum L.) plants were infected with either a TMV strain inducing chlorosis or with a strain inducing mild symptoms, and both the accumulation pattern of TMV-CP inside chloroplasts as well as the rates of photosynthetic electron transport were followed. The CP of the TMV strain inducing chlorosis was detected inside chloroplasts 3 days after infection, and thereafter accumulated at a rapid rate, first in the stroma and then in the thylakoid membranes. On the other hand, the CP of the TMV strain that caused only mild symptoms accumulated in chloroplasts to lower levels and little CP was associated with the thylakoids. In vivo and in vitro measurements of electron transport revealed that photosystem II activity was inhibited in plants infected with the aggressive TMV strain while no reduction was observed in plants infected with the mild strain. The capacity of chloroplasts to synthesize proteins was equivalent in organelles isolated from healthy and virus-infected leaves. The possibility that a large accumulation of TMV-CP inside chloroplasts may affect photosynthesis in virus-infected plants by inhibiting photosystem II activity is discussed.     

Presence of Plant Viruses in some Rivers of Southern Italy

Water samples were collected from some rivers of Southern Italy whose water is normally used for irrigation and checked for the presence of plant viruses. The assayes were carried out by centrifuging at 5000x g one liter of water, resuspending the sediments in phosphate buffer and testing their infectivities on Chenopodium quinoa Willd. It was possible to isolate tobacco mosaic virus, cucumber mosaic virus and two more viruses not identified yet.     

The electron-microscopic proof of tobacco necrosis virus in hop plant

Following a previous biological and serological evidence of tobacco necrosis virus by means of an electron microscope, isometric virus particles of an average size of 26 ± 0.5 nm were found, in ultrathin sections prepared from the tissue of sprouting buds of underground stalks and from that of runners butts with symptoms of mosaic line pattern, as well as from the tissue of the indicator plantChenopodium quinoa.     

Czech and scandinavian isolates resembling dandelion yellow mosaic virus

Virus isolates resembling the dandelion yellow mosaic virus (synonym: lettuce necrosis virus) were obtained from dandelion plants in twenty five localities of Bohemia and also of Norway, Sweden and Finland. All isolates were sap transmissible merely to lettuce, but some of them also toChenopodium quinoa; other test plants could not be infected. Attempted serological and biological identification of the isolates with some viruses presumed to be able to infect spontaneously dandelion plants have failed.     

Changes in phosphoenolpyruvate carboxylase and ribulosebisphosphate carboxylase activities in tobacco plants infected with tobacco mosaic virus

Considerable changes in the activities of phosphoenolpyruvate carboxylase and ribulosebisphosphate carboxylase were found inNicotiana tabacum cv. Sarasun plants infected with TMV. Ribulosebisphosphate carboxylase is inhibited at the time of maximum TMV reproduction, but its decreased activity is at the same time partly compensated by phosphoenol-pyruvate carboxylase in the shoots of infected plants. The pattern of activity of this enzyme nearly exactly reflects the pattern of reproduction of the tobacco mosaic virus.     

Purification and conservation of infective sugar beet mosaic virus

Sugar beet mosaic virus (SBMV) was precipitated by polyethyleneglycol (PEG) 6000 from the cell sap of infected sugar beet leaves. After centrifugation and addition of dextrane T 10 the virus was lyophilized. Its infectious activity was demonstrated by mechanical transmission toChenopodium quinoa Willd. and to sugar beet. Stability of infectious activity of the lyophilized virus was verified.     

Vergleich des Eiweissanteiles des Wildstammes vom Tabakmosaikvirus mit dem einer Temperaturmutante

Summary The protein of the common strainvulgare of tobacco mosaic virus was compared analytically with the protein of a temperature-mutantthermophilum. The latter was isolated from tobacco plants which were infected with common TMV and cultured at 35°C. Per polypeptid chain of 17 500 molecular weight the protein of the mutation strain contained one threonine residue more and one isoleucine (or leucine) residue less than common TMV protein.     

3. Isolation of Viruses by Electro-Extraction of Infected Plants

ABSTRACT

The isolation of viruses from infected plant material by a process termed electro-extraction appeared to be a convenient and simple method of obtaining viruses in a fair state of purity. The method has the advantage over the conventional methods of virus purification that the infected plant tissue is not disintegrated and that organic solvents such as chloroform and butanol are avoided. The procedure used was demonstrated on the extraction of tobacco mosaic virus (TMV) from infected tobacco and turnip yellow mosaic virus (TYMV) from Chinese cabbage plants. To obtain the virus it was found advisable to freeze and thaw the plants prior to extraction.     

22. Purification of PVY° and Its Soluble Antigen by Physico Chemical Means

ABSTRACT

Potato virus Y° was purified by centrifugation of infected and minced plant tissue in the virus extraction rotor. As the initial seeding material was heavily contaminated with tobacco mosaic virus (TMV) this virus was isolated and antibody was elicited in chickens. The chicken antibody (IgY) against TMV was used for removing this extraneous virus from the original PVY° seeding material prior to propagating PVY° in tobacco plants, CV Glutinosa.     

Isolation and Characterization of Two Protein Isoforms with Antiviral Activity from Chenopodium album L Leaves

Two antiviral proteins (AVPs) named CAP-I and CAP-II isolated and purified from the leaves of Chenopodium album cv Pusa Bathua 1 were found to inhibit tobacco mosaic virus (TMV) and sunnhemp rosette virus (SRV) infection on their respective host plants. The molecular weight of both the AVPs was found to be 24 kD. They were devoid of carbohydrate moiety and were highly basic with pI ～10.2. However, they differed with respect to amino acid composition and N-terminal sequence. They also differed with respect to IC₅₀ values, and CAP-I was found to be 2.5 fold more effective than CAP-II in inhibiting viral infection.