Correlation of viral RNA biosynthesis with glucose-6-phosphate dehydrogenase activity and host resistance

Changes in glucose-6-phosphate dehydrogenase (G6P DH; EC 1.1.1.49) activity caused by infection of tobacco ( Nicotiana tabacum L.) leaves with potato virus Y (PVY), cucumber mosaic virus, potato virus X, tobacco rattle virus and turnip mosaic virus, the subcellular localisation of G6P DH isozymes in mesophyll protoplasts derived from healthy and PVY-infected tobacco leaves, as well as G6P DH control and the relationship of its isozymes with the degree of tobacco resistance to PVY multiplication, were studied. The activities of G6P DH were markedly increased in locally and systemically infected leaves and the time courses of the activity linearly correlated with those of virus multiplication. In leaves infected with PVY, the activity time courses of the crude and the partially purified G6P DH were coincident. This probably indicates the involvement of coarse regulation of the enzyme. PVY content linearly correlated with enhanced G6P DH activity in leaf discs derived from susceptible, tolerant and resistant cultivars of tobacco. The increased activity of the enzyme in infected protoplasts and plant tissues was predominantly caused by the increased activity of chloroplastic isozymes. This was confirmed by the specific staining of isozymes after electrophoretic separation of chloroplastic proteins of tobacco leaves. These findings enable the degree of resistance to virus multiplication to be quantified for the use of gene manipulation and breeding.     

THE DISTRIBUTION OF VIRUSES IN LEAVES AND THEIR INACTIVATION BY ULTRA-VIOLET IRRADIATION

The infectivity of sap expressed from the lower epidermis stripped from leaves systemically infected with potato virus Y , henbane mosaic virus or tobacco mosaic virus was compared with that of sap from the underlying mesophyll. Results suggested that the concentration of virus in each of the two tissues was about the same.
Ultra-violet irradiation of leaves infected with potato virus Y or henbane mosaic virus greatly reduced the infectivity of sap expressed from subepidermal tissues.     

Plant Viruses Affecting Tomatoes in Jordan. Identification and Prevalence

A survey was conducted during the period 1979–1982 to identify viruses affecting tomatoes in the Jordan Valley. Results indicated the rare occurrence of mosaic diseases caused by tobacco mosaic virus, cucumber mosaic virus, or potato virus Y. However, tomato yellow leaf curl virus was predominant.     

HEAT-THERAPY OF VIRUS-INFECTED PLANTS

Virus-free plants were produced from parents systemically infected with the following five viruses: tomato bushy stunt, carnation ring spot, cucumber mosaic, tomato aspermy and Abutilon variegation. The leaves formed while the infected plants were kept at 36°C. were free from symptoms, and test plants inoculated from these remained uninfected. When cuttings were taken from the infected plants at the end of the treatment most grew into healthy plants. The treated plants themselves usually developed symptoms after varying lengths of time at 20°C, but some that before treatment were infected with tomato aspermy, cucumber mosaic or Abutilon variegation viruses, remained permanently healthy.
The same method failed to cure plants infected with tomato spotted wilt, potato virus X and tobacco mosaic virus, although it decreased their virus content. Heat-therapy seems not to be correlated with the thermal inactivation end point of the virus in vitro.     

THE CORRELATION OF APHID NUMBERS WITH THE SPREAD OF LEAF ROLL AND RUGOSE MOSAIC IN POTATO CROPS

An analysis of the results of experiments in different parts of England and Wales from 1941 to 1947 on the spread of potato leaf roll and rugose mosaic showed that leaf roll spread was correlated with the number of alate Myzus persicae (Sulzer) caught on sticky traps throughout the potato-growing season; there was some correlation with the maximum count of M. persicae per 100 leaves, but this possibly results from the correlation between trapped aphids and the number per 100 leaves. Spread of rugose mosaic (potato virus Y) was correlated to a lesser degree with number of M, persicae , perhaps because other aphid species are often vectors. With both diseases higher correlations were obtained when the infected plants were dispersed among the healthy crop than when they were placed together in a row. It is concluded that it is possible to predict the average health of potato stocks in the following year from average trap data; further work may enable the health of individual stocks to be predicted.     

The relationships of some viruses causing necrotic diseases of the potato

Potato virus B , and some other viruses with reactions in potato varieties different from any previously described, are strains of virus X . All produce intracellular inclusions which vary with different hosts and virus strains. Except with virus B, the inclusions are larger and more frequent in potato than in tobacco or tomato. All give systemic infection when inoculated to tobacco, tomato and potato varieties in which they are carried or cause mosaic symptoms; some give systemic infection when inoculated to varieties in which they cause top-necrosis, whereas others give only local lesions.
Potato virus C is a strain of virus Y: in tobacco and a few potato varieties both produce similar symptoms, but in those varieties in which Y causes leaf-drop streak, C causes top-necrosis. C causes systemic infection when inoculated to tobacco and to potato varieties in which it causes mosaic symptoms, but not when inoculated to potato varieties in which it causes top-necrosis. Virus C was not transmitted by M . persicae. Viruses C and Y produce a few small intracellular inclusions in potato and tobacco.
Virus A is not related to Y or X : no inclusions were found in plants infected with A alone.     

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.     

Antiviral RNA silencing is restricted to the marginal region of the dark green tissue in the mosaic leaves of tomato mosaic virus-infected tobacco plants

Mosaic is a common disease symptom caused by virus infection in plants. Mosaic leaves of Tomato mosaic virus (ToMV)-infected tobacco plants consist of yellow-green and dark green tissues that contain large and small numbers of virions, respectively. Although the involvement of RNA silencing in mosaic development has been suggested, its role in the process that results in an uneven distribution of the virus is unknown. Here, we investigated whether and where ToMV-directed RNA silencing was established in tobacco mosaic leaves. When transgenic tobaccos defective in RNA silencing were infected with ToMV, little or no dark green tissue appeared, implying the involvement of RNA silencing in mosaic development. ToMV-related small interfering RNAs were rarely detected in the dark green areas of the first mosaic leaves, and their interior portions were susceptible to infection. Thus, ToMV-directed RNA silencing was not effective there. By visualizing the cells where ToMV-directed RNA silencing was active, it was found that the effective silencing occurs only in the marginal regions of the dark green tissue ( approximately 0.5 mm in width) and along the major veins. Further, the cells in the margins were resistant against recombinant potato virus X carrying a ToMV-derived sequence. These findings demonstrate that RNA silencing against ToMV is established in the cells located at the margins of the dark green areas, restricting the expansion of yellow-green areas, and consequently defines the mosaic pattern. The mechanism of mosaic symptom development is discussed in relation to the systemic spread of the virus and RNA silencing.     

The effects of the repellents dodecanoic acid and polygodial on the acquisition of non-, semi- and persistent plant viruses by the aphid Myzus persicae

Few Myzus persicae settled on polygodial (1 g litre^-1) or dodecanoic acid (5 g litre^-1)-treated leaves and few nymphs were deposited. Polygodial decreased acquisition of the semi-persistent beet yellows virus and the non-persistent potato virus Y and dodecanoic acid that of the persistent potato leafroll and beet mild yellowing viruses, of beet yellows virus and of the bimodally-transmitted cauliflower mosaic virus. However dodecanoic acid increased the acquisition of potato virus Y.     

Effects of Salicylate on Systemic Invasion of Tobacco Plants by Various Viruses

Salicylate watered onto soil in which White Burley tobacco plants were grown represents a reversible stress characterized by stomatal closure, slight slackening of plant growth and low chlorophyll loss. Salicylate affected viral pathogenesis in opposite ways. It had no effect against local and systemic infections by potato virus X (PVX), potato virus Y⁰ (PVY⁰) or tobacco mosaic virus (TMV), whereas it completely prevented systemic infection by alfalfa mosaic virus (AIMV) or tobacco, rattle virus (TRV) in a high proportion of treated plants. When infection moved from leaves inoculated with AIMV or TRV, the tendency to limit systemic spread was shown by the restriction of systemic infection to very limited areas erratically distributed in some uninoculated leaves. The salicylate-induced restriction of AIMV or TRV infectivity to inoculated leaves did not appear due to inhibition of virus multiplication because the inoculation of potentially resistant leaves of salicylate-reated plants resulted in virus antigen accumulation comparable to that of untreated controls. Salicylate may therefore inhibit some long distance virus transport function. Salicylate appears able to evoke true hypersensitivity only against systemic viruses able to induce local necrotic lesions, probably by activating some genetic information for resistance that is normally not expressed.     

THE REACTION OF VIRUS-INFECTED POTATO PLANTS TO PHYTOPHTHORA INFESTANS

The growth of Phytophthora infestans was retarded on leaves of potato plants that had been artificially inoculated with virus X or with virus Y.
Using different virus strains and potato varieties, the effect of virus infection on blight development was found to be greater, the more severe the systemic virus symptoms exhibited on the infected leaves before P. infestans inoculation.
The development of the fungus was never increased by virus infection.
The reduced blight development on virus-infected leaves is partially caused by an increase of resistance to infection. It is also suggested that virus infection alters the nutritional status of leaves to one less favourable for the development of P. infestans.     

Experiments on the spread of rugose mosaic and leaf roll in potato crops in 1946

A series of experiments on the spread of potato rugose mosaic (virus Y ), and leaf roll, which has been in progress on a uniform plan since 1943, was ended in 1946. Mean values for thirteen centres in England and Wales showed that in 1946 69% of the infections with virus Y and 48 % of those with leaf-roll virus reached the tubers of Majestic potatoes by the beginning of August. There was usually little subsequent increase of rugose mosaic, but a late increase of leaf roll was associated with a relatively high initial spread. Three-quarters of the virus Y and over half the leaf-roll infections occurred within five plants distance of the source. There was no close correlation between the spread of either virus and the maximum number of Myzus persicae , either apterous forms on the plants or alate forms caught on adhesive traps, but centres with high trap catches in July and August showed pronounced late season spread of leaf roll. There were marked differences at different centres in the relative spread of the two viruses. The amount of spread and the gradients from source of infection of the two viruses are compared over the period 1943–6.     

Hypersensitive response in cucumber mosaic virus-inoculated Arabidopsis thaliana

The responses of 12 Arabidopsis ecotypes to cucumber mosaic virus strain Y (CMV(Y)) or strain O (CMV(O)) were characterized. Except for ecotype C24, all ecotypes were susceptible to both strains of CMV. In C24, CMV(O) multiplied systemically, but CMV(Y) did not spread systemically and induced only local necrotic spots in virus-inoculated leaves 21–27 h after inoculation. In CMV(Y)-inoculated C24 leaves, virus was confined to the inoculated leaves, and the amount of the pathogenesis-related-1 protein increased during the progress of local necrotic spot formation. These results indicate that C24 mounts a hypersensitive response (HR) to CMV(Y). By genetic analysis of crosses between C24 and ecotype Columbia or Landsberg (erecta) which are susceptible to CMV(Y) infection, the HR to CMV(Y) in C24 was found to be determined by a single major dominant gene whose function was influenced by a modifier gene from the Landsberg ecotype. Comparison of the responses between C24 leaves inoculated with pseudorecombinants of both strains of CMV suggested that the HR was controlled by CMV RNA3. The molecular interaction between the single major gene and CMV(Y) RNA3 is likely to induce the HR in CMV(Y)-inoculated C24.     

The shallot aphis,Myzus ascalonicus Doncaster,and its behaviour as a vector of plant viruses

A new species of aphis, Myzus ascalonicus Doncaster, is briefly described, and compared with Myzus persicae Sulz., which it resembles superficially. It has been found on shallots in storage and on onions and other species of plants both in glasshouses and in the open between October and June. Its summer habits and hosts are unknown. In comparative virus transmission tests with M. persicae it was found that Myzus ascalonicus transmits dandelion yellow mosaic virus, which is not transmitted by Myzus persicae ; and also cucumber virus I, Hyoscyamus virus III and sugar-beet yellows virus, all of which are transmitted by M. persicae. Myzus ascalonicus does not transmit the viruses of potato Y severe etch, lettuce mosaic and sugar-beet mosaic which are transmitted by Myzus persicae.     

Mapping the virus and host genes involved in the resistance response in cucumber mosaic virus-Infected Arabidopsis thaliana

A yellow strain of cucumber mosaic virus (CMV) [CMV(Y)] induces a resistance response characterized by inhibition of virus systemic movement with development of necrotic local lesions in the virus-inoculated leaves of Arabidopsis thaliana ecotype C24. In this report, the avirulence determinant in the virus genome was defined and the resistance gene (RCY1) of C24 was genetically mapped. The response of C24 to CMV containing the chimeric RNA3 between CMV(Y) and a virulent strain of CMV indicated that the coat protein gene of CMV(Y) determined the localization of the virus in the inoculated leaves of C24. The RCY1 locus was mapped between two CAPS markers, DFR and T43968, which were located in the region containing genetically defined disease resistance genes and their homologues. These results indicate that the resistance response to CMV(Y) in C24 is determined by the combination of the coat protein gene and RCY1 on chromosome 5.     

Movement of plant viruses is delayed in a beta-1,3-glucanase-deficient mutant showing a reduced plasmodesmatal size exclusion limit and enhanced callose deposition

Susceptibility to virus infection is decreased in a class I beta-1,3-glucanase (GLU I)-deficient mutant (TAG4.4) of tobacco generated by antisense transformation. TAG4.4 exhibited delayed intercellular trafficking via plasmodesmata of a tobamovirus (tobacco mosaic virus), of a potexvirus (recombinant potato virus X expressing GFP), and of the movement protein (MP) 3a of a cucumovirus (cucumber mosaic virus). Monitoring the cell-to-cell movement of dextrans and peptides by a novel biolistic method revealed that the plasmodesmatal size exclusion limit (SEL) of TAG4.4 was also reduced from 1.0 to 0.85 nm. Therefore, GLU I-deficiency has a broad effect on plasmodesmatal movement, which is not limited to a particular virus type. Deposition of callose, a substrate for beta-1,3-glucanases, was increased in TAG4.4 in response to 32 degrees C treatment, treatment with the fungal elicitor xylanase, and wounding, suggesting that GLU I has an important function in regulating callose metabolism. Callose turnover is thought to regulate plasmodesmatal SEL. We propose that GLU I induction in response to infection may help promote MP-driven virus spread by degrading callose.     

Experiments on the Spread of Potato Virus X Between Plants in Contact

Experiments on the spread of five strains of potato virus X were made with seven potato varieties and with tomato plants both under glass and in the field. Spread by leaf contact between healthy and infected plants was confirmed, and it was also found that spread could occur between plants whose only contact was below ground.
The rate of spread was much greater in tomato than in potato plants, and virulent strains of the virus, which achieve a high concentration in infected plants, spread more rapidly than avirulent strains. In only one experiment with potatoes did more than 10% of the healthy potato plants exposed to infection become infected during one season.
Datura stramonium and tomato plants became infected when growing in soil containing sap or residues from X -infected plants.
It was common in the field for potato plants whose foliage gave no reaction for virus X at the end of the season to yield a mixed progeny of healthy and infected tubers. Such infections are thought to result from underground spread.
Attempts to transmit virus X from infected to healthy potatoes by means of Rhizoctonia solani failed. No examples of infection were found except when healthy plants came into direct contact with sources of the virus.