The effect of inoculation with an attenuated mutant strain of tobacco mosaic virus on the growth and yield of early glasshouse tomato crops

In trials in 1973-5 at the Glasshouse Investigational Unit for Scotland, the yield of fruit from tomato cv. Eurocross BB inoculated at the seedling stage with the Mil-16 attenuated strain of tobacco mosaic virus was 5–8-9-4% greater than that from uninoculated plants which became naturally infected with a severe indigenous strain of the virus within 7–8 wk of planting. The increase in fruit yield, particularly of better grades, resulted in higher gross financial returns (up to 25p/plant) from inoculated plants. The yields from the Mil-16 protected plants were up to 14% greater than those from plants artificially inoculated at the seedling stage with the indigenous severe virus. Inoculation with Mil-16 had little adverse effect on early growth or the rate of fruit development on the first five trusses, but in 1973 the final yield of inoculated plants was depressed c. 5% compared with that from plants substantially free from infection for 14 wk after planting. In 1 year's test no benefit from inoculation with Mil-16 was recorded in cv. Cudlow Cross.     

The establishment of a new strain of tobacco mosaic virus resulting from the use of resistant varieties of tomato

Tomato varieties with monogenic resistance (tolerance) to tobacco mosaic virus (TMV) were used by British growers for the first time in 1966. Samples of TMV collected before 1966, and in 1967 and 1968, were tested for their ability to produce virus symptoms on a series of isogenic tomato differential hosts differing in three factors for TMV resistance. Samples collected before 1966 yielded only strain o of TMV, which was unable to overcome any of the three factors. Strain 1, able to infect the newly-introduced resistant varieties, was found with increased frequency after 1966. It was found less frequently on nurseries which had resumed growing susceptible varieties, suggesting that in these it may be unable to compete successfully with strain o. The dangers of indiscriminate release of varieties protected from disease by only one gene are discussed.     

Nucleotide sequence of the tobacco mosaic virus (tomato strain) genome and comparison with the common strain genome

The sequence of about 4,500 nucleotides of the internal part of tobacco mosaic virus (TMV)-tomato strain (L) RNA has been newly determined using cloned cDNAs. Together with the previously determined partial sequences at both ends, the entire sequence of the 6,384 nucleotide genome has been completed. The 130K (1,115 amino acids), 180K (1,615 amino acids), 30K (263 amino acids) and coat protein (158 amino acids) cistrons are located at residues 72-3442, 72-4922, 4906-5700, and 5703-6182 on the genome, respectively. Sequence polymorphism was not observed except for heterogeneity in the length of the A cluster near the 3' end. The homology of the nucleotide sequences of TMV-L and TMV-vulgare, a common strain, is about 80% on average. Remarkable differences between them were found in a part of the N-terminal portion of the 130K/180K protein and the C-terminal portion of the 30K protein. A new method for cDNA cloning was developed by which the cDNA of the 5'-terminus of viral RNA can be cloned efficiently.     

Strain-genotype interaction of tobacco mosaic virus in tomato

The symptoms and virus content of isogenic tomato genotypes differing by three tobacco mosaic virus (TMV) resistance factors, Tm-I, Tm-2 and Tm-2², were studied in relation to various isolates of TMV and four strains were identified. The common strain induced no symptoms on plants with any of the factors for resistance, one strain caused symptoms on Tm-I plants, one on Tm-2 plants and one on both Tm-I and Tm-2 plants and also on Tm-I Tm-2 plants. No strain induced symptoms on Tm-2² plants. The gene, Tm-I, was found to be dominant or incompletely dominant for preventing symptom development but was recessive or intermediate for limiting virus multiplication of the common strain. Both Tm-2 and Tm-2² were dominant for a hypersensitive response to the common strain. Virus multiplication was temperature-dependent. The background or varietal genotype did not affect virus multiplication. A systemic necrosis of Tm-2² plants occurred only when heterozygous Tm-2² was not protected by other factors against specific strains of TMV. The complexity of the host genotype, pathogen genotype and environment interactions are outlined and the exploitation of the resistance factors in tomato breeding discussed.     

Biological and physico-chemical properties of an atypical mutant of tobacco mosaic virus

Summary A mutant of tobacco mosaic virus (TMV) was isolated from a population of TMV-RNA molecules inactivated to 0.1% level of survival by treatment with nitrous acid This mutant is poorly transported in Samsun tobacco and induces chlorotic spots, oak-leaf patterns, ringspots and necrosis. Rate zonal sucrose density-gradient and equilibrium sedimentations in sucrose and CsCl indicate that the purified mutant preparations contain a heterogenous population of particles including some free coat protein. These vary in length, but approximately 5–10% are infectious rods of standard (15×300 nm) dimensions. Purified mutant RNA contain an overabundance of fragmented and a few infectious molecules. Compared to the parent strain, the mutant capsid protein contains two additional residues of arginine and one of glycine, but lacks one residue each of glutamic acid (or glutamine), serine and threonine. These observations and other data based on temperature sensitivity, exposure to urea and serology indicate that the mutant particles are inherently unstable and break during the purification procedure.     

Unique nature of an attenuated strain of tobacco mosaic virus: autoregulation

An attenuated strain L11A of tobacco mosaic virus (TMV) multiplied like wild type strain L at an early stage of infection in tomato leaves. Four days after inoculation, however, multiplication of L11A was drastically reduced (autoregulation) compared with the constant multiplication of L. In mixed infections, L11A strongly inhibited the multiplication of homologous strain L. Experiments with cucumber mosaic virus (CMV) or tobacco plants revealed that the inhibitory mechanism of L11A is not host-specific but virus-specific, and the autoregulatory mechanism is effective only for TMV. RNA synthesis in L11A infected leaves 4 days after inoculation was studied by polyacrylamide gel electrophoresis. Synthesis of TMV-RNA and its replicative intermediate were strongly inhibited, whereas the replicative form of TMV-RNA and ribosomal RNA were synthesized as in the case of L infection. Synthesis of non-coat-protein was studied by the incorporation of radioactive histidine into subcellular fractions derived from leaves infected with L or L11A for 4 days. Different patterns of the two strains in protein synthesis were noted. At least three proteins were predominantly synthesized in L11A infection. One of them was observed in the mitochondria fraction. From its position in polyacrylamide gel, it could be viral coded 165K protein which is considered to be involved in viral RNA replication. These results suggest that the unique nature of attenuated virus L11A, i.e. autoregulation, resulted from the inhibitory mechanism of viral RNA synthesis due to overproduction of 165K protein and is quite distinct from interferon, intrinsic interference or interference by defective virus.     

Cross protection in transgenic tobacco plants expressing a mild strain of tobacco mosaic virus

Summary Cross protection of plant viruses is a phenomenon in which plants infected with one strain of a virus are protected from the effects of superinfection by other related strains. Recently, we have succeeded in the introduction and expression of a cDNA copy of the tobacco mosaic virus (TMV) genomic RNA in transgenic tobacco plants. Using this system, we introduced a cDNA copy of a mild strain of TMV into tobacco plants. The transgenic plants did not develop any severe symptoms upon inoculation with a virulent TMV strain, indicating that these transgenic plants were cross protected against TMV infection. The system described here can be a useful model system to study the mechanism(s) of cross protection.     

Strain changes in populations of tobacco mosaic virus from tomato crops

The incidence of tobacco mosaic virus (TMV) Strain 1 in tomato crops of TMV-susceptible cultivars increased from 12% in 1971 to 64% in 1974. This change coincided with the introduction of cross protection to reduce the effects of tomato mosaic. Of the isolates collected from nurseries where the technique was used in 1974, 94% were of Strain 1 compared with 39 % from the other nurseries. No new strains were found and all the isolates were tomato forms of TMV.     

Interaction of tomato mosaic virus movement protein with tobacco RIO kinase

Tomato mosaic virus (ToMV) has a regulatory gene encoding a movement protein (MP) that is involved in the cell-to-cell movement of viral RNA through plasmodesmata. To identify the host cell factors interacting with ToMV MP, we used a recombinant MP probe to isolate cDNA clones from a phage expression library of Nicotiana tabacum by a far-Western screening method. One of the cDNA clones encoded an MP-interacting protein, MIP-T7, homologous to the yeast novel protein kinase, Rio1p. We isolated a full-length cDNA by RT-PCR. The putative gene product was designated NtRIO, and shared 33 and 73% amino acid identity with yeast and Arabidopsis RIO kinases, respectively. In vitro analyses using recombinant proteins showed that NtRIO also interacted with a different MP derived from Cucumber mosaic virus. NtRIO had autophosphorylation activity and phosphorylated ToMV MP. Addition of recombinant tobacco casein kinase 2 resulted in a marked increase in the phosphorylation of NtRIO. The interaction between NtRIO and ToMV MP was inhibited by phosphorylation of NtRIO.     

The movement of tobacco mosaic viruses and potato virus X through tomato plants

Tomato aucuba mosaic virus, tobacco mosaic virus and potato virus X took 3'5-4, 5 and 3 days respectively to move from inoculated tomato leaflets into the petioles and stems
On reaching the stem each virus usually first moved downward, but in some plants both upward and downward movement occurred simultaneously and in a few
upward movement occurred first.
All three viruses travelled through the stem at approximately the same rate. Each was capable of travelling more than 80 cm. during the first 12 hr. after entering the stem, giving a minimal average rate of about 8 cm. per hr.
Uninfected pieces of stem invariably occurred between infected pieces. Maximum lengths of stem through which virus particles had apparently passed without causing infection, were 44.5, 49 and 39 cm. for the three viruses.     

Detection of tobacco mosaic virus and tomato mosaic virus in pepper seeds by enzyme linked immunosorbent assay (ELISA)

Pepper seed samples were tested for the infection of tobacco mosaic virus (TMV) and tomato mosaic virus (ToMV) by enzyme linked immunosorbent assay (ELISA). Out of 26 pepper seed samples tested, 17 were infected with TMV and ToMV in ELISA. About 34.7% of pepper seed samples were found to be healthy. Infections of TMV or ToMV were recorded to be 61.53% and 11.5%, respectively of the total tested seed samples.