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At least 19 recordsLinked to original sources
PubMed2002

In situ localization and tissue distribution of the replication-associated proteins of Cucumber mosaic virus in tobacco and cucumber.

The replication-associated proteins encoded by Cucumber mosaic virus (CMV), the 1a and 2a proteins, were detected by immunogold labeling in two host species of this virus, tobacco (Nicotiana tabacum) and cucumber (Cucumis sativus). In both hosts, the 1a and 2a proteins colocalized predominantly to the vacuolar membranes, the tonoplast. While plus-strand CMV RNAs were found distributed throughout the cytoplasm by in situ hybridization, minus-strand CMV RNAs were barely detectable but were found associated with the tonoplast. In both cucumber and tobacco, 2a protein was detected at higher densities than 1a protein. The 1a and 2a proteins also showed quantitative differences with regard to tissue distributions in tobacco and cucumber. About three times as much 2a protein was detected in CMV-infected cucumber tissues as in CMV-infected tobacco tissues. In tobacco, high densities of these proteins were observed only in vascular bundle cells of minor veins. In contrast, in cucumber, high densities of 1a and 2a proteins were observed in mesophyll cells, followed by epidermis cells, with only low levels being observed in vascular bundle cells. Differences were also observed in the distributions of 2a protein and capsid protein in vascular bundle cells of the two host species. These observations may represent differences in the relative rates of tissue infection in different hosts or differences in the extent of virus replication in vascular tissues of different hosts.

Cucumis sativus↗
PubMed2005

Transgenic cucumbers harboring the 54-kDa putative gene of Cucumber fruit mottle mosaic tobamovirus are highly resistant to viral infection and protect non-transgenic scions from soil infection.

Cucumber fruit mottle mosaic tobamovirus (CFMMV) causes severe mosaic symptoms and yellow mottling on leaves and fruits and, occasionally, severe wilting of cucumber (Cucumis sativus L.) plants. No genetic source of resistance against this virus has been identified in cucumber. The gene coding for the putative 54-kDa replicase gene of CFMMV was cloned into an Agrobacterium tumefaciens binary vector, and transformation was performed on cotyledon explants of a parthenocarpic cucumber cultivar. R1 seedlings were screened for resistance to CFMMV by symptom expression, back inoculation on an alternative host and ELISA. From a total of 14 replicase-containing R1 lines, eight resistant lines were identified. Line 144--homozygous for the putative 54-kDa replicase gene--was immune to CFMMV infection by mechanical and graft inoculation, and to root infection following planting in CFMMV-infested soil. A substantial delay of symptom appearance was observed following infection by three additional cucurbit-infecting tobamoviruses. When used as a rootstock, line I44 protected susceptible cucumber scions from soil infection by CFMMV. This paper is the first report on protection of a susceptible cultivar against a soil-borne viral pathogen, by grafting onto a transgenic rootstock.

Base Sequence↗
PubMed1982

Comparative studies on ribonucleic acid dependent RNA polymerases in cucumber mosaic virus infected cucumber and tobacco and uninfected tobacco plants.

RNA-dependent RNA polymerases have been isolated in almost pure form from cucumber mosaic virus (CMV) infected cucumber cotyledons and from tobacco leaves and were compared with the less pure enzyme from uninfected tobacco. The purified polymerase from cucumber shows on sodium dodecyl sulfate gels two peptide chains of about 100 and 112 kdaltons. The enzyme from tobacco shows a close doublet of about 125 kdaltons, which is also present in the less pure preparation from healthy tobacco. While both the cucumber and tobacco enzymes can use many polynucleotides and RNAs as templates, considerable quantitative differences exist, poly(C) being by far the most effective template for the cucumber enzyme but of low activity with the tobacco enzyme and poly(UG) being highly active with the latter but not the former. Poly(A) and poly(G) are inactive. Different viral RNAs, including CMV RNA, show smaller differences. The sedimentation rates of the enzyme from both sources are the same as that of gamma-globulin. A uridine 5'-triphosphate (UTP) terminal transferase also present in both plants sediments much more slowly and can be completely removed from the RNA polymerases. However, slight nucleolytic activity remains associated with the purified polymerases and appears to be proportional to the polymerase activity. The conclusion derived from these data is that the RNA-dependent RNA polymerases of different plants differ and are not detectably affected by virus infection in qualitative terms while being produced in greatly increased amounts upon some virus infections. Similar conclusions were previously reached with less purified enzyme preparations from tobacco as compared to cowpea, infected with different viruses, if any.

Kinetics↗
PubMed2003

Tyrosinase inhibitory activity of cucumber compounds: enzymes responsible for browning in cucumber.

The inhibition of mushroom tyrosinase by cucumber extracts was evaluated. The inhibitory effect was measured by both polarographic and spectrophotometric methods. The commercial aldehyde, trans,cis-2,6-nonadienal, described as a major volatile compound of cucumber, was characterized as a noncompetitive inhibitor against 4-tert-butylcatechol oxidation by mushroom tyrosinase. The K(I) obtained was 3.4 mM. Polyphenol oxidase (PPO) activity was not detected in cucumber skin extracts. However, the presence of PPO was revealed by Western blot; a single band was found with a M(r) of 53 kDa. These results support the assumption that the enzyme PPO is present in the cucumber skin, but its activity is inhibited. Peroxidase (PO) was also found in cucumber skin extracts. This enzyme was detected in the soluble fraction but not in the membrane fraction. The kinetic characterization of PO was carried out. Native isoelectric focusing revealed several acidic PO isoenzymes with a pI in the range between 5 and 6, a basic isoenzyme, and one principal neutral isoenzyme of pI = 7.2.

Catechol Oxidase↗
PubMed2026

Strain Diversity and Resistance to Cucumber Green Mottle Mosaic Virus (CGMMV) in Cucumber.

Cucumber green mottle mosaic virus (CGMMV) is a tobamovirus that causes disease in cucumber crops worldwide, leading to significant economic losses. To study the variability of CGMMV in southeastern Spain, partial genome sequences were obtained from isolates collected in 2017 and 2020 from cucumber crops. Phylogenetic analyses revealed that isolates clustered into two major groups, Asian (AS)-like and European (EU)-like CGMMV isolates. These two groups coexisted in the same area, crops, and even individual plants, although the AS type predominated. The accumulation and symptom expression of molecularly cloned isolates from these two groups were assessed in two cucumber cultivars (resistant and susceptible) under both summer and winter conditions. An in planta antagonistic interaction was detected between the AS and EU isolates, in which the accumulation of CGMMV-EU was suppressed during mixed infections. A multivariate analysis did not identify statistically significant differences because of variations in environmental conditions. Unlike CGMMV-EU, CGMMV-AS did not show significant differences in accumulation based on the plant genotype. To further investigate this, the response to CGMMV-AS infection was analyzed in additional susceptible and resistant cultivars. All cultivars appeared to be similarly susceptible to CGMMV-AS, in contrast to CGMMV-EU, which accumulated to a much lower extent in resistant compared with susceptible plants. These results reinforce the need to continue epidemiological surveillance, identify new sources of resistance, and implement strict control of infected seed trade, given the growing threat that CGMMV-AS isolates pose to cucumber cultivation.

agroinfectious clone↗
PubMed2004

Isolation of cucumber CsARF cDNAs and expression of the corresponding mRNAs during gravity-regulated morphogenesis of cucumber seedlings.

Cucumber seedlings show positive gravitropism and bend in the transition zone between the hypocotyl and the root. The peg, a specialized protuberance, develops on the concave side of the bending transition zone. Auxin and the mRNA of an auxin-inducible gene (CsIAA1) isolated from cucumber are differentially accumulated across the transition zone during the gravity-regulated peg formation. In this study, five cDNAs of Auxin Response Factors (ARFs) from cucumber were isolated and their mRNA accumulation was compared with that of CsIAA1. The tissue specificity of CsARF2 mRNA accumulation was similar to that of CsIAA1. Because the structural character of CsARF2 predicts that it is a transcriptional activator, CsARF2 may be involved in the activation of CsIAA1 transcription, which plays a role in gravity-regulated peg formation. Neither gravity nor auxin affected mRNA accumulation of five CsARFs including CsARF2, suggesting that CsARF2 may be regulated at a post-transcriptional level to induce the asymmetric expression of the CsIAA1 gene in response to gravistimulation and auxin in cucumber seedlings.

Amino Acid Sequence↗
PubMed2006

Potential involvement of a cucumber homolog of phloem protein 1 in the long-distance movement of Cucumber mosaic virus particles.

The systemic movement of Cucumber mosaic virus (CMV) in cucumber plants was analyzed. The structure that is translocated and its putative interactions with phloem components were analyzed in phloem exudate (PE) samples, which reflect sieve tubes stream composition. Rate zonal centrifugation and electron-microscopy analyses of PE from CMV-infected plants showed that CMV moves through sieve tubes as virus particles. Gel overlay assays revealed that CMV particles interact with a PE protein, p48. The amino-acid sequence of several tryptic peptides of p48 was determined. Partial amino-acid sequence of p48 showed it was a cucumber homolog of phloem protein 1 (PP1) from pumpkin, with which p48 also shares several chemical properties. PP1 from pumpkin has plasmodesmata-gating ability and translocates in sieve tubes. Encapsidated CMV RNA in PE samples from infected plants was less accessible to digestion by RNase A than RNA in purified CMV particles, a property that was reconstituted by the in vitro interaction of purified CMV particles and protein p48. These results indicate that the interaction with p48 modifies CMV particle structure and suggest that CMV particles interact with the cucumber homolog of PP1 during translocation in the sieve tubes.

Amino Acid Sequence↗
PubMed2004

Analysis of the systemic colonization of cucumber plants by Cucumber green mottle mosaic virus.

Systemic movement of Cucumber green mottle mosaic virus (CGMMV) in cucumber plants was shown to be from photoassimilate source to sink, thus indicating phloem transport. Nevertheless, CGMMV was not detected by immunocytochemical procedures in the intermediary cell-sieve element complex in inoculated cotyledons, where photoassimilate loading occurs. In stem internodes, CGMMV was first localized in the companion cells of the external phloem and subsequently in all tissues except the medulla, therefore suggesting leakage of the virus from, and reloading into, the transport phloem during systemic movement. In systemically infected sink leaves, CGMMV was simultaneously detected in the xylem and phloem. Interestingly, CGMMV accumulated to high levels in the differentiating tracheids of young leaves implying that the xylem could be involved in the systemic movement of CGMMV. This possibility was tested using plants in which cell death was induced in a portion of the stem by steam treatment. At 24 degrees C, steam treatment effectively prevented the systemic movement of CGMMV, even though viral RNA was detected in washes of the xylem above the steamed internode suggesting that xylem circulation occurred. At 29 degrees C, CGMMV systemically infected steam-treated cucumber plants, indicating that CGMMV can move systemically via the xylem. Xylem transport of CGMMV was, however, less efficient than phloem transport in terms of the time required for systemic infection and the percentage of plants infected.

Cotyledon↗
PubMed1976

Investigation on the infection of cucumber mesophyll protoplasts with cucumber mosaic virus.

Isolated protoplasts from the first leaf mesophyll of cucumber plants have been successfully infected in vitro with cucumber mosaic virus (CMV). Virus instability before, during and after inoculation of the protoplasts resulted in low infectivities when extracts were assayed on cowpea; however, viral RNA extraction improved the bioassay technique. Attempts to optimize inoculation and incubation of protoplasts are outlined, incorporating the improved assay.

Culture Techniques↗
PubMed1999

Virus particles of cucumber green mottle mosaic tobamovirus move systemically in the phloem of infected cucumber plants.

Systemic movement through the phloem of infected host plants is a key process in the life cycle of plant viruses, knowledge of which is scant. A main point to be elucidated is the structural form in which virus infection moves within the phloem. Indirect evidence suggests that virions might be the viral structure that moves in the phloem, but data from direct analysis in phloem sap have not been reported. We have done such analysis in the system cucumber (from which phloem exudate can be collected)/cucumber green mottle mosaic tobamovirus (CGMMV). CGMMV has structurally well-characterized particles. Both CGMMV coat protein and RNA were found in phloem exudate from infected cucumbers. Analysis of the accessibility of CGMMV RNA in phloem exudate to RNase A indicates that it is protected within a ribonucleoprotein structure. The accessibility to RNase A of the RNA in these structures was as in virus particles. Centrifugation analyses showed that the ribonucleoprotein structures in the phloem exudate have the same mass and isopycnic density as virions. Virus particles indistinguishable from purified virions were detected by electron microscopy in phloem exudate. No evidence of free RNA or other CGMMV-related structure was found in phloem exudate of infected plants. These results indicate that CGMMV movement in the phloem occurs mainly, if not exclusively, in the form of virus particles.

Capsid↗
PubMed1996

Cucumber chloroplast trnL(CAA) gene: nucleotide sequence and in vivo expression analysis in etiolated cucumber seedlings treated with benzyladenine and light.

The nucleotide sequence of a 714 bp BamHI-EcoRI fragment of cucumber chloroplast DNA was determined. The fragment contained a gene for tRNA(Leu) together with its flanking regions. The trnL(CAA) gene sequence is about 99% in similarity to broad bean, cauliflower, maize, spinach and tobacco corresponding genes. The relative expression level of the gene was determined by Northern (tRNA) gel blot and Northern (total cellular RNA) slot-blot analyses using the trnL gene probe in 6-day old etiolated cucumber seedlings and the seedlings that had been kept in the dark (dark-grown), treated with benzyladenine (BA) and kept in the dark (BA-treated dark-grown), illuminated (light-grown), and treated with BA and illuminated (BA-treated light-grown), for additional 4, 8 or 12 hr. The trnL transcripts and tRNA(Leu) levels in BA-treated dark-grown seedlings were 5 and 3 times higher, respectively after 4 hr BA treatment, while in the BA treated light-grown seedlings the level of trnL transcripts was only 3 times higher and had no detectable effect on mature tRNA(Leu) when compared to the time-4 hr dark-grown seedlings. However, the level of mature tRNA(Leu) did not show marked changes in the light-grown seedlings, whereas the level of trnL transcripts increases 3 times after 8 hr illumination of dark-grown seedlings. These data indicate that both light and cytokinin can signal changes in plastid tRNA gene expression. The possible regulatory mechanisms for such changes are discussed.

Base Sequence↗
PubMed1994

Mapping helper virus functions for cucumber mosaic virus satellite RNA with pseudorecombinants derived from cucumber mosaic and tomato aspermy viruses.

P-TAV is a strain of tomato aspermy virus (TAV) able to efficiently support the systemic accumulation of some (i.e., B2-satRNA) but not of other (i.e., Ix-satRNA) strains of the satellite RNA (satRNA) of cucumber mosaic virus (CMV) in both tobacco and in tomato. As reported for V-TAV, the failure to support the systemic accumulation of Ix-satRNA seems to be due to an inefficient support of its systemic movement. Pseudorecombinants obtained by the exchange of RNAs 1 + 2 between P-TAV and Trk7-CMV, an efficient helper for the systemic accumulation of Ix-satRNA, were assayed for their ability to support the accumulation of CMV-satRNAs in tobacco plants and protoplasts. Pseudorecombinants having RNAs 1 + 2 from CMV supported the systemic movement and accumulation of CMV-satRNA as efficiently as CMV, whereas pseudorecombinants having RNAs 1 + 2 from TAV supported the CMV-satRNA very poorly. Thus, the ability to support the systemic movement and accumulation of CMV-satRNA is determined primarily by RNAs 1 + 2 and not by RNA 3, which is presumed to encode movement functions in the cucumoviruses and only has a minor, modulating effect on the systemic accumulation of satRNA. This suggests that for systemic movement CMV-satRNA has to interact with (the gene products of) RNAs 1 and/or 2 or that these viral RNAs compete with the satRNA for interaction with the coat or other movement proteins.

Cucumovirus↗
PubMed1997

Characterization of cucumber mosaic virus. IV. Movement protein and coat protein are both essential for cell-to-cell movement of cucumber mosaic virus.

cDNA clones of cucumber mosaic virus (CMV) RNA 3 were modified to express the jellyfish green fluorescent protein (GFP) in place of the 3a movement protein (MP) or coat protein (CP), as fusions to the N (GFP-3a) or C (3a-GFP) terminus of the MP or from a separate open reading frame as part of tricistronic RNAs 3. CMV RNA transcripts containing the individual modified RNAs 3 were unable to infect either Nicotiana tabacum or Nicotiana benthamiana systemically. Infection, as measured by confocal microscopy of GFP fluorescence, generally was limited to one to three epidermal cells at each inoculation site. Limited cell-to-cell movement, but not systemic movement, could be detected by complementation involving expression of MP and CP from two different RNA 3 constructs, each also expressing GFP. Infection involving RNA 3 expressing the GFP-3a fusion showed bright granules of variable size distributed predominantly and nonuniformly throughout the cytoplasm and, to a lesser extent, associated with the cell wall in single fluorescent cells, while infections expressing the 3a-GFP fusion showed bright, punctate fluorescence associated only with the cell wall. Infected cells expressing either 3a-GFP or free GFP showed a halo of less bright, fluorescent, neighboring cells, indicating limited movement of GFP. The initially infected cells also allowed movement of 10-kDa fluorescent dextran to the neighboring halo cells, while infection did not spread, suggesting different requirements for movement of either MP or dextran versus RNA.

Capsid↗
PubMed1998

Transgenically expressed cucumber mosaic virus RNA 1 simultaneously complements replication of cucumber mosaic virus RNAs 2 and 3 and confers resistance to systemic infection.

Tobacco plants transformed with a cDNA copy of RNA 1 of the Fny strain of cucumber mosaic virus (CMV) promoted the asymptomatic accumulation of inoculated viral RNAs 2 and 3, which could be detected in noninoculated leaves, suggesting that the transgene also permitted viral long-distance movement. Typical symptoms of infection appeared later and correlated with the appearance of viral RNA 1 regenerated from the transgenic mRNA. Although all R0-generation plants were susceptible to Fny-CMV, one line displaying variable susceptibility to the virus in R1-and R2-generations led to selected R3-generation lines with systemic resistance to Fny-CMV. In the inoculated leaves of resistant plants, a dramatic decrease in the accumulation of viral RNA 1 was observed, relative to susceptible plants. No viral RNAs were detected in noninoculated leaves of the resistant plants, but such leaves were susceptible to infection. Furthermore, these leaves could sustain replication of inoculated CMV RNAs 2 and 3, indicating that a complete transgene-silencing had not been induced. Although a transgene-mediated, CMV RNA 1-suppression occurred in the inoculated leaf of resistant plants, the absence of a complete systemically acquired silencing suggests the existence of additional interferences with viral infection that prevented systemic infection by viral RNAs 2 and 3.

Cucumovirus↗