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nucleotide sequence of a cucumber mosaic virus satellite rna associated with a tomato top stunting. 19921282706
a single nucleotide change within a plant virus satellite rna alters the host specificity of disease induction.some rna plant viruses contain satellite rnas which are dependent upon their associated virus for replication and encapsidation. some cucumber mosaic virus satellite rnas induce chlorosis on any of several host plants, including either tobacco or tomato. the exchange of sequence domains between cdna clones of chlorosis-inducing and non-pathogenic satellite rnas delimited the chlorosis domain for both tobacco and tomato plants to the same region. site-directed mutagenesis of one nucleotide (149) ...19921284656
differential interactions among strains of tomato aspermy virus and satellite rnas of cucumber mosaic virus.tomato and tobacco plants were inoculated with either of two strains of tomato aspermy virus, 1-tav or v-tav, and each of six isolates of cucumber mosaic virus satellite rna (cmv-satrna), b1, b2, b3, ix, or wl2. ribonuclease protection assays, used to detect total satrna and encapsidated satrna, revealed that g-satrna generated new satellite rna not of the inoculated sequence. the other cmv-satrnas were compared for their ability (1) to replicate, (2) to modulate symptoms, (3) to reduce tav accu ...19921370738
cucumber mosaic virus satellite rna (y strain): analysis of sequences which affect yellow mosaic symptoms on tobacco.plants infected with cucumber mosaic virus (cmv) (kin strain) produce a mild mosaic disease on tobacco whereas infections of cmv with satellite rna (strain y) cause a severe yellow mosaic. analysis of recombinant and mutant forms of satellite rna identified a site (nucleotides 185/186) in the y satellite rna that affects the ability to induce the yellow mosaic in combination with cmv but not with tomato aspermy virus. the location of this site with respect to other mutations in the satellite rna ...19901698919
differential replication in zucchini squash of a cucumber mosaic virus satellite rna maps to rna 1 of the helper virus.cucumber mosaic virus (cmv) supports the replication and encapsidation of its satellite rna, both in solanaceous and cucurbit host plants; however, different strains of cmv support the replication of satellite rnas with different efficiency. in addition, replication of satellite rna is very efficient in solanaceous host plants and generally poor in cucurbit host plants. the wl1-satellite (wl1-sat) rna is an exception, and replicates to high levels in both solanaceous and curcubit host plants wit ...19911704659
replication of cucumber mosaic virus satellite rna in vitro by an rna-dependent rna polymerase from virus-infected tobacco.an rna-dependent rna polymerase purified from tobacco infected with cucumber mosaic virus catalyzes the synthesis of (-) and (+) strands of the viral satellite rna, carna 5, but fails to replicate the satellite rna of peanut stunt virus (psv). the enzyme replicates the genomic rnas of the three principal cucumoviruses cmv, psv and tomato aspermy virus (tav) with varying efficiencies. the specificity with which cmv rdrp replicates different sequence-unrelated rna templates suggests that the site ...19911720399
serial passage of infectious transcripts of a cucumber mosaic virus satellite rna clone results in sequence heterogeneity.populations of the d-satellite rna (d-sat) of cucumber mosaic virus (cmv) have a major site of heterogeneity at which a significant proportion of the molecules vary from each other. the generation of this heterogeneity was investigated by using infectious transcript rna of a prototype d-sat clone as an inoculum of uniform sequence to initiate serial passages in five different host plant species. rnase protection assays indicated that heterogeneity at the same site seen in natural d-sat rna popul ...19902158699
induction of tobacco chlorosis by certain cucumber mosaic virus satellite rnas is specific to subgroup ii helper strains.two satellite (sat) rnas of cucumber mosaic virus (cmv), b2- and wl3-sat rnas, which induce systemic chlorosis on tobacco, were inoculated onto tobacco with a number of cmv strains. systemic chlorosis was observed only when these satellite rnas were associated with subgroup ii cmv strains. infection of tobacco with various pseudorecombinants of subgroup i and ii cmv strains, together with wl3- or b2-sat rna, suggests that chlorosis is associated with rna 2 of subgroup ii cmv strains. chlorosis w ...19902330677
cucumber mosaic virus satellite rna (strain y): analysis of sequences which affect systemic necrosis on tomato.the location of a sequence within the y satellite rna of cucumber mosaic virus (cmv) that confers the ability to induce necrosis on tomato plants has been analysed using chimeric satellite rnas. these recombinant rna molecules contained parts of the y (necrogenic) and ra (benign) satellite rnas and were inoculated into tomato plants together with cmv helper virus. from the composition of the recombinant satellite rnas that induced necrosis it was concluded that, of the nucleotides which differ b ...19902374005
a gene coding for a monomeric form of cucumber mosaic virus satellite rna confers tolerance to cmv.a gene whose transcript bears a monomeric form of cucumber mosaic virus (cmv) satellite rna was introduced into tobacco (nicotiana tabacum 'xanthi' nc) plants by using an ri plasmid-based vector system. on cmv infection, the transcript of the satellite rna gene was used as a template to yield unit-length satellite rna, which was efficiently amplified by the virus. plants bearing the satellite rna gene displayed long-term tolerance to cmv infection and were also tolerant to cmv infection by aphid ...19882485143
determination of sequence and structural requirements for pathogenicity of a cucumber mosaic virus satellite rna (y-satrna).we describe the use of biologically active cdna clones to investigate genetic determinants of a satellite rna that modulates symptoms normally induced by its helper virus, cucumber mosaic virus (cmv). for this purpose, we have investigated a cmv satellite rna (y-satrna) that induces bright yellow symptoms on tobacco and necrosis on tomato. to determine the pathogenicity-modulating domain of y-satrna, several insertion and deletion mutants were created by using various restriction sites in the cd ...19892485234
requirement of 3'-terminal guanosine in (-)-stranded rna for in vitro replication of cucumber mosaic virus satellite rna by viral rna-dependent rna polymerase.the 3' terminus of the (-) rna strand in the replicative forms of several (+)-stranded rna viruses possesses an unpaired guanosine with unknown function. this unpaired guanosine is also found at the 3' terminus of the (-) strand in the double-stranded form of two cucumoviral satellite rnas. using a cucumber mosaic virus (cmv) rna-dependent rna polymerase capable of replicating the satellite rna in vitro, the 3'-terminal guanosine of the satellite (-) strand was shown to be an absolute requiremen ...19947514230
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 ...19947526544
the replication of a necrogenic cucumber mosaic virus satellite is temperature-sensitive in tomato.lethal necrosis development in tomato plants infected with cucumber mosaic virus (cmv) strain d containing the necrogenic satellite d-carna 5 and held at 32 degrees c is shown to be impaired. carna 5 accumulation in tomato at 32 degrees c is reduced about 100-fold compared to accumulation in plants held at 24 degrees c, while viral rna accumulation is reduced about 5-fold. cmv-infected tomato held for 3 days at 24 degrees c prior to shift to 32 degrees c do not develop lethal necrosis. longer in ...19957544109
effect of temperature on cucumber mosaic virus satellite-induced lethal tomato necrosis is helper virus strain dependent.the effect of temperature on the response of tomato (lycopersicon esculentum mill. cv. rutgers) to infections with the necrogenic cucumber mosaic virus (cmv) satellite d-carna 5 was investigated with each of four cmv strains d, 1, y and s functioning as helper virus. at 24 degrees c lethal necrosis was observed in all infections. however, at 32 degrees c the response varied from total absence or reduction of necrosis with some strains to accelerated lethal necrosis with others. the total lack of ...19957544110
support of a cucumber mosaic virus satellite rna maps to a single amino acid proximal to the helicase domain of the helper virus.cucumber mosaic virus (cmv) is a tripartite rna virus that can support the replication of satellite rnas, small molecular parasites of the virus. satellite rnas can have a dramatic effect on the helper virus and the host plant in a manner specific to the helper, satellite, and host. previously, we showed that the sny-cmv strain is not able to support the replication of the wl1 satellite rna in zucchini squash and that this phenotype maps to rna 1. in the present study, we use recombinant cdna cl ...19978985390
evolutionary dynamics of cucumber mosaic virus satellite rna during natural epidemics in italy.the evolutionary dynamics of 22 variants of cucumber mosaic virus satellite rna (cmv satrna) isolated in italy during virus epidemics from 1988 to 1993 were investigated on the basis of their primary structure and biological properties. most of the variants were amplified from total nucleic acid preparations extracted from field-infected plants, thus representing wild isolates of cmv satrna. eleven variants were associated with subgroup ii cmv strains, 10 with subgroup i and 1 with a mixed infec ...19979123858
formation of multimers of cucumber mosaic virus satellite rna.double-stranded rna multimers of cucumber mosaic virus (cmv) satellite rna were detected in cmv-infected plants. rt-pcr showed that plus-sense and minus-sense monomers and plus-sense multimers of satellite rna were present. multimeric minus-sense rna was not present except in the form of multimeric dsrna. sequence analysis of 52 cloned junction regions in head-to-tail repeats of unit-length satellite rna indicated that about 35% of the junction sequences were precise fusions of monomer units, 56 ...19979129669
[studies on construction of artificial mutants of cucumber mosaic virus satellite rna and their biological activity].based on the full length cdna clone of a cucumber mosaic virus satellite rna, which was 369nt in size, artificial mutants were developed by the method of error-prone pcr and dna shuffling. the new satellite cdnas were transcribed in vitro into ssrna and pseudo-recombined with a helper cucumber mosaic virus, which contains no satellite rna. sequence analysis showed that a to t/g or g to a replacement all the four mutants, named ms1, ms5, ms6 and ms11 respectively, and there is no c to g or g to c ...200516245882
[construction of infectious cdna clone of cucumber mosaic virus satellite rna xjs1 and preliminary study on its biological function].cucumber mosaic virus (cmv) sugar beet isolate caused yellow mosaic, leaf distortion, crinkle and stunt symptoms on sugar beet in nature. it exhibited some special biological properties with narrower host range and had no symptom on nicotiana glutinosa l. and nicotiana tobacum l. cv. nc-89. a new satellite rna, xjs1 was found to be associated with the helper virus. in order to know the cause of the special pathogenicity of the cmv isolate. full-length infectious cdna clone of cmv satellite rna x ...200616736580
dcl4 targets cucumber mosaic virus satellite rna at novel secondary structures.it has been reported that plant virus-derived small interfering rnas (vsirnas) originated predominantly from structured single-stranded viral rna of a positive single-stranded rna virus replicating in the cytoplasm and from the nuclear stem-loop 35s leader rna of a double-stranded dna (dsdna) virus. increasing lines of evidence have also shown that hierarchical actions of plant dicer-like (dcl) proteins are required in the biogenesis process of small rnas, and dcl4 is the primary producer of vsi ...200717609283
in vitro translation of cucumoviral satellites. i. purification and nucleotide sequence of cucumber mosaic virus-associated rna 5 from cucumber mosaic virus strain s.the satellite cucumber mosaic virus (cmv)-associated rna 5 (carna 5) of cmv strain s (cmv-s) which previously had been assigned the capability both to direct the synthesis of two small proteins in vitro (r. a. owens and j. m. kaper, 1977, virology, 80, 196-203) and to induce the tomato necrosis disease in the presence of its helper virus (j. m. kaper and h. e. waterworth, 1977, science, 196, 429-431), has been reinvestigated. polyacrylamide gel electrophoretic analyses under partially denaturing ...198618640643
characterizing multiple exogenous and endogenous small rna populations in parallel with subfemtomolar sensitivity using a streptavidin gel-shift assay.here we present a simple and inexpensive gel-shift assay for the detection and quantification of small rnas. the assay is at least 5-10 times more sensitive than a conventional northern, and is highly scalable. total rna is first size purified to enrich the desired size range, phosphatase treated, and then radiolabeled to high specific activity using polynucleotide kinase. the resulting rna stock is then hybridized to an excess of biotinylated dna probe oligonucleotide, prior to mixing with stre ...200919237462
oligonucleotide microarray-based detection and identification of 10 major tomato viruses.a dna microarray chip was developed for screening 10 major economically important tomato viruses from infected plants using "combimatrix" platform 40-mer oligonucleotide probes. a total of 279 oligonucleotide virus probes were specific for simultaneous multiple detection, identification, differentiation and/or genotyping of each of the following tomato rna viruses and/or strains and a virus satellite: cucumber mosaic virus, cucumber mosaic virus satellite rna, tomatoinfectiouschlorosisvirus, tom ...201020470828
cucumber mosaic virus satellite rnas that induce similar symptoms in melon plants show large differences in fitness.two groups of cmv-satrnas, necrogenic and non-necrogenic, can be differentiated according to the symptoms they cause in tomato plants, a host in which they also differ in fitness. in most other cmv hosts these cmv-satrna cause similar symptoms. here we analyse if in melon plants, in which the two groups of cmv-satrnas cause similar symptoms, they differ in traits determining their relative fitness. for this, 10 necrogenic and 10 non-necrogenic field satrna genotypes were assayed with fny-cmv as ...201121562122
differential effects of cucumber mosaic virus satellite rnas in the perturbation of microrna-regulated gene expression in tomato.viral infections generally cause disease symptoms by interfering with the microrna (mirna)-mediated regulation of gene expression of host plants. in tomato leaves, the accumulation levels of eleven mirnas and ten target mrnas were enhanced by different degrees upon cucumber mosaic virus (cmv)-fny and tomato aspermy virus (tav)-bj infections. the ability of cmv-fny to interfere with mirna pathway was dramatically suppressed in the addition of the benign satellite (sat) rna variant (satyn12), but ...201121590278
heterologous replicase driven 3' end repair of cucumber mosaic virus satellite rna.to investigate the extent of the 3' end repair in a satellite rna of cucumber mosaic virus (cmv) strain q (q(sat)) by a heterologous tomato aspermy virus (tav), a set of biologically active agrotransformants corresponding to the three genomic rnas of tav was developed. analysis of nicotiana benthamiana plants agroinfiltrated with tav and either wild type or each of the six 3' deletion mutants of q(sat) revealed that (i) heterologous replicase failed to generate q(sat) multimers, a hallmark featu ...201525705791
nicotiana small rna sequences support a host genome origin of cucumber mosaic virus satellite rna.satellite rnas (satrnas) are small noncoding subviral rna pathogens in plants that depend on helper viruses for replication and spread. despite many decades of research, the origin of satrnas remains unknown. in this study we show that a β-glucuronidase (gus) transgene fused with a cucumber mosaic virus (cmv) y satellite rna (y-sat) sequence (35s-gus:sat) was transcriptionally repressed in n. tabacum in comparison to a 35s-gus transgene that did not contain the y-sat sequence. this repression wa ...201525568943
cloning and profiling of small rnas from cucumber mosaic virus satellite rna.rna silencing is not only a gene regulation mechanism that is conserved in a broad range of eukaryotes but also an adaptive immune response against foreign nucleic acids including viruses in plants. a major feature of rna silencing is the production of small rna (srna) of 21-24 nucleotides (nt) in length from double-stranded (ds) or hairpin-like (hp) rna by dicer-like (dcl) proteins. these srnas guide the binding and cleavage of cognate single-stranded (ss) rna by an rna silencing complex. like ...201525287499
functional significance of a hepta nucleotide motif present at the junction of cucumber mosaic virus satellite rna multimers in helper-virus dependent replication.satellite rnas (satrna) associated with cucumber mosaic virus (cmv) have been shown to generate multimers during replication. we have discovered that multimers of a cmv satrna generated in the absence of its helper virus (hv) are characterized by the addition of a hepta nucleotide motif (hnm) at the monomer junctions. here, we evaluated the functional significance of hnm in hv-dependent replication by ectopically expressing wild type and mutant forms of satrna multimers in planta either in (+) o ...201323146208
correction: nicotiana small rna sequences support a host genome origin of cucumber mosaic virus satellite rna.[this corrects the article doi: 10.1371/journal.pgen.1004906.].201525915543
analysis of mechanisms involved in the cucumber mosaic virus satellite rna-mediated transgenic resistance in tomato plants.transgenic tomato (lycopersicon esculentum mill. cv. uc82) plants expressing a benign variant of cucumber mosaic virus satellite rna (cmv tfn-satrna) were generated. the transformed plants did not produce symptoms when challenged with a satrna-free strain of cmv (cmv-fl). the same plant lines initially were susceptible to necrosis elicited by a cmv strain supporting a necrogenic variant of satrna (cmv-77), but a phenotype of total recovery from the necrosis was observed in the newly developing l ...200414714873
template-independent repair of the 3' end of cucumber mosaic virus satellite rna controlled by rnas 1 and 2 of helper virus.rna viruses which do not have a poly(a) tail or a trna-like structure for the protection of their vulnerable 3' termini may have developed a different strategy to maintain their genome integrity. we provide evidence that deletions of up to 7 nucleotides from the 3' terminus of cucumber mosaic cucumovirus (cmv) satellite rna (satrna) were repaired in planta in the presence of the helper virus (hv) cmv. sequence comparison of 3'-end-repaired satrna progenies, and of satrna and hv rna, suggested th ...19989573276
analysis of the in vitro secondary structure of cucumber mosaic virus satellite rna.the solution conformation of two variants of cucumber mosaic virus satellite rna (cmv satrna) was analyzed using several enzymatic and chemical probes. ribonuclease t1 and nuclease s1 were used to map unpaired nucleotides, and nuclease v1 was used to detect double-stranded, or stacked, bases. chemical probing with dimethylsulphate and diethylpyrocarbonate also identified unpaired and unstacked nucleotides, respectively. modified or cleaved positions were identified by direct gel electrophoresis ...19979292503
variation in the hypervariable region of cucumber mosaic virus satellite rnas is affected by the helper virus and the initial sequence context.the d satellite rna (sat rna) of cucumber mosaic virus (cmv) was previously shown to contain a region of hypervariability around nucleotide 230, in wild-type populations and in cdna clones and progeny of one such clone (pdsat4) after passage with the subgroup i strain fny-cmv. this hypervariable region (hvr) consists of a series of consecutive a and/or u residues. we found that variability is also generated in the hvr of transcript derived from pdsat4 after passage with the subgroup ii strain ls ...19957530401
replication of cucumber mosaic virus satellite rna from negative-sense transcripts produced either in vitro or in transgenic plants.both positive [(+)] and negative [(-)] sense versions of two satellite rna (satrna) genes from cucumber mosaic virus (cmv), the necrogenic i17n and the nonnecrogenic r, have been introduced into the genome of tobacco plants. on infection with satrna-free cmv, satrna was amplified in plants expressing each of the four genes. all four genes confer protection against cmv infection. however, co-inoculation of plants with viral rna and cmv satrna transcripts synthesized in vitro showed that (-) sense ...19947513743
genetic analysis of helper virus-specific selective amplification of cucumber mosaic virus satellite rnas.satellite rnas (sat-rnas) are small molecular parasites associated with a number of plant rna viruses. the cucumber mosaic virus (cmv) sat-rnas are ca. 335 nucleotides and have evolved to produce a large number of closely related sat-rnas. different cucumoviruses can act as helper viruses in the amplification of cmv sat-rnas. we have found that different helper viruses show a preference for a particular sat-rna in a mixed infection. in this study the specificity of wl47 sat-rna amplification by ...19957536251
in vitro synthesis of infectious rnas from cdnas of cucumber mosaic virus satellite rna (strain y) after removal of non-viral bases with ribonuclease h. 19872827104
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