| immunodetection of grapevine fanleaf virus satellite rna-encoded protein in infected chenopodium quinoa. | an antiserum was raised against a fusion protein containing the c-terminal half of the protein (p3) encoded by the satellite rna of grapevine fanleaf virus (gflv; f13 isolate) and the n-terminal portion of the ci repressor of phage lambda. this antiserum specifically recognized p3 synthesized in the in vitro wheatgerm translation system and also in infected chenopodium quinoa plants. in these plants, the amount of virus increased for 10 days, then remained constant for up to 21 days, whereas p3 ... | 1992 | 1279107 |
| replication of grapevine fanleaf virus satellite rna transcripts in chenopodium quinoa protoplasts. | a set of full-length cdna clones of the satellite rna of grapevine fanleaf nepovirus isolate f13 (gflv-f13) was constructed with a variable number of additional, non-viral nucleotides at the 5' and 3' ends. the biological activity of the rnas transcribed from these constructs was tested in chenopodium quinoa protoplasts using a helper virus. when inoculated with arabis mosaic virus s (armv-s) rna as helper, transcripts with 33 non-viral nucleotides at the 5' end (tr45p4) did not replicate, where ... | 1992 | 1383395 |
| cross-reacting and heterospecific monoclonal antibodies produced against arabis mosaic nepovirus. | monoclonal antibodies (mabs) were produced against arabis mosaic nepovirus (amv). a hybridoma screening procedure was applied which involved the testing of culture supernatants, before the hybridomas were cloned to single cell lines, for their reaction with eight nepoviruses [amv, cherry leafroll virus (clrv), grapevine fanleaf virus (gflv), peach rosette mosaic virus, raspberry ringspot virus (rrsv), tobacco ringspot virus, tomato black ring virus (tbrv) and tomato ringspot virus]. in addition ... | 1992 | 1402798 |
| effects of site-directed mutagenesis on the presumed catalytic triad and substrate-binding pocket of grapevine fanleaf nepovirus 24-kda proteinase. | grapevine fanleaf nepovirus (gflv) has a bipartite plus-sense rna genome. its structural and functional proteins originate from polyprotein maturation by at least one virus-encoded proteinase. here we describe the cloning of the 24-kda proteinase cistron located between the virus-linked protein (vpg) and the rna-dependent rna polymerase cistron in gflv rna1 (nucleotides 3966 to 4622). proteinase expressed from this clone is able to cleave gflv polyprotein p2 in order to produce the coat protein ... | 1992 | 1519363 |
| amino-acid sequence comparison of nepovirus coat proteins. | the amino acid sequence of the coat proteins of several nepoviruses was determined by a combination of peptide and nucleic acid sequencing (grapevine fanleaf virus, arabis mosaic virus, tomato blackring virus, grapevine chrome mosaic virus). these sequences were compared and showed homologies ranging from 10% to 69%, and 96.7% for the two arabis mosaic virus strains. 10% homology does not reflect any relationship between viruses, and our results implicate, that nepoviruses, considering the homol ... | 1992 | 1589965 |
| complete nucleotide sequence and genetic organization of grapevine fanleaf nepovirus rna1. | the nucleotide sequence of the genomic rna1, 7342 nucleotides (nt) of grapevine fanleaf virus strain f13 (gflv-f13) has been determined from cdna clones. the complete sequence contained only one long open reading frame (orf) of 6852 nucleotides extending from nucleotide 243 to 7101. the putative polyprotein encoded by this orf is 2284 amino acids in length with an mr of 253k. the location of genome-linked protein and comparison of the primary structure of the 253k polyprotein to that of other cl ... | 1991 | 1655953 |
| the nucleotide sequence of a satellite rna associated with arabis mosaic nepovirus. | the nucleotide sequence of a satellite rna (satrna) associated with a lilac isolate of arabis mosaic virus (armv) was determined from cdna copies. the sequence was 1104 nucleotides in length excluding the poly(a) tail, contained a long open reading frame which encodes a polypeptide of 360 amino acids, with an mr of 39k. nucleotide sequence comparisons revealed that the armv-associated satrna shared 83% nucleotide identity with a satrna from grapevine fanleaf nepovirus, but no extensive sequence ... | 1990 | 1693660 |
| cdna sequence of the capsid protein gene and 3' untranslated region of a fanleaf isolate of grapevine fanleaf virus. | | 1991 | 1840668 |
| nucleotide sequence of tomato ringspot virus rna-2. | the sequence of tomato ringspot virus (tomrsv) rna-2 has been determined. it is 7273 nucleotides in length excluding the 3' poly(a) tail and contains a single long open reading frame (orf) of 5646 nucleotides in the positive sense beginning at position 78 and terminating at position 5723. a second in-frame aug at position 441 is in a more favourable context for initiation of translation and may act as a site for initiation of translation. the tomrsv rna-2 3' noncoding region is 1550 nucleotides ... | 1991 | 1856689 |
| transgenic plants and insect cells expressing the coat protein of arabis mosaic virus produce empty virus-like particles. | the 3' end of the rna-2 of arabis mosaic virus (armv) was cloned and sequenced. the n-terminal amino acid sequence of the virion coat protein was determined by edman degradation and the corresponding coding region identified. this gene was modified at the 5' and 3' ends by use of mismatched primers in the polymerase chain reaction (pcr), in order to facilitate the cloning of the gene, and to provide it with a methionine initiation codon. the modified cloned gene was expressed in transgenic plant ... | 1991 | 1875193 |
| cloning and in vitro characterization of the grapevine fanleaf virus proteinase cistron. | the region of the genomic rna-1 from grapevine fanleaf virus isolate f13 (gflv-f13), containing the proteinase cistron and flanking sequences (nucleotides 3894 to 4789) of the gflv polyprotein, was modified by pcr mutagenesis to create a start codon and cloned in a transcription vector. the transcripts from the resulting clone (pvp7) produced, upon translation in rabbit reticulocyte lysate, a 37.8-kda protein which was subsequently cleaved to a stable 28-kda product. autocleavage was maximal at ... | 1991 | 1962449 |
| in vitro expression of a chimeric coat protein gene from grapevine fanleaf virus (strain f 13). | the coat protein (cp) cistron of grapevine fanleaf virus strain f13 (gflv-f13) has been located in the c-terminal region of the 122k polyprotein encoded by the genomic rna 2 [serghini et al. (1990) j. gen. virol. 71: 1433-1441]. a chimeric cp gene of gflv-f13 including a short sequence corresponding to 3 restriction sites, the leader sequence of the gflv-f13 satellite rna and an initiation codon was constructed. transcripts from this construct were translated in wheat germ extract with equal eff ... | 1991 | 2018431 |
| primary structure and location of the genome-linked protein (vpg) of grapevine fanleaf nepovirus. | the genome linked protein vpg covalently linked to the rnas of grapevine fanleaf nepovirus has been sequenced. the vpg (mr = 2931) composed of 24 residues is linked by its n-terminal ser beta-oh group to the viral rnas. the vpg mapped from residues 1218 to 1241 of the 253k polyprotein encoded by gflv rna1. | 1991 | 2060618 |
| rna2 of grapevine fanleaf virus: sequence analysis and coat protein cistron location. | the nucleotide sequence of the genomic rna2 (3774 nucleotides) of grapevine fanleaf virus strain f13 was determined from overlapping cdna clones and its genetic organization was deduced. two rapid and efficient methods were used for cdna cloning of the 5' region of rna2. the complete sequence contained only one long open reading frame of 3555 nucleotides (1184 codons, 131k product). the analysis of the n-terminal sequence of purified coat protein (cp) and identification of its c-terminal residue ... | 1990 | 2374004 |
| the nucleotide sequence of satellite rna in grapevine fanleaf virus, strain f13. | the nucleotide sequence of cdna copies of grapevine fanleaf virus (strain f13) satellite rna has been determined. the primary structure obtained was 1114 nucleotides in length, excluding the poly(a) tail, and contained only one long open reading frame encoding a 341 residue, highly hydrophilic polypeptide of mr37275. the coding sequence was bordered by a leader of 14 nucleotides and a 3'-terminal non-coding region of 74 nucleotides. no homology has been found with small satellite rnas associated ... | 1989 | 2471799 |
| the use of peroxidase anti-peroxidase (pap) complexes in the detection of plant viruses by elisa. | an elisa test system for detection of plant viruses in field samples is described, based on the unlabelled antibody method using a peroxidase-antiperoxidase (pap) complex. novel features of the system include the use of acid-treated naked bacteria as combined carrier-adjuvants for the production of rabbit antiviral antibodies, and the use of acid-treated chicken antibodies (igy) for antigen trapping in the elisa. systems for detection of potato virus y (pvy), potato leafroll virus (plrv), grapev ... | 1989 | 2685004 |
| characterization of antigenic structures on arabis mosaic virus with monoclonal antibodies. | seven different epitopes on arabis mosaic virus (armv) were discerned. neo-, crypto-, and epitopes exposed on the virion and isolated coat protein were differentiated by their reactivity with monoclonal antibodies in an indirect enzyme-linked immunosorbent assay. the monoclonal antibodies, produced in vitro and in vivo, were of iggl, igm and iga isotype. no epitope exclusively specific for one isolate was found. one epitope was specific for armv isolates only. with the common epitopes an operati ... | 1986 | 3753200 |
| location of the replication determinants of the satellite rna associated with grapevine fanleaf nepovirus (strain f13). | a large satellite rna of 1114 nucleotides, named rna3, is always found associated with the genomic rnas of grapevine fanleaf virus, isolate f13 (gflv-f13). rna3 encodes a non-structural protein (p3) of m(r) 37k to which no function has previously been assigned. full-length cdna clones of rna3 were mutated in the 5' and 3' non-coding regions and in the 37k open reading frame. the ability of transcripts obtained from these clones to be replicated was investigated by protoplast infection in the pre ... | 1993 | 7505623 |
| coat protein gene sequence of an austrian isolate of grapevine fanleaf virus. | the nucleotide sequence of the coat protein cistron of an austrian isolate of grapevine fanleaf virus (gflv-fc) from vitis vinifera cv. french colombard was determined. it shows small differences at the rna level as well as at the protein level compared to the sequences of already published grapevine fanleaf virus strains. the differences may be a result of the natural variation among virus populations or a consequence of selection in a special host plant. as the virus rna sequence reported here ... | 1995 | 7646340 |
| a method combining immunocapture and pcr amplification in a microtiter plate for the detection of plant viruses and subviral pathogens. | a method for the detection of rna viral and subviral plant pathogens was developed that combines pathogen partial purification by solid-phase adsorbed antibodies, reverse transcriptional-polymerase chain reaction (rt-pcr) and quantitation of the amplified products by fluorescence. the reverse transcription of the rna is performed directly on the retained material without any previous thermal or chemical disruption of the virus particles. the whole procedure can be carried out in a microtiter pla ... | 1993 | 8113346 |
| differential proteolytic activities of precursor and mature forms of the 24k proteinase of grapevine fanleaf nepovirus. | the presence of a genome-linked protein (vpg) at the rna 5'-end of the genome is a characteristic of different groups of animal and plant positive-sense single-stranded rna viruses. these viruses express their structural and functional proteins from polyproteins that are sequentially processed by at least one viral proteinase. the grapevine fanleaf nepovirus 24k chymotrypsin-like cysteine proteinase, located between the vpg and the rna polymerase in the rna-1 encoded polyprotein p1, is active in ... | 1994 | 8128640 |
| vpg northern-immunoblots as a means for detection of viral rnas in protoplasts or plants infected with grapevine fanleaf nepovirus. | anti-genome-linked viral protein (anti-vpg) antibodies were produced from a synthetic peptide corresponding to the integral vpg sequence of grapevine fanleaf nepovirus-f13. these antibodies allowed detection of viral vpg-linked proteins which occur during the processing of viral polyproteins and of viral rnas in total rna extracts from infected protoplasts or plants after northern blotting. these highly specific antibodies recognised rnas from two grapevine fanleaf virus strains but not from ara ... | 1993 | 8328916 |
| genome organization of grapevine fanleaf nepovirus rna2 deduced from the 122k polyprotein p2 in vitro cleavage products. | the full-length transcript of grapevine fanleaf virus (gflv) rna2 produces a primary product of 122k when translated in the rabbit reticulocyte system. this 122k polyprotein is completely processed in vitro by the rna1-encoded 24k proteinase. the positions of the cleavage sites within the polyprotein have been mapped and the genome organization of gflv-f13 rna2 has been established. the order of mature proteins in the 122k polyprotein is the amino-terminal 28k protein, the 38k protein followed b ... | 1993 | 8376967 |
| biologically active transcripts from cloned cdna of genomic grapevine fanleaf nepovirus rnas. | transcripts were produced in vitro by run-off transcription from full-length cdna of rna1 and rna2 of grapevine fanleaf nepovirus (gflv; isolate f13) cloned downstream from a bacteriophage rna polymerase promoter. these transcripts, which possess a 5' terminal cap structure and a non-viral g residue instead of the naturally occurring genome-linked viral protein (vpg), are infectious to chenopodium quinoa protoplasts when inoculated by electroporation. synthetic rna1 alone replicated in protoplas ... | 1993 | 8429300 |
| the cloning, sequence analysis, and expression in e. coli of coat protein gene of grapevine fanleaf virus gh. | fragments of 5' end and 3' end of the coat protein gene of a chinese strain grapevine fanleaf virus (gflv-gh) were separately amplified through rt-pcr. the two products were cloned and ligated into a complete full-length coat protein gene via an inherent bgiii restriction site. the entire gene contains 1512 nucleotides which encode a 504 amino acids coat protein, molecular weight approximately 56 kda. comparison with gflv f-13 shows 88.4% homology in nucleotides and 95.8% similarity in deduced a ... | 1996 | 8988353 |
| the 119 kda and 124 kda polyproteins of arabis mosaic nepovirus (isolate s) are encoded by two distinct rna2 species. | arabis mosaic virus (armv) is a nepovirus that is serologically distantly related to grapevine fanleaf virus (gflv). both armv and gflv induce grapevine degeneration disease. several armv isolates, unlike isolates of gflv, produce upon in vitro translation of rna2 a polyprotein (p2) that forms a double band in polyacrylamide-sds gels. cloning of full-length copies of rna2 of an armv isolate from grapevine (armv-s) revealed that this isolate contained two rna2s of different length, called rna2-u ... | 1995 | 9049336 |
| grapevine fanleaf nepovirus p38 putative movement protein is not transiently expressed and is a stable final maturation product in vivo. | the putative 38 kda movement protein (p38) gene, located on the rna2 of grapevine fanleaf nepovirus (gflv), was cloned in escherichia coli and expressed as a fusion protein fused to six histidines (6hisp38). this protein was purified and used to produce a specific polyclonal antiserum from which immunoglobulins were isolated by immunoaffinity against the recombinant 6hisp38. western immunoblot analyses on gflv rna2 in vitro maturation products showed that the antibodies were specific for p38 pro ... | 1995 | 9049337 |
| cell-to-cell movement of beet necrotic yellow vein virus: i. heterologous complementation experiments provide evidence for specific interactions among the triple gene block proteins. | cell-to-cell movement of beet necrotic yellow vein virus (bnyvv) requires three proteins encoded by a triple gene block (tgb) on viral rna 2. a bnyvv rna 3-derived replicon was used to express movement proteins to functionally substitute for the bnyvv tgb proteins was tested by coinoculation of tgb-defective bnyvv with the various replicons to chenopodium quinoa. trans-heterocomplementation was successful with the movement protein (p30) of tobacco mosaic virus but not with the tubule-forming mov ... | 1998 | 9650295 |
| the nine c-terminal residues of the grapevine fanleaf nepovirus movement protein are critical for systemic virus spread. | the grapevine fanleaf virus (gflv) rna2-encoded polyprotein p2 is proteolytically cleaved by the rna1-encoded proteinase to yield protein 2a, 2b(mp) movement protein and 2c(cp) coat protein. to further investigate the role of the 2b(mp) and 2c(cp) proteins in virus movement, rna2 was engineered by alternatively replacing the gflv 2b(mp) and 2c(cp) genes with their counterparts from the closely related arabis mosaic virus (armv). transcripts of all chimeric rna2s were able to replicate in chenopo ... | 1999 | 10374950 |
| protein 2a of grapevine fanleaf nepovirus is implicated in rna2 replication and colocalizes to the replication site. | rna2 of grapevine fanleaf virus is replicated in trans by the rna1-encoded replication machinery. full processing of the rna2-encoded polyprotein p2 yields protein 2a of unknown function, the movement protein 2b(mp), and the coat protein 2c(cp). analysis of a set of deletion mutants in the p2-coding sequence revealed that protein 2a is necessary but not sufficient for rna2 replication. in addition to the 5' and 3' noncoding sequences and the 2a-coding sequence, an additional sequence coding for ... | 1999 | 10544127 |
| gflv replication in electroporated grapevine protoplasts. | grapevine fanleaf virus (gflv), responsible for the economically important court-noué disease, is exclusively transmitted to its natural host in the vineyards through xiphinema nematodes. we have developed direct inoculation of gflv into grapevine through protoplast electroporation. protoplasts were isolated from mesophyll of in vitro-grown plants and from embryogenic cell suspensions. permeation conditions were determined by monitoring calcein uptake. low salt poration medium was selected. elec ... | 2000 | 10814824 |
| [purification, serology and detection of grapevine leaf roll virus]. | closterovirus-like particles associated with grapevine leaf roll disease were purified from stem phloem tissue by differential centrifugation and cesium sulphate-sucrose density gradient centrifugation. the particles were between 660-2000 nm long. most of them were about 1400 nm long and also decorated by the antiserum against glrv ny-1 isolate. the purified preparation was tested for their serological relatedness in indirect elisa. the results indicated that these closterovirus-like particles r ... | 1997 | 11189365 |
| complete nucleotide sequences of the rnas 2 of german isolates of grapevine fanleaf and arabis mosaic nepoviruses. | the rnas 2 of an arabis mosaic virus (armv) and a grapevine fanleaf virus (gflv) isolate, originating from south west of germany near neustadt an der weinstrasse (nw), were sequenced. they are 3820 and 3775 nucleotides long respectively, and both contain one open reading frame encoding a polypeptide of 1110 amino acids. their 5' non-coding regions contain conserved and repeated sequences, which are able to form stem-loop structures. nucleotide sequence comparisons between the full-length rnas 2 ... | 2001 | 11325468 |
| quasispecies nature of the genome of grapevine fanleaf virus. | genetic diversity was characterized in 14 isolates of grapevine fanleaf virus (gflv) recovered from grapevine (vitis vinifera). virions were collected by immunocapture, and a 1557 bp fragment containing part of the viral coat protein gene and part of the untranslated region to its 3' side was amplified by rt-pcr. sequence variation among isolates was characterized by restriction fragment length polymorphism (rflp) analysis and by sequencing. the avaii-generated rflp patterns from the various iso ... | 2001 | 11413392 |
| involvement of rna2-encoded proteins in the specific transmission of grapevine fanleaf virus by its nematode vector xiphinema index. | the nepovirus grapevine fanleaf virus (gflv) is specifically transmitted by the nematode xiphinema index. to identify the rna2-encoded proteins involved in x. index-mediated spread of gflv, chimeric rna2 constructs were engineered by replacing the 2a, 2b(mp), and/or 2c(cp) sequences of gflv with their counterparts in arabis mosaic virus (armv), a closely related nepovirus which is transmitted by xiphinema diversicaudatum but not by x. index. among the recombinant viruses obtained from transcript ... | 2001 | 11878885 |
| grapevine fanleaf virus replication occurs on endoplasmic reticulum-derived membranes. | infection by grapevine fanleaf nepovirus (gflv), a bipartite rna virus of positive polarity belonging to the comoviridae family, causes extensive cytopathic modifications of the host endomembrane system that eventually culminate in the formation of a perinuclear "viral compartment." we identified by immunoconfocal microscopy this compartment as the site of virus replication since it contained the rna1-encoded proteins necessary for replication, newly synthesized viral rna, and double-stranded re ... | 2002 | 12163601 |
| preliminary attempts to biolistic inoculation of grapevine fanleaf virus. | biolistics has been studied to inoculate grapevine fanleaf virus (gflv), a nepovirus, to its natural woody host, vitis sp., and its herbaceous host, chenopodium quinoa. at first, bombardment conditions for in vitro and greenhouse grown plants were set using the uida reporter gene. the infectious feature of the cartridges was then evaluated by studying infection of c. quinoa plants. systemic infection was obtained with either gflv particles or rna extracts in experimental conditions which gave al ... | 2003 | 12565151 |
| involvement of the secretory pathway and the cytoskeleton in intracellular targeting and tubule assembly of grapevine fanleaf virus movement protein in tobacco by-2 cells. | grapevine fanleaf virus (gflv) is one of a large class of plant viruses whose cell-to-cell transport involves the passage of virions through tubules composed of virus-encoded movement protein (mp). the tubules are embedded within modified plasmodesmata, but the mechanism of targeting of mp to these sites is unknown. to study intracellular gflv mp trafficking, a green fluorescent protein-mp fusion (gfp:mp) was expressed in transgenic tobacco by-2 suspension cells under the control of an inducible ... | 2003 | 12953111 |
| the specific transmission of grapevine fanleaf virus by its nematode vector xiphinema index is solely determined by the viral coat protein. | the viral determinants involved in the specific transmission of grapevine fanleaf virus (gflv) by its nematode vector xiphinema index are located within the 513 c-terminal residues of the rna2-encoded polyprotein, that is, the 9 c-terminal amino acids of the movement protein (2bmp) and contiguous 504 amino acids of the coat protein (2ccp) [virology 291 (2001) 161]. to further delineate the viral determinants responsible for the specific spread, the four amino acids that are different within the ... | 2004 | 15003859 |
| complete nucleotide sequence of the rna 1 of a grapevine isolate of arabis mosaic virus. | the complete nucleotide sequence of the genomic rna 1 of the grapevine isolate nw (neustadt an der weinstrasse) of arabis mosaic virus (armv) was determined. it is 7334 nucleotides long excluding the poly(a) tail, and contains one long open reading frame encoding a polypeptide of 2284 amino acids. the 5' and 3' non-coding regions were 227 and 252 nucleotides long respectively, and showed stretches of high identity with the corresponding 5' and 3' non-coding regions of armv-nw rna 2. the analysis ... | 2004 | 15098112 |
| field safety assessment of recombination in transgenic grapevines expressing the coat protein gene of grapevine fanleaf virus. | one of the major environmental safety issues over transgenic crops containing virus-derived genes relates to the outcome of recombination events between viral transgene transcripts and rnas from indigenous virus populations. we addressed this issue by assessing the emergence of viable grapevine fanleaf virus (gflv) recombinants in transgenic grapevines expressing the gflv coat protein (cp) gene. test plants consisted of nontransgenic scions grafted onto transgenic and nontransgenic rootstocks th ... | 2004 | 15198204 |
| population structure and genetic variability within isolates of grapevine fanleaf virus from a naturally infected vineyard in france: evidence for mixed infection and recombination. | the nematode-borne grapevine fanleaf virus, from the genus nepovirus in the family comoviridae, causes severe degeneration of grapevines in most vineyards worldwide. we characterized 347 isolates from transgenic and conventional grapevines from two vineyard sites in the champagne region of france for their molecular variant composition. the population structure and genetic diversity were examined in the coat protein gene by ic-rt-pcr-rflp analysis with ecori and styi, and nucleotide sequencing, ... | 2004 | 15269386 |
| sensitive and reliable detection of grapevine fanleaf virus in a single xiphinema index nematode vector. | grapevine fanleaf virus (gflv) is specifically transmitted from plant to plant by the ectoparasitic nematode xiphinema index. a sensitive and reliable procedure was developed to readily detect gflv in a single viruliferous x. index, regardless of the nematode origin, i.e. greenhouse rearings or vineyard soils. the assay is based on bead milling to disrupt nematodes extracted from soil samples, solid-phase extraction of total nematode rnas, and amplification of a 555bp fragment of the coat protei ... | 2004 | 15488624 |
| complete nucleotide sequence of the rna-2 of grapevine deformation and grapevine anatolian ringspot viruses. | the nucleotide sequence of rna-2 of grapevine anatolian ringspot virus (garsv) and grapevine deformation virus (gdefv), two recently described nepoviruses, has been determined. these rnas are 3753 nt (gdefv) and 4607 nt (garsv) in size and contain a single open reading frame encoding a polyprotein of 122 kda (gdefv) and 150 kda (garsv). full-length nucleotide sequence comparison disclosed 71-73% homology between gdefv rna-2 and that of grapevine fanleaf virus (gflv) and arabis mosaic virus (armv ... | 2005 | 15830151 |
| rt-pcr-rflp for genetic diversity analysis of tunisian grapevine fanleaf virus isolates in their natural host plants. | genetic diversity was characterized in 20 isolates of grapevine fanleaf virus (gflv) recovered from naturally infected grapevine plants (vitis vinifera) in the north of tunisia. viral rnas were isolated by oligoprobe capture, and a 605 bp fragment containing a part of the viral coat protein gene was amplified by rt-pcr. sequence variation among isolates was characterized by restriction fragment length polymorphism (rflp) analysis and confirmed by sequencing. the gflv infections are found as a co ... | 2005 | 15967236 |
| characterization of a naturally occurring recombinant isolate of grapevine fanleaf virus. | the naturally occurring grapevine fanleaf virus (gflv) recombinant isolate a17b was recovered from its grapevine host by sap inoculation and serial passages onto gomphrena globosa, a pseudo local lesion herbaceous host, and chenopodium quinoa, a systemic herbaceous host, to characterize some of its biological properties. sequence analysis of the cp gene, in which a recombinational event was previously detected, demonstrated the genetic stability of recombinant isolate a17b over a 5-year period i ... | 2005 | 15968475 |
| molecular characterization of grapevine plants transformed with gflv resistance genes: i. | the grapevine fanleaf virus-coat protein (gflv cp) gene was inserted through agrobacterium-mediated transformation in vitis vinifera "nebbiolo", "lumassina" and "blaufränkisch". two plasmids were used: pga-cp+ (full-length gflv cp gene with an introduced start codon) and pga-as (same gene in antisense orientation). forty-three transgenic lines were regenerated. as several lines in southern blots share same hybridization patterns, eight independent line groups resulted for "nebbiolo", one for "lu ... | 2005 | 16240119 |
| molecular characterization of grapevine plants transformed with gflv resistance genes: ii. | a collection of 127 putatively transgenic individuals of vitis vinifera cv. russalka was characterized by pcr and southern hybridization. six different constructs containing the neomycin phosphotransferase (nptii) marker gene and sequences of the grapevine fanleaf virus coat protein (gflv cp) gene including non-translatable and truncated forms were transferred via agrobacterium-mediated transformation. detection of transgenic sequences by pcr was positive in all lines. southern blot analysis rev ... | 2006 | 16408176 |
| efficient methods for sample processing and cdna synthesis by rt-pcr for the detection of grapevine viruses and viroids. | template preparation is important in reverse-transcription polymerase chain reaction (rt-pcr)-based detection methods. a tissuelyser with tungsten carbide beads was used for simultaneous processing of up to 48 samples under the same conditions in the development of a simple and rapid procedure to prepare a plant extract for rt reaction. a sandpaper method was also developed by which wood tissue of dormant cuttings could be macerated easily to process with minimal time and effort. it was also dem ... | 2006 | 16510197 |
| partial molecular characterization of a mild isolate of grapevine fanleaf virus from south moravia, czech republic. | an atypical mild isolate hv5 of grapevine fanleaf virus (gflv) was found in a south moravian viticulture region in czech republic. partial sequence of its rna2 was determined and compared with available sequences of typical gflv isolates. two genomic regions, namely a 814 nt-long one spanning the movement protein (mp) gene and a 5'-part of the coat protein (cp) gene. and a 1426 nt-long one covering a part of the cp gene and the adjacent 3'-non-coding region (3'-ncr) were analyzed. although no hv ... | 2006 | 17131940 |
| development of degenerate and species-specific primers for the differential and simultaneous rt-pcr detection of grapevine-infecting nepoviruses of subgroups a, b and c. | based on the nucleotide sequence homology of rna-1 and rna-2 of nepoviruses isolated from grapevines, three sets of degenerate primers, one for each of the three subgroups of the genus (a, b and c), were designed and proved effective for rt-pcr detection of subgroups in infected grapevines and herbaceous hosts. primers designed specifically for detecting subgroup a species amplified a fragment of 255 bp from samples infected by grapevine fanleaf virus (gflv), arabis mosaic virus (armv), tobacco ... | 2007 | 17187868 |
| diversity of grapevine fanleaf virus isolates from iran. | enzyme-linked immunosorbent assay (elisa) testing of 126 grapevine samples, from vineyards in the northwest region of iran, detected grapevine fanleaf virus (gflv) in 33 samples. total rna from eight of the infected samples were subjected to reverse transcription polymerase chain reaction (rt-pcr) analysis using primers which corresponded to the virus coat protein and 3' non coding region of rna 2. an expected 1620 bp dna fragment was amplified from all the tested samples. pcr products from isol ... | 2007 | 17521761 |
| genetic and qtl analysis of resistance to xiphinema index in a grapevine cross. | resistance to the dagger nematode xiphinema index has been an important objective in grape rootstock breeding programs. this nematode not only causes severe feeding damage to the root system, but it also vectors grapevine fanleaf virus (gflv), the causal agent of fanleaf degeneration and one of the most severe viral diseases of grape. the established screening procedures for dagger nematode resistance are time consuming and can produce inconsistent results. a fast and reliable greenhouse-based s ... | 2008 | 18004541 |
| comparison of low-density arrays, rt-pcr and real-time taqman rt-pcr in detection of grapevine viruses. | low-density arrays (lda) have been designed based on the real-time rt-pcr (taqman) assays for the specific detection of 13 viruses that infect grapevines in addition to the housekeeping gene 18s rrna. the viruses included in the study are grapevine leafroll associated viruses 1, 2, 3, 4, 5, and 9, grapevine leafroll associated virus-2 redglobe (glrav-2rg) strain, ruspestris stem pitting associated virus, grapevine vitivirus a, grapevine vitivirus b, grapevine fanleaf virus, tomato ringspot virus ... | 2008 | 18329731 |
| the synthesis and processing of the nepovirus grapevine fanleaf virus proteins in rabbit reticulocyte lysate. | translation of a mixture of the two grapevine fanleaf virus (gflv) rnas in reticulocyte lysates in the presence of the amino acid analogs canavanine, s-aminoethyl-cysteine, and p-fluorophenylalanine gave two products of molecular weights (mr) 220,000 (from rna-1) and 125,000 (from rna-2). in the absence of these analogs a protease apparently induced by rna-1 catalyzed cleavage of the rna-2 product (mr 125,000) into two proteins of mr 68,000 and 58,000. the 58,000 mr protein was shown to be the l ... | 1983 | 18639155 |
| multiple interspecies recombination events within rna2 of grapevine fanleaf virus and arabis mosaic virus. | sequence alignments and siscan analyses inferred multiple interspecies recombination events within rna2 of strains ghu of grapevine fanleaf virus (gflv) and ta of arabis mosaic virus (armv), two closely related subgroup a nepoviruses in the family comoviridae. interspecies recombination events were identified in the 5' untranslated region, the putative homing protein and movement protein genes but not in the coat protein gene and 3' untranslated region. these findings suggest a dynamic relations ... | 2008 | 18695933 |
| isolate-specific detection of grapevine fanleaf virus from xiphinema index through dna-based molecular probes. | abstract tests with a real-time reverse transcription-polymerase chain reaction (rt-pcr) were performed on specimens of xiphinema index collected from the rhizosphere of grapevine fanleaf virus (gflv)-infected grapevines at palagiano, italy. a 1,157-bp fragment of the gflv rna-2 coat protein (cp) gene was amplified and sequenced. a fluorescent scorpion probe was designed to detect a highly conserved cp region. a second region with isolate-specific multiple nucleotide polymorphisms was used to de ... | 2005 | 18943119 |
| multiplex polymerase chain reaction identification of single individuals of the longidorid nematodes xiphinema index, x. diversicaudatum, x. vuittenezi, and x. italiae using specific primers from ribosomal genes. | abstract the species x. index, x. diversicaudatum, x. vuittenezi, and x. italiae are established (e) or putative (p) vectors of grapevine fanleaf virus (gflv) (e), arabis mosaic virus (e), grapevine chrome mosaic virus (p), and gflv (p) nepoviruses of grapevine, respectively. all four species are very closely related taxonomically and their low field densities make them difficult to identify from morphological and morphometrical diagnostic characters when only single or few individuals are detec ... | 2003 | 18943130 |
| spatial distribution of the dagger nematode xiphinema index and its associated grapevine fanleaf virus in french vineyard. | the nematode xiphinema index is, economically, the major virus vector in viticulture, transmitting specifically the grapevine fanleaf virus (gflv), the most severe grapevine virus disease worldwide. increased knowledge of the spatial distribution of this nematode, both horizontally and vertically, and of correlative gflv plant infections, is essential to efficiently control the disease. in two infested blocks of the bordeaux vineyard, vertical distribution data showed that the highest numbers of ... | 2008 | 18943213 |
| survival of xiphinema index in vineyard soil and retention of grapevine fanleaf virus over extended time in the absence of host plants. | abstract grapevine fanleaf virus (gflv) is transmitted specifically from grapevine to grapevine by the ectoparasitic root-feeding nematode xiphinema index. limited information is available on the survival of x. index in vineyard soil and on the retention of gflv by x. index over extended periods of time. we addressed these two issues by quantifying the numbers of living x. index recovered from soil samples that were collected in three naturally gflv-infected vineyards in france and subsequently ... | 2005 | 18943467 |
| simultaneous detection of nine grapevine viruses by multiplex reverse transcription-polymerase chain reaction with coamplification of a plant rna as internal control. | abstract a multiplex reverse transcription-polymerase chain reaction (mrt-pcr) was developed for simultaneous detection of nine grapevine viruses: arabis mosaic virus, grapevine fanleaf virus, grapevine virus a, grapevine virus b, rupestris stem pitting-associated virus, grapevine fleck virus, grapevine leafroll-associated virus-1, -2, and -3, in combination with a plant rna internal control used as an indicator of the effectiveness of rna extraction and rt-pcr. primers were designed from conser ... | 2006 | 18943959 |
| grapevine fanleaf virus (gflv)-specific antibodies confer gflv and arabis mosaic virus (armv) resistance in nicotiana benthamiana. | grapevine fanleaf virus (gflv) is one of the most destructive pathogens of grapevine. in this study, we generated monoclonal antibodies binding specifically to the coat protein of gflv. antibody fl(3), which bound most strongly to gflv and showed cross-reactivity to arabis mosaic virus (armv), was used to construct the single-chain antibody fragment scfvgflvcp-55. to evaluate the potential of this single-chain variable fragment (scfv) to confer antibody-mediated virus resistance, transgenic nico ... | 2009 | 19161351 |
| biotin-avidin elisa detection of grapevine fanleaf virus in the vector nematode xiphinema index. | the value of biotin-avidin (b-a) elisa for the detection of grapevine fanleaf virus (gflv) in xiphinema was estimated with field populations and greenhouse subpopulations. samples consisted of increasing numbers of adults ranging from 1 to 64 in multiples of two. tests with virus-free x. index populations reared on grapevine and fig plants as negative controls did not reveal a noticeable effect of the host plant. elisa absorbances of virus-free x. index samples were greater than corresponding ab ... | 1993 | 19279786 |
| effect of grapevine fanleaf virus on the reproduction and survival of its nematode vector, xiphinema index thorne &allen. | studies on the virus-vector interaction between the grapevine fanleaf virus (gfv) and its nematode vector, xiphinema index, indicate the virus had no measurable effect on the rate of reproduction of its vector, but significantly influenced survnal of the nematodes. | 1969 | 19325663 |
| high-sensitivity detection of fruit tree viruses using bacterial magnetic particles. | prunus necrotic ring spot virus (pnrsv) and grapevine fanleaf virus (gflv) were detected by fluoroimmunoassay using bacterial magnetic particles (bmps), and a double antibody sandwich enzyme linked immunosorbent assay (das-elisa). for the fluoroimmunoassay, fluorescein isothiocyanate labeled anti-pnrsv antibody or anti-gflv antibody was conjugated onto bmps of magnetospirillum gryphiswaldense msr-1. with this method, a very low minimum antigen concentration (1 x 10(6) dilution of the original sa ... | 2009 | 19341408 |
| genetic structure and variability of virus populations in cross-protected grapevines superinfected by grapevine fanleaf virus. | recombination was assessed in a vineyard site in which grapevines cross-protected with mild strains ghu of grapevine fanleaf virus (gflv) or ta of arabis mosaic virus (armv) were superinfected with gflv field isolates following transmission by the nematode vector xiphinema index. the genetic structure and variability within rna2 of isolates from grapevines co-infected with gflv field isolates and either gflv-ghu or armv-ta were characterized to identify intra- and interspecies recombinants. sequ ... | 2009 | 19409944 |
| microarray immunoassay for the detection of grapevine and tree fruit viruses. | development and application of dna microarrays for plant disease diagnosis has to date been limited, and for antibody arrays even more so. in this work, an antibody microarray procedure was developed and its usefulness for the detection of plant viruses demonstrated. using the conventional monoplex immunoassay elisa technique as a benchmark, the procedure was used to detect several grapevine and tree fruit viruses. in a direct labelling approach, arabis mosaic virus (armv), and grapevine fanleaf ... | 2009 | 19410603 |
| gene silencing and virus resistance based on defective interfering constructs in transgenic nicotiana benthamiana is not linked to accumulation of sirna. | rna interference (rnai) was established in nicotiana benthamiana plants by introducing constructs containing a defective interfering (di) sequence from tomato bushy stunt virus (tbsv) in combination with a conserved sense-sequence from the target grapevine fanleaf virus (gflv). silencing in plants was confirmed by agrobacterium-mediated infiltration of a gfp-sensor containing the gflv-derived target sequence. the transgene-induced rnai led to silencing of the gfp-sensor and gfp fluorescence was ... | 2009 | 19419883 |
| genetic variability within the coat protein gene of grapevine fanleaf virus isolates from south africa and the evaluation of rt-pcr, das-elisa and immunostrips as virus diagnostic assays. | grapevine fanleaf virus (gflv) is responsible for severe fanleaf degeneration in grapevines of all major wine producing regions of the world, including south africa. in order to successfully control the spread of the virus, specific and reliable diagnostic assays are necessary. the genetic variability of 12 gflv isolates recovered from naturally infected grapevine plants in the western cape region of south africa were characterised. these samples were subjected to rna extraction, rt-pcr analysis ... | 2009 | 19428739 |
| transgene silencing in grapevines transformed with gflv resistance genes: analysis of variable expression of transgene, sirnas production and cytosine methylation. | eight transgenic grapevine lines transformed with the coat protein gene of grapevine fanleaf virus (gflv-cp) were analyzed for a correlation between transgene expression, sirnas production and dna methylation. bisulphite genome sequencing was used for a comprehensive analysis of dna methylation. methylated cytosine residues of cpg and cpnpg sites were detected in the gflv-cp transgene, in the t7 terminator and in the 35s promoter of three grapevines without transgene expression, but no detectabl ... | 2010 | 19507046 |
| genome diversity and intra- and interspecies recombination events in grapevine fanleaf virus. | abstract grapevine fanleaf virus (gflv) was documented in self-rooted vines of four grapevine (vitis vinifera) cultivars in eastern washington. gflv was found as mixed infection in cvs. pinot noir, chardonnay, and cabernet franc and as single infections in cv. merlot. fanleaf disease symptoms were only observed in the first two cultivars. the spatial distribution of gflv-infected grapevines was random, suggesting primary spread through planting virus-infected cuttings rather than infield transmi ... | 2009 | 19900006 |
| a diagnostic oligonucleotide microarray for simultaneous detection of grapevine viruses. | at least 58 viruses have been reported to infect grapevines causing economic damage globally. conventional detection strategies based on serological assays, biological indexing and rt-pcr targeting one or few viruses in each assay are widely used. grapevines are prone to contain mixed infections of several viruses, making the use of these techniques time-consuming. a 70-mer oligonucleotide microarray able to detect simultaneously a broad spectrum of known viruses as well as new viruses by cross- ... | 2010 | 19914293 |
| transmission competency of single-female xiphinema index lines for grapevine fanleaf virus. | grapevine fanleaf virus (gflv) is vectored specifically from grapevine to grapevine by the ectoparasitic nematode xiphinema index. limited information is available on the vector competency of x. index populations from diverse geographical origins. we determined the transmissibility of two gflv strains showing 4.6% amino acid divergence within their coat protein (e.g., strains f13 and ghu) by seven clonal lines of x. index developed from seven distinct populations from the mediterranean basin (cy ... | 2010 | 20205542 |
| cloning and characterization of xir1, a locus responsible for dagger nematode resistance in grape. | the dagger nematode, xiphinema index, feeds aggressively on grape roots and in the process, vectors grapevine fanleaf virus (gflv) leading to the severe viral disease known as fanleaf degeneration. resistance to x. index and gflv has been the key objective of grape rootstock breeding programs. a previous study found that resistance to x. index derived from vitis arizonica was largely controlled by a major quantitative trait locus, xir1 (x. index resistance 1), located on chromosome 19. the study ... | 2010 | 20490447 |
| a stretch of 11 amino acids in the betab-betac loop of the coat protein of grapevine fanleaf virus is essential for transmission by the nematode xiphinema index. | grapevine fanleaf virus (gflv) and arabis mosaic virus (armv) from the genus nepovirus, family secoviridae, cause a severe degeneration of grapevines. gflv and armv have a bipartite rna genome and are transmitted specifically by the ectoparasitic nematodes xiphinema index and xiphinema diversicaudatum, respectively. the transmission specificity of both viruses maps to their respective rna2-encoded coat protein (cp). to further delineate the gflv cp determinants of transmission specificity, three ... | 2010 | 20519403 |
| genetic structure and molecular variability of grapevine fanleaf virus populations. | to gain insights into the evolutionary mechanisms of grapevine fanleaf virus (gflv) from the genus nepovirus, family secoviridae, the sequences of the complete coding region of rna2, including genes 2a(hp), 2b(mp) and 2c(cp), and partial sequence from the rna1-encoded gene 1e(pol) of 14 gflv isolates from three naturally infected california vineyards were characterized. phylogenetic analyses suggested two to three evolutionarily divergent lineages that did not reflect the vineyard origin of the ... | 2010 | 20540977 |
| a one-step reverse transcription real-time pcr assay for the detection and quantitation of grapevine fanleaf virus. | a taqman(®) one-step reverse transcription real-time pcr (rt-qpcr) assay was developed for the specific detection and relative quantitation of grapevine fanleaf virus (gflv), the causal agent of grapevine fanleaf degeneration disease. the assay was targeted to a conservative region located in the 2a(hp) gene of the gflv rna2 molecule. the assay specificity was evaluated on gflv isolates from a wide range of geographical regions and on other viruses infecting grapevines. the sensitivity of the de ... | 2010 | 20804787 |
| quantitation of grapevine leafroll associated virus-1 and -3, grapevine virus a, grapevine fanleaf virus and grapevine fleck virus in field-collected vitis vinifera l. 'nebbiolo' by real-time reverse transcription-pcr. | taqman one-step real-time qrt-pcr assays were developed for the quantitation of grapevine leafroll associated virus-1 and -3 (glrav-1 and -3), grapevine virus a (gva), grapevine fanleaf virus (gflv) and grapevine fleck virus (gfkv) in vitis vinifera l. virus load in the progenies of three 'nebbiolo' clones planted in two experimental vineyards in piemonte (northwestern italy), and carrying the viruses in different combinations, was evaluated. quantitation primers were designed on the rna-depende ... | 2010 | 21167205 |
| strategies for the crystallization of viruses: using phase diagrams and gels to produce 3d crystals of grapevine fanleaf virus. | the small icosahedral plant rna nepovirus grapevine fanleaf virus (gflv) is specifically transmitted by a nematode and causes major damage to vineyards worldwide. to elucidate the molecular mechanisms underlying the recognition between the surface of its protein capsid and cellular components of its vector, host and viral proteins synthesized upon infection, the wild type gflv strain f13 and a natural mutant (gflv-td) carrying a gly297asp mutation were purified, characterized and crystallized. s ... | 2011 | 21352920 |
| development of nine polymorphic microsatellite markers for the phytoparasitic nematode xiphinema index, the vector of the grapevine fanleaf virus. | we report isolation, characterization and cross-species amplification of nine microsatellite loci from the phytoparasitic nematode xiphinema index, the vector of grapevine fanleaf virus. levels of polymorphism were evaluated in 62 individuals from two x. index populations. the number of alleles varies between two and 10 depending on locus and population. observed heterozygosity on loci across both populations varied from 0.32 to 0.857 (mean 0.545). the primers were tested for cross-species ampli ... | 2009 | 21564611 |
| structural insights into viral determinants of nematode mediated grapevine fanleaf virus transmission. | many animal and plant viruses rely on vectors for their transmission from host to host. grapevine fanleaf virus (gflv), a picorna-like virus from plants, is transmitted specifically by the ectoparasitic nematode xiphinema index. the icosahedral capsid of gflv, which consists of 60 identical coat protein subunits (cp), carries the determinants of this specificity. here, we provide novel insight into gflv transmission by nematodes through a comparative structural and functional analysis of two gfl ... | 2011 | 21625570 |
| tubule-guided cell-to-cell movement of a plant virus requires class xi myosin motors. | cell-to-cell movement of plant viruses occurs via plasmodesmata (pd), organelles that evolved to facilitate intercellular communications. viral movement proteins (mp) modify pd to allow passage of the virus particles or nucleoproteins. this passage occurs via several distinct mechanisms one of which is mp-dependent formation of the tubules that traverse pd and provide a conduit for virion translocation. the mp of tubule-forming viruses including grapevine fanleaf virus (gflv) recruit the plant p ... | 2011 | 22046131 |
| molecular characterization of phylogenetically distinct isolates of grapevine fanleaf virus from iran based on 2a(hp) gene. | movement and coat protein genes from grapevine fanleaf virus (gflv) isolates have been characterized previously from iran. in this study, an optimized reverse transcription polymerase chain reaction protocol was established to amplify rna2 genomic segment corresponding to the hypothetical protein (2a(hp)). the sequence of 2a(hp) was compared with that of previously reported gflv strains/isolates from other countries which showed 82-86% sequence identities. the 2a(hp) gene from iran appeared to b ... | 2012 | 23730004 |
| transient expression of artificial micrornas targeting grapevine fanleaf virus and evidence for rna silencing in grapevine somatic embryos. | grapevines are affected worldwide by viruses that compromise fruit yield and quality. grapevine fanleaf virus (gflv) causes fanleaf degeneration disease, a major threat to grapevine production. transgenic approaches exploiting the rna silencing machinery have proven suitable for engineering viral resistance in several crop species. however, the artificial microrna (amirna)-based strategy has not yet been reported in grapevine. we developed two amirna precursors (pre-amirnas) targeting the coat p ... | 2012 | 22427113 |
| transformation of grapevine rootstocks with the coat protein gene of grapevine fanleaf nepovirus. | control of fanleaf disease induced by the grapevine fanleaf nepovirus (gflv) today is based on sanitary selection and soil disinfection with nematicides. this way of control is not always efficient and nematicides can be dangerous pollutants. coat protein (cp) mediated protection could be an attractive alternative. we have transferred a chimeric cp gene of gflv-f13 via agrobacterium tumefaciens lba4404 into two rootstock varieties: vitis rupestris and 110 richter (v. rupestris x v. berlandieri). ... | 1995 | 24185595 |
| protection against virus infection in tobacco plants expressing the coat protein of grapevine fanleaf nepovirus. | grapevine fanleaf nepovirus (gflv) is responsible for the economically significant "court-noué" disease in vineyards. its genome is made up of two single-stranded rna molecules (rna1 and rna2) which direct the synthesis of polyproteins p1 and p2 respectively. a chimeric coat protein gene derived from the c-terminal part of p2 was constructed and subsequently introduced into a binary transformation vector. transgenic nicotiana benthamiana plants expressing the coat protein under the control of th ... | 1994 | 24193837 |
| comparative detection of a large population of grapevine viruses by taqman(®) rt-qpcr and elisa. | grapevine (vitis spp.) can be infected by numerous viruses that are often widespread and of great economic importance. reliable detection methods are necessary for sanitary selection which is the only way to partly control grapevine virus diseases. biological indexing and elisa are currently the standard methods for screening propagation material, and pcr-methods are becoming increasingly popular. due to the diversity of virus isolates, it is essential to verify that the tests allow the detectio ... | 2017 | 27923589 |
| detection of nepovirus vector and nonvector xiphinema species in grapevine. | fanleaf degeneration is considered the most damaging viral disease of grapevine. the two major nepoviruses involved are grapevine fanleaf virus (gflv) and arabis mosaic virus (armv) which are respectively and specifically transmitted by the dagger nematodes xiphinema index and x. diversicaudatum. the methods described below are aimed at detecting four prevalent grapevine xiphinema species: the vector species previously mentioned and two nonvector species x. vuittenezi and x. italiae. | 2015 | 25981253 |
| multiplex rt-pcr method for the simultaneous detection of nine grapevine viruses. | viral diseases are a serious pathological problem for grapevines, and in recent years the need for increasingly specific and rapid diagnostic methods for the selection of propagation materials has grown. arabis mosaic virus, grapevine fanleaf virus, grapevine virus a, grapevine virus b, grapevine rupestris stem pitting-associated virus, grapevine fleck virus, and grapevine leafroll-associated viruses 1, 2, and 3 are nine of the most widespread viruses that naturally infect grapevines. a multiple ... | 2015 | 25287494 |
| phylogenetic and recombination analysis of the homing protein domain of grapevine fanleaf virus (gflv) isolates associated with 'yellow mosaic' and 'infectious malformation' syndromes in grapevine. | the rna2 of seven grapevine fanleaf virus (gflv) isolates from vines with yellow mosaic (ym) symptoms from different origin were sequenced. these sequences showed a high variability in the homing protein (2a(hp)) and, in five of them, a putative recombination with arabis mosaic virus (armv) was detected. to investigate recombination frequency, the partial sequences of the 2a(hp) of 28 additional gflv isolates from nine different countries, showing either ym or infectious malformations (mf) sympt ... | 2014 | 24916055 |
| complete nucleotide sequence of a south african isolate of grapevine fanleaf virus and its associated satellite rna. | the complete sequences of rna1, rna2 and satellite rna have been determined for a south african isolate of grapevine fanleaf virus (gflv-sach44). the two rnas of gflv-sach44 are 7,341 nucleotides (nt) and 3,816 nt in length, respectively, and its satellite rna (satrna) is 1,104 nt in length, all excluding the poly(a) tail. multiple sequence alignment of these sequences showed that gflv-sach44 rna1 and rna2 were the closest to the south african isolate, gflv-sapcs3 (98.2% and 98.6% nt identity, r ... | 2013 | 23867805 |
| genetic variability, evolution, and biological effects of grapevine fanleaf virus satellite rnas. | large satellite rnas (type b satrnas) of grapevine fanleaf virus (gflv) from the genus nepovirus, family secoviridae were identified in a naturally infected vineyard and a grapevine germplasm collection. these gflv satrna variants had a higher nucleotide sequence identity with satrnas of arabis mosaic virus (armv) strains nw and j86 (93.8 to 94.6%) than with the satrna of gflv strain f13 and those of other armv strains (68.3 to 75.0%). phylogenetic analyses showed no distinction of gflv and armv ... | 2013 | 23718838 |
| the backbone model of the arabis mosaic virus reveals new insights into functional domains of nepovirus capsid. | arabis mosaic virus (armv) and grapevine fanleaf virus (gflv) are two picorna-like viruses from the genus nepovirus, consisting in a bipartite rna genome encapsidated into a 30 nm icosahedral viral particle formed by 60 copies of a single capsid protein (cp). they are responsible for a severe degeneration of grapevines that occurs in most vineyards worldwide. although sharing a high level of sequence identity between their cp, armv is transmitted exclusively by the ectoparasitic nematode xiphine ... | 2013 | 23376736 |
| real-time multiplex rt-pcr for the simultaneous detection of the five main grapevine viruses. | a real-time multiplex rt-pcr has been developed for the simultaneous detection and identification of the major rna viruses that infect grapevines (grapevine fanleaf virus, arabis mosaic virus, grapevine leafroll-associated virus 1, grapevine leafroll-associated virus 3 and grapevine fleck virus). serial dilutions of infected plant extracts were tested using the new method, and the results were compared with those obtained using a commercially available elisa and real-time singleplex rt-pcr. the ... | 2013 | 23219809 |
| complete nucleotide sequence of a south african isolate of grapevine fanleaf virus. | the complete sequences of rna1 and rna2 have been determined for a south african isolate of grapevine fanleaf virus (gflv-sapcs3). the two rnas are, respectively, 7,342 and 3,817 nucleotides in length, excluding the poly(a) tails. rna1 has a large open reading frame (orf) of 6,852 nucleotides and a 5'-utr and a 3'-utr of 243 and 244 nucleotides, respectively. rna2 encodes for an orf of 3,330 nucleotides and has the highest nucleotide identity (90.4 %) with gflv-f13. the full length nucleotide se ... | 2012 | 22669541 |
| grapevine deformation virus: completion of the sequence and evidence on its origin from recombination events between grapevine fanleaf virus and arabis mosaic virus. | the complete nucleotide (nt) sequence of grapevine deformation virus (gdefv) rna-1 has been determined. it consists of 7386 nt, excluding the poly(a) tail, and contains a single open reading frame (orf) encoding a polyprotein (p1) of 252 kda. p1 comprises the 1a(pro-cof) proteinase cofactor, the 1b(hel) ntp-binding protein, the 1c(vpg) viral protein genome-linked, the 1d(prot) proteinase and the 1e(pol) rna-dependent rna polymerase, all of which are conserved domains in polyproteins of different ... | 2012 | 22480575 |
| the role of plasmodesma-located proteins in tubule-guided virus transport is limited to the plasmodesmata. | intercellular spread of plant viruses involves passage of the viral genome or virion through a plasmodesma (pd). some viruses severely modify the pd structure, as they assemble a virion carrying tubule composed of the viral movement protein (mp) inside the pd channel. successful modulation of the host plant to allow infection requires an intimate interaction between viral proteins and both structural and regulatory host proteins. to date, however, very few host proteins are known to promote viru ... | 2016 | 27339685 |
| a leafhopper-transmissible dna virus with novel evolutionary lineage in the family geminiviridae implicated in grapevine redleaf disease by next-generation sequencing. | a graft-transmissible disease displaying red veins, red blotches and total reddening of leaves in red-berried wine grape (vitis vinifera l.) cultivars was observed in commercial vineyards. next-generation sequencing technology was used to identify etiological agent(s) associated with this emerging disease, designated as grapevine redleaf disease (grd). high quality rna extracted from leaves of grape cultivars merlot and cabernet franc with and without grd symptoms was used to prepare cdna librar ... | 2013 | 23755117 |
| production of polyclonal antibody against grapevine fanleaf virus movement protein expressed in escherichia coli. | the genomic region of grapevine fanleaf virus (gflv) encoding the movement protein (mp) was cloned into pet21a and transformed into escherichia coli strain bl21 (de3) to express the protein. induction was made with a wide range of isopropyl-β-d-thiogalactopyranoside (iptg) concentrations (1, 1.5, and 2 mm) each for duration of 4, 6, or 16 h. however, the highest expression level was achieved with 1 mm iptg for 4 h. identity of the expressed protein was confirmed by sodium dodecyl sulphate polyac ... | 2016 | 27721695 |
| the fifty distal amino acids of the 2a(hp) homing protein of grapevine fanleaf virus elicit a hypersensitive reaction on nicotiana occidentalis. | avirulence factors are critical for the arm's race between a virus and its host in determining incompatible reactions. the response of plants to viruses from the genus nepovirus in the family secoviridae, including grapevine fanleaf virus (gflv), is well characterized although the nature and characteristics of a viral avirulence factor remains elusive. by using infectious clones of gflv strains f13 and ghu in a reverse genetics approach with wild-type, assortant and chimeric viruses, the determi ... | 2017 | 28387986 |