| analysis of the dsrnas of apple chlorotic leaf spot virus. | double-stranded rnas were isolated from plants infected with five different isolates of apple chlorotic leaf spot virus (aclsv). analysis by page and by northern blot hybridization showed that six major species of viral dsrna of approximately 7.5, 6.4, 5.4, 2.2, 1.1 and 1.0 kbp can be detected in infected plants, irrespective of the aclsv isolate used. in addition to the dsrna of 7.5 kbp corresponding to the full-length genome, the size and position on the genome of the 2.2 and 1.1 kbp species i ... | 1992 | 1634872 |
| conservation of the putative methyltransferase domain: a hallmark of the 'sindbis-like' supergroup of positive-strand rna viruses. | computer-assisted comparisons of the large proteins involved in the replication of viral rna have revealed a novel domain located near the n termini of these proteins and conserved throughout the so-called 'sindbis-like' supergroup of positive-strand rna viruses. this domain encompasses four distinct conserved motifs, with motifs i, ii and iv containing an invariant his residue, the aspxxarg signature and an invariant tyr residue, respectively. each of the two large groups of viruses within this ... | 1992 | 1645151 |
| diverse groups of plant rna and dna viruses share related movement proteins that may possess chaperone-like activity. | amino acid sequences of plant virus proteins mediating cell-to-cell movement were compared to each other and to protein sequences in databases. two families of movement proteins have been identified, the members of which show statistically significant sequence similarity. the first, larger family (i) encompasses the movement proteins of tobamo-, tobra-, caulimo- and comoviruses, apple chlorotic leaf spot virus (aclsv) and geminiviruses with bipartite genomes. thus this family includes viruses wh ... | 1991 | 1684985 |
| use of monoclonal antibodies for the identification of different antigenic domains in apple chlorotic leaf spot virus. | thirteen monoclonal antibodies (mabs) specific for apple chlorotic leaf spot virus (aclsv), produced by the somatic cell hybridization technique, were used to investigate the antigenic structure of the virus. epitope specificity studies showed that these mabs defined in aclsv particles seven independent antigenic domains, representing at least eight distinct epitopes. one of them was present only in virions and not in dissociated subunits. it appeared that the interaction between a mab and the v ... | 1990 | 1700692 |
| phylogeny of capsid proteins of rod-shaped and filamentous rna plant viruses: two families with distinct patterns of sequence and probably structure conservation. | computer-assisted comparative analysis of all available amino acid sequences of the capsid proteins of positive strand rna plant viruses with helical capsids is described. two distinct families of homologous proteins were delineated through statistically significant sequence similarities, one including the capsid proteins of rod-shaped viruses (tobamo-, tobra-, hordei-, and furoviruses) and the other those of filamentous viruses (poty-, bymo-, potex-, carla-, and closteroviruses). it was conclud ... | 1991 | 1871982 |
| the phylogeny of rna-dependent rna polymerases of positive-strand rna viruses. | representative amino acid sequences of the rna-dependent rna polymerases of all groups of positive-strand rna viruses were aligned hierarchiacally, starting with the most closely related ones. this resulted in delineation of three large supergroups. within each of the supergroups, the sequences of segments of approximately 300 amino acid residues originating from the central and/or c-terminal portions of the polymerases could be aligned with statistically significant scores. specific consensus p ... | 1991 | 1895057 |
| nucleotide sequence and genomic organization of apple chlorotic leaf spot closterovirus. | the nucleotide sequence of apple chlorotic leaf spot closterovirus (aclsv) genomic rna has been determined from cdna clones. it is 7555 nucleotides in length excluding the 3' terminal poly(a) tail and contains three putative open reading frames capable of encoding proteins of 216.5, 50, and 28 kda. aclsv rna has untranslated regions of 151 and 190 nucleotides at its 5' and 3' termini, respectively. the 216.5-kda orf encodes a protein which contains the conserved "signature" sequences and has sig ... | 1990 | 2219716 |
| production and use of monoclonal antibodies for the detection of apple chlorotic leaf spot virus. | stable hybridoma cell lines secreting antibodies specific for the apple chlorotic leaf spot virus (clsv) were produced by fusing spleen cells of a biozzi mouse immunized with clsv p863 strain, with the non-secretory p3 x63 ag8.653 myeloma cell line. two hybridoma clones producing monoclonal antibodies of the igg1 subclass were obtained. these monoclonal antibodies were used for virus detection by enzyme-linked immunosorbent assay (elisa). in contrast to polyclonal antisera to clsv, which always ... | 1989 | 2476459 |
| some properties of apple chlorotic leaf spot virus and their relation to purification problems. | | 1971 | 4106981 |
| soluble antigens associated with infection with apple chlorotic leaf spot virus. | | 1973 | 4199139 |
| cherry virus a: cdna cloning of dsrna, nucleotide sequence analysis and serology reveal a new plant capillovirus in sweet cherry. | the nucleotide sequence (7383 nucleotides) of a newly identified member of the genus capillovirus, cherry virus a (cva), was obtained from cdna clones. the cdna was generated from dsrna extracted from plant tissue infected with little cherry virus (lcv). small amounts of lcv dsrna served as template nucleic acid and enabled the construction of a library of which, unexpectedly, 7.5% of the recombinant plasmids were specific for cva. the genome organization of cva resembles that of apple stem groo ... | 1995 | 7636482 |
| papain-like proteinase of turnip yellow mosaic virus: a prototype of a new viral proteinase group. | sequence comparisons predicted a potential papain-like proteinase domain in the n-terminal cleavage product (nrp) of the large nonstructural replicase polyprotein (rp) of turnip yellow mosaic virus (tymv). replacement of the predicted catalytic amino acids, cys-783 by ser, or of his-869 by glu, abolished cleavage of the 206k rp into a approximately 150 k nrp and a approximately 78 k c-terminal product in reticulocyte lysates, while other substitutions exerted no apparent influence on proteolysis ... | 1995 | 7710355 |
| expression, subcellular location and modification of the 50 kda protein encoded by orf2 of the apple chlorotic leaf spot trichovirus genome. | a putative movement protein of molecular mass 50 kda encoded by the orf2 of the apple chlorotic leaf spot virus (aclsv) genome was expressed in escherichia coli using an expression vector and was then used to produce an antiserum. immunoblot analysis using an antiserum raised against this protein showed that the orf2 protein of aclsv was detected in both cell wall and cell membrane fractions prepared from infected chenopodium quinoa tissues. the orf2 protein from infected tissues had a molecular ... | 1995 | 7782779 |
| progress towards a higher taxonomy of viruses. | the current consensus view is that a higher hierarchical taxonomy of viruses cannot be established for two reasons. firstly, viruses appear to be polyphyletic in origin, with several sets of viruses arising by different, independent routes at different times. secondly, subsequent virus adaptation for survival in different host/vector combinations has involved the selective acquisition of additional genes by a process of cassette or modular evolution, with these additional gene modules coming fro ... | 1993 | 8140287 |
| complete nucleotide sequence of the genome of an apple isolate of apple chlorotic leaf spot virus. | the complete nucleotide sequence of the genome of an apple isolate of apple chlorotic leaf spot virus (aclsva) was determined. the genome is 7552 nucleotides excluding the poly(a) tail and contains three open reading frames (orfs 1, 2 and 3), encoding proteins with m(r) values of 216,503 (216.5k), 50,453 (50.4k) and 21,394 (21.4k), respectively. nucleotide sequence comparisons between aclsv-a and the previously sequenced aclsv from plum (aclsv-p) showed that the sequence identity at the nucleoti ... | 1993 | 8376968 |
| localization of fruit tree viruses by immuno-tissue printing in infected shoots of malus sp. and prunus sp. | immuno-tissue printing protocols for the localization of apple chlorotic leaf spot virus (aclsv), stem grooving virus (sgv) and plum pox virus (ppv) in shoots of prunus and malus in vitro have been established for routine diagnosis in a virus elimination program. since these viruses belong to different virus genera, the protocols were adapted according to the properties of the virus under investigation. accumulation of aclsv was highest in the base of the stem and decreased towards the apex of t ... | 1995 | 8537455 |
| the nucleotide sequence and genomic organization of grapevine virus b. | grapevine virus b (gvb) is a tentative member of the genus trichovirus. the 5'-terminal region of the rna genome of gvb comprises 5437 nucleotides and has been sequenced by the dideoxynucleotide chain termination method. evidence was obtained that the rna is capped. two putative open reading frames (orfs) were identified. orf 1 coded for a 194.7 kda polypeptide with conserved motifs of replication-related proteins of positive-strand rna viruses (i.e. methyltransferase, helicase and rna-dependent ... | 1996 | 8887502 |
| in vitro translation of apple chlorotic leaf spot virus (aclsv) rna. | the genomic rna of apple chlorotic leaf spot virus was translated in a rabbit reticulocyte lysate system, yielding a large, 190 k product as well as several other polypeptides of smaller size (60, 56, 22 and 15 kda). the 22 kda product was immunoprecipitated by an anti-aclsv serum and comigrated with purified aclsv coat protein. in vitro translation of rna transcripts prepared from cloned aclsv cdnas demonstrates that the coat protein is synthesised by initiation on the second in frame aug codon ... | 1996 | 8973521 |
| grapevine virus a: nucleotide sequence, genome organization, and relationship in the trichovirus genus. | the 5' terminal region of the genomic rna of grapevine virus a (gva), a tentative member of the trichovirus genus, encompassing 5466 nucleotides, was sequenced. evidence was obtained that the rna is capped. two putative open reading frames (orf) were identified: orf 1 that codes for a 194 kda polypeptide with conserved motifs of replication-related proteins of positive-strand rna viruses, and orf 2 that encodes a 19 kda polypeptide with no significant homology with protein sequences from databas ... | 1997 | 9125055 |
| complete nucleotide sequence of the genome of a severe cherry isolate of apple chlorotic leaf spot trichovirus (aclsv). | the genome of the balaton 1 severe cherry isolate of apple chlorotic leaf spot trichovirus (aclsv-bal1) has been cloned and sequenced. the genomic rna is 7549 nucleotide long, excluding the poly a tail. the genomic organization, with three overlapping open reading frames (orf), is similar to that of the other sequenced aclsv isolates. sequence comparisons indicate a high variability between aclsv isolates, with overall nucleotide sequence homology levels between 76 and 82%. the coat protein, enc ... | 1997 | 9170508 |
| grapevine berry inner necrosis, a new trichovirus: comparative studies with several known trichoviruses. | biological, morphological and serological properties of grapevine berry inner necrosis virus (ginv), the casual virus of grapevine berry inner necrosis disease occurring in japan, were compared with those of several known trichoviruses. host range and particle length of ginv were quite similar to those of apple chlorotic leaf spot trichovirus (aclsv). in ultrathin sections of the infected tissues, ginv particles existed in aggregated masses in the cytoplasm of vascular parenchyma and mesophyll c ... | 1997 | 9267448 |
| apple chlorotic leaf spot virus 50 kda protein is targeted to plasmodesmata and accumulates in sieve elements in transgenic plant leaves. | we investigated the in situ localization of the 50 kda protein encoded by orf2 of apple chlorotic leaf spot virus (aclsv) genome which is thought to be a movement protein. in immunogold electron microscopy of aclsv-infected chenopodium quinoa leaves, the 50 kda protein was localized on plasmodesmata and nearby cytoplasm. observation of transgenic nicotiana occidentalis leaves expressing the 50 kda protein fused to enhanced green fluorescent protein (egfp) by fluorescence and confocal laser scann ... | 1999 | 10664400 |
| nucleotide sequence and genome organisation of cherry mottle leaf virus and its relationship to members of the trichovirus genus. | the nucleotide sequence of cherry mottle leaf virus (cmlv) was determined and compared to sequences of a number of plant viruses including the type member of the trichovirus genus (apple chlorotic leafspot virus, aclsv), and members of the vitivirus genus including grapevine virus b, (gvb). the cmlv genome was determined to consist of 8003 nt excluding the poly(a) tail at the 3' end of the genome. the overall a + u content of cmlv genomic rna was 59.1%, which is similar to aclsv, but significant ... | 2000 | 10881685 |
| intracellular distribution, cell-to-cell trafficking and tubule-inducing activity of the 50 kda movement protein of apple chlorotic leaf spot virus fused to green fluorescent protein. | the 50 kda protein (50kp) encoded by orf2 of apple chlorotic leaf spot virus (aclsv) fused to green fluorescent protein (gfp) was expressed transiently in cells of nicotiana occidentalis and chenopodium quinoa leaves. its intracellular distribution, cell-to-cell trafficking in leaf epidermis and tubule formation on the surface of protoplasts were analysed. the 50kp-gfp fluorescence was distributed as small irregular spots or a fibrous network structure on the periphery of epidermal cells and pro ... | 2000 | 10900048 |
| detection of four apple viruses by multiplex rt-pcr assays with coamplification of plant mrna as internal control. | two multiplex rt-pcr assays with specific coamplification of plant mrna as an internal control from total nucleic acids are described for the parallel detection of apple chlorotic leaf spot virus, apple stem pitting virus, apple mosaic virus and apple stem grooving virus. all are important economically and common pathogens in commercial apple and pear cultivars, except for apple mosaic virus. four virus specific primer pairs and one primer pair which allows the specific amplification of mrna of ... | 2002 | 11684306 |
| detection of apple chlorotic leaf spot virus using a 5' nuclease assay with a fluorescent 3' minor groove binder-dna probe. | the development of a real-time 5' nuclease rt-pcr assay for the detection of apple chlorotic leaf spot virus (aclsv) from infected plant material is described. a short fluorogenic 3' minor groove binder-dna hydrolysis probe was used to circumvent genome variability between isolates and target a short conserved sequence. the covalent attachment of the minor groove binder moiety at the 3' end of the probe increased the probe/target duplex stability and raised the melting temperature to a range sui ... | 2002 | 12020797 |
| the 50-kda protein of apple chlorotic leaf spot virus interferes with intracellular and intercellular targeting and tubule-inducing activity of the 39-kda protein of grapevine berry inner necrosis virus. | to understand why transgenic nicotiana occidentalis plants expressing a functional movement protein (mp) of apple chlorotic leaf spot virus (aclsv) show specific resistance to grapevine berry inner necrosis virus (ginv), the mps of aclsv (50kp) and ginv (39kp) were fused to green, yellow, or cyan fluorescent proteins (gfp, yfp, or cfp). these fusion proteins were transiently expressed in leaf cells of both transgenic (50kp) and nontransgenic (nt) plants, and the intracellular and intercellular t ... | 2003 | 12650450 |
| multiplex rt-pcr-elisa compared with bioassay for the detection of four apple viruses. | a sensitive and reliable multiplex rt-pcr-elisa technique for the detection of apple chlorotic leaf spot virus, apple stem pitting virus, apple mosaic virus and apple stem grooving virus was developed. this technique is compared with the method used commonly for indexing by woody indicators, which is time consuming and expensive. for the rt-pcr-elisa technique, the amplified products were labeled with digoxigenin during the rt-pcr by incorporation of a digoxigenin labeled primer. after hybridiza ... | 2003 | 12798242 |
| stable expression of foreign proteins in herbaceous and apple plants using apple latent spherical virus rna2 vectors. | infectious cdna clones of apple latent spherical virus (alsv)-rna1 (pealsr1) and -rna2 (pealsr2) were constructed using an enhanced 35s promoter. a viral vector was constructed from pealsr2 by creating artificial protease processing sites by duplicating the q/g protease cleavage site between 42kp and vp25. eight rna2-derived vectors expressing gfp with varied sizes of duplications around the 42kp/vp25 junction were constructed and tested for infectivity in chenopodium quinoa. the results indicat ... | 2004 | 15290379 |
| mapping the rna-binding domain on the apple chlorotic leaf spot virus movement protein. | the rna-binding properties of the cell-to-cell movement protein (mp) of apple chlorotic leaf spot virus were analysed. mp was expressed in escherichia coli and was used in uv-crosslinking analysis, using a digoxigenin-utp-labelled rna probe and gel-retardation analysis. the analyses demonstrated that mp bound cooperatively to single-stranded rna (ssrna). when analysed for nacl dependence of the rna-binding activity, the majority of the mp could bind ssrna even in binding buffer with 1 m nacl. fu ... | 2005 | 15604450 |
| simultaneous detection of six stone fruit viruses by non-isotopic molecular hybridization using a unique riboprobe or 'polyprobe'. | a new strategy for the simultaneous detection of plant viruses by molecular hybridization has been developed. two, four or six viral sequences were fused in tandem and transcribed to render unique riboprobes and designated as 'polyprobes'. the 'polyprobe four' (poly 4) covered the four ilarviruses affecting stone fruit trees including apple mosaic virus (apmv), prunus necrotic ringspot virus (pnrsv), prune dwarf virus (pdv), and american plum line pattern virus (aplpv) whereas the 'polyprobe two ... | 2004 | 15664050 |
| simultaneous detection and identification of four pome fruit viruses by one-tube pentaplex rt-pcr. | a pentaplex reverse-transcription polymerase chain reaction (pentaplex rt-pcr) in a single tube was developed for the simultaneous detection of the pome fruit viruses: apple stem pitting virus (aspv), apple stem grooving virus (asgv), apple chlorotic leaf spot virus (aclsv) and apple mosaic virus (apmv). this is the first report of the simultaneous detection of all four viruses and host mrna as an internal specific control. pentaplex rt-pcr was applied successfully throughout the year, using dif ... | 2006 | 16337013 |
| apple chlorotic leaf spot virus 50 kda movement protein acts as a suppressor of systemic silencing without interfering with local silencing in nicotiana benthamiana. | apple chlorotic leaf spot virus (aclsv) is the type species of the genus trichovirus and its single-stranded, plus-sense rna genome encodes a 216 kda protein (p216) involved in replication, a 50 kda movement protein (p50) and a 21 kda coat protein (cp). in this study, it was investigated whether these proteins might have rna silencing-suppressor activities by agrobacterium-mediated transient assay in the green fluorescent protein-expressing nicotiana benthamiana line 16c. the results indicated t ... | 2007 | 17170464 |
| combinations of two amino acids (ala40 and phe75 or ser40 and tyr75) in the coat protein of apple chlorotic leaf spot virus are crucial for infectivity. | amino acid sequences of apple chlorotic leaf spot virus (aclsv) coat protein (cp) were compared between 12 isolates from apple, plum and cherry, and 109 cdna clones that were amplified directly from infected apple tissues. phylogenetic analysis based on the amino acid sequences of cp showed that the isolates and cdna clones were separated into two major clusters in which the combinations of the five amino acids at positions 40, 59, 75, 130 and 184 (ala(40)-val(59)-phe(75)-ser(130)-met(184) or se ... | 2007 | 17698674 |
| the complete nucleotide sequence of an isolate of apple chlorotic leaf spot virus from peach (prunus persica (l.) batch). | | 2008 | 18392552 |
| characterization of apricot pseudo-chlorotic leaf spot virus, a novel trichovirus isolated from stone fruit trees. | abstract a trichovirus closely related to apple chlorotic leaf spot virus (aclsv) was detected in symptomatic apricot and japanese plum from italy. the sus2 isolate of this agent cross-reacted with anti-aclsv polyclonal reagents but was not detected by broad-specificity anti- aclsv monoclonal antibodies. it had particles with typical trichovirus morphology but, contrary to aclsv, was unable to infect chenopodium quinoa and c. amaranticolor. the sequence of its genome (7,494 nucleotides [nt], mis ... | 2005 | 18943045 |
| interference of long-distance movement of grapevine berry inner necrosis virus in transgenic plants expressing a defective movement protein of apple chlorotic leaf spot virus. | abstract transgenic nicotiana occidentalis plants expressing a movement protein (p50) and partially functional deletion mutants (deltaa and deltac) of the apple chlorotic leaf spot virus (aclsv) showed resistance to grapevine berry inner necrosis virus (ginv). the resistance is highly effective and ginv was below the level of detection in both inoculated and uninoculated upper leaves. in contrast, ginv accumulated in inoculated and uninoculated leaves of nontransgenic (nt) plants and transgenic ... | 2006 | 18943419 |
| polyvalent degenerate oligonucleotides reverse transcription-polymerase chain reaction: a polyvalent detection and characterization tool for trichoviruses, capilloviruses, and foveaviruses. | abstract a polyvalent nested reverse transcription-polymerase chain reaction (rt-pcr) test using degenerate primers containing inosine (polyvalent degenerate oligonucleotides [pdo]) was developed for filamentous fruit tree viruses belonging to the genera trichovirus, capillovirus, and foveavirus. the 362-bp product was amplified from nucleic acid extracts obtained from prunus and malus leaf samples. all the viruses targeted were detected, demonstrating the polyvalence of the test. the variabilit ... | 2005 | 18943777 |
| transgenic nicotiana occidentalis plants expressing the 50-kda protein of apple chlorotic leaf spot virus display increased susceptibility to homologous virus, but strong resistance to grapevine berry inner necrosis virus. | abstract the 50-kda protein (p50) encoded by the open reading frame 2 of apple chlorotic leaf spot virus (aclsv), a putative movement protein, was expressed in transgenic nicotiana occidentalis plants. p50 in transgenic plants was mainly detected in a modified form in the cell wall fraction, similar to that in infected leaves. the p50-expressing plants (p50 plants) complemented the systemic spread of the p50-defective mutants of an infectious cdna clone of aclsv (pclsf), indicating that p50 in t ... | 2000 | 18944625 |
| inhibition of long-distance movement of rna silencing signals in nicotiana benthamiana by apple chlorotic leaf spot virus 50 kda movement protein. | apple chlorotic leaf spot virus 50 kda movement protein (p50) acts as a suppressor of systemic silencing in nicotiana benthamiana. here, we investigate the mode of action of p50 suppressor. an agroinfiltration assay in gfp-expressing n. benthamiana line16c (gfp-plant) showed that p50 could not prevent the short-distance spread of silencing. in grafting experiments, the systemic silencing was inhibited in gfp-plants (scion) grafted on p50-expressing n. benthamiana (p50-plant; rootstock) when gfp ... | 2008 | 18954886 |
| molecular variability analyses of apple chlorotic leaf spot virus capsid protein. | the complete sequences of the coat protein (cp) gene of 26 isolates of apple chlorotic leaf spot virus (aclsv) from india were determined. the isolates were obtained from various pome (apple, pear and quince) and stone (plum, peach, apricot, almond and wild himalayan cherry) fruit trees. other previously characterized aclsv isolates and trichoviruses were used for comparative analysis. indian aclsv isolates among themselves and with isolates from elsewhere in the world shared 91-100% and 70-98% ... | 2010 | 21289443 |
| Strategies to facilitate the development of uncloned or cloned infectious full-length viral cDNAs: Apple chlorotic leaf spot virus as a case study. | Approaches to simplify and streamline the construction of full-length infectious cDNA clones (FL-cDNAs) are needed. Among desirable improvements are the ability to use total nucleic acids (TNA) extracts from infected hosts (to bypass viral purification limitations) for the direct one-step amplification of large FL-cDNAs, the possibility to inoculate plants with uncloned FL-cDNAs and the simplified cloning of these large molecules. | 2011 | 22040379 |
| determination of major viral and sub viral pathogens incidence in apple orchards in himachal pradesh. | apple is the major commercial horticulture crop in himachal pradesh and other hill states of jammu & kashmir, uttarakhand and some parts of northeastern states of india. in order to gather data on health status and incidence of virus and virus-like pathogens in apple orchards, survey was conducted in the month of june and september, 2010 in hatkoti, rohru, kuthara, jubbal and khadapathar areas of major apple producing shimla district of himachal pradesh. a total of 250 samples were collected and ... | 2011 | 23730008 |
| characterization of apple stem grooving virus and apple chlorotic leaf spot virus identified in a crab apple tree. | apple stem grooving virus (asgv), apple chlorotic leaf spot virus (aclsv), and prunus necrotic ringspot virus (pnrsv) were identified in a crab apple tree by small rna deep sequencing. the complete genome sequence of aclsv isolate bj (aclsv-bj) was 7554 nucleotides and shared 67.0%-83.0% nucleotide sequence identity with other aclsv isolates. a phylogenetic tree based on the complete genome sequence of all available aclsv isolates showed that aclsv-bj clustered with the isolates sy01 from hawtho ... | 2017 | 27990565 |
| deep sequencing analysis of apple infecting viruses in korea. | deep sequencing has generated 52 contigs derived from five viruses; apple chlorotic leaf spot virus (aclsv), apple stem grooving virus (asgv), apple stem pitting virus (aspv), apple green crinkle associated virus (agcav), and apricot latent virus (aplv) were identified from eight apple samples showing small leaves and/or growth retardation. nucleotide (nt) sequence identity of the assembled contigs was from 68% to 99% compared to the reference sequences of the five respective viral genomes. sequ ... | 2016 | 27721694 |
| genome sequences of three apple chlorotic leaf spot virus isolates from hawthorns in china. | the genome sequences of apple chlorotic leaf spot virus (aclsv) isolates from three accessions of hawthorns (crataegus pinnatifida) grown at shenyang agricultural university were determined using illumina rna-seq. to confirm the assembly data from the de novo sequencing, two aclsv genomic sequences (sy01 and sy02) were sequenced using the sanger method. the sy01 and sy02 sequences obtained with the sanger method showed 99.5% and 99.7% nucleotide identity with the transcriptome data, respectively ... | 2016 | 27519059 |
| a multiple rt-pcr assay for simultaneous detection and differentiation of latent viruses and apscarviroids in apple trees. | apple chlorotic leaf spot virus (aclsv), apple stem grooving virus (asgv), and apple stem pitting virus (aspv) are three latent viruses frequently occurring in apple trees worldwide. in field orchards, these viruses are frequently found in a mixed infection with viroids in the genus apscarviroid, including apple scar skin viroid, and apple dimple fruit viroid. together these viruses and viroids could cause serious damage to apple fruit production worldwide. rapid and efficient detection methods ... | 2016 | 27054889 |
| cryotherapy as a method for reducing the virus infection of apples (malus sp.). | there is an urgent need in kazakhstan for virus-free nursery stock to reinvigorate the industry and preserve historic cultivars. an in vitro collection of apples could be used for virus testing and elimination and to provide virus-free elite stock plants to nurseries. | 2016 | 26964019 |
| [identification of host factors interacting with the movement protein of apple chlorotic leaf spot virus by yeast two-hybrid system]. | in order to identify host factors which interact with the movement protein (mp) of apple chlorotic leaf spot virus (aclsv), aclsv mp was cloned into the bait vector pgbkt7 and used to screen a cdna library of malus sylvestris cv. r12740-7a, which had previously been constructed by yeast two-hybrid sequencing transformation. the protein functions of the identified host factors were determined according to their gene annotations in genbank. the result showed that the bait plasmid pgbkt7-mp showed ... | 2015 | 26164936 |
| a simplified strategy for studying the etiology of viral diseases: apple stem grooving virus as a case study. | a simple method to amplify infective, complete genomes of single stranded rna viruses by long distance pcr (ld pcr) from woody plant tissues is described in detail. the present protocol eliminates partial purification of viral particles and the amplification is achieved in three steps: (i) easy preparation of template rna by incorporating a pre processing step before loading onto the column (ii) reverse transcription by amv or superscript reverse transcriptase and (iii) amplification of cdna by ... | 2015 | 25486082 |
| simultaneous detection of major pome fruit viruses and a viroid. | a rapid and sensitive two-step rt-pcr protocol for simultaneous detection of major apple viruses, namely apple mosaic virus (apmv), apple stem pitting virus (aspv), apple stem grooving virus (asgv), apple chlorotic leaf spot virus (aclsv) and apple scar skin viroid (assvd), was developed. five specific primer pairs were tested and confirmed for these viruses and viroid together in a single tube, giving amplicons of ~198, ~330, ~370, ~547 and ~645 bp corresponding to asgv, assvd, aspv, apmv and a ... | 2014 | 25320423 |
| the rt-pcr identification and sequence analysis of apple chlorotic leaf spot virus from apple cultivars in jiaodong peninsula, china. | a set of specific primer pairs was utilized to detect apple chlorotic leaf spot virus (aclsv) from seven different apple cultivars in jiaodong peninsula via reverse transcription polymerase chain reaction (rt-pcr), and the sequence of aclsv genome was analysed. the results indicate that: (1) high-purity total rna could be successfully isolated using plant rna rapid extraction kit. the ratios of a260/a280 varied between 1.8 and 2.1. the fragmentation in agarose gel was good and the 28s and 16s ba ... | 2014 | 26019509 |
| genetic variation analysis of apple chlorotic leaf spot virus coat protein reveals a new phylogenetic type and two recombinants in china. | in this study, we generated sequences of the apple chlorotic leaf spot virus (aclsv) coat protein (cp) gene. genetic variation and phylogenetic analyses were carried out on these sequences along with others reported previously. aclsv populations clustered into four types: in three of the four types, combinations of three amino acids at positions of 40, 75 and 79 were conserved. the fourth phylogenetic type, newly identified here, was characterized by co-variation of ser(40)-tyr(75)-ser(79). stat ... | 2014 | 24318575 |
| multiplex rt-pcr detection and distribution of four apple viruses in china. | apple trees are natural hosts of four economically important virus species in china. we used a simple, sensitive multiplex rt-pcr protocol with an internal control to simultaneously detect and differentiate four apple viruses: apple chlorotic leaf spot virus (aclsv), apple mosaic virus (apmv), apple stem pitting virus (aspv), and apple stem grooving virus (asgv). this multiplex rt-pcr could be used as an alternative to other routinely used detection methods. we used this protocol to evaluate the ... | 2013 | 24294957 |
| molecular detection of latent apple chlorotic leaf spot virus in elite mother plants of apple. | apple chlorotic leaf spot virus (aclsv; family betaflexiviridae genus trichovirus) is one of the economically important latent virus infecting apple (malus × domestica borkh.). reverse transcriptase polymerase chain reaction (rt-pcr) procedures were used to amplify coat protein gene of aclsv. among 5 primer sets used, two primer sets (1f1r and 1f2r) amplified fragments of expected size (432 bp). products visible on agarose gel were produced using templates extracted from apple leaves. the result ... | 2012 | 24293825 |
| simultaneous detection and differentiation of three viruses in pear plants by a multiplex rt-pcr. | a multiplex rt-pcr (mrt-pcr) assay was developed for detection and differentiation of the apple stem pitting virus (aspv), apple stem grooving virus (asgv) and apple chlorotic leaf spot virus (aclsv), which are viruses frequently occurring in pear trees. different combinations of mixed primer pairs were tested for their specificity and sensitivity for the simultaneous detection of the three viruses. three primer pairs were used to amplify their fragments of 247bp, 358bp and 500bp, respectively. ... | 2014 | 24269332 |
| a duplex, sybr green i-based rt-qpcr assay for the simultaneous detection of apple chlorotic leaf spot virus and cherry green ring mottle virus in peach. | co-infections of apple chlorotic leaf spot virus (aclsv) and cherry green ring mottle virus (cgrmv) in peach is common in china and have resulted in significant yield reductions. a reliable, sensitive and quantitive method is needed to detect and distinguish between aclsv and cgrmv in peach. | 2013 | 23938201 |
| a one-step multiplex rt-pcr assay for simultaneous detection of four viruses that infect peach. | a multiplex reverse transcription polymerase chain reaction (mrt-pcr) assay was developed to enable the simultaneous detection and differentiation of four viruses that infect peach, namely apple chlorotic leaf spot virus (aclsv), cherry green ring mottle virus (cgrmv), prunus necrotic ringspot virus (pnrsv) and apricot pseudo-chlorotic leaf spot virus (apclsv). in this study, four pairs of primers, one specific for each virus, were designed; the corresponding pcr products were 632, 439, 346 and ... | 2013 | 23777367 |
| characterization of plant virus-encoded gene silencing suppressors. | agroinfiltration assay using green fluorescent protein (gfp)-expressing nicotiana benthamiana line 16c is a powerful method for screening of putative plant virus-encoded gene silencing suppressors. this method allows the investigator to know whether the putative viral suppressor inhibits silencing in a cell (local silencing) and/or spreading of silencing throughout a plant (systemic silencing). additionally, grafting experiments using transgenic plants expressing the suppressor and the gfp will ... | 2012 | 22678576 |
| complete nucleotide sequences of the genomes of two isolates of apple chlorotic leaf spot virus from peach (prunus persica) in china. | the complete nucleotide sequences of two isolates of apple chlorotic leaf spot virus (z1 and z3) collected from peach in henan province, china, were determined. the genomes of both z1 and z3 were found to contain three open reading frames (orfs). sequence analysis showed that genomic sequences of z1 and z3 isolates shared 67.4%-82.9% and 67.2%-82.6% identity, respectively, with the other eight isolates of aclsv that have been reported previously. based on the putative amino acid sequences of the ... | 2012 | 22278708 |