| characterization of cymbidium mosaic virus coat protein gene and its expression in transgenic tobacco plants. | cymbidium mosaic virus (cymv) is the most prevalent virus infecting orchids. here, we report the isolation of partial cdna clones encoding the genomic rna of cymv. like most of the polyadenylated monopartite positive-strand rna viruses, the open reading frame (orf) coding for the viral coat protein (cp) is located at the 3' end. the orf predicts a polypeptide chain of 220 amino acids with a molecular weight of 23,600. sequence comparison of this orf to the cp sequences of potato virus x(pvx) and ... | 1992 | 1600145 |
| ubiquitinated conjugates are found in preparations of several plant viruses. | recently d.d. dunigan, r.g. dietzgen, j.e. schoelz, and m. zaitlin (virology 165, 310-312, 1988) demonstrated that a small proportion of the subunits of tobacco mosaic virus particles were conjugated with the small protein ubiquitin. we have now detected ubiquitinated conjugates in immunoblots of virion preparations of several other plant viruses, using anti-human ubiquitin antiserum. based on their polyacrylamide gel migrations, plant virus-associated ubiquitin-immunoreactive proteins were cons ... | 1990 | 2162109 |
| rapid detection of cymbidium mosaic virus by the polymerase chain reaction (pcr). | a direct, simple and sensitive method was developed for the detection of cymbidium mosaic virus (cymv), based on the polymerase chain reaction (pcr). two oligonucleotide primers were selected from regions that are homologous to potexviruses and cymv, and used to hybridize with purified viral rna and particles. this resulted in the amplification of a 313 bp fragment after 30 cycles of pcr in all samples. a less prominent fragment of 227 bp was also obtained due to mispriming of the second primer. ... | 1993 | 8432762 |
| rapid detection and identification of odontoglossum ringspot virus by polymerase chain reaction amplification. | the odontoglossum ringspot tobamovirus (orsv) movement and coat proteins genes were selected for the design of oligonucleotide primers for amplification of a 1,085 bp fragment. a combined assay of reverse transcription and the polymerase chain reaction (rt-pcr) was performed with 20-mer orsv-specific primers and crude nucleic acid extracts from virus-infected orchids for rapid detection of the virus. the lowest concentration of template viral rna required for detection was 10 fg. the rt-pcr is a ... | 1995 | 8522141 |
| an appraisal of the banded and paracrystalline cytoplasmic inclusions induced in cymbidium mosaic potexvirus- and odontoglossum ringspot tobamovirus infected orchid cells using confocal laser scanning microscropy. | the cymbidium mosaic potexvirus (cymv) banded inclusions and the odontoglossum ringspot tobamovirus (orsv) paracrystalline inclusions were studied in flowers and leaves of nine orchid cultivars using the confocal laser scanning microscope. the inclusions varied in length and width and were mostly located adjacent to the cell walls. some orsv inclusions fully extended across entire infected cells. we propose that the cymv banded inclusion was formed from virus aggregates which aligned as periodic ... | 1996 | 8634017 |
| coat protein gene and 3'-noncoding region of a new korean isolate of cymbidium mosaic virus. | a new korean isolate of cymbidium mosaic virus (denoted as cymmv-k2), a member of potexviruses, was identified and isolated from a korean cultivar of cymbidium species when the nucleotide sequence of the 3'-terminal region of the viral rna was compared with those of the corresponding regions of two singaporean isolates (denoted as cymmv-s1 and cymmv-s2) and a korean isolate (denoted as cymmv-k1), the nucleotide sequence of the coat protein gene of cymmv-k2 was highly homologous to other cymbidiu ... | 1997 | 9085262 |
| the use of dig-labelled crna probes for the detection of cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) in orchids. | dig-labelled sense and antisense crna probes were synthesized from cdna clones of cymmv and orsv for virus detection in infected plants. a slot-blot hybridization assay was developed using either crude leaf extracts or total rna from infected leaves. the assay could detect 50 and 250 pg of purified cymmv and orsv rna, respectively. as little as 30 mg of nicotiana benthamiana infected leaves was sufficient to provide positive detection. cymmv and orsv were detected at 3125 and 625 times dilution ... | 1998 | 9562413 |
| phylogenetic analysis of triple gene block viruses based on the tgb 1 homolog gene indicates a convergent evolution. | the complete nucleotide sequence of the triple gene block one (tgb 1) of cymbidium mosaic potexvirus (cymmv) was compared to those from other potex-, carla-, furo- and hordeiviruses. seven conserved motifs in the tgb 1, including the atp-gtp binding domain (p-loop) consensus gxxgktsts, were found in all four virus genera. we propose that all tgbv can be classified into phylogenetic clusters based on their tgb 1 homolog genes. these clusters can be further delineated to form subgroups. the first ... | 1998 | 9654683 |
| simultaneous td/rt-pcr detection of cymbidium mosaic potexvirus and odontoglossum ringspot tobamovirus with a single pair of primers. | cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) are the two most prevalent and economically important orchid viruses which have attained a world-wide distribution. we have designed one pair of common primers, which were able to detect, simultaneously, both viruses by a single polymerase chain reaction (pcr). sequence alignment and primer analyses were employed in primer design. the effect of touch-down pcr, primer annealing temperature, number of thermal cycles, ... | 1998 | 9694327 |
| synthesis of biologically active cdna clones of cymbidium mosaic potexvirus using a population cloning strategy. | biologically active cdna clones of cymbidium mosaic potexvirus (cymmv) were synthesized using a population cloning strategy. three populations of overlapping rt-pcr products encompassing the entire viral rna of cymmv were ligated into pbluescriptks+ with t7 rna polymerase promoter fused to the 5' extreme of the viral cdna. capped-rna in vitro transcripts were infectious. this is the first report of successful synthesis of biologically active cymmv clones. unlike the conventional methods, populat ... | 1998 | 9739338 |
| rapid simultaneous detection of two orchid viruses using lc- and/or maldi-mass spectrometry. | liquid chromatography/mass spectrometry (lc/ms) and matrix-assisted laser desorption-ionization (maldi) mass spectrometry are capable of providing molecular mass information on biological samples with high speed, accuracy and sensitivity. with mass spectrometry, identifying a virus based on the molecular weight of its coat protein is relatively simple and accurate. the technique can be applied to all viruses with known coat protein molecular weights. using the lc/ms and/or maldi, this paper desc ... | 2000 | 10716342 |
| genetic variability in the coat protein genes of two orchid viruses: cymbidium mosaic virus and odontoglossum ringspot virus. | the variability in coat protein gene sequences of cymbidium mosaic virus (cymmv) and odontoglossum ringspot virus (orsv) that naturally infect orchids worldwide was investigated. samples were collected from korea, singapore and taiwan. the sequence data were compared with available published coat protein gene sequences of cymmv and orsv, including those from japan and thailand. among cymmv isolates, the homology was 89.1%-99.7% and 93.2%-100% at the nucleotide and amino acid levels, respectively ... | 2002 | 12376755 |
| complete nucleotide sequence of the genomic rna of a french isolate of pepino mosaic virus (pepmv). brief report. | the complete nucleotide sequence of the genomic rna of a french isolate of pepino mosaic virus (pepmv) was determined. with a length of 6,425 nucleotides excluding the poly(a) tail at the 3' terminus, the pepmv genome contains five major open reading frames encoding a 164 kda replicase, triple gene block proteins of 26 kda, 14 kda and 9 kda, and a 25 kda coat protein. this genome organization, as well as characteristic consensus motifs and high degrees of similarity of pepmv proteins with those ... | 2002 | 12417957 |
| [viruses of orchids of the natural flora in ukraine]. | orchids from biogeocenosis of natural landscapes of ukraine (the crimea, carpathians) were analyzed for presence of viral pathogens. it is demonstrated that tobacco rattle virus (trv), tomato aspermy virus (tav), cymbidium mosaic virus (cymmv) and arabis mosaic virus (armv) are the most spreaded pathogens of orchid plants. there are orchid species distinguished in natural flora of ukraine with characteristic high frequency of viral infection. | 2004 | 15208857 |
| transcription analysis of peloric mutants of phalaenopsis orchids derived from tissue culture. | tissue culture has been widely used for mass propagation of phalaenopsis. however, somaclonal variation occurred during micropropagation process posed a severe problem by affecting product quality. in this study, wild type and peloric flower buds of phalaenopsis hybrids derived from flower stalk nodal culture were used for cdna-rapd and cdna suppression subtractive hybridization analyses in order to study their genetic difference in terms of expressed sequence tags. a total of 209 ests from norm ... | 2005 | 16117854 |
| gene stacking in phalaenopsis orchid enhances dual tolerance to pathogen attack. | cymbidium mosaic virus (cymmv) and erwinia carotovora have been reported to cause severe damage to orchid plants. to enhance the resistance of orchids to both viral and bacterial phytopathogens, gene stacking was applied on phalaenopsis orchid by double transformation. plbs originally transformed with cymmv coat protein cdna (cp) were then re-transformed with sweet pepper ferredoxin-like protein cdna (pflp) by agrobacterium tumefaciens, to enable expression of dual (viral and bacterial) disease ... | 2005 | 16145836 |
| evidence that cymbidium mosaic virus (cymmv) isolates divide into two subgroups based on nucleotide diversity of coat protein and replicase genes. | the genetic diversity of cymbidium mosaic virus (cymmv, family flexiviridae) was assessed by analysing the nucleotide sequences of coat protein (cp) and partial rna-dependent rna polymerase (rdrp) genes. thirty cymmv sequences from vanilla isolates, obtained in this work by direct sequencing of rt-pcr products, were compared to the sequences from ornamental orchid isolates available in genbank. the cymmv population exhibited overall low genetic diversity (pi=0.054 and pi=0.053 for cp and rdrp ge ... | 2007 | 17216134 |
| molecular beacons: a new approach to plant virus detection. | abstract molecular beacons are single-stranded nucleic acid molecules with a stem-loop conformation. the stem portion consists of complementary sequences at the 5' and 3' terminals of the molecule, while the loop portion consists of probe sequences that are complementary to the target sequences of choice. a fluorescent moiety is attached to one end, while a quenching moiety is attached to the opposite end. reverse transcription-polymerase chain reactions are carried out with primers that amplify ... | 2000 | 18944619 |
| detection of cymbidium mosaic potexvirus and odontoglossum ringspot tobamovirus using immuno-capillary zone electrophoresis. | abstract immuno-capillary zone electrophoresis (i-cze) is a technique that combines the specificity afforded by serological assays with the sensitivity, rapidity, and automation in detection provided by capillary zone electrophoresis. cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) were detected in their purified forms as well as in the crude saps of infected nicotiana benthamiana leaves and oncidium orchid flowers. the two orchid virus-antibody complexes were r ... | 1999 | 18944725 |
| cymbidium mosaic virus coat protein gene in antisense confers resistance to transgenic nicotiana occidentalis. | the nucleotide sequence of the 3'-terminal region of the korean isolate of cymbidium mosaic virus (cymv-ca) from a naturally infected cattleya was determined. the sequence contains an open reading frame (orf) coding for the viral coat protein (cp) at the 3'-end and three other orfs (triple gene block or movement protein) of cymv. the cp gene encodes a polypeptide chain of 220 amino acids with a molecular mass of 23,760 da. the deduced cp sequence showed a strong homology with those of two cymvs ... | 1999 | 10672926 |
| the gprime package: computer programs for identifying the best regions of aligned genes to target in nucleic acid hybridisation-based diagnostic tests, and their use with plant viruses. | the gprime (group primer design) programs examine aligned sets of gene sequences to discover homologous regions to be targeted in diagnostic tests. the core program moves a 'window' over the aligned sequences and calculates, at each window position, a 'redundancy value', namely the number of sequences that would represent all permutations of the variable sequence positions within that window. regions with minimal redundancy values may then be targeted in diagnostic tests based on oligonucleotide ... | 1998 | 9763130 |
| synergism in replication of cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) rna in orchid protoplasts. | an in vitro orchid protoplast isolation method to study replication kinetics of cymmv and orsv was developed. this method allows the isolation of viable and raphid-free petal protoplasts from an orchid hybrid, dendrobium sonia (dendrobium caesar x dendrobium tomie drake). the optimum field strength for both viral rna to achieve good efficiency of electroporation was 750 v/cm and the optimum viral rna concentration required was 1 microgram and 4 micrograms per 2 x 10(6) protoplasts for cymmv and ... | 1998 | 9722873 |
| genome characterization of a korean isolate of cymbidium mosaic virus. | the complete nucleotide sequence of the genomic rna of a korean isolate of cymbidium mosaic virus (cymmv-k2) was determined. the genomic rna is 6227 nucleotides in length, excluding the poly(a) tail. it contains a 5'-noncoding region (ncr) of 73 nucleotides, five open reading frames (orfs 1 to 5) which encode proteins with m(r) 160 kda rna-dependent rna polymerase (orf1), 26 kda movement protein 1 (orf2), 13 kda movement protein 2 (orf3), 10 kda movement protein 3 (orf4), 24 kda coat protein (of ... | 1998 | 9638650 |
| cymbidium mosaic potexvirus rna: complete nucleotide sequence and phylogenetic analysis. | the complete nucleotide sequence of the genomic rna of cymbidium mosaic potexvirus (cymmv) was determined to be 6227 nucleotides in length, excluding the poly (a) tail at the 3' terminus. similar to other potexviruses, its genome organisation is comprised of five major open reading frames (orfs 1 to 5), encoding a mr 160 kda putative rna-dependent rna polymerase (rdrp); a mr 26kda/13kda/10kda triple-gene-block (tgb) and a mr 24 kda coat protein. the cymmv encoded proteins shared a high degree of ... | 1997 | 9125051 |
| use of reverse transcription-polymerase chain reaction for cloning of coat protein-encoding genes of cymbidium mosaic virus. | a reverse transcription-polymerase chain reaction (rt-pcr) has been developed for the cloning of coat protein-encoding genes (cp) of cymbidium mosaic virus (cymv) isolated from three different infected species of thai orchid: cattleya, mokara and oncidium. the analysis of the compiled sequences of the cdna clones shows a single open reading frame which encoded a cymv cp gene. the gene is 669 nucleotides (nt) long and codes for a 23 761-da protein. the nt sequences of cymv cp from the thai isolat ... | 1996 | 8955635 |
| cymbidium mosaic virus rna synthesis in isolated orchid protoplasts. | an in vitro protoplast host system has been developed to facilitate the study of cymbidium mosaic virus (cymv) replication mechanisms. cymv, a member of the potexvirus group of plant viruses, is among the most significant pathogens of cultivated orchid plants, but a convenient system for studying its molecular genetics has not been available previously. a cdna probe representing a segment of the cymv genome was used to detect viral rna following inoculation of vanda floral protoplasts with virus ... | 1993 | 8249447 |
| nucleotide sequence and in vitro translation of the coat protein gene of cymbidium mosaic virus. | the nucleotide sequence of the coat protein gene of cymbidium mosaic virus (cymv) has been determined from a cdna clone that encompassed the 3'-region of the virus genome. the 0.75 kb coat protein open reading frame (orf) codes for a 24kd polypeptide. the coat protein coding region was separated from the 3'-poly(a) terminal of the viral genome by an 80 base noncoding region. the derived amino acid sequence of this orf showed considerable homology with other potexvirus coat protein sequences, not ... | 1993 | 8103614 |
| cloning and sequencing of a cdna encoding the coat protein of a korean isolate of cymbidium mosaic virus. | the 3'-terminal nucleotide (nt) sequence of the coat protein (cp)-encoding gene of the korean isolate of cymbidium mosaic potexvirus (cymv-k) has been determined. the sequence contained a full-length orf coding for the viral cp. it comprises 663 nt and a polypeptide chain of 220 amino acids with a m(r) of 23,718. the stop codon (tga) is followed by a 3'-untranslated region (3'-utr) of 79 nt and a poly(a) tract. the putative polyadenylation signal, aataaa motif, is found in the terminus of the 3' ... | 1995 | 7758973 |
| virus resistance in orchids. | orchid plants, phalaenopsis and dendrobium in particular, are commercially valuable ornamental plants sold worldwide. unfortunately, orchid plants are highly susceptible to viral infection by cymbidium mosaic virus (cymmv) and odotoglossum ringspot virus (orsv), posing a major threat and serious economic loss to the orchid industry worldwide. a major challenge is to generate an effective method to overcome plant viral infection. with the development of optimized orchid transformation biotechnolo ... | 2014 | 25438783 |
| multiplex rt-pcr detection of three common viruses infecting orchids. | a multiplex reverse transcription polymerase chain reaction (rt-pcr) assay was developed for simultaneous detection of three orchid viruses: cymbidium mosaic virus (cymmv), odontoglossum ringspot virus (orsv), and orchid fleck virus (ofv). primers were used to amplify nucleocapsid protein gene fragments of 845 bp (orsv), 505 bp (cymmv) and 160 bp (ofv). a 60-bp amplicon of plant glyceraldehyde-3-phophate dehydrogenase mrna was included as an internal control against false negatives. the assay wa ... | 2014 | 24980395 |
| nucleotide sequences of the two orfs upstream to the coat protein gene of cymbidium mosaic virus. | | 1992 | 1581564 |
| cymbidium mosaic potexvirus isolate-dependent host movement systems reveal two movement control determinants and the coat protein is the dominant. | little is known about how plant viruses of a single species exhibit different movement behavior in different host species. two cymbidium mosaic potexvirus (cymmv) isolates, m1 and m2, were studied. both can infect phalaenopsis orchids, but only m1 can systemically infect nicotiana benthamiana plants. protoplast inoculation and whole-mount in situ hybridization revealed that both isolates can replicate in n. benthamiana; however, m2 was restricted to the initially infected cells. genome shuffling ... | 2009 | 19345971 |
| detection of two orchid viruses using quartz crystal microbalance-based dna biosensors. | abstract we have developed a piezoelectric dna-sensor based on dna-rna hybridization for the detection of two orchid viruses, cymbidium mosaic virus (cymmv) and odontoglossum ringspot virus (orsv). specific oligonucleotide probes modified with a mercaptohexyl group at the 5'-phosphate end were directly immobilized onto 10-mhz at-cut quartz crystal microbalance (qcm). qcms coated with such oligonucleotide probes were exposed to test solutions containing viral rna for hybridization. various experi ... | 2002 | 18944263 |
| the rx gene confers resistance to a range of potexviruses in transgenic nicotiana plants. | rx-mediated resistance was analyzed in rx-expressing transgenic nicotiana plants. the infection outcome of nine potato virus x isolates mutated at amino acid positions 121 and 127 of the coat protein (cp) confirmed the key role of these amino acids but provided a more complex picture than previously reported. in particular, in rx-expressing nicotiana spp., eliciting activity modulated by amino acid 121 was conditioned by the nature of amino acid 127. these results suggest that the specificity of ... | 2008 | 18700820 |
| characterization of hawaiian isolates of cymbidium mosaic virus (cymmv) co-infecting dendrobium orchid. | we report here the isolation and characterization of three distinct isolates of cymbidium mosaic virus (cymmv) co-infecting dendrobium orchid in hawaii. isolates 1 and 2 were phylogenetically distinct from previously reported cymmv isolates. however, isolate 3 was highly similar to previously reported cymmv sequences and could be localised to cymmv subgroup a. isolate 2 localised to cymmv subgroup b. thus, we report here the first full-length cymmv subgroup b isolate. isolate 1 represents a reco ... | 2008 | 18458814 |
| [production of monoclonal antibodies to cymbidium mosaic virus and application in orchids virus detection]. | three hybridoma cell lines, 2c6, 5b7 and 12g9, secreting monoclonal antibodies (mcabs) against cymbidium mosaic virus (cymmv) were produced by fusing mouse myeloma cells (sp2/0) with spleen cells from balb/ c immunized by the cymmv particles. the three mcabs could specifically react with cymmv. the titres of ascitic fluids of two mcabs are up to 10(-6) in i-elisa. isotypes and subclasses of the the three mcabs belong to igg1. isotypes of light strains of the three mcabs all belong to kappa. they ... | 2007 | 18062276 |
| complete nucleotide sequence analysis of cymbidium mosaic virus indian isolate: further evidence for natural recombination among potexviruses. | the complete nucleotide sequence of an indian strain of cymbidium mosaic virus (cymmv)was determined and compared with other potexviruses. phylogenetic analyses on the basis of rna-dependent rna polymerase (rdrp), triple gene block protein and coat protein (cp) amino acid sequences revealed that cymmv is closely related to the narcissus mosaic virus (nmv), scallion virus x (svx), pepino mosaic virus (pepmv)and potato aucuba mosaic virus (pamv). different sets of primers were used for the amplifi ... | 2007 | 17762139 |
| [molecular identification of cymbidium mosaic potexvirus and odontoglossum ringspot tobamovirus complex infected phalaenopsis and its pathological ultrastructural alteration]. | filamentous and rod-shaped virions, and aggregated crystals were observed in infected leaves of by negative staining and ultramicrotomy. histologic study synchronously showed typical crystal forms of the two virions: the crystals from filamentous particles congregated in strips, arrayed in multilayer and piled in certain angles or helix between layers; while the crystals from rod-shaped particles arrayed in parallel, angle-layer or helix. the two kinds of crystals both presented in parenchyma ce ... | 2007 | 17580669 |
| strategies for functional validation of genes involved in reproductive stages of orchids. | plants in the largest family of angiosperms, orchidaceae, are diverse in both specialized pollination and ecological strategies and provide a rich source for investigating evolutionary relationships and developmental biology. however, studies in orchids have been hindered by several challenges that include low transformation efficiency and long regeneration time. to overcome such obstacles, we selected a symptomless cymbidium mosaic virus (cymmv) isolate for constructing virus-induced gene-silen ... | 2007 | 17189336 |
| use of superparamagnetic beads for the isolation of a peptide with specificity to cymbidium mosaic virus. | a modified method for the rapid isolation of specific ligands to whole virus particles is described. biopanning against cymbidium mosaic virus was carried out with a commercial 12-mer random peptide display library. a solution phase panning method was devised using streptavidin-coated superparamagnetic beads. the solution based panning method was more efficient than conventional immobilized target panning when using whole viral particles of cymbidium mosaic virus as a target. enzyme-linked immun ... | 2006 | 16781785 |
| detection and serological relationships of cymbidium mosaic potexvirus isolates. | twenty-two isolates of cymbidium mosaic virus (cymv) were isolated from 35 orchid plants suspected of being infected with cymv. among the three methods used for detecting cymv, immunoelectron microscopy (iem-1) was shown to be the most sensitive method, being able to detect the virus in 71.43% of suspected cymv-infected plants while the electron microscopic method and the indexing plant method could detect 51.43 and 42.86%, respectively. out of 12 symptomless plants investigated, 25% were found ... | 1999 | 16232444 |
| transgenic resistance to cymbidium mosaic virus in dendrobium expressing the viral capsid protein gene. | a taiwan isolate of cymbidium mosaic virus (cymmv-cs) was isolated from infected cymbidium sinesis willd. the cdna of the capsid protein (cp) gene was synthesized and sequenced. alignment of this cp gene with other reported cps revealed homologies of 92-98% at the nucleotide level and 98-99% at the amino acid level. to generate virus-resistant varieties, the cymmv-cs cp gene was transformed into dendrobium protocorms through particle bombardment. transformants were selected on medium supplemente ... | 2005 | 15865047 |
| reciprocal function of movement proteins and complementation of long-distance movement of cymbidium mosaic virus rna by odontoglossum ringspot virus coat protein. | complementation of movement and coat proteins of the orchid-infecting potexvirus cymbidium mosaic virus (cymmv) and tobamovirus odontoglossum ringspot virus (orsv) was investigated. nicotiana benthamiana, which is susceptible to both cymmv and orsv, was used as a model system. four transgenic lines, each harbouring one of the movement protein (mp) or coat protein (cp) genes of cymmv or orsv, were constructed. the mp of cymmv consists of three overlapping open reading frames, together called the ... | 2005 | 15831968 |
| the complete sequence of the genomic rna of an isolate of lily virus x (genus potexvirus). | the complete sequence of the genomic rna of an isolate of lily virus x (lvx) has been determined for the first time. the isolate from the netherlands was 5823 nucleotide (nt) long excluding the 3'-poly(a) tail, making it the shortest reported potexvirus sequence. the 5'-non-coding region begins with ggaaaa like that of scallion virus x (scavx) and some isolates of cymbidium mosaic virus (cymmv), whereas those of other sequenced potexviruses probably all begin with gaaaa. the genome organisation ... | 2004 | 15578239 |
| detection of two orchid viruses using quartz crystal microbalance (qcm) immunosensors. | quartz crystal microbalance (qcm) immunosensors are based on the principle that adsorption of substances on the surface of a quartz crystal changes its resonance oscillation frequency. a qcm immunosensor was developed for the detection of both cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) by pre-coating the qcms with virus-specific antibodies. upon binding of virions in either purified form or crude sap of infected orchids with the immobilised virus antibodies ... | 2002 | 11684305 |
| simultaneous quantitation of two orchid viruses by the taqman real-time rt-pcr. | simultaneous quantitation of two orchid viruses, cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv), were carried out using the taqman((r)) real-time rt-pcr, a novel detection technique that combines rt-pcr with the power of fluorescent detection. four taqman((r)) probes were synthesized, targeting at the rna-dependent rna polymerase (rdrp) and coat protein (cp) genes of both viruses. the reporter dye fam (6-carboxyfluorescein) was used to label the 5' terminus of ... | 2000 | 10856762 |
| one-step reverse transcription loop-mediated isothermal amplification assay for rapid detection of cymbidium mosaic virus. | cymbidium mosaic virus (cymmv) is the most prevalent orchid virus. a single-tube one-step betaine-free reverse transcription (rt) loop-mediated isothermal amplification (lamp) assay was developed for the rapid and easy detection of orchid-infecting cymmv. five sets of primers were designed based on the conserved regions among various virus isolates. the specificity and the sensitivity of the assay were then evaluated using the rt-lamp reaction. within 1h under isothermal conditions at 60°c the t ... | 2011 | 21237208 |
| the complete sequence of cymbidium mosaic virus from vanilla fragrans in hainan, china. | the complete nucleotide sequence of cymbidium mosaic virus (cymmv) isolated from vanilla in hainan province, china was determined for the first time. it comprised 6,224 nucleotides; sequence analysis suggested that the isolate we obtained was a member of the genus potexvirus, and its sequence shared 86.67-96.61% identities with previously reported sequences. phylogenetic analysis suggested that cymmv from vanilla fragrans was clustered into subgroup a and the isolates in this subgroup displayed ... | 2011 | 21442302 |
| genetic variability in the coat protein genes of cymbidium mosaic virus isolates from orchids. | the variability in the nucleotide (nt) and amino acid (aa) sequences of the coat protein (cp) of cymbidium mosaic virus (cymmv), which naturally infects orchids worldwide, was investigated. the cp genes of 55 cymmv isolates originating from different locations in korea were amplified using rt-pcr and sequenced. the encoded cp consists of 223 aa. the cp sequences of the korean isolates were compared with those of previously published cymmv isolates originating from different countries at both nt ... | 2011 | 22015427 |
| in vitro expression and production of antibody against cymbidium mosaic virus coat protein. | polyclonal rabbit antisera were produced using coat protein of cymbidium mosaic virus (cymmv) indian isolate expressed in e. coli as gst fusion. the expressed protein was purified by gst-fusion protein purification kit for use as an immunogen in rabbits. antisera prepared in this manner reacted in double antibody sandwich enzyme-linked immunosorbent assay (das elisa) with extract from cymmv-infected tissue. the results indicate that polyclonal antisera prepared from expressed cymmv coat proteins ... | 2012 | 23730003 |
| optimizing virus-induced gene silencing efficiency with cymbidium mosaic virus in phalaenopsis flower. | virus-induced gene silencing (vigs) is a good way to study floral gene functions of orchids, especially those with a long life cycle. to explore the applicability and improve viral silencing efficiency for application of cymbidium mosaic virus (cymmv)-induced gene silencing, we examined several variables, including the optimal length of the dna fragment, the effect of developmental maturation status of inflorescence, and suitable inoculation sites. a cymmv-based vigs system can be used with orch ... | 2012 | 23352400 |
| production of virus-free orchid cymbidium aloifolium (l.) sw. by various tissue culture techniques. | orchids are affected by many viruses resulting in poor growth, yield and quality, and an overall decline in population. cymbidium mosaic virus (cymmv) is one of the common orchid viruses found in cymbidium species but it infects different orchid genera. in this study cymbidium aloifolium was propagated in vitro using ms medium at different strength (1.0, ½, and ¼) with or without 0.5 mg/l bap (6-benzylaminopurine) and 0.5 mg/l naa (naphthalene acetic acid). to provide disease-free planting mater ... | 2016 | 27812549 |
| differentially expressed genes of chenopodium amaranticolor in response to cymbidium mosaic virus infection. | cymbidium mosaic virus (cymmv)-induced expressed sequence tag (est) clones from chenopodium amaranticolor were identified. cymmv was mechanically inoculated onto c. amaranticolor, and local lesion symptoms were observed. inoculated leaves were collected on serial days post inoculation (dpi) to identify activated or suppressed genes. mrna isolation and suppression subtractive hybridization (ssh) were then performed to identify differentially expressed genes related to the local lesion response. f ... | 2016 | 27364083 |
| simultaneous detection of cymbidium mosaic virus and odontoglossum ringspot virus in orchids using multiplex rt-pcr. | a system for simultaneous detection of two orchid-infecting viruses was developed and applied to several orchid species. the detection system involved multiplex reverse transcription-polymerase chain reaction (rt-pcr) and could simultaneously identify cymbidium mosaic virus (cymmv) and odontoglossum ringspot virus (orsv) from the orchid species studied. multiplex rt-pcr was conducted using two virus-specific primer pairs and an internal control pair of primers to amplify the cymmv and orsv coat ... | 2015 | 26542829 |
| detection of viruses directly from the fresh leaves of a phalaenopsis orchid using a microfluidic system. | early detection of pathogens is crucial for the effective surveillance of diseases. many efforts have been made to explore methods which can detect these pathogens within a short period of time without requiring a tedious protocol. however, these developed methods have disadvantages such as they are relatively time-consuming or require specialized laboratory facilities. in this work, we have developed an integrated microfluidic system for rapid and automatic detection of viruses by direct analys ... | 2013 | 23751373 |
| metabolome of vanilla planifolia (orchidaceae) and related species under cymbidium mosaic virus (cymmv) infection. | the genus vanilla which belongs to the orchidaceae family comprises more than 110 species of which two are commercially cultivated (vanilla planifolia and vanilla xtahitensis). the cured pods of these species are the source of natural vanilla flavor. in intensive cultivation systems the vines are threatened by viruses such as cymbidium mosaic virus (cymmv). in order to investigate the effect of cymmv on the growth and metabolome of vanilla plants, four accessions grown in intensive cultivation s ... | 2012 | 22902551 |
| a high-throughput virus-induced gene-silencing vector for screening transcription factors in virus-induced plant defense response in orchid. | the large number of species and worldwide spread of species of orchidaceae indicates their successful adaptation to environmental stresses. thus, orchids provide rich resources to study how plants have evolved to cope with stresses. this report describes our improvement of our previously reported orchid virus-induced gene silencing vector, pcymmv-pro60, with a modified gateway cloning system which requires only one recombination and can be inoculated by agroinfiltration. we cloned 1,700 dna frag ... | 2012 | 22397405 |
| the npr1 ortholog phanpr1 is required for the induction of phapr1 in phalaenopsis aphrodite. | systematic acquired resistance (sar) is an effective broad-spectrum defense mechanism that confers long-lasting protection against biotrophic pathogens trough defense related salicylic acid (sa) signaling. gene(s) involved in sar have been extensively studied in dicot plants; however, remains largely unresolved in monocot plants. npr1, an evolutionary conserved gene, plays a central role in sar, and pr-1 is widely used as a marker for effective sa signaling. | 2013 | 28510874 |
| direct detection of orchid viruses using nanorod-based fiber optic particle plasmon resonance immunosensor. | a fiber optic particle plasmon resonance (foppr) immunosensor is developed for label-free detection of orchid viruses that use gold nanorods (aunrs) as the sensing material. the aunrs are employed to create a near-infrared sensing window to solve the color interference problem of sample matrix for direct sensing of target analyte. this work cannot be achieved using gold nanospheres (aunss) because the signal of sample color absorption largely overlaps the signal of molecular recognition events i ... | 2014 | 24001513 |
| virus-induced gene silencing unravels multiple transcription factors involved in floral growth and development in phalaenopsis orchids. | orchidaceae, one of the largest angiosperm families, has significant commercial value. isolation of genes involved in orchid floral development and morphogenesis, scent production, and colouration will advance knowledge of orchid flower formation and facilitate breeding new varieties to increase the commercial value. with high-throughput virus-induced gene silencing (vigs), this study identified five transcription factors involved in various aspects of flower morphogenesis in the orchid phalaeno ... | 2013 | 23956416 |