| comparative oligonucleotide fingerprints of three plant viroids. | 5' phosphorylation in vitro with gamma-32p-atp and t4 phage induced polynucleotide kinase was used to obtain rnaase a and rnaase t1 fingerprints of three plant viroids: potato spindle tuber viroid from tomato (pstv-tom), chrysanthemum stunt viroid from cineraria (chsv-cin) and citrus exocortis viroid from gynura aurantiaca (cev-gyn). these three viroids differ significantly from each other as judged from their oligonucleotide patterns. this supports the concept of individual viroid species. | 1977 | 896482 |
| 32p- and biotin-labelled in vitro transcribed crna probes for the detection of potato spindle tuber viroid and chrysanthemum stunt viroid. | replacing nick-translated dna probes by in vitro transcribed complementary rna (crna) probes considerably increased the sensitivity of dot-blot detection tests of potato spindle tuber viroid and chrysanthemum stunt viroid. as compared to the limit of detection of 5-10 pg of viroid obtained with 32p-labelled dna probes, crna probes allow the detection of less than 1 pg of pure viroid. when labelled with biotin by incorporation of biotin-labelled ribonucleotides, the crna probes have a limit of de ... | 1990 | 1691524 |
| nonradioactive, photobiotin-labelled dna probes for routine diagnosis of viroids in plant extracts. | avocado sunblotch viroid (asbv), coconut cadang cadang viroid (cccv), chrysanthemum stunt viroid (csv) and potato spindle tuber viroid (pstv) were detected in plant extracts by dot-blot hybridization using nonradioactive photobiotin-labelled nucleic acid probes. recombinant dna probes, containing full-length monomer viroid inserts in the plasmid vectors psp64 or puc9, were biotinylated with photobiotin and used as sonicated double-stranded dna fragments. using fresh leaf material, a general meth ... | 1989 | 2654169 |
| synthetic oligonucleotide hybridization probes to diagnose hop stunt viroid strains and citrus exocortis viroid. | four species of synthetic oligonucleotide probes for the diagnosis of hop stunt viroid (hsv) and citrus exocortis viroid (cev) were devised. probe hsv-1 detected all the members of hsv group, such as hsv-hop, hsv-grapevine, hsv-cucumber, hsv-citrus and a viroid-like rna isolated from plum trees affected by plum dapple fruit disease. probe hsv-2 discriminated hsv-grapevine from the other members of hsv group. hsv-hop and hsv-grapevine consist of the same numbers of nucleotides, with only one nucl ... | 1988 | 3366851 |
| nucleotide sequence and secondary structure of citrus exocortis and chrysanthemum stunt viroid. | the complete nucleotide sequence of citrus exocortis viroid (cev, propagated in gymura) and chrysanthemum stunt viroid (csv, propagated in cineraria) has been established, using labelling in vitro and direct rna sequencing methods and a new screening procedure for the rapid selection of suitable rna fragments from limited digests. the covalently closed circular single-stranded viroid rnas consist of 371 (cev) and 354 (csv) nucleotides, respectively. as previously shown for potato spindle tuber v ... | 1982 | 7060550 |
| chrysanthemum stunt viroid: primary sequence and secondary structure. | the sequence of the 356 nucleotide residues of chrysanthemum stunt viroid (csv) has been determined. overlapping linear viroid fragments were obtained by partial ribonuclease digestion, radiolabelled in vitro at their 5'-ends, and sequenced using partial enzymic cleavage methods. of the csv sequence, 69% is contained in the published sequence of potato spindle tuber viroid (pstv). differences in the primary sequence of csv and pstv suggest that neither the positive nor putative negative strands ... | 1981 | 7279660 |
| avocado sunblotch viroid: primary sequence and proposed secondary structure. | the sequence of the 247 nucleotide residues of the single strand circular rna of avocado sunblotch viroid (asbv) was determined using partial enzymic cleavage methods on overlapping viroid fragments obtained by partial ribonuclease digestion followed by 32p-labelling in vitro at their 5'-ends. asbv is much smaller than potato spindle tuber viroid (pstv; 359 residues) and chrysanthemum stunt viroid (csv; 356 residues). a secondary structure model for asbv is proposed and contains 67% of its resid ... | 1981 | 7322921 |
| [design of a universal biotin-containing oligonucleotide diagnostic kit for detecting viroid plant diseases]. | a non-radioactive diagnosticum for plant viroid diseases has been designed, based on hybridization of a biotin-labeled 26-member oligonucleotide probe with the viroid rna site identical for potato spindle tuber viroid and chrysanthemum stunt viroid. the biotin label has been introduced into the synthetic oligonucleotide probe by the high-yield acylation of the oligonucleotide aminoalkylamide with the biotin imidazolide or n-hydroxysuccinimide ester. hybridization techniques have been elaborated ... | 1993 | 8250981 |
| a simple, rapid method of nucleic acid extraction without tissue homogenization for detecting viroids by hybridization and rt-pcr. | a simple, rapid method of nucleic acid extraction on a microcentrifuge tube scale for detecting viroids is presented. five distinct citrus viroids (cvds), chrysanthemum stunt viroid (csvd), hop stunt viroid (hsvd), hop latent viroid (hlvd) and potato spindle tuber viroid (pstvd) were detected in their natural host plants by hybridization using crna probes and reverse transcription-polymerase chain reaction (rt-pcr). nucleic acids (na) were liberated from tissues by incubation in a buffer contain ... | 1999 | 10029324 |
| restoration of secondary hairpin ii is associated with restoration of infectivity of a non-viable recombinant viroid. | mutagenesis and/or construction of recombinants by exchange of genomic regions between parental molecules constitute powerful tools for the study of viroids. however, a large proportion of such modifications results in molecules, which have lost their infectivity. such is the case for a recombinant viroid named cecs, obtained by replacing the right half of a citrus exocortis viroid (cevd) by the same region from chrysanthemum stunt viroid (csvd). in an effort to recover viable infectious progeny ... | 2001 | 11311425 |
| production of anti-virus, viroid plants by genetic manipulations. | many pathogenic plant viruses are rna viruses, which initiate production of double-stranded rna intermediates when they replicate in host plant cells. introduction of double-stranded rna-specific ribonucleases such as the schizosaccharomyces pombe derived pac i protein and animal cell derived interferon-induced 2',5'-oligoadenylate synthetase (2-5 aase)/ribonuclease l (rnase l) system into various plants may make plants resistant to various pathogenic viruses and viroids. we have demonstrated th ... | 2002 | 12449532 |
| an rt-pcr primer pair for the detection of pospiviroid and its application in surveying ornamental plants for viroids. | a primer pair for reverse transcription-polymerase chain reaction (rt-pcr), based on the conserved sequences of the members of genus pospiviroid was designed to yield a fragment of about 200 base pairs (bp). since pospiviroids infect a large number of plants species and a few members of the genus pospiviroid have been already detected in some ornamental plants, the primer pair was evaluated for its efficacy using ornamental plants. the method of return-polyacrylamide gel electrophoresis (r-page) ... | 2004 | 14738987 |
| development of a one tube-one step rt-pcr protocol for the detection of seven viroids in four genera: apscaviroid, hostuviroid, pelamoviroid and pospiviroid. | a one tube-one step rt-pcr was developed for the detection of seven viroids (apple scar skin viroid, apple dimple fruit viroid, pear blister canker viroid, hop stunt viroid, chrysanthemum stunt viroid, citrus exocortis viroid and peach latent mosaic viroid) in four genera that infect eight plant species. the efficiency and specificity of this method were optimized by the use of moloney-murine leukemia virus (m-mlv) reverse-transcriptase and hotstartaq dna polymerase which allowed increase sensit ... | 2004 | 15350729 |
| measurement of viroid sequence homology by hybridization with complementary dna prepared in vitro. | dna complementary to potato spindle tuber viroid (ystv cdna) has been used in rna dna hybridization experiments to study pstv replication in a variety of hosts and to measure the amount of sequence homology between ystv and several independently isolated viroids. when pstv cdna was hybridized with low molecular weight rna isolated from pstv-infected tomato at different times after inoculation, pstv . pstv cdna hybrids were detectable before symptoms began to appear. low molecular weight rna was ... | 1978 | 18627881 |
| hybridization analysis of chrysanthemum stunt viroid with complementary dna and the quantitation of viroid rna sequences in extracts of infected plants. | in order to develop a method for the quantitation of viroids in extracts of infected plants by hybridization analysis with labeled complementary dna (cdna), the kinetics of hybridization of chrysanthemum stunt viroid (csv) with [32p]cdna were investigated. two buffer systems were used, each with a total cation concentration of 0.19 m, with one containing no formamide (buffer a) and the other 40% (v/v) formamide (buffer b). initial experiments with buffer a at 60 degrees showed that the rate of h ... | 1979 | 18631615 |
| characterization of a viroid associated with avocado sunblotch disease. | a viroid has been purified from avocado leaves infected by sunblotch disease and designated the avocado sunblotch viroid. it is a covalently closed circular rna molecule with a molecular weight lower than that of chrysanthemum stunt viroid and citrus exocortis viroid while hybridization analysis with 32p-labeled complementary dna indicated that it is a single rna species. it could be detected as a stainable rna band on polyacrylamide tube gel electrophoresis of partially purified extracts of onl ... | 1979 | 18631620 |
| comparative sequence and structure of different isolates of citrus exocortis viroid. | the nucleotide sequences of two new australian isolates of citrus exocortis viroid (cev), cev-de25 and cev-de26, have been determined and compared with the previously sequenced australian isolate, cev-a (j. e. visvader, a. r. gould, g. e. breuning, and r. h. symons. febs lett.137, 288-292 (1982).) and a californian isolate, cev-c (h. j. gross, g. krupp, h. domdey, m. raba, p. jank, c. lossow, h. alberty, k. ramm, and h. l. sanger. eur. j. biochem.121, 249-257 (1982).). sequencing of recombinant ... | 1983 | 18639139 |
| sequence variability of chrysanthemum stunt viroid in different chrysanthemum cultivars. | viroids are the smallest infectious agents, and their genomes consist of a short single strand of rna that does not encode any protein. chrysanthemum stunt viroid (csvd), a member of the family pospiviroidae, causes chrysanthemum stunt disease. here, we report the genomic variations of csvd to understand the sequence variability of csvd in different chrysanthemum cultivars. we randomly sampled 36 different chrysanthemum cultivars and examined the infection of csvd in each cultivar by reverse tra ... | 2017 | 28149699 |
| agrobacterium-mediated inoculation of chrysanthemum (chrysanthemum morifolium) plants with chrysanthemum stunt viroid. | agroinfiltration was tested as a method of inoculation of chrysanthemum plants with chrysanthemum stunt viroid (csvd). binary vectors harboring dimeric csvd sequences in sense and antisense orientations were constructed, and agrobacterium transfected with these binary vectors was infiltrated into chrysanthemum leaves. northern blotting and reverse transcription polymerase chain reaction analysis showed that local infection was established within 7 days and systemic infection within 20 days. csvd ... | 2016 | 27155239 |
| digestion of chrysanthemum stunt viroid by leaf extracts of capsicum chinense indicates strong rna-digesting activity. | csvd could not infect nicotiana benthamiana when the plants were pretreated with crude leaf extract of capsicum chinense 'sy-2'. c. chinense leaves were revealed to contain strong rna-digesting activity. several studies have identified active antiviral and antiviroid agents in plants. capsicum plants are known to contain antiviral agents, but the mechanism of their activity has not been determined. we aimed to elucidate the mechanism of capsicum extract's antiviroid activity. chrysanthemum stunt ... | 2016 | 27053224 |
| low temperature treatment affects concentration and distribution of chrysanthemum stunt viroid in argyranthemum. | chrysanthemum stunt viroid (csvd) can infect argyranthemum and cause serious economic loss. low temperature treatment combined with meristem culture has been applied to eradicate viroids from their hosts, but without success in eliminating csvd from diseased argyranthemum. the objectives of this work were to investigate (1) the effect of low temperature treatment combined with meristem culture on elimination of csvd, (2) the effect of low temperature treatment on csvd distribution pattern in sho ... | 2016 | 26973607 |
| callose deposition in plasmodesmata and viroid invasion of the shoot apical meristem. | | 2016 | 26870015 |
| elimination of chrysanthemum stunt viroid and chrysanthemum chlorotic mottle viroid from infected chrysanthemum by cryopreservation. | chrysanthemum morifolium 'borami' and 'secret pink' showing symptoms of stunt disease caused by chrysanthemum stunt viroid (csvd) and 'yellow cap' showing chlorotic mottle disease caused by chrysanthemum chlorotic mottle viroid (cchmvd) were confirmed to be infected by the respective viroids by using reverse transcription polymerase chain reaction (rt-pcr). real-time pcr results showed that the viroid concentrations in the infected cultivars varied between the different regions of origin (chilgo ... | 2016 | 26315819 |
| complete genome sequences of chrysanthemum stunt viroid from a single chrysanthemum cultivar. | the chrysanthemum stunt viroid (csvd), a member of the genus pospiviroid with a single circular rna genome, infects many chrysanthemum species. here, we report 25 complete genome sequences of csvd in a single chrysanthemum cultivar, revealing 20 variants. | 2015 | 26251489 |
| simultaneous detection of tomato spotted wilt virus, dahlia mosaic virus and chrysanthemum stunt viroid by multiplex rt-pcr in dahlias and their distribution in japanese dahlias. | tomato spotted wilt virus (tswv), dahlia mosaic virus (dmv) and chrysanthemum stunt viroid (csvd) are economically important viruses and viroid that infect cultivated dahlias. prior to this investigation, no multiplex rt-pcr assay for the detection of dahlia virus and viroid infections existed. in this study, we report the development of a multiplex rt-pcr that simultaneously detects tswv, dmv and csvd infections in dahlias. in addition, a simple rt-pcr method that does not require rna extractio ... | 2015 | 25976592 |
| invasion of shoot apical meristems by chrysanthemum stunt viroid differs among argyranthemum cultivars. | chrysanthemum stunt viroid (csvd) is a damaging pathogen attacking argyranthemum plants. our study attempted to reveal distribution patterns of csvd in shoot apical meristems (sam) and to explore reasons for differential ability of csvd to invade sam of selected argyranthemum cultivars. symptom development was also observed on greenhouse-grown argyranthemum plants. viroid localization using in situ hybridization revealed that the ability of csvd to invade sam differed among cultivars. in disease ... | 2015 | 25763000 |
| a multiplex rt-pcr for simultaneous detection and identification of five viruses and two viroids infecting chrysanthemum. | pathogens causing significant economic losses in chrysanthemum include tomato aspermy virus (tav), chrysanthemum virus b (cvb), cucumber mosaic virus (cmv), tobacco mosaic virus (tmv), potato virus y (pvy), chrysanthemum stunt viroid (csvd) and chrysanthemum chlorotic mottle viroid (cchmvd). a multiplex reverse transcription polymerase chain reaction (rt-pcr) method, using specific primer sets for each virus or viroid, was developed for simultaneous detection and differentiation of tav, cvb, cmv ... | 2015 | 25698104 |
| construction of infectious cdna clone of a chrysanthemum stunt viroid korean isolate. | chrysanthemum stunt viroid (csvd), a noncoding infectious rna molecule, causes seriously economic losses of chrysanthemum for 3 or 4 years after its first infection. monomeric cdna clones of csvd isolate sk1 (csvd-sk1) were constructed in the plasmids pgem-t easy vector and puc19 vector. linear positive-sense transcripts synthesized in vitro from the full-length monomeric cdna clones of csvd-sk1 could infect systemically tomato seedlings and chrysanthemum plants, suggesting that the linear csvd ... | 2014 | 25288987 |
| multiplex reverse transcription loop-mediated isothermal amplification for the simultaneous detection of cvb and csvd in chrysanthemum. | a multiplex reverse transcription loop-mediated isothermal amplification (mrt-lamp) assay was developed for the simultaneous detection of chrysanthemum virus b (cvb) and chrysanthemum stunt viroid (csvd), which are the major viral pathogens of chrysanthemum worldwide. two sets of mrt-lamp primers were designed for the coat protein gene of cvb and the complete nucleotide sequence of csvd, and a restriction enzyme cleavage site was inserted into two pairs of species-specific primers. the mrt-lamp ... | 2014 | 25241144 |
| [application of screening microarray technology in genus level for detection of pospiviroid]. | the aim was to establish an effective screening microarray at genus level for pospiviroid. we analyzed nucleotide sequences from pospiviroid viroid and designed 19 probes with genus identification characteristics. the standards of these probes included the characters of (i) a gc content between 40 and 60%, (ii) less than 50% of single nucleotide, (iii) less than 4 continuous mononucleotides, and (iv) less than 6 nucleotides in the inner hairpin. we synthesized microarrays by using these probes o ... | 2014 | 25009845 |
| a current overview of two viroids that infect chrysanthemums: chrysanthemum stunt viroid and chrysanthemum chlorotic mottle viroid. | the chrysanthemum (dendranthema x grandiflorum) belongs to the family asteraceae and it is one of the most popular flowers in the world. viroids are the smallest known plant pathogens. they consist of a circular, single-stranded rna, which does not encode a protein. chrysanthemums are a common host for two different viroids, the chrysanthemum stunt viroid (csvd) and the chrysanthemum chlorotic mottle viroid (cchmvd). these viroids are quite different from each other in structure and function. he ... | 2013 | 23594461 |
| dahlia latent viroid: a recombinant new species of the family pospiviroidae posing intriguing questions about its origin and classification. | a viroid-like rna has been detected in two asymptomatic dahlia accessions by return and double page. it appeared smaller than chrysanthemum stunt viroid and potato spindle tuber viroid, the two members of the genus pospiviroid, family pospiviroidae, reported in this ornamental previously. rt-pcr with primers designed for amplifying all pospiviroids produced no amplicons, but rt-pcr with random primers revealed a 342 nt rna. the sequence of this rna was confirmed with specific primers, which addi ... | 2013 | 23255620 |
| a multiplex rt-pcr for rapid and simultaneous detection of viruses and viroids in chrysanthemum. | chrysanthemum plants are subject to serious virus diseases, so detection and identification of virus pathogens is important to prevent the virus spread. a reliable one-step multiplex rt-pcr was developed to simultaneously detect two viruses and two viriods: chrysanthemum virus b, tomato aspermy virus, chrysanthemum stunt viroid and chrysanthemum chlorotic mottle viroid. in addition, we investigated the detection limit and the efficiency of single and multiplex rt-pcr assays. the results showed t ... | 2013 | 23020260 |
| sequence comparisons of global chrysanthemum stunt viroid variants: multiple polymorphic positions scattered through the viroid genome. | the nucleotide sequences of cdna clones of three chrysanthemum stunt viroid (csvd) isolates (one each from the usa, china, and australia) were determined and analyzed. the sequences of csvd cdna clones of the us and australian isolates were both quasi-species, while the cdna clones of the chinese isolate contained only a single variant. a comparison of the nucleotide sequences of 117 isolates and cdna clones obtained from 16 countries showed that in some cases identical csvd isolates were found ... | 2013 | 22961404 |
| a single polyprobe for detecting simultaneously eight pospiviroids infecting ornamentals and vegetables. | the spread of viroids belonging to the genus pospiviroid (family pospiviroidae), recorded recently in ornamentals and vegetables in several european countries, calls for fast, efficient and sensitive detection methods. based on bioinformatics analyses of sequence identity among all pospiviroids, a digoxigenin-labeled polyprobe (pospiprobe) was developed that, when tested by dot-blot and northern-blot hybridization, detected potato spindle tuber viroid, citrus exocortis viroid, columnea latent vi ... | 2012 | 22935607 |
| in vitro-transcribed chrysanthemum stunt viroid rna is infectious to chrysanthemum and other plants. | chrysanthemum stunt viroid (csvd), a noncoding rna, is known to cause chrysanthemum stunt disease, which affects the yield of flowers. to gain insights into csvd replication, infection, and the reasons for the spreading of csvd disease in chrysanthemum plants, we prepared linear csvd rna and analyzed its ability to cause disease in chrysanthemum plants. we found that linear csvd replicated as efficiently as csvd rna isolated from the infected chrysanthemum plants. additionally, the linear csvd r ... | 2009 | 19055435 |
| one-step site-directed mutagenesis of viroid dimeric cdna. | the study of viroids (plant pathogens constituted by small non-coding rna) has greatly benefited from the use of site-directed mutagenesis tools. however, compared to viral systems, this technique is complicated by the fact that, usually, infectious cdnas carry two copies of the viroid genome. a simple method for a one-step site-directed mutagenesis of viroids is described and tested by estimating the rate of mutation incorporation of three random nucleotide substitutions in each chrysanthemum s ... | 2007 | 17604131 |
| direct rt-pcr method for detecting two chrysanthemum viroids using minimal amounts of plant tissue. | a direct reverse transcription-polymerase chain reaction (rt-pcr) method for detecting the chrysanthemum stunt viroid (csvd) and chrysanthemum chlorotic mottle viroid (cchmvd) to screen for a viroid-free chrysanthemum plant at a small plant size was established and named microtissue direct rt-pcr. a razor or syringe needle was used for rna template preparations. under a stereoscopic microscope, a razor or syringe needle was used to pierce, a tissue sample to a depth of 0.1-0.2mm, and the sample ... | 2006 | 16102850 |
| chrysanthemum stunt viroid disturbs the photoperiodic response for flowering of chrysanthemum plants. | chrysanthemum ( dendranthema grandiflorum kitam.) is one of the qualitative short-day flowering plants. therefore, the flowering of chrysanthemum can usually be controlled by photoperiod. however, it was noted that 'piato' plants infected by the chrysanthemum stunt viroid (csvd) flowered autonomously even under long-day conditions. in this study, csvd-free and csvd-infected plants were prepared by culturing different-sized dissected shoot apical meristems (sams) of 'piato'. using these csvd-free ... | 2004 | 15549375 |
| elimination of chrysanthemum stunt viroid from an infected chrysanthemum cultivar by shoot regeneration from a leaf primordium-free shoot apical meristem dome attached to a root tip. | in this research we eliminated chrysanthemum stunt viroid (csvd) from a highly infected chrysanthemum cultivar using a newly established method. 'piato' is one of the most difficult cultivars in which to obtain csvd-free plants by conventional methods. leaf primordium-free shoot apical meristems (lp-free sams) of 'piato' plants were dissected and attached to csvd-free chrysanthemum or cabbage root tips. as shown by nested-pcr, csvd was not detected in some shoots regenerated on both types of roo ... | 2004 | 14986059 |
| detection of chrysanthemum stunt viroid (csv) using nick translated probes in a dot-blot hybridization assay. | we developed a dot-blot hydridization assay for the detection of chrysanthemum stunt viroid (csv) in chrysanthemum plant samples. the probe, a recombinant plasmid containing a full-length monomeric cdna copy of csv, is labelled with (32p) by nick-translation. the influence of the hybridization conditions, of the sample denaturation technique and of the plant sap components on the final sensitivity has been studied. the optimized system, involving a formaldehyde denaturation step, allows the dete ... | 1988 | 3410928 |
| graphics of rna secondary structure; towards an object-oriented algorithm. | we present a new algorithm for the display of rna secondary structure. the principle of the algorithm is entirely different from those currently in use in that our algorithm is 'object oriented' while current algorithms are 'procedural'. the circular rna molecule of chrysanthemum stunt viroid was used as input data for demonstrating the operation of the program. the major interest of this method will be found in its potential use in simulation graphics of rna folding processes. | 1987 | 2455587 |
| computer program for prediction of the optimal and suboptimal secondary structures of long rna molecules. | we present an algorithm for prediction of rna secondary structures. the program consists of three parts: the first computes location and free energy of every possible stem-loop structure, the second computes probability of its formation, and the third lists the positions and free energies of all the stem-loops in the order of their probability sizes. the circular rna molecule of chrysanthemum stunt viroid was used as an input data for demonstrating the operation of the program. | 1985 | 2468406 |