| partial characterisation of citrus leaf blotch virus, a new virus from nagami kumquat. | citrus leaf blotch virus (clbv) was purified from leaves of nagami kumquat sra-153 that showed bud union crease when propagated on troyer citrange. virions were filamentous particles (960 x 14 nm) containing a 42 kda protein and a single-stranded rna (ssrna) of about 9,000 nt (mr 3 x 10(6)). infected tissue contained three species of double-stranded rna (dsrna) of mr 6, 4.5 and 3.4 x 10(6). the nucleotide sequence of several complementary dna (cdna) clones showed significant similarities with re ... | 2001 | 11315643 |
| the nucleotide sequence and genomic organization of citrus leaf blotch virus: candidate type species for a new virus genus. | the complete nucleotide sequence of citrus leaf blotch virus (clbv) was determined. clbv genomic rna (grna) has 8747 nt, excluding the 3'-terminal poly(a) tail, and contains three open reading frames (orfs) and untranslated regions (utr) of 73 and 541 nucleotides at the 5' and 3' termini, respectively. orf1 potentially encodes a 227.4-kda polypeptide, which has methyltransferase, papain-like protease, helicase, and rna-dependent rna polymerase motifs. orf2 encodes a 40.2-kda polypeptide containi ... | 2001 | 11504557 |
| characterization of two kinds of subgenomic rnas produced by citrus leaf blotch virus. | citrus leaf blotch virus (clbv) has a single-stranded, positive-sense, genomic rna (grna) organized in three orfs, which encode a polyprotein involved in replication (rp), a potential movement protein (mp), and coat protein (cp). northern blot hybridization of total, virion, or double-stranded rna with probes of different grna regions revealed that clbv produces two 3'-coterminal and two 5'-coterminal subgenomic rnas (sgrnas). the 3'-coterminal sgrnas contain the mp (3'mp sgrna) and cp (3'cp sgr ... | 2002 | 12033792 |
| low genetic variation between isolates of citrus leaf blotch virus from different host species and of different geographical origins. | the population structure and genetic diversity of citrus leaf blotch virus (clbv) were estimated by single-strand conformation polymorphism and nucleotide sequence analyses of two genomic regions located within the replicase (r) and the coat protein (c) genes. analysis of 30 cdna clones of each genomic region from two clbv isolates showed that both isolates contained a predominant haplotype and others closely related. analysis of 37 clbv spanish field isolates showed low genetic diversity (0.004 ... | 2002 | 12237442 |
| development of a full-genome cdna clone of citrus leaf blotch virus and infection of citrus plants. | citrus leaf blotch virus (clbv), a member of the family flexiviridae, has a ~9-kb single-stranded, positive-sense genomic rna encapsidated by a 41-kda coat protein. clbv isolates are associated with symptom production in citrus including leaf blotching of dweet tangor and stem pitting in etrog citron (dweet mottle disease), and some isolates are associated with bud union crease on trifoliate rootstocks, but koch's postulates for this virus were not fulfilled. a full-genome cdna of clbv isolate s ... | 2008 | 19019007 |
| detection and quantitation of citrus leaf blotch virus by taqman real-time rt-pcr. | a real-time rt-pcr assay based on the taqman chemistry was developed for reliable detection and quantitation of citrus leaf blotch virus (clbv) in citrus plants. detection by this method was highly specific and about one thousand times more sensitive than detection by conventional rt-pcr. an external standard curve using in vitro synthesized rna transcripts of the selected target allowed a reproducible quantitative assay, with a wide dynamic range (seven logarithmic units of concentration) and v ... | 2009 | 19406167 |
| a generic method to identify plant viruses by high-resolution tandem mass spectrometry of their coat proteins. | although a number of protocols have been developed for detection of viruses at the genus or family level, universal approaches to detect and identify unknown viruses are still required. high-resolution tandem mass spectrometry was used to identify accurately peptide masses and their constituent sequences from partially purified plant virus preparations. analysis of the peptide fragment masses against a virus database using pattern-matching algorithms identified sequences with homology to known v ... | 2010 | 19712699 |
| nucleotide sequence and genome organization of dweet mottle virus and its relationship to members of the family betaflexiviridae. | the nucleotide sequence of dweet mottle virus (dmv) was determined and compared to sequences of members of the families alphaflexiviridae and betaflexiviridae. the dmv genome has 8,747 nucleotides (nt) excluding the 3' poly-(a) tail. dmv genomic rna contains three putative open reading frames (orfs) and untranslated regions of 73 nt at the 5' and 541 nt at 3' termini. orf1 potentially encoding a 227.48-kda polyprotein, which has methyltransferase, oxygenase, endopeptidase, helicase, and rna-depe ... | 2010 | 20644968 |
| mapping the subgenomic rna promoter of the citrus leaf blotch virus coat protein gene by agrobacterium-mediated inoculation. | citrus leaf blotch virus has a single-stranded positive-sense genomic rna (grna) of 8747 nt organized in three open reading frames (orfs). the orf1, encoding a polyprotein involved in replication, is translated directly from the grna, whereas orfs encoding the movement (mp) and coat (cp) proteins are expressed via 3' coterminal subgenomic rnas (sgrnas). we characterized the minimal promoter region critical for the cp-sgrna expression in infected cells by deletion analyses using agrobacterium-med ... | 2010 | 20708769 |
| The Citrus leaf blotch virus movement protein acts as silencing suppressor. | To counteract plant antiviral defense based on RNA silencing, many viruses express proteins that inhibit this mechanism at different levels. The genome of Citrus leaf blotch virus (CLBV) encodes a 227-kDa protein involved in replication, a 40-kDa movement protein (MP), and a 41-kDa coat protein (CP). To determine if any of these proteins might have RNA silencing suppressor activities, we have used Agrobacterium-mediated transient assays in the green fluorescent protein (GFP)-expressing Nicotiana ... | 2011 | 21948005 |
| the resistance of sour orange to citrus tristeza virus is mediated by both the salicylic acid and rna silencing defence pathways. | citrus tristeza virus (ctv) induces in the field the decline and death of citrus varieties grafted on sour orange (so) rootstock, which has forced the use of alternative decline-tolerant rootstocks in affected countries, despite the highly desirable agronomic features of the so rootstock. declining citrus plants display phloem necrosis below the bud union. in addition, so is minimally susceptible to ctv compared with other citrus varieties, suggesting partial resistance of so to ctv. here, by si ... | 2016 | 27588892 |
| precocious flowering of juvenile citrus induced by a viral vector based on citrus leaf blotch virus: a new tool for genetics and breeding. | the long juvenile period of citrus trees (often more than 6 years) has hindered genetic improvement by traditional breeding methods and genetic studies. in this work, we have developed a biotechnology tool to promote transition from the vegetative to the reproductive phase in juvenile citrus plants by expression of the arabidopsis thaliana or citrus flowering locus t (ft) genes using a citrus leaf blotch virus-based vector (clbvinpr-atft and clbvinpr-cift, respectively). citrus plants of differe ... | 2016 | 26920394 |
| characterization of new isolates of apricot vein clearing-associated virus and of a new prunus-infecting virus: evidence for recombination as a driving force in betaflexiviridae evolution. | double stranded rnas from prunus samples gathered from various surveys were analyzed by a deep-sequencing approach. contig annotations revealed the presence of a potential new viral species in an azerbaijani almond tree (prunus amygdalus) and its genome sequence was completed. its genomic organization is similar to that of the recently described apricot vein clearing associated virus (avcav) for which two new isolates were also characterized, in a similar fashion, from two japanese plums (prunus ... | 2015 | 26086395 |
| development and validation of a multiplex reverse transcription quantitative pcr (rt-qpcr) assay for the rapid detection of citrus tristeza virus, citrus psorosis virus, and citrus leaf blotch virus. | a single real-time multiplex reverse transcription quantitative polymerase chain reaction (rt-qpcr) assay for the simultaneous detection of citrus tristeza virus (ctv), citrus psorosis virus (cpsv), and citrus leaf blotch virus (clbv) was developed and validated using three different fluorescently labeled minor groove binding qpcr probes. to increase the detection reliability, coat protein (cp) genes from large number of different isolates of ctv, cpsv and clbv were sequenced and a multiple sequ ... | 2015 | 25907469 |
| effectiveness of gene silencing induced by viral vectors based on citrus leaf blotch virus is different in nicotiana benthamiana and citrus plants. | virus induced gene silencing (vigs) is an effective technology for gene function analysis in plants. we assessed the vigs effectiveness in nicotiana benthamiana and citrus plants of different citrus leaf blotch virus (clbv)-based vectors, using insets of the phytoene desaturase (pds) gene. while in n. benthamiana the silencing phenotype was induced only by the construct carrying a 58-nt pds hairpin, in citrus plants all the constructs induced the silencing phenotype. differences in the generatio ... | 2014 | 25010281 |
| deep-sequencing analysis of an apricot tree with vein clearing symptoms reveals the presence of a novel betaflexivirus. | deep-sequencing technology applied on double stranded rna recovered from an apricot tree with vein clearing symptoms allowed the identification of a novel virus with a single-stranded rna genome, for which the provisional name apricot vein clearing-associated virus (avcav) is proposed. its genome comprises 7315nt, excluding the poly(a) tail, covering four open reading frames (orfs). the putative virus-encoded proteins, i.e., replicase (rep), movement protein (mp), coat protein (cp) and nucleic a ... | 2014 | 24389094 |
| citrus leaf blotch virus invades meristematic regions in nicotiana benthamiana and citrus. | to invade systemically host plants, viruses need to replicate in the infected cells, spread to neighbouring cells through plasmodesmata and move to distal parts of the plant via sieve tubes to start new infection foci. to monitor the infection of nicotiana benthamiana plants by citrus leaf blotch virus (clbv), leaves were agroinoculated with an infectious cdna clone of the clbv genomic rna expressing green fluorescent protein (gfp) under the transcriptional control of a duplicate promoter of the ... | 2013 | 23560714 |
| characterization of the complete genome of a novel citrivirus infecting actinidia chinensis. | a ssrna virus from kiwifruit (actinidia spp.) was identified as a member of the family betaflexiviridae. it was mechanically transmitted to the herbaceous indicators nicotiana benthamiana, n. clevelandii, n. glutinosa and n. occidentalis. the complete genome was comprised of three orfs and a 3'poly (a) tail. phylogenetic analysis of the entire genome indicated it was a novel member of the genus citrivirus (family betaflexiviridae). the complete nucleotide sequence differed from that of citrus le ... | 2013 | 23494225 |
| development of viral vectors based on citrus leaf blotch virus to express foreign proteins or analyze gene function in citrus plants. | viral vectors have been used to express foreign proteins in plants or to silence endogenous genes. this methodology could be appropriate for citrus plants that have long juvenile periods and adult plants that are difficult to transform. we developed viral vectors based on citrus leaf blotch virus (clbv) by duplicating a minimum promoter (92 bp) either at the 3' untranslated region (clbv3'pr vector) or at the intergenic region between the movement and coat protein (cp) genes (clbvinpr vector). th ... | 2012 | 22670755 |
| tepovirus, a novel genus in the family betaflexiviridae. | tepovirus is a new monotypic genus of plant viruses typified by potato virus t (pvt), a virus with helically constructed filamentous particles that are 640 nm long, previously classified as unassigned species in the family betaflexiviridae. virions have a single-stranded positive-sense polyadenylated rna genome that is 6.5 kb in size, and a single type of coat protein with a size of 24 kda. the viral genome contains three slightly overlapping orfs encoding, respectively, the replication-related ... | 2012 | 22592959 |