kinetics of interaction of cotton leaf curl kokhran virus-dabawali (clcukv-dab) coat protein and its mutants with ssdna.gemini viral assembly and transport of viral dna into nucleus for replication, essentially involve dna-coat protein interactions. the kinetics of interaction of cotton leaf curl kokhran virus-dabawali recombinant coat protein (rcp) with dna was studied by electrophoretic mobility shift assay (emsa) and surface plasmon resonance (spr). the rcp interacted with ssdna with a k(a), of 2.6+/-0.29 x 10(8) m(-1) in a sequence non-specific manner. the cp has a conserved c2h2 type zinc finger motif compos ...200919237182
the hypersensitive response induced by the v2 protein of a monopartite begomovirus is countered by the c2 protein.a functional analysis of the v2 protein of two monopartite begomoviruses, papaya leaf curl virus (palcuv) and cotton leaf curl kokhran virus (clcukov), has been performed. expression of the v2 gene from a potato virus x (pvx) vector resulted in severe leaf curling followed by a hypersensitive response (hr) in nicotiana benthamiana and n. tabacum, demonstrating that the v2 protein is a pathogenicity determinant and a target of host defence responses. agroinfiltration of a pvx vector expressing th ...201020447273
virus-induced gene silencing using a modified betasatellite: a potential candidate for functional genomics of crops.betasatellites are geminivirus-associated single-stranded dna molecules that play an important role in symptom modulation. a vigs vector was developed by modifying cotton leaf curl multan betasatellite (clcumb). clcumb dna was modified by replacing the βc1 gene with a multiple cloning site. the silencing ability of the modified clcumb was investigated by cloning a fragment of a host gene (su) or a reporter transgene (uida) into the modified clcumb and co-agroinoculation with cotton leaf curl mul ...201424610555
amplicon-based rna interference targeting v2 gene of cotton leaf curl kokhran virus-burewala strain can provide resistance in transgenic cotton plants.the conserved coat or v2 gene of begomoviruses is responsible for viral movement in the plant cells. rnai technology was used to silence v2 gene for resistance against these viruses in transgenic plants. the transformation of the rnai-based gene construct targeting v2 gene of clcukov-bur, cloned under 35s promoter, was done in two elite cotton varieties mnh-786 and vh-289 using shoot apex cut method of gene transformation. the transformation efficiency was found to be 3.75 and 2.88 % in mnh-786 ...201627757798
g5, a phage single-stranded dna-binding protein, fused with a nuclear localization signal, attenuates symptoms and reduces begomovirus-betasatellite accumulation in transgenic plants.cotton leaf curl disease is caused by several monopartite begomoviruses and is the major threat to cotton production in the indian subcontinent. the disease has been shown to be associated with four distinct species, including cotton leaf curl kokhran virus (clcukov), and a specific betasatellite-cotton leaf curl multan betasatellite (clcumub). transgenic nicotiana benthamiana plants were produced which constitutively express the escherichia coli phage m13 encoded, sequence nonspecific single-st ...201627364491
frequent occurrence of tomato leaf curl new delhi virus in cotton leaf curl disease affected cotton in pakistan.cotton leaf curl disease (clcud) is the major biotic constraint to cotton production on the indian subcontinent, and is caused by monopartite begomoviruses accompanied by a specific dna satellite, cotton leaf curl multan betasatellite (clcumb). since the breakdown of resistance against clcud in 2001/2002, only one virus, the "burewala" strain of cotton leaf curl kokhran virus (clcukov-bur), and a recombinant form of clcumb have consistently been identified in cotton across the major cotton growi ...201627213535
the 35-amino acid c2 protein of cotton leaf curl kokhran virus, burewala, implicated in resistance breaking in cotton, retains some activities of the full-length protein.with one exception, all the begomoviruses characterized so far encode an ~134-amino acid (aa) (a)c2 protein. the exception is the "burewala" strain of cotton leaf curl kokhran virus (clcukov-bu), associated with resistance breaking in cotton across pakistan and northwestern india, that encodes a truncated 35-aa c2. the c2 protein encoded by begomoviruses performs multiple functions including suppression of post-transcriptional gene silencing (ptgs), modulating microrna (mirna) expression and may ...201627209537
three gene products of a begomovirus-betasatellite complex restore expression of a transcriptionally silenced green fluorescent protein transgene in nicotiana benthamiana.single-stranded dna geminiviruses replicate via double-stranded dna intermediates forming mini-chromosomes that are targets for transcriptional gene silencing (tgs) in plants. the ability of the cotton leaf curl kokhran virus (clcukov)-cotton leaf curl multan betasatellite (clcumub) proteins, replication-associated protein (rep), transcriptional activator protein (trap), c4, v2 and βc1, to suppress tgs was investigated by using the nicotiana benthamiana line 16-tgs (16-tgs) harbouring a transcri ...201525537949
biochemical characterization of c4 protein of cotton leaf curl kokhran virus-dabawali.cotton leaf curl kokhran virus-dabawali (clcukv-dab) is a monopartite begomovirus encoding two proteins v1 and v2 in the virion sense and four proteins c1, c2, c3 and c4 in the complementary sense. the c4 protein of monopartite begomoviruses has been implicated to play a role in symptom determination and virus movement. the present work aims at the biochemical characterization of this protein.201323500017
multiple begomoviruses found associated with cotton leaf curl disease in pakistan in early 1990 are back in cultivated cotton.the first epidemic of cotton leaf curl disease (clcud) in early 1990's in the indian subcontinent was associated with several distinct begomoviruses along with a disease-specific betasatellite. resistant cotton varieties were introduced in late 1990's but soon resistance was broken and was associated with a single recombinant begomovirus named burewala strain of cotton leaf curl kokhran virus that lacks a full complement of a gene encoding a transcription activator protein (trap). in order to un ...201728386113
further changes in the cotton leaf curl disease complex: an indication of things to come?cotton leaf curl disease (clcud) has been a problem for cotton production in pakistan and india since the early 1990s. the disease is caused by begomoviruses associated with a specific satellite, the cotton leaf curl multan betasatellite (clcumb). in 2001, resistance introduced into cotton was broken by a recombinant begomovirus, cotton leaf curl kokhran virus strain burewala (clcukov-bur). unusually, in resistant cotton, this virus lacked an intact transcriptional activator protein (trap) gene, ...201728721488
molecular diagnostic development for begomovirus-betasatellite complexes undergoing diversification: a case least five begomoviral species that cause leaf curl disease of cotton have emerged recently in asia and africa, reducing fiber quality and yield. the potential for the spread of these viruses to other cotton-vegetable growing regions throughout the world is extensive, owing to routine, global transport of alternative hosts of the leaf curl viruses, especially ornamentals. the research reported here describes the design and validation of polymerase chain reaction (pcr) primers undertaken to fa ...201728438632
cotton leaf curl burewala virus with intact or mutant transcriptional activator proteins: complexity of cotton leaf curl disease.cotton leaf curl disease (clcud) is a serious disease of cotton on the indian subcontinent. in the present study, three cotton leaf curl viruses, cotton leaf curl burewala virus (clcubuv), cotton leaf curl kokhran virus (clcukov) and cotton leaf curl multan virus (clcumv), and their associated satellites, cotton leaf curl multan betasatellite (clcumb) and cotton leaf curl multan alphasatellite (clcuma), were detected. clcubuv with either intact (clcubuv-1) or mutant (clcubuv-2) transcriptional a ...201525772572
diversity, mutation and recombination analysis of cotton leaf curl geminiviruses.the spread of cotton leaf curl disease in china, india and pakistan is a recent phenomenon. analysis of available sequence data determined that there is a substantial diversity of cotton-infecting geminiviruses in pakistan. phylogenetic analyses indicated that recombination between two major groups of viruses, cotton leaf curl multan virus (clcumuv) and cotton leaf curl kokhran virus (clcukov), led to the emergence of several new viruses. recombination detection programs and phylogenetic analyse ...201626963635
effects of the mutation of selected genes of cotton leaf curl kokhran virus on infectivity, symptoms and the maintenance of cotton leaf curl multan betasatellite.cotton leaf curl kokhran virus (clcukov) is a cotton-infecting monopartite begomovirus (family geminiviridae). the effects of mutation of the coat protein (cp), v2, c2 and c4 genes of clcukov on infectivity and symptoms in nicotiana benthamiana were investigated. each mutation introduced a premature stop codon which would lead to premature termination of translation of the gene. mutation of the cp gene abolished infectivity. however, transient expression of the clcukov cp gene under the control ...201222871297
functional analysis of cotton leaf curl kokhran virus/cotton leaf curl multan betasatellite rna silencing south asia, cotton leaf curl disease (clcud) is caused by a complex of phylogenetically-related begomovirus species and a specific betasatellite, cotton leaf curl multan betasatellite (clcumub). the post-transcriptional gene silencing (ptgs) suppression activities of the transcriptional activator protein (trap), c4, v2 and βc1 proteins encoded by cotton leaf curl kokhran virus (clcukov)/clcumub were assessed in nicotiana benthamiana. a variable degree of local silencing suppression was observ ...201526512705
functional characterization of coat protein and v2 involved in cell to cell movement of cotton leaf curl kokhran virus-dabawali.the functional attributes of coat protein (cp) and v2 of the monopartite begomovirus, cotton leaf curl kokhran virus- dabawali were analyzed in vitro and in vivo by their overexpression in e. coli, insect cells and transient expression in the plant system. purified recombinant v2 and cp proteins were shown to interact with each other using elisa and surface plasmon resonance. confocal microscopy of sf21 cells expressing v2 and cp proteins revealed that v2 localized to the cell periphery and cp t ...201122110597
cotton leaf curl disease in sindh province of pakistan is associated with recombinant begomovirus components.cotton leaf curl disease (clcud) is a devastating disease of cotton causing severe losses to cotton across the punjab province of pakistan and northeastern india. although the disease has been reported as occurring sporadically in sindh province, pakistan, this has not caused significant losses. however, in the last few years the disease has become more significant in sindh province. clcud is caused by begomoviruses in association with a disease-specific symptom determining satellite (cotton lea ...201020621137
genetic variability of begomoviruses associated with cotton leaf curl disease originating from india.cotton leaf curl disease (clcud) causing viruses belong to the begomovirus genus of the family geminiviridae. most begomoviruses are bipartite with two molecules of circular single stranded dna (a and b) encapsidated in icosahedral geminate particles. however, the begomoviruses associated with clcud have dna-beta instead of dna-b. in this communication we report the complete genomic sequence of dna-a component of two clcud-causing begomoviruses, cotton leaf curl kokhran virus-dabawali (clcukv-da ...200415669112
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