ABSTRACT
The genus Dichorhavirus includes plant-infecting rhabdoviruses with bisegmented genomes that are horizontally transmitted by false spider mites of the genus Brevipalpus. The complete genome sequences of three isolates of the putative dichorhavirus clerodendrum chlorotic spot virus were determined using next-generation sequencing (Illumina) and traditional RT-PCR. Their genome organization, sequence similarity and phylogenetic relationship to other viruses, and transmissibility by Brevipalpus yothersi mites support the assignment of these viruses to a new species of dichorhavirus, as suggested previously. New data are discussed stressing the reliability of the current rules for species demarcation and taxonomic status criteria within the genus Dichorhavirus.
Subject(s)
Clerodendrum/virology , Genome, Viral , Hibiscus/virology , Plant Diseases/virology , RNA, Viral/genetics , Rhabdoviridae/genetics , Animals , Arachnid Vectors/virology , High-Throughput Nucleotide Sequencing , Mites/virology , Phylogeny , Plant Leaves/virology , Rhabdoviridae/classification , Rhabdoviridae/isolation & purification , Whole Genome SequencingABSTRACT
Clerodendron inerme, a common hedge plant grown in India, is affected by a yellow mosaic disease caused by a begomovirus. In the present study, the complete genome (DNA A) of this virus was cloned and sequenced. The total size of DNA A is 2760 nucleotides. The genome of this virus contains six open reading frames and a non-coding intergenic region of 293 nucleotides. Nucleotide sequence comparison analysis revealed maximum sequence identity with Papaya leaf curl virus-Pakistan [Pakistan:Cotton:2002] (73.9%). As this virus had less than 89% identity with other begomoviruses, it was identified as a new begomovirus species and tentatively, named as Clerodendron yellow mosaic virus-[India:New Delhi:2007] (ClYMV-[IN:ND:07]).
Subject(s)
Begomovirus/classification , Begomovirus/genetics , Clerodendrum/virology , Genome, Viral/genetics , Plant Diseases/virology , Sequence Analysis, DNA , Begomovirus/isolation & purification , India , Molecular Sequence Data , PhylogenyABSTRACT
Two begomovirus isolates (YX2-I and YX2-II) were identified from Clerodendrum cyrtophyllum showing yellow mosaic symptoms collected in Jiangsu province of China. Sequence analysis reveals that YX2-I is a distinct begomovirus species for which the name Clerodendrum golden mosaic Jiangsu virus (ClGMJSV-[CN:YX2:08]) is proposed. YX2-II is an isolate of bipartite begomovirus Clerodendrum golden mosaic China virus (ClGMCNV-[CN:YX2:08]). Infectious clones of the two viruses were constructed and agroinoculated into Nicotiana benthamiana, N. glutinosa, N. tabacum Samsun, Petunia hybrida, Solanum lycopersicum, Capsicum annuum, S. melongena, Glycine max and Gossypium hirsutum plants. ClGMJSV induced leaf curling and stunting symptoms in N. benthamiana, N. glutinosa, N. tabacum Samsun, and P. hybrida, and ClGMCNV infected N. benthamiana, N. glutinosa, and N. tabacum Samsun with severe symptoms and P. hybrida without obvious symptom. Latent infection in N. benthamiana, N. glutinosa, N. tabacum Samsun, and P. hybrida plants was observed when plants were inoculated with ClGMCNV DNA-A alone. In addition, we illustrated that ClGMCNV DNA-A was capable of interacting with the betasatellite associated with Tobacco curly shoot virus (TbCSB) to produce symptoms in N. benthamiana and N. glutinosa plants, and ClGMJSV could interact with TbCSB but not with ClGMCNV DNA-B in N. benthamiana plants.
Subject(s)
Begomovirus/growth & development , Begomovirus/genetics , Clerodendrum/virology , Host Specificity , Plant Diseases/virology , Begomovirus/isolation & purification , Begomovirus/physiology , Capsicum/virology , China , Cluster Analysis , DNA, Viral/chemistry , DNA, Viral/genetics , Gossypium/virology , Solanum lycopersicum/virology , Petunia/virology , Phylogeny , Sequence Analysis, DNA , Glycine max/virology , Nicotiana/virologyABSTRACT
Rapid micropropagation through adventitious shoot induction from in vitro raised leaf explants of Clerodendrum aculeatum (Verbenaceae), was successfully achieved for the first time. Basal portion of the leaves showed highest regeneration potential when grown on MS medium supplemented with BA (5.0 mg/l) and NAA and IBA (0.5 mg/l of each). Shoots after elongation in growth regulator-free medium, were rooted in MS medium containing 0.5 mg/l of NAA and IBA. Aqueous leaf extract of in vitro raised plants, induced high degree of resistance against viruses in susceptible healthy hosts when applied prior to virus inoculation. Upon purification from leaves of cultured plants, the resistance inducing protein, showed molecular mass of 34 kDa. Amount of resistance inducing protein obtained from leaves of cultured plants, was consistent throughout the year, as compared to the protein isolated from leaves of field grown plants, which showed marked seasonal fluctuation. The purified 34 kDa protein from in vitro raised plants, was serologically related to field grown plants and possessed similar characteristics. The micropropagated plants were successfully established in earthen pots under greenhouse conditions.
