Hibiscus Chlorotic Ringspot, a Widespread Virus Disease in the Ornamental Hibiscus Rosa-Sinensis.

Jones, Dennis; Behncken, G. M.

doi:10.1071/app9800004

Cited by 22 publications

(7 citation statements)

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“…The virus is found worldwide where hibiscus is cultivated (16,41,42). The symptoms on HCRSV-infected plants range from generalized mottle to chlorotic ringspots and vein-banding patterns, severe stunting, and flower distortion (42,45).…”

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A Six-Nucleotide Segment within the 3′ Untranslated Region of Hibiscus Chlorotic Ringspot Virus Plays an Essential Role in Translational Enhancement

Koh

Liu

Wong

2002

J Virol

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RNA plant viruses use various translational regulatory mechanisms to control their gene expression.Translational enhancement of viral mRNAs that leads to higher levels of protein synthesis from specific genes may be essential for the virus to successfully compete for cellular translational machinery. The control elements have yet to be analyzed for members of the genus Carmovirus, a small group of plant viruses with positive-sense RNA genomes. In this study, we examined the 3 untranslated region (UTR) of hibiscus chlorotic ringspot virus (HCRSV) genomic RNA (gRNA) and subgenomic RNA (sgRNA) for its role in the translational regulation of viral gene expression. The results showed that the 3 UTR of HCRSV significantly enhanced the translation of several open reading frames on gRNA and sgRNA and a viral gene in a bicistronic construct with an inserted internal ribosome entry site. Through deletion and mutagenesis studies of both the bicistronic construct and full-length gRNA, we demonstrated that a six-nucleotide sequence, GGGCAG, that is complementary to the 3 region of the 18S rRNA and a minimal length of 180 nucleotides are required for the enhancement of translation induced by the 3 UTR.The 3Ј untranslated regions (UTRs) of plant virus RNAs are likely to function in a manner similar to that of the corresponding regions of cellular mRNA in regulating translation and maintaining RNA stability. Due to their great variety in terminal structures, the 3Ј UTRs of plus-strand viral RNAs play varied roles in replication and translation (5). For example, the turnip yellow mosaic virus RNA contains a 3Ј tRNA mimic believed to regulate minus-strand RNA synthesis (6). For brome mosaic virus, the 3Ј UTR harbors a unique promoter element that directs minus-strand RNA synthesis (23). The 3Ј UTR of tobacco mosaic virus contains a translation enhancer (8).Hibiscus chlorotic ringspot virus (HCRSV), a member of the genus Carmovirus, is an isometric monopartite plant virus which measures 28 nm in diameter. The virus is found worldwide where hibiscus is cultivated (16,41,42). The symptoms on HCRSV-infected plants range from generalized mottle to chlorotic ringspots and vein-banding patterns, severe stunting, and flower distortion (42,45). HCRSV possesses a singlestranded positive-sense RNA that is not polyadenylated at the 3Ј terminus. The genomic RNA (gRNA) is 3,911 nucleotides (nt) long and has the potential to encode seven open reading frames (ORFs), including two novel ORFs, p23 and p25 (12). Two 3Ј-coterminated subgenomic RNA (sgRNA) species have been identified.In this study, we examined the 3Ј UTR of HCRSV gRNA and sgRNA for its role in the regulation of translation. Through deletion studies and elimination of putative translation enhancer elements, we were able to identify the prerequisites required for the translational enhancement induced by the 3Ј UTR of HCRSV. The results showed that the 3Ј UTR of HCRSV differentially enhanced the translation of ORFs on gRNA and sgRNA and an HCRSV gene in a bicistronic construct by 1.5-t...

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confidence: 99%

A Six-Nucleotide Segment within the 3′ Untranslated Region of Hibiscus Chlorotic Ringspot Virus Plays an Essential Role in Translational Enhancement

Koh

Liu

Wong

2002

J Virol

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“…It was first identified in a hibiscus cultivar imported to the United States from El Salvador. The virus is found worldwide where hibiscus is cultivated (15,36,37). The symptoms on HCRSVinfected plants range from a generalized mottle to chlorotic ring spots and vein-banding patterns (37).…”

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confidence: 99%

Complete Nucleotide Sequence and Genome Organization of Hibiscus Chlorotic Ringspot Virus, a New Member of the Genus Carmovirus : Evidence for the Presence and Expression of Two Novel Open Reading Frames

Huang

Koh

Weng

et al. 2000

J Virol

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The complete nucleotide sequence of hibiscus chlorotic ringspot virus (HCRSV) was determined. The genomic RNA (gRNA) is 3,911 nucleotides long and has the potential to encode seven viral proteins in the order of 28 (p28), 23 (p23), 81 (p81), 8 (p8), 9 (p9), 38 (p38), and 25 (p25) kDa. Excluding two unique open reading frames (ORFs) encoding p23 and p25, the ORFs encode proteins with high amino acid similarity to those of carmoviruses. In addition to gRNA, two 3-coterminated subgenomic RNA (sgRNA) species were identified. Full-length cDNA clones derived from gRNA and sgRNA were constructed under the control of a T7 promoter. Both capped and uncapped transcripts derived from the full-length genomic cDNA clone were infectious. In vitro translation and mutagenesis assays confirmed that all the predicted ORFs except the ORF encoding p8 are translatable, and the two novel ORFs (those encoding p23 and p25) may be functionally indispensable for the viral infection cycle. Based on virion morphology and genome organization, we propose that HCRSV be classified as a new member of the genus Carmovirus in family Tombusviridae.Hibiscus chlorotic ringspot virus (HCRSV) is an isometric monopartite plant virus which measures 28 nm in diameter. It was first identified in a hibiscus cultivar imported to the United States from El Salvador. The virus is found worldwide where hibiscus is cultivated (15,36,37). The symptoms on HCRSVinfected plants range from a generalized mottle to chlorotic ring spots and vein-banding patterns (37). Many hibiscus hybrids grown in the tropics as ornamental plants showed severe stunting and flower distortion when infected by HCRSV (41).Serological evidence has demonstrated that HCRSV belongs to the genus Carmovirus (13). So far, the complete nucleotide sequences for seven carmoviruses have been determined. These include the type member carnation mottle virus (CarMV) (9), turnip crinkle virus (TCV) (3), melon necrotic spot virus (MNSV) (27), cardamine chlorotic fleck virus (CCFV) (33), cowpea mottle virus (CPMoV) (42), saguaro cactus virus (SCV) (38), and galinsoga mosaic virus (GaMV) (6).All carmoviruses have a genome organization similar to that of CarMV (Fig. 1). These viruses possess two open reading frames (ORFs), one of which results from an in-frame readthrough mechanism, with the potential to encode two polypeptides starting from the first AUG (5,8,22). Both proteins are putative subunits of the viral replicase. Two centrally located small ORFs encode proteins p8 and p9, which are required for virus movement (10, 17). The coat protein gene is located in the 3Ј region of the genome. CPMoV also has a unique ORF that encodes a 28-kDa protein by in-frame readthrough of the ORF encoding p9 [ORF(p9)] (42). The function of p28 of CPMoV is still unknown.Here we report the complete nucleotide sequence, genome organization, construction of a cDNA clone from which infectious transcripts can be derived, and expression of HCRSV in vitro. In addition to the five ORFs found in other carmoviruses, the HCRSV geno...

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“…Virus-infected H. rosa-sinensis can display various symptoms. Infection with Hibiscus chlorotic ringspot virus (HCRSV) exhibits mottling or chlorotic spots, rings, or vein-banding symptoms under natural conditions, which may disappear during the summer months (Jones and Behncken, 1980). Infection with Cotton leaf curl Multan virus (CLCuMuV), exhibits leaf curling, vein swelling, vein darkening and enations on the veins on the undersides of the leaves (She et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

High throughput RNA sequencing discovers symptomatic and latent viruses: an example from ornamental Hibiscus

Zhou

Richert-Poeggeler

Wang

et al. 2022

Preprint

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Hibiscus rosa-sinensis L. (Hibiscus, Malvaceae) is an ornamental species grown widely in amenity plantings. We collected leaves on an urban roadside pavement (sidewalk) near a market in Guangzhou which showed multiple symptoms of leaf rolling, deformation and chlorosis. Initial evaluation by electron microscopy using negative staining of drip preparations revealed the presence of tobamovirus-like particles. Total RNA was extracted, and, unusually, without any RNA selection based on sequence, was used for cDNA library construction and high-throughput survey sequencing. From the 814 Mb of clean sequence data (from 2,712,161 paired reads of 150 bp) reads representing chloroplast, ribosomal, and mitochondrial genes were filtered out, eliminating 79.1% of reads. 1,135,848 x 150 bp of the sequence was retained and screened for viral sequences. Assembly of these sequences detected nine virus species from seven virus genera comprising three tobamoviruses, namely, Tobacco mosaic virus, Tobacco mild green mosaic virus and Hibiscus latent Singapore virus, Turnip mosaic virus (Potyvirus), Potato virus M (Carlavirus), Hibiscus chlorotic ringspot virus (Betacarmovirus), Fabavirus sp (Fabavirus), Cotton leaf curl Multan virus (Begomovirus) and a putative mitoviruses replicating in mitochondria, Chenopodium quinoa mitovirus 1. Mapping the reads to complete virus reference sequences showed high and uniform coverage of the genomes from 3,729x coverage for Turnip mosaic virus to 22x for Cotton leaf curl Multan virus. By comparison, nuclear reference genes actin showed 14x coverage and polyubiquitin 27x. Notable variants from reference sequences (SNPs) were identified. With the low cost of sequencing and potential for semi-automated bioinformatic pipelines, the whole-RNA approach has huge potential for identifying multiple undiagnosed viruses in ornamental plants, resulting in the ability to take preventive measures in production facilities against spread and to product quality for the mutual benefit of producers and consumers.

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Hibiscus Chlorotic Ringspot, a Widespread Virus Disease in the Ornamental Hibiscus Rosa-Sinensis.

Cited by 22 publications

References 1 publication

A Six-Nucleotide Segment within the 3′ Untranslated Region of Hibiscus Chlorotic Ringspot Virus Plays an Essential Role in Translational Enhancement

A Six-Nucleotide Segment within the 3′ Untranslated Region of Hibiscus Chlorotic Ringspot Virus Plays an Essential Role in Translational Enhancement

Complete Nucleotide Sequence and Genome Organization of Hibiscus Chlorotic Ringspot Virus, a New Member of the Genus Carmovirus : Evidence for the Presence and Expression of Two Novel Open Reading Frames

High throughput RNA sequencing discovers symptomatic and latent viruses: an example from ornamental Hibiscus

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