Characterization of a virus infecting Citrus volkameriana with citrus leprosis-like symptoms.

A Citrus volkameriana tree displaying symptoms similar to citrus leprosis on its leaves and bark was found in Hawaii. Citrus leprosis virus C (CiLV-C)-specific detection assays, however, were negative for all tissues tested. Short, bacilliform virus-like particles were observed by transmission electron microscopy in the cytoplasm of symptomatic leaves but not in healthy controls. Double-stranded (ds) RNAs ≈8 and 3 kbp in size were present in symptomatic leaf tissue but not in healthy controls. Excluding poly(A) tails, the largest molecule, RNA1, was 8,354 bp in length. The ≈3 kbp dsRNA band was found to be composed of two distinct molecules, RNA2 and RNA3, which were 3,169 and 3,113 bp, respectively. Phylogenetic analyses indicated that the RNA-dependent RNA polymerase (RdRp) domain located in RNA1 was most closely related to the RdRp domain of CiLV-C. A reverse-transcription polymerase chain reaction assay developed for the detection of this virus was used to screen nearby citrus trees as well as Hibiscus arnottianus plants with symptoms of hibiscus green spot, a disease associated with infection by Hibiscus green spot virus (HGSV). All nearby citrus trees tested negative with the assay; however, symptomatic H. arnottianus plants were positive. All three RNAs were present in symptomatic H. arnottianus and were >98% identical to the RNAs isolated from C. volkameriana. We contend that the virus described in this study is HGSV, and propose that it be the type member of a new virus genus, Higrevirus.

An assemblage of closteroviruses infects Hawaiian ti (Cordyline fruticosa L.).

The ti plant (Cordyline fruticosa L.) is culturally important throughout most of Polynesia and has considerable economic importance in Hawai'i where the foliage is commonly used in cultural ceremonies as well as food and ornamental industries. In Hawai'i, ringspot symptoms were recently observed on leaves of the common green variety of ti growing in Kahalu'u on the island of O'ahu, and Wailuku and Hana on the island of Maui. High molecular weight double-stranded (ds)RNAs were isolated from the leaves of symptomatic plants as well as plants without symptoms. A cDNA library derived from the dsRNAs present in symptomatic plants was generated and sequenced. These sequences indicated at least four distinct clostero-like viruses were present in the plants, and phylogenetic analyses suggested they were most closely related to Little cherry virus 1, an unassigned member of the family Closteroviridae. The 16,883 nucleotide genome of one of these viruses was determined and predicted to contain ten open reading frames with an organization typical of closteroviruses. Reverse-transcription PCR revealed this virus was present in both symptomatic and asymptomatic ti plants, making it unlikely to be responsible for the observed ringspot symptoms. We propose the name Cordyline virus 1 (CoV-1) for this virus and include it as a new, unassigned member of the family Closteroviridae.

Genetic diversity and evidence for recent modular recombination in Hawaiian Citrus tristeza virus.

The Hawaiian Islands are home to a widespread and diverse population of Citrus tristeza virus (CTV), an economically important pathogen of citrus. In this study, we quantified the genetic diversity of two CTV genes and determined the complete genomic sequence for two strains of Hawaiian CTV. The nucleotide diversity was estimated to be 0.0565 + or - 0.0022 for the coat protein (CP) gene (n = 137) and 0.0822 + or - 0.0033 for the p23 gene (n = 30). The genome size and organization of CTV strains HA18-9 and HA16-5 were similar to other fully sequenced strains of CTV. The 3'-terminal halves of their genomes were nearly identical (98.5% nucleotide identity), whereas the 5'-terminal halves were more distantly related (72.3% nucleotide identity), suggesting a possible recombination event. Closer examination of strain HA16-5 indicated that it arose through recent recombination between the movement module of an HA18-9 genotype, and the replication module of an undescribed CTV genotype.

Genome organization and phylogenetic relationship of Pineapple mealybug wilt associated virus-3 with family Closteroviridae members.

The nucleotide sequence of Pineapple mealybug wilt associated virus-3 (PMWaV-3) (Closteroviridae: Ampelovirus), spanning seven open reading frames (ORFs) and the untranslatable region of the 3' end was determined. Based on the amino acid identities with orthologous ORFs of PMWaV-1 (54%-73%) and PMWaV-2 (13%-35%), we propose PMWaV-3 is a new species in the PMWaV complex. PMWaV-3 lacks an intergenic region between ORF1b and ORF2, encodes a relatively small, 28.8 kDa, coat protein, and lacks a coat protein duplicate. Phylogenetic analyses were used to analyze seven different domains and ORFs from members of the family Closteroviridae. Two distinct clades within the recognized genus Ampelovirus were observed; one that includes PMWaV-3 and PMWaV-1 and several GLRaVs and another that includes PMWaV-2 and GLRaV-3, the type member of the genus Ampelovirus.