Genomic characterization of Calla lily chlorotic spot virus and design of broad-spectrum primers for detection of tospoviruses

Local chlorotic spots resembling early lesions characteristic of citrus leprosis (CL) were observed in leaves of two sweet orange (Citrus sinensis L.) trees in Teresina, State of Piauí, Brazil, in early 2017. However, despite the similarities, these spots were generally larger than those of a typical CL and showed rare or no necrosis symptoms. In symptomatic tissues, transmission electron microscopy revealed the presence of viroplasms in the nuclei of the infected parenchymal cells and rod-shaped particles with an average size of approximately 40 × 100 nm, resembling those typically observed during infection by dichorhaviruses. A bipartite genome of the putative novel virus, tentatively named citrus chlorotic spot virus (CiCSV) (RNA1 = 6,518 nucleotides [nt] and RNA2 = 5,987 nt), revealed the highest nucleotide sequence identity values with the dichorhaviruses coffee ringspot virus strain Lavras (73.8%), citrus leprosis virus N strain Ibi1 (58.6%), and orchid fleck virus strain So (56.9%). In addition to citrus, CiCSV was also found in local chlorotic lesions on leaves of the ornamental plant beach hibiscus (Talipariti tiliaceum (L.) Fryxell). Morphological characterization of mites recovered from the infected plants revealed at least two different types of Brevipalpus. One of them corresponds to Brevipalpus yothersi. The other is slightly different from B. yothersi mites but comprises traits that possibly place it as another species. A mix of the two mite types collected on beach hibiscus successfully transmitted CiCSV to arabidopsis plants but additional work is required to verify whether both types of flat mite may act as viral vectors. The current study reveals a newly described dichorhavirus associated with a citrus disease in the northeastern region of Brazil.

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Complete genome sequence of a distinct calla lily chlorotic spot virus isolated in mainland China

Archives of Virology ◽

10.1007/s00705-015-2630-9 ◽

2015 ◽

Vol 161 (1) ◽

pp. 219-222 ◽

Cited By ~ 4

Author(s):

Ye Xu ◽

Si-bo Wang ◽

Yong-zhong Li ◽

Hong-zheng Tao ◽

Ya-ning Huang ◽

...

Keyword(s):

Genome Sequence ◽

Complete Genome Sequence ◽

Complete Genome ◽

Mainland China ◽

Spot Virus ◽

Chlorotic Spot ◽

Calla Lily

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Biological and Genomic Characterization of a Novel Tobamovirus Infecting Hoya spp.

Plant Disease ◽

10.1094/pdis-04-18-0667-re ◽

2018 ◽

Vol 102 (12) ◽

pp. 2571-2577 ◽

Cited By ~ 3

Author(s):

Scott Adkins ◽

Tom D’Elia ◽

Kornelia Fillmer ◽

Patchara Pongam ◽

Carlye A. Baker

Keyword(s):

Virus Infection ◽

Chenopodium Quinoa ◽

Amino Acid Sequences ◽

Chlorotic Spot ◽

Individual Gene ◽

Genome Nucleotide Sequence ◽

Experimental Host Range ◽

Genomic Characterization ◽

Systemic Infections

Foliar symptoms suggestive of virus infection were observed on the ornamental plant hoya (Hoya spp.; commonly known as waxflower) in Florida. An agent that reacted with commercially available tobamovirus detection reagents was mechanically transmitted to Chenopodium quinoa and Nicotiana benthamiana. Rod-shaped particles ∼300 nm in length and typical of tobamoviruses were observed in partially purified virion preparations by electron microscopy. An experimental host range was determined by mechanical inoculation with virions, and systemic infections were observed in plants in the Asclepiadaceae, Apocynaceae, and Solanaceae families. Some species in the Solanaceae and Chenopodiaceae families allowed virus replication only in inoculated leaves, and were thus only local hosts for the virus. Tested plants in the Amaranthaceae, Apiaceae, Brassicaceae, Cucurbitaceae, Fabaceae, and Malvaceae did not support either local or systemic virus infection. The complete genome for the virus was sequenced and shown to have a typical tobamovirus organization. Comparisons of genome nucleotide sequence and individual gene deduced amino acid sequences indicate that it is a novel tobamovirus sharing the highest level of sequence identity with Streptocarpus flower break virus and members of the Brassicaceae-infecting subgroup of tobamoviruses. The virus, for which the name Hoya chlorotic spot virus (HoCSV) is proposed, was detected in multiple hoya plants from different locations in Florida.

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SEQUENCE ANALYSIS OF S RNA OF CALLA LILY CHLOROTIC SPOT VIRUS, A NEW TOSPOVIRUS

Acta Horticulturae ◽

10.17660/actahortic.2006.722.13 ◽

2006 ◽

pp. 103-110

Author(s):

Y.H. Lin ◽

T.C. Chen ◽

M.H. Chung ◽

C.C. Chen ◽

S.D. Yeh ◽

...

Keyword(s):

Sequence Analysis ◽

Spot Virus ◽

Chlorotic Spot ◽

Calla Lily

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Serological Comparison and Molecular Characterization for Verification of Calla lily chlorotic spot virus as a New Tospovirus Species Belonging to Watermelon silver mottle virus Serogroup

Phytopathology ◽

10.1094/phyto-95-1482 ◽

2005 ◽

Vol 95 (12) ◽

pp. 1482-1488 ◽

Cited By ~ 38

Author(s):

Yu-Hsuan Lin ◽

Tsung-Chi Chen ◽

Hei-Ti Hsu ◽

Fang-Lin Liu ◽

Fang-Hua Chu ◽

...

Keyword(s):

Open Reading Frames ◽

Mottle Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Spot Virus ◽

Heterologous Antigen ◽

Chlorotic Spot ◽

Calla Lily ◽

Homologous Antigen ◽

Distinct Member ◽

Watermelon Silver Mottle Virus

Calla lily chlorotic spot virus (CCSV) isolated from central Taiwan was recently identified as a tospovirus serologically but distantly related to Watermelon silver mottle virus (WSMoV). To clarify the serological relationship between the two viruses, rabbit polyclonal antibody (PAb) to CCSV and mouse monoclonal antibodies (MAbs) to WSMoV NP or CCSV NP were produced in this investigation, using purified nucleocapsid protein (NP) as immunogens. The PAb to CCSV NP reacted stronger with the homologous antigen than with the heterologous antigen, with much lower A405 readings in indirect enzyme-linked immunosorbent assay (ELISA) and low-intensity banding in immunoblotting. MAbs produced to CCSV NP or WSMoV NP reacted specifically with the homologous antigens but not with the heterologous antigens in both ELISA and immunoblot analyses. The CCSV S RNA was determined to be 3,172 nucleotides in length, with an inverted repeat at the 5′ and 3′ ends and two open reading frames encoding the NP and a nonstructural (NSs) protein in an ambisense arrangement. A typical 3′-terminal sequence (5′-AUUGCUCU-3′) that is shared by all members of the genus Tospovirus also is present in the CCSV S RNA. The CCSV NP and NSs protein share low amino acid identities of 20.1 to 65.1% and 19.9 to 66.1%, respectively, with those of reported tospoviruses. Phylogenetic dendrogram analysis indicates that CCSV is a distinct member in the genus Tospovirus. The results provide evidence that CCSV is a new species in the genus Tospovirus and belongs to WSMoV serogroup.

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First Report of Capsicum chlorosis virus Causing Yellow Stripes on Calla Lilies in Taiwan

Plant Disease ◽

10.1094/pdis-91-9-1201c ◽

2007 ◽

Vol 91 (9) ◽

pp. 1201-1201 ◽

Cited By ~ 9

Author(s):

C. C. Chen ◽

C. H. Huang ◽

T. C. Chen ◽

S. D. Yeh ◽

Y. H. Cheng ◽

...

Keyword(s):

Chenopodium Quinoa ◽

Amino Acid Sequences ◽

N Gene ◽

Impatiens Necrotic Spot Virus ◽

Leaf Extracts ◽

Spot Virus ◽

Chlorotic Spot ◽

Calla Lily ◽

Capsicum Chlorosis Virus ◽

Virus Isolates

Tomato spotted wilt virus (TSWV) and Calla lily chlorotic spot virus (CCSV) are two recognized species of the Tospovirus genus in the family Bunyaviridae infecting calla lily (Zantedeschia spp.). During 2005, 15 virus isolates were collected from different calla lily plants exhibiting yellow stripes on their leaves in Ho-Li, a major calla lily-production township in Taiwan. After three successive local lesion passages on Chenopodium quinoa Willd., diseased leaf tissues individually infected by these isolates were preserved in liquid nitrogen and used for subsequent identification studies. Using the tospovirus genus-specific primers gL3637 and gL4435c designed from the L RNA, an 800-bp DNA fragment was amplified in reverse transcription-PCR from all 15 isolates. Moreover, leaf extracts of the diseased calla lilies and the C. quinoa plants inoculated with the 15 virus isolates reacted with antisera against the nucleocapsid proteins (NP) of Capsicum chlorosis virus (CaCV)-gloxinia and Watermelon silver mottle virus (WSMoV), but not to monoclonal antibodies against the NP of TSWV, CCSV, Peanut chlorotic fan-spot virus (PCFV), or Impatiens necrotic spot virus (INSV) in indirect ELISA. These results indicate that the 15 virus isolates are tospoviruses belonging to the WSMoV serogroup. Additionally, we amplified and sequenced the full-length N gene from these tospovirus isolates using primers WN2328 (5′-CCATTGGTTTGCCTCCG-3′) and WN3534 (5′-CGTCGACAGAGCAATCGAGGC-3′) designed from the S RNA of WSMoV. The deduced amino acid sequences of the N protein from these 15 tospovirus isolates showed a greater than 92% identity to that of CaCV (GenBank Accession No. NC-008301). Furthermore, results of phylogenetic analysis of the 15 isolates on the basis of amino acids sequences, both genetic distance and parsimony trees indicated that they were all genetically clustered within CaCV using INSV, TSWV, and WSMoV as outgroups. The results indicate that the virus causing yellow stripes in calla lilies is a strain of CaCV. To our knowledge, this is the first evidence that CaCV can naturally infect calla lilies and cause yellow stripe symptoms. Reference: (1) F.-H. Chu et al. Phytopathology 91:361, 2001.

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Molecular Characterization of Sorghum Chlorotic Spot Virus, a Proposed Furovirus

Journal of General Virology ◽

10.1099/0022-1317-69-9-2335 ◽

1988 ◽

Vol 69 (9) ◽

pp. 2335-2345 ◽

Cited By ~ 10

Author(s):

T. L. Kendall ◽

W. G. Langenberg ◽

S. A. Lommel

Keyword(s):

Molecular Characterization ◽

Spot Virus ◽

Chlorotic Spot

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Calla lily chlorotic spot virus from Spider Lily (Hymenocallis litteralis) and Tobacco (Nicotiana tabacum) in the South-west of China

Journal of Phytopathology ◽

10.1111/j.1439-0434.2011.01873.x ◽

2012 ◽

Vol 160 (4) ◽

pp. 201-205 ◽

Cited By ~ 7

Author(s):

Yating Liu ◽

Xiuping Lu ◽

Long Zhi ◽

Yuanxian Zheng ◽

Xuejiao Chen ◽

...

Keyword(s):

Nicotiana Tabacum ◽

The South ◽

Spot Virus ◽

Chlorotic Spot ◽

Calla Lily ◽

South West

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Biological and Molecular Characterization of Apple chlorotic leaf spot virus Causing Chlorotic Leaf Spot on Pear (Pyrus pyrifolia) in Taiwan

HortScience ◽

10.21273/hortsci.45.7.1073 ◽

2010 ◽

Vol 45 (7) ◽

pp. 1073-1078 ◽

Cited By ~ 2

Author(s):

Zhong-Bin Wu ◽

Hsin-Mei Ku ◽

Yuh-Kun Chen ◽

Chung-Jan Chang ◽

Fuh-Jyh Jan

Keyword(s):

Amino Acid ◽

Leaf Spot ◽

Phylogenetic Analyses ◽

Amino Acid Sequences ◽

Pyrus Pyrifolia ◽

Spot Virus ◽

Chlorotic Spot ◽

Cp Gene ◽

Chlorotic Leaf

Pear plants (Pyrus pyrifolia var. Hengshen) showing symptoms of chlorotic spots on leaves were observed in orchards in central Taiwan in 2004. The sap of diseased leaves reacted positively to Apple chlorotic leaf spot virus (ACLSV) antiserum. A purified virus isolate (LTS1) from pear was characterized by host range, electron microscopy, phylogenetic analyses, serological property, and back-inoculation experiments to pear. Fifteen of 28 species of tested plants were susceptible to this virus after mechanical inoculation. Pathogenicity of ACLSV isolate LTS1 was verified by back-inoculating to pear seedlings. Filamentous virions of ≈12 × 750 nm were observed in the preparations of purified virus. Virus particles accumulated in the cytoplasm were observed in the ultrathin sections of LTS1-infected pear leaf tissue. Sequence analyses of the coat protein (CP) gene of LTS1 and the CP gene of LTS2, which originated from a distinct symptomatic pear sample, shared 81.4% to 92.6% nucleotide and 87.6% to 98.4% amino acid identities with those of the CP of 35 ACLSV isolates available in GenBank. ACLSV isolates were grouped into four clusters, i.e., Asia I, II, III, and Europe, and isolates LTS1 and LTS2 were classified as members of cluster Asia II and Asia I, respectively, based on phylogenetic data. Moreover, the variability of amino acid sequences of the CP gene of 37 ACLSV isolates showed geographically associated clustering in the phylogenetic tree. To our knowledge, this is the first study on the characterization of ACLSV causing the leaf chlorotic spot disease of pear in Taiwan. This study also provides the phylogenetic relationships among ACLSV populations based on amino acid sequences of CPs, which are correlated with their geographic origins.

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