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2021
Chabi-Jesus, Camila; Ramos-González, Pedro L.; Postclam-Barro, Matheus; Fontenele, Rafaela Salgado; Harakava, Ricardo; Bassanezi, Renato B.; Moreira, Alecio S.; Kitajima, Elliot W.; Varsani, Arvind; Freitas-Astúa, Juliana
Molecular Epidemiology of Citrus Leprosis Virus C: A New Viral Lineage and Phylodynamic of the Main Viral Subpopulations in the Americas Journal Article
Em: Frontiers in Microbiology, vol. 12, pp. 986, 2021, ISSN: 1664302X.
Resumo | Links | BibTeX | Tags: Brevipalpus mites, Cilevirus, citrus leprosis disease, Kitaviridae, virus evolution
@article{nokey,
title = {Molecular Epidemiology of Citrus Leprosis Virus C: A New Viral Lineage and Phylodynamic of the Main Viral Subpopulations in the Americas},
author = {Camila Chabi-Jesus and Pedro L. Ramos-González and Matheus Postclam-Barro and Rafaela Salgado Fontenele and Ricardo Harakava and Renato B. Bassanezi and Alecio S. Moreira and Elliot W. Kitajima and Arvind Varsani and Juliana Freitas-Astúa},
doi = {10.3389/FMICB.2021.641252/BIBTEX},
issn = {1664302X},
year = {2021},
date = {2021-01-01},
journal = {Frontiers in Microbiology},
volume = {12},
pages = {986},
publisher = {Frontiers Media S.A.},
abstract = {Despite the importance of viral strains/variants as agents of emerging diseases, genetic and evolutionary processes affecting their ecology are not fully understood. To get insight into this topic, we assessed the population and spatial dynamic parameters of citrus leprosis virus C (CiLV-C, genus Cilevirus, family Kitaviridae). CiLV-C is the etiological agent of citrus leprosis disease, a non-systemic infection considered the main viral disorder affecting citrus orchards in Brazil. Overall, we obtained 18 complete or near-complete viral genomes, 123 complete nucleotide sequences of the open reading frame (ORF) encoding the putative coat protein, and 204 partial nucleotide sequences of the ORF encoding the movement protein, from 430 infected Citrus spp. samples collected between 1932 and 2020. A thorough examination of the collected dataset suggested that the CiLV-C population consists of the major lineages CRD and SJP, unevenly distributed, plus a third one called ASU identified in this work, which is represented by a single isolate found in an herbarium sample collected in Asuncion, Paraguay, in 1937. Viruses from the three lineages share about 85% nucleotide sequence identity and show signs of inter-clade recombination events. Members of the lineage CRD were identified both in commercial and non-commercial citrus orchards. However, those of the lineages SJP were exclusively detected in samples collected in the citrus belt of São Paulo and Minas Gerais, the leading Brazilian citrus production region, after 2015. The most recent common ancestor of viruses of the three lineages dates back to, at least, ∼1500 years ago. Since citrus plants were introduced in the Americas by the Portuguese around the 1520s, the Bayesian phylodynamic analysis suggested that the ancestors of the main CiLV-C lineages likely originated in contact with native vegetation of South America. The intensive expansion of CRD and SJP lineages in Brazil started probably linked to the beginning of the local citrus industry. The high prevalence of CiLV-C in the citrus belt of Brazil likely ensues from the intensive connectivity between orchards, which represents a potential risk toward pathogen saturation across the region.},
keywords = {Brevipalpus mites, Cilevirus, citrus leprosis disease, Kitaviridae, virus evolution},
pubstate = {published},
tppubtype = {article}
}
Despite the importance of viral strains/variants as agents of emerging diseases, genetic and evolutionary processes affecting their ecology are not fully understood. To get insight into this topic, we assessed the population and spatial dynamic parameters of citrus leprosis virus C (CiLV-C, genus Cilevirus, family Kitaviridae). CiLV-C is the etiological agent of citrus leprosis disease, a non-systemic infection considered the main viral disorder affecting citrus orchards in Brazil. Overall, we obtained 18 complete or near-complete viral genomes, 123 complete nucleotide sequences of the open reading frame (ORF) encoding the putative coat protein, and 204 partial nucleotide sequences of the ORF encoding the movement protein, from 430 infected Citrus spp. samples collected between 1932 and 2020. A thorough examination of the collected dataset suggested that the CiLV-C population consists of the major lineages CRD and SJP, unevenly distributed, plus a third one called ASU identified in this work, which is represented by a single isolate found in an herbarium sample collected in Asuncion, Paraguay, in 1937. Viruses from the three lineages share about 85% nucleotide sequence identity and show signs of inter-clade recombination events. Members of the lineage CRD were identified both in commercial and non-commercial citrus orchards. However, those of the lineages SJP were exclusively detected in samples collected in the citrus belt of São Paulo and Minas Gerais, the leading Brazilian citrus production region, after 2015. The most recent common ancestor of viruses of the three lineages dates back to, at least, ∼1500 years ago. Since citrus plants were introduced in the Americas by the Portuguese around the 1520s, the Bayesian phylodynamic analysis suggested that the ancestors of the main CiLV-C lineages likely originated in contact with native vegetation of South America. The intensive expansion of CRD and SJP lineages in Brazil started probably linked to the beginning of the local citrus industry. The high prevalence of CiLV-C in the citrus belt of Brazil likely ensues from the intensive connectivity between orchards, which represents a potential risk toward pathogen saturation across the region.
Chabi-Jesus, Camila; Ramos-González, Pedro L.; Postclam-Barro, Matheus; Fontenele, Rafaela Salgado; Harakava, Ricardo; Bassanezi, Renato B.; Moreira, Alecio S.; Kitajima, Elliot W.; Varsani, Arvind; Freitas-Astúa, Juliana
Molecular Epidemiology of Citrus Leprosis Virus C: A New Viral Lineage and Phylodynamic of the Main Viral Subpopulations in the Americas Journal Article
Em: Frontiers in Microbiology, vol. 12, pp. 986, 2021, ISSN: 1664302X.
Resumo | Links | BibTeX | Tags: Brevipalpus mites, Cilevirus, citrus leprosis disease, Kitaviridae, virus evolution
@article{nokey,
title = {Molecular Epidemiology of Citrus Leprosis Virus C: A New Viral Lineage and Phylodynamic of the Main Viral Subpopulations in the Americas},
author = {Camila Chabi-Jesus and Pedro L. Ramos-González and Matheus Postclam-Barro and Rafaela Salgado Fontenele and Ricardo Harakava and Renato B. Bassanezi and Alecio S. Moreira and Elliot W. Kitajima and Arvind Varsani and Juliana Freitas-Astúa},
doi = {10.3389/FMICB.2021.641252/BIBTEX},
issn = {1664302X},
year = {2021},
date = {2021-01-01},
journal = {Frontiers in Microbiology},
volume = {12},
pages = {986},
publisher = {Frontiers Media S.A.},
abstract = {Despite the importance of viral strains/variants as agents of emerging diseases, genetic and evolutionary processes affecting their ecology are not fully understood. To get insight into this topic, we assessed the population and spatial dynamic parameters of citrus leprosis virus C (CiLV-C, genus Cilevirus, family Kitaviridae). CiLV-C is the etiological agent of citrus leprosis disease, a non-systemic infection considered the main viral disorder affecting citrus orchards in Brazil. Overall, we obtained 18 complete or near-complete viral genomes, 123 complete nucleotide sequences of the open reading frame (ORF) encoding the putative coat protein, and 204 partial nucleotide sequences of the ORF encoding the movement protein, from 430 infected Citrus spp. samples collected between 1932 and 2020. A thorough examination of the collected dataset suggested that the CiLV-C population consists of the major lineages CRD and SJP, unevenly distributed, plus a third one called ASU identified in this work, which is represented by a single isolate found in an herbarium sample collected in Asuncion, Paraguay, in 1937. Viruses from the three lineages share about 85% nucleotide sequence identity and show signs of inter-clade recombination events. Members of the lineage CRD were identified both in commercial and non-commercial citrus orchards. However, those of the lineages SJP were exclusively detected in samples collected in the citrus belt of São Paulo and Minas Gerais, the leading Brazilian citrus production region, after 2015. The most recent common ancestor of viruses of the three lineages dates back to, at least, ∼1500 years ago. Since citrus plants were introduced in the Americas by the Portuguese around the 1520s, the Bayesian phylodynamic analysis suggested that the ancestors of the main CiLV-C lineages likely originated in contact with native vegetation of South America. The intensive expansion of CRD and SJP lineages in Brazil started probably linked to the beginning of the local citrus industry. The high prevalence of CiLV-C in the citrus belt of Brazil likely ensues from the intensive connectivity between orchards, which represents a potential risk toward pathogen saturation across the region.},
keywords = {Brevipalpus mites, Cilevirus, citrus leprosis disease, Kitaviridae, virus evolution},
pubstate = {published},
tppubtype = {article}
}
Despite the importance of viral strains/variants as agents of emerging diseases, genetic and evolutionary processes affecting their ecology are not fully understood. To get insight into this topic, we assessed the population and spatial dynamic parameters of citrus leprosis virus C (CiLV-C, genus Cilevirus, family Kitaviridae). CiLV-C is the etiological agent of citrus leprosis disease, a non-systemic infection considered the main viral disorder affecting citrus orchards in Brazil. Overall, we obtained 18 complete or near-complete viral genomes, 123 complete nucleotide sequences of the open reading frame (ORF) encoding the putative coat protein, and 204 partial nucleotide sequences of the ORF encoding the movement protein, from 430 infected Citrus spp. samples collected between 1932 and 2020. A thorough examination of the collected dataset suggested that the CiLV-C population consists of the major lineages CRD and SJP, unevenly distributed, plus a third one called ASU identified in this work, which is represented by a single isolate found in an herbarium sample collected in Asuncion, Paraguay, in 1937. Viruses from the three lineages share about 85% nucleotide sequence identity and show signs of inter-clade recombination events. Members of the lineage CRD were identified both in commercial and non-commercial citrus orchards. However, those of the lineages SJP were exclusively detected in samples collected in the citrus belt of São Paulo and Minas Gerais, the leading Brazilian citrus production region, after 2015. The most recent common ancestor of viruses of the three lineages dates back to, at least, ∼1500 years ago. Since citrus plants were introduced in the Americas by the Portuguese around the 1520s, the Bayesian phylodynamic analysis suggested that the ancestors of the main CiLV-C lineages likely originated in contact with native vegetation of South America. The intensive expansion of CRD and SJP lineages in Brazil started probably linked to the beginning of the local citrus industry. The high prevalence of CiLV-C in the citrus belt of Brazil likely ensues from the intensive connectivity between orchards, which represents a potential risk toward pathogen saturation across the region.
Leastro, Mikhail Oliveira; Freitas-Astúa, Juliana; Kitajima, Elliot Watanabe; Pallás, Vicente; Sánchez-Navarro, Jesús
Membrane Association and Topology of Citrus Leprosis Virus C2 Movement and Capsid Proteins Journal Article
Em: Microorganisms 2021, Vol. 9, Page 418, vol. 9, iss. 2, pp. 418, 2021, ISSN: 2076-2607.
Resumo | Links | BibTeX | Tags: Cilevirus, membrane association, Movement protein, p29 capsid protein, topology
@article{Leastro2021,
title = {Membrane Association and Topology of Citrus Leprosis Virus C2 Movement and Capsid Proteins},
author = {Mikhail Oliveira Leastro and Juliana Freitas-Astúa and Elliot Watanabe Kitajima and Vicente Pallás and Jesús Sánchez-Navarro},
url = {https://www.mdpi.com/2076-2607/9/2/418/htm https://www.mdpi.com/2076-2607/9/2/418},
doi = {10.3390/MICROORGANISMS9020418},
issn = {2076-2607},
year = {2021},
date = {2021-01-01},
journal = {Microorganisms 2021, Vol. 9, Page 418},
volume = {9},
issue = {2},
pages = {418},
publisher = {Multidisciplinary Digital Publishing Institute},
abstract = {Although citrus leprosis disease has been known for more than a hundred years, one of its causal agents, citrus leprosis virus C2 (CiLV-C2), is poorly characterized. This study described the association of CiLV-C2 movement protein (MP) and capsid protein (p29) with biological membranes. Our findings obtained by computer predictions, chemical treatments after membrane fractionation, and biomolecular fluorescence complementation assays revealed that p29 is peripherally associated, while the MP is integrally bound to the cell membranes. Topological analyses revealed that both the p29 and MP expose their N- and C-termini to the cell cytoplasmic compartment. The implications of these results in the intracellular movement of the virus were discussed.},
keywords = {Cilevirus, membrane association, Movement protein, p29 capsid protein, topology},
pubstate = {published},
tppubtype = {article}
}
Although citrus leprosis disease has been known for more than a hundred years, one of its causal agents, citrus leprosis virus C2 (CiLV-C2), is poorly characterized. This study described the association of CiLV-C2 movement protein (MP) and capsid protein (p29) with biological membranes. Our findings obtained by computer predictions, chemical treatments after membrane fractionation, and biomolecular fluorescence complementation assays revealed that p29 is peripherally associated, while the MP is integrally bound to the cell membranes. Topological analyses revealed that both the p29 and MP expose their N- and C-termini to the cell cytoplasmic compartment. The implications of these results in the intracellular movement of the virus were discussed.
2020
Arena, Gabriella D.; Ramos-González, Pedro Luis; Falk, Bryce W.; Casteel, Clare L.; Freitas-Astúa, Juliana; Machado, Marcos A.
Plant Immune System Activation Upon Citrus Leprosis Virus C Infection Is Mimicked by the Ectopic Expression of the P61 Viral Protein Journal Article
Em: Frontiers in Plant Science, vol. 11, pp. 1188, 2020, ISSN: 1664462X.
Resumo | Links | BibTeX | Tags: Arabidopsis thaliana, Cilevirus, hypersensitive response, Jasmonic acid, Nicotiana benthamiana, plant–virus interaction, RNA-Seq, Salicylic acid
@article{Arena2020,
title = {Plant Immune System Activation Upon Citrus Leprosis Virus C Infection Is Mimicked by the Ectopic Expression of the P61 Viral Protein},
author = {Gabriella D. Arena and Pedro Luis Ramos-González and Bryce W. Falk and Clare L. Casteel and Juliana Freitas-Astúa and Marcos A. Machado},
doi = {10.3389/FPLS.2020.01188/BIBTEX},
issn = {1664462X},
year = {2020},
date = {2020-01-01},
journal = {Frontiers in Plant Science},
volume = {11},
pages = {1188},
publisher = {Frontiers Media S.A.},
abstract = {Citrus leprosis virus C (CiLV-C, genus Cilevirus, family Kitaviridae) is an atypical virus that does not spread systemically in its plant hosts. Upon its inoculation by Brevipalpus mites, only localized lesions occur, and the infection remains limited to cells around mite feeding sites. Here, we aimed to gain insights into the putative causes of viral unfitness in plants by expanding the limited knowledge of the molecular mechanisms underlying plant/kitavirid interactions. Firstly, we quantified the CiLV-C viral RNAs during the infection in Arabidopsis thaliana plants using RT-qPCR and systematized it by defining three stages of distinguishing subgenomic and genomic RNA accumulation: i) 0–24 h after infestation, ii) 2–4 days after infestation (dai), and iii) 6–10 dai. Accordingly, the global plant response to CiLV-C infection was assessed by RNA-Seq at each period. Results indicated a progressive reprogramming of the plant transcriptome in parallel to the increasing viral loads. Gene ontology enrichment analysis revealed the induction of cell growth-related processes at the early stages of the infection and the triggering of the SA-mediated pathway, ROS burst and hypersensitive response (HR) at the presymptomatic stage. Conversely, infected plants downregulated JA/ET-mediated pathways and processes involved in the primary metabolism including photosynthesis. Marker genes of unfolded protein response were also induced, suggesting a contribution of the endoplasmic reticulum stress to the cell death caused by the viral infection. Finally, we transiently expressed CiLV-C proteins in Nicotiana benthamiana plants to undertake their roles in the elicited plant responses. Expression of the CiLV-C P61 protein consistently triggered ROS burst, upregulated SA- and HR-related genes, increased SA levels, reduced JA levels, and caused cell death. Mimicry of responses typically observed during CiLV-C–plant interaction indicates P61 as a putative viral effector causing the HR-like symptoms associated with the infection. Our data strengthen the hypothesis that symptoms of CiLV-C infection might be the outcome of a hypersensitive-like response during an incompatible interaction. Consequently, the locally restricted infection of CiLV-C, commonly observed across infections by kitavirids, supports the thesis that these viruses, likely arising from an ancestral arthropod-infecting virus, are unable to fully circumvent plant defenses.},
keywords = {Arabidopsis thaliana, Cilevirus, hypersensitive response, Jasmonic acid, Nicotiana benthamiana, plant–virus interaction, RNA-Seq, Salicylic acid},
pubstate = {published},
tppubtype = {article}
}
Citrus leprosis virus C (CiLV-C, genus Cilevirus, family Kitaviridae) is an atypical virus that does not spread systemically in its plant hosts. Upon its inoculation by Brevipalpus mites, only localized lesions occur, and the infection remains limited to cells around mite feeding sites. Here, we aimed to gain insights into the putative causes of viral unfitness in plants by expanding the limited knowledge of the molecular mechanisms underlying plant/kitavirid interactions. Firstly, we quantified the CiLV-C viral RNAs during the infection in Arabidopsis thaliana plants using RT-qPCR and systematized it by defining three stages of distinguishing subgenomic and genomic RNA accumulation: i) 0–24 h after infestation, ii) 2–4 days after infestation (dai), and iii) 6–10 dai. Accordingly, the global plant response to CiLV-C infection was assessed by RNA-Seq at each period. Results indicated a progressive reprogramming of the plant transcriptome in parallel to the increasing viral loads. Gene ontology enrichment analysis revealed the induction of cell growth-related processes at the early stages of the infection and the triggering of the SA-mediated pathway, ROS burst and hypersensitive response (HR) at the presymptomatic stage. Conversely, infected plants downregulated JA/ET-mediated pathways and processes involved in the primary metabolism including photosynthesis. Marker genes of unfolded protein response were also induced, suggesting a contribution of the endoplasmic reticulum stress to the cell death caused by the viral infection. Finally, we transiently expressed CiLV-C proteins in Nicotiana benthamiana plants to undertake their roles in the elicited plant responses. Expression of the CiLV-C P61 protein consistently triggered ROS burst, upregulated SA- and HR-related genes, increased SA levels, reduced JA levels, and caused cell death. Mimicry of responses typically observed during CiLV-C–plant interaction indicates P61 as a putative viral effector causing the HR-like symptoms associated with the infection. Our data strengthen the hypothesis that symptoms of CiLV-C infection might be the outcome of a hypersensitive-like response during an incompatible interaction. Consequently, the locally restricted infection of CiLV-C, commonly observed across infections by kitavirids, supports the thesis that these viruses, likely arising from an ancestral arthropod-infecting virus, are unable to fully circumvent plant defenses.
2017
Arena, Gabriella Dias; Ramos-González, Pedro Luis; Nunes, Maria Andréia; Jesus, Camila Chabi; Calegario, Renata Faier; Kitajima, Elliot Watanabe; Novelli, Valdenice Moreira; Freitas-Astúa, Juliana
Arabidopsis thaliana as a model host for Brevipalpus mite-transmitted viruses Journal Article
Em: Scientia Agricola, vol. 74, iss. 1, pp. 85-89, 2017, ISSN: 1678-992X.
Resumo | Links | BibTeX | Tags: Cilevirus, Clerodendrum chlorotic spot virus, Coffee ringspot virus, Dichorhavirus, Solanum violaefolium ringspot virus
@article{Arena2017,
title = {\textit{Arabidopsis thaliana} as a model host for \textit{Brevipalpus} mite-transmitted viruses},
author = {Gabriella Dias Arena and Pedro Luis Ramos-González and Maria Andréia Nunes and Camila Chabi Jesus and Renata Faier Calegario and Elliot Watanabe Kitajima and Valdenice Moreira Novelli and Juliana Freitas-Astúa},
url = {http://www.scielo.br/j/sa/a/mLKYNzqSqGsJZwmP5XM5LbH/?lang=en},
doi = {10.1590/1678-992X-2015-0380},
issn = {1678-992X},
year = {2017},
date = {2017-01-01},
journal = {Scientia Agricola},
volume = {74},
issue = {1},
pages = {85-89},
publisher = {Escola Superior de Agricultura "Luiz de Queiroz"},
abstract = {ABSTRACT: Brevipalpus-transmitted viruses (BTV) are a taxonomically diverse group of plant viruses which severely affect a number of major crops. Members of the group can be sub-classified into cytoplasmic (BTV-C) or nuclear type (BTV-N) according to the accumulation sites of virions in the infected plant cells. Both types of BTV produce only local infections near the point of inoculation by viruliferous mites. Features of BTV-plant interactions such as the failure of systemic spread in their natural hosts are poorly understood. In this study we evaluated Arabidopsis thaliana, a model plant commonly used for the study of plant-virus interactions, as an alternative host for BTV. Infection of Arabidopsis with the BTV-N Coffee ringspot virus and Clerodendrum chlorotic spot virus, and the BTV-C Solanum violaefolium ringspot virus, were mediated by viruliferous Brevipalpus mites collected in the wild. Upon infestation, local lesions appeared in 7 to 10 days on leaves of, at least, 80 % of the assayed plants. Presence of viral particles and characteristic cytopathic effects were detected by transmission electron microscopy (TEM) and the viral identities confirmed by specific reverse-transcriptase polymerase chain reaction (RT-PCR) and further amplicon sequencing. The high infection rate and reproducibility of symptoms of the three different viruses assayed validate A. thaliana as a feasible alternative experimental host for BTV.},
keywords = {Cilevirus, Clerodendrum chlorotic spot virus, Coffee ringspot virus, Dichorhavirus, Solanum violaefolium ringspot virus},
pubstate = {published},
tppubtype = {article}
}
ABSTRACT: Brevipalpus-transmitted viruses (BTV) are a taxonomically diverse group of plant viruses which severely affect a number of major crops. Members of the group can be sub-classified into cytoplasmic (BTV-C) or nuclear type (BTV-N) according to the accumulation sites of virions in the infected plant cells. Both types of BTV produce only local infections near the point of inoculation by viruliferous mites. Features of BTV-plant interactions such as the failure of systemic spread in their natural hosts are poorly understood. In this study we evaluated Arabidopsis thaliana, a model plant commonly used for the study of plant-virus interactions, as an alternative host for BTV. Infection of Arabidopsis with the BTV-N Coffee ringspot virus and Clerodendrum chlorotic spot virus, and the BTV-C Solanum violaefolium ringspot virus, were mediated by viruliferous Brevipalpus mites collected in the wild. Upon infestation, local lesions appeared in 7 to 10 days on leaves of, at least, 80 % of the assayed plants. Presence of viral particles and characteristic cytopathic effects were detected by transmission electron microscopy (TEM) and the viral identities confirmed by specific reverse-transcriptase polymerase chain reaction (RT-PCR) and further amplicon sequencing. The high infection rate and reproducibility of symptoms of the three different viruses assayed validate A. thaliana as a feasible alternative experimental host for BTV.