Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures

Autores
Samarfard, S.; Bejerman, Nicolas; Sharman, M.; Trucco, Verónica Milagros; Giolitti, Fabián José; Dietzgen, R. G.
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In 2010, a severe disease of lucerne (Medicago sativa L.) named alfalfa dwarf disease (ADD) was reported in Argentina. High throughput sequencing of diseased plants indicated the presence of five viruses, the (−) ssRNA virus, alfalfa dwarf virus (ADV), the (+) RNA viruses, alfalfa mosaic virus (AMV), bean leafroll virus (BLRV) and alfalfa enamovirus 1 (AEV-1) and the ssDNA virus, alfalfa leaf curl virus (ALCV). In this study, we determined which ADD-associated viruses are present in Australian lucerne that showed diverse virus-like symptoms. A duplex RT-PCR was developed for simultaneous detection of ADV and AMV using a cloned non-infectious ADV RNA fragment as positive control. Similarly, the presence of BLRV and AEV-1 was determined by duplex RT-PCR, and ALCV by PCR. Only AMV and BLRV that are endemic to Australia were detected. None of the novel exotic viruses ADV, AEV-1 and ALCV were detected in lucerne samples collected between 2015 and 2017. However, AMV and BLRV were detected in 78% and 70% of tested samples, respectively. Based on analysis of coat protein (CP) nucleotide sequences, Australian BLRV isolates are closely related to each other and to the Argentine Manfredi isolate. Phylogenetic analyses based on CP gene nucleotide sequences confirmed separation of AMV isolates into two subgroups. The majority of AMV isolates, including all those from Argentina and Australia, clustered in subgroup I with isolates from various hosts and geographic origins. All Australian AMV isolates were closely related to AMV isolated from ADD-affected lucerne in Argentina.
Fil: Samarfard, S.. The University Of Queensland; Australia
Fil: Bejerman, Nicolas. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet. Córdoba. Unidad de Fitopatología y Modernización Agrícola (UFYMA); Argentina
Fil: Sharman, M.. The University Of Queensland; Australia
Fil: Trucco, Verónica Milagros. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; Argentina
Fil: Giolitti, Fabián José. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; Argentina
Fil: Dietzgen, R. G.. The University Of Queensland; Australia
Materia
Biosecurity
Diagnostics
Geminivirus
Genetic Diversity
Next Generation Sequencing
Rhabdovirus
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/69501
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/69501
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pasturesSamarfard, S.Bejerman, NicolasSharman, M.Trucco, Verónica MilagrosGiolitti, Fabián JoséDietzgen, R. G.BiosecurityDiagnosticsGeminivirusGenetic DiversityNext Generation SequencingRhabdovirushttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1In 2010, a severe disease of lucerne (Medicago sativa L.) named alfalfa dwarf disease (ADD) was reported in Argentina. High throughput sequencing of diseased plants indicated the presence of five viruses, the (−) ssRNA virus, alfalfa dwarf virus (ADV), the (+) RNA viruses, alfalfa mosaic virus (AMV), bean leafroll virus (BLRV) and alfalfa enamovirus 1 (AEV-1) and the ssDNA virus, alfalfa leaf curl virus (ALCV). In this study, we determined which ADD-associated viruses are present in Australian lucerne that showed diverse virus-like symptoms. A duplex RT-PCR was developed for simultaneous detection of ADV and AMV using a cloned non-infectious ADV RNA fragment as positive control. Similarly, the presence of BLRV and AEV-1 was determined by duplex RT-PCR, and ALCV by PCR. Only AMV and BLRV that are endemic to Australia were detected. None of the novel exotic viruses ADV, AEV-1 and ALCV were detected in lucerne samples collected between 2015 and 2017. However, AMV and BLRV were detected in 78% and 70% of tested samples, respectively. Based on analysis of coat protein (CP) nucleotide sequences, Australian BLRV isolates are closely related to each other and to the Argentine Manfredi isolate. Phylogenetic analyses based on CP gene nucleotide sequences confirmed separation of AMV isolates into two subgroups. The majority of AMV isolates, including all those from Argentina and Australia, clustered in subgroup I with isolates from various hosts and geographic origins. All Australian AMV isolates were closely related to AMV isolated from ADD-affected lucerne in Argentina.Fil: Samarfard, S.. The University Of Queensland; AustraliaFil: Bejerman, Nicolas. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet. Córdoba. Unidad de Fitopatología y Modernización Agrícola (UFYMA); ArgentinaFil: Sharman, M.. The University Of Queensland; AustraliaFil: Trucco, Verónica Milagros. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; ArgentinaFil: Giolitti, Fabián José. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; ArgentinaFil: Dietzgen, R. G.. The University Of Queensland; AustraliaCsiro Publishing2018-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/69501Samarfard, S.; Bejerman, Nicolas; Sharman, M.; Trucco, Verónica Milagros; Giolitti, Fabián José; et al.; Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures; Csiro Publishing; Australasian Plant Pathology; 47; 2; 3-2018; 215-2250815-3191CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://link.springer.com/10.1007/s13313-017-0533-9info:eu-repo/semantics/altIdentifier/doi/10.1007/s13313-017-0533-9info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:39:58Zoai:ri.conicet.gov.ar:11336/69501instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:39:59.109CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures
title Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures
spellingShingle Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures
Samarfard, S.
Biosecurity
Diagnostics
Geminivirus
Genetic Diversity
Next Generation Sequencing
Rhabdovirus
title_short Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures
title_full Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures
title_fullStr Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures
title_full_unstemmed Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures
title_sort Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures
dc.creator.none.fl_str_mv Samarfard, S.
Bejerman, Nicolas
Sharman, M.
Trucco, Verónica Milagros
Giolitti, Fabián José
Dietzgen, R. G.
author Samarfard, S.
author_facet Samarfard, S.
Bejerman, Nicolas
Sharman, M.
Trucco, Verónica Milagros
Giolitti, Fabián José
Dietzgen, R. G.
author_role author
author2 Bejerman, Nicolas
Sharman, M.
Trucco, Verónica Milagros
Giolitti, Fabián José
Dietzgen, R. G.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Biosecurity
Diagnostics
Geminivirus
Genetic Diversity
Next Generation Sequencing
Rhabdovirus
topic Biosecurity
Diagnostics
Geminivirus
Genetic Diversity
Next Generation Sequencing
Rhabdovirus
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv In 2010, a severe disease of lucerne (Medicago sativa L.) named alfalfa dwarf disease (ADD) was reported in Argentina. High throughput sequencing of diseased plants indicated the presence of five viruses, the (−) ssRNA virus, alfalfa dwarf virus (ADV), the (+) RNA viruses, alfalfa mosaic virus (AMV), bean leafroll virus (BLRV) and alfalfa enamovirus 1 (AEV-1) and the ssDNA virus, alfalfa leaf curl virus (ALCV). In this study, we determined which ADD-associated viruses are present in Australian lucerne that showed diverse virus-like symptoms. A duplex RT-PCR was developed for simultaneous detection of ADV and AMV using a cloned non-infectious ADV RNA fragment as positive control. Similarly, the presence of BLRV and AEV-1 was determined by duplex RT-PCR, and ALCV by PCR. Only AMV and BLRV that are endemic to Australia were detected. None of the novel exotic viruses ADV, AEV-1 and ALCV were detected in lucerne samples collected between 2015 and 2017. However, AMV and BLRV were detected in 78% and 70% of tested samples, respectively. Based on analysis of coat protein (CP) nucleotide sequences, Australian BLRV isolates are closely related to each other and to the Argentine Manfredi isolate. Phylogenetic analyses based on CP gene nucleotide sequences confirmed separation of AMV isolates into two subgroups. The majority of AMV isolates, including all those from Argentina and Australia, clustered in subgroup I with isolates from various hosts and geographic origins. All Australian AMV isolates were closely related to AMV isolated from ADD-affected lucerne in Argentina.
Fil: Samarfard, S.. The University Of Queensland; Australia
Fil: Bejerman, Nicolas. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet. Córdoba. Unidad de Fitopatología y Modernización Agrícola (UFYMA); Argentina
Fil: Sharman, M.. The University Of Queensland; Australia
Fil: Trucco, Verónica Milagros. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; Argentina
Fil: Giolitti, Fabián José. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; Argentina
Fil: Dietzgen, R. G.. The University Of Queensland; Australia
description In 2010, a severe disease of lucerne (Medicago sativa L.) named alfalfa dwarf disease (ADD) was reported in Argentina. High throughput sequencing of diseased plants indicated the presence of five viruses, the (−) ssRNA virus, alfalfa dwarf virus (ADV), the (+) RNA viruses, alfalfa mosaic virus (AMV), bean leafroll virus (BLRV) and alfalfa enamovirus 1 (AEV-1) and the ssDNA virus, alfalfa leaf curl virus (ALCV). In this study, we determined which ADD-associated viruses are present in Australian lucerne that showed diverse virus-like symptoms. A duplex RT-PCR was developed for simultaneous detection of ADV and AMV using a cloned non-infectious ADV RNA fragment as positive control. Similarly, the presence of BLRV and AEV-1 was determined by duplex RT-PCR, and ALCV by PCR. Only AMV and BLRV that are endemic to Australia were detected. None of the novel exotic viruses ADV, AEV-1 and ALCV were detected in lucerne samples collected between 2015 and 2017. However, AMV and BLRV were detected in 78% and 70% of tested samples, respectively. Based on analysis of coat protein (CP) nucleotide sequences, Australian BLRV isolates are closely related to each other and to the Argentine Manfredi isolate. Phylogenetic analyses based on CP gene nucleotide sequences confirmed separation of AMV isolates into two subgroups. The majority of AMV isolates, including all those from Argentina and Australia, clustered in subgroup I with isolates from various hosts and geographic origins. All Australian AMV isolates were closely related to AMV isolated from ADD-affected lucerne in Argentina.
publishDate 2018
dc.date.none.fl_str_mv 2018-03
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/69501
Samarfard, S.; Bejerman, Nicolas; Sharman, M.; Trucco, Verónica Milagros; Giolitti, Fabián José; et al.; Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures; Csiro Publishing; Australasian Plant Pathology; 47; 2; 3-2018; 215-225
0815-3191
CONICET Digital
CONICET
url http://hdl.handle.net/11336/69501
identifier_str_mv Samarfard, S.; Bejerman, Nicolas; Sharman, M.; Trucco, Verónica Milagros; Giolitti, Fabián José; et al.; Development and validation of PCR assays for detection of alfalfa dwarf disease-associated viruses in Australian lucerne pastures; Csiro Publishing; Australasian Plant Pathology; 47; 2; 3-2018; 215-225
0815-3191
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://link.springer.com/10.1007/s13313-017-0533-9
info:eu-repo/semantics/altIdentifier/doi/10.1007/s13313-017-0533-9
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Csiro Publishing
publisher.none.fl_str_mv Csiro Publishing
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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