Development of a molecular tool for the diagnosis of leprosis, a major threat to citrus production in the Americas
Identifieur interne : 002B63 ( Main/Exploration ); précédent : 002B62; suivant : 002B64Development of a molecular tool for the diagnosis of leprosis, a major threat to citrus production in the Americas
Auteurs : Eliane Cristina Locali [Brésil] ; Juliana Freitas-Astua [Brésil] ; Alessandra Alves De Souza [Brésil] ; Marco Aurélio Takita [Brésil] ; Gustavo Astua-Monge [Brésil] ; Renata Antonioli [Brésil] ; Elliot W. Kitajima [Brésil] ; Marcos Antonio Machado [Brésil]Source :
- Plant disease [ 0191-2917 ] ; 2003.
Descripteurs français
- Pascal (Inist)
English descriptors
- KwdEn :
Abstract
Citrus leprosis virus (CiLV), a tentative member of the Rhabdoviridae family, affects citrus trees in Brazil, where it is transmitted by mites Brevipalpus spp. It also occurs in other South American countries and was recently identified in Central America. This northbound spread of CiLV is being considered a serious threat to the citrus industry of the United States. However, despite its importance, difficulties related to the biology of CiLV have hindered much of the progress regarding its accurate detection, leaving both the analyses of symptoms and electron microscopy as the only tools available. An attempt to overcome this problem was made by constructing a cDNA library from double-stranded RNA extracted from leprosis lesions of infected Citrus sinensis (sweet orange) leaves. After cloning and sequencing, specific primers were designed to amplify putative CiLV genome regions with similarity to genes encoding the movement protein and replicase of other plant viruses. RNA from infected citrus plants corresponding to different varieties and locations were amplified by reverse transcription-polymerase chain reaction (RT-PCR) using the two pairs of primers. Amplified products were purified, cloned in pGEM-T, and sequenced. The sequences confirmed the genomic regions previously associated with CiLV. The results demonstrate that RT-PCR was specific, accurate, rapid, and reliable for the detection of CiLV.
Affiliations:
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<front><div type="abstract" xml:lang="en">Citrus leprosis virus (CiLV), a tentative member of the Rhabdoviridae family, affects citrus trees in Brazil, where it is transmitted by mites Brevipalpus spp. It also occurs in other South American countries and was recently identified in Central America. This northbound spread of CiLV is being considered a serious threat to the citrus industry of the United States. However, despite its importance, difficulties related to the biology of CiLV have hindered much of the progress regarding its accurate detection, leaving both the analyses of symptoms and electron microscopy as the only tools available. An attempt to overcome this problem was made by constructing a cDNA library from double-stranded RNA extracted from leprosis lesions of infected Citrus sinensis (sweet orange) leaves. After cloning and sequencing, specific primers were designed to amplify putative CiLV genome regions with similarity to genes encoding the movement protein and replicase of other plant viruses. RNA from infected citrus plants corresponding to different varieties and locations were amplified by reverse transcription-polymerase chain reaction (RT-PCR) using the two pairs of primers. Amplified products were purified, cloned in pGEM-T, and sequenced. The sequences confirmed the genomic regions previously associated with CiLV. The results demonstrate that RT-PCR was specific, accurate, rapid, and reliable for the detection of CiLV.</div>
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<tree><country name="Brésil"><region name="État de São Paulo"><name sortKey="Locali, Eliane Cristina" sort="Locali, Eliane Cristina" uniqKey="Locali E" first="Eliane Cristina" last="Locali">Eliane Cristina Locali</name>
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<name sortKey="Antonioli, Renata" sort="Antonioli, Renata" uniqKey="Antonioli R" first="Renata" last="Antonioli">Renata Antonioli</name>
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<name sortKey="Kitajima, Elliot W" sort="Kitajima, Elliot W" uniqKey="Kitajima E" first="Elliot W." last="Kitajima">Elliot W. Kitajima</name>
<name sortKey="Machado, Marcos Antonio" sort="Machado, Marcos Antonio" uniqKey="Machado M" first="Marcos Antonio" last="Machado">Marcos Antonio Machado</name>
<name sortKey="Takita, Marco Aurelio" sort="Takita, Marco Aurelio" uniqKey="Takita M" first="Marco Aurélio" last="Takita">Marco Aurélio Takita</name>
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