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 | Acesso ao texto completo restrito à biblioteca da Embrapa Mandioca e Fruticultura. Para informações adicionais entre em contato com cnpmf.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Mandioca e Fruticultura. |
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Data corrente: |
14/11/2022 |
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Data da última atualização: |
14/11/2022 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
ARENA, G. D.; RAMOS-GONZÁLEZ, P. L.; TASSI, A. D.; MACHADO, M. A.; ASTUA, J. de F. |
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Afiliação: |
GABRIELLA D. ARENA, Instituto Biológico; PEDRO L. RAMOS GONZÁLEZ, Instituto Biológico; ALINE D. TASSI, Instituto Biológico; MARCOS A. MACHADO, Centro de Citricultura Sylvio Moreira; JULIANA DE FREITAS ASTUA, CNPMF. |
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Título: |
A TaqMan RT qPCR assay for absolute quantifcation of citrus leprosis virus C lineage SJP: disclosing the subgenomic/genomic ratio in plant and mite vector, plant organ specifc viral loads, and the kinetics of viral accumulation in plants. |
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Ano de publicação: |
2022 |
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Fonte/Imprenta: |
Tropical Plant Pathology, November, 2022. |
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DOI: |
https://doi.org/10.1007/s40858-022-00539-4 |
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Idioma: |
Inglês |
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Conteúdo: |
Citrus leprosis virus C (CiLV-C) causes citrus leprosis, a re-emergent viral disease afecting citrus production in Latin America. Here, we developed two TaqMan RT-qPCR assays to detect and quantify CiLV-C lineage SJP, prevalent in the Brazilian citrus belt and the world?s main sweet orange production area. Assays targeted sequences within the genes p29 and RdRp. ORF p29 is transcribed in a subgenomic RNA (sgRNA) and codes for the putative capsid protein. ORF RdRp, coding for the replicase, is directly translated from the genomic RNA (gRNA). After assessing the efciency and sensitivity of the assays, the targets were quantifed in (i) symptomatic tissues of feld-collected sweet orange (Citrus sinensis) samples, in a time course after infection in both (ii) Arabidopsis thaliana and (iii) sweet orange plants, and in (iv) the mite vector Brevipalpus yothersi. Sweet orange fruits support higher quantities of CiLV-C molecules than stems and leaves. Amounts of viral molecules in late lesions of diferent developmental stages collected in the feld remain stable, but CiLV-C quantities progressively increase from the early stages of the infection to the appearance of symptoms in both A. thaliana and C. sinensis. The high sensibility of the assays allowed the quantifcation of CiLV-C in early infection periods, even during the asymptomatic period of plants, and in scant amounts of B. yothersi individuals. In plants, a higher accumulation of p29 than RdRp was reported (sgRNA/gRNA up to 95), refecting the transcription of the p29 sgRNA. In mites, p29 quantities were only slightly higher than RdRp (sgRNA/gRNA of 2), adding a new tool to evaluate the putative replication of CiLV-C in its vector, a challenging aspect of the study of mite-virus interplay. The methods developed here contribute to a more accurate analysis of citrus leprosis epidemiology and shed light on unknown features of the virus-vector interaction MenosCitrus leprosis virus C (CiLV-C) causes citrus leprosis, a re-emergent viral disease afecting citrus production in Latin America. Here, we developed two TaqMan RT-qPCR assays to detect and quantify CiLV-C lineage SJP, prevalent in the Brazilian citrus belt and the world?s main sweet orange production area. Assays targeted sequences within the genes p29 and RdRp. ORF p29 is transcribed in a subgenomic RNA (sgRNA) and codes for the putative capsid protein. ORF RdRp, coding for the replicase, is directly translated from the genomic RNA (gRNA). After assessing the efciency and sensitivity of the assays, the targets were quantifed in (i) symptomatic tissues of feld-collected sweet orange (Citrus sinensis) samples, in a time course after infection in both (ii) Arabidopsis thaliana and (iii) sweet orange plants, and in (iv) the mite vector Brevipalpus yothersi. Sweet orange fruits support higher quantities of CiLV-C molecules than stems and leaves. Amounts of viral molecules in late lesions of diferent developmental stages collected in the feld remain stable, but CiLV-C quantities progressively increase from the early stages of the infection to the appearance of symptoms in both A. thaliana and C. sinensis. The high sensibility of the assays allowed the quantifcation of CiLV-C in early infection periods, even during the asymptomatic period of plants, and in scant amounts of B. yothersi individuals. In plants, a higher accumulation of p29 than RdRp was reported (sgRNA/gRNA up to 95)... Mostrar Tudo |
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Palavras-Chave: |
Kitaviridae; Real-time PC. |
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Thesagro: |
Doença de Planta; Fruta Cítrica; Leprose Cítrica. |
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Thesaurus Nal: |
Arabidopsis; Brevipalpus; Cilevirus; Citrus. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 02910naa a2200289 a 4500
001 2148257
005 2022-11-14
008 2022 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.1007/s40858-022-00539-4$2DOI
100 1 $aARENA, G. D.
245 $aA TaqMan RT qPCR assay for absolute quantifcation of citrus leprosis virus C lineage SJP$bdisclosing the subgenomic/genomic ratio in plant and mite vector, plant organ specifc viral loads, and the kinetics of viral accumulation in plants.$h[electronic resource]
260 $c2022
520 $aCitrus leprosis virus C (CiLV-C) causes citrus leprosis, a re-emergent viral disease afecting citrus production in Latin America. Here, we developed two TaqMan RT-qPCR assays to detect and quantify CiLV-C lineage SJP, prevalent in the Brazilian citrus belt and the world?s main sweet orange production area. Assays targeted sequences within the genes p29 and RdRp. ORF p29 is transcribed in a subgenomic RNA (sgRNA) and codes for the putative capsid protein. ORF RdRp, coding for the replicase, is directly translated from the genomic RNA (gRNA). After assessing the efciency and sensitivity of the assays, the targets were quantifed in (i) symptomatic tissues of feld-collected sweet orange (Citrus sinensis) samples, in a time course after infection in both (ii) Arabidopsis thaliana and (iii) sweet orange plants, and in (iv) the mite vector Brevipalpus yothersi. Sweet orange fruits support higher quantities of CiLV-C molecules than stems and leaves. Amounts of viral molecules in late lesions of diferent developmental stages collected in the feld remain stable, but CiLV-C quantities progressively increase from the early stages of the infection to the appearance of symptoms in both A. thaliana and C. sinensis. The high sensibility of the assays allowed the quantifcation of CiLV-C in early infection periods, even during the asymptomatic period of plants, and in scant amounts of B. yothersi individuals. In plants, a higher accumulation of p29 than RdRp was reported (sgRNA/gRNA up to 95), refecting the transcription of the p29 sgRNA. In mites, p29 quantities were only slightly higher than RdRp (sgRNA/gRNA of 2), adding a new tool to evaluate the putative replication of CiLV-C in its vector, a challenging aspect of the study of mite-virus interplay. The methods developed here contribute to a more accurate analysis of citrus leprosis epidemiology and shed light on unknown features of the virus-vector interaction
650 $aArabidopsis
650 $aBrevipalpus
650 $aCilevirus
650 $aCitrus
650 $aDoença de Planta
650 $aFruta Cítrica
650 $aLeprose Cítrica
653 $aKitaviridae
653 $aReal-time PC
700 1 $aRAMOS-GONZÁLEZ, P. L.
700 1 $aTASSI, A. D.
700 1 $aMACHADO, M. A.
700 1 $aASTUA, J. de F.
773 $tTropical Plant Pathology, November, 2022.
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