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A nested-polymerase chain reaction protocol for the detection of Mycosphaerella nawae in persimmon

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A nested-polymerase chain reaction protocol for the detection of Mycosphaerella nawae in persimmon

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dc.contributor.author Berbegal Martínez, Mónica es_ES
dc.contributor.author Mora Sala, Beatriz es_ES
dc.contributor.author García Jiménez, José es_ES
dc.date.accessioned 2016-09-19T14:44:51Z
dc.date.available 2016-09-19T14:44:51Z
dc.date.issued 2013-10
dc.identifier.issn 0929-1873
dc.identifier.uri http://hdl.handle.net/10251/70111
dc.description.abstract Mycosphaerella nawae is the causal agent of circular leaf spot of persimmon. A polymerase chain reaction (PCR) based protocol was developed for M. nawae-specific identification from pure culture, or infected symptomatic and asymptomatic persimmon tissues. Variation among the internal transcribed spacer regions (ITS) of the ribosomal DNA (rDNA) sequences of potentially related fungal species in persimmon orchards was analyzed for a primer pair design. Specificity was confirmed using multiple isolates of these species, other fungal pathogens that cause foliar diseases in persimmon and contaminants commonly obtained in the isolation process. The detection threshold for M. nawae DNA was lowered from 50 pg to 500 fg when nested-PCR was evaluated instead of single PCR. The nested-PCR protocol developed in this study showed its suitability to be applied for the specific detection of M. nawae from three types of naturally infected persimmon tissues: from lesions in fresh leaves, from pseudothecia present in lesions in leaf litter, and from infected asymptomatic leaves. The protocol can be useful for routine diagnosis, disease monitoring programs and for epidemiological research. es_ES
dc.description.sponsorship M. Berbegal was a contract holder of the "Campus de Excelencia Internacional" program of the Universitat Politecnica de Valencia. This research was financially supported by Fundacion Agroalimed (Conselleria de Agricultura, Pesca i Alimentacio, Generalitat Valenciana). We thank J. Armengol for critically reading the manuscript prior to submission. en_EN
dc.language Inglés es_ES
dc.publisher Springer Verlag (Germany) es_ES
dc.relation.ispartof European Journal of Plant Pathology es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject Circular leaf spot es_ES
dc.subject Diagnosis es_ES
dc.subject Diospyros kaki es_ES
dc.subject Nested-PCR es_ES
dc.subject.classification BOTANICA es_ES
dc.subject.classification PRODUCCION VEGETAL es_ES
dc.subject.classification ECOSISTEMAS AGROFORESTALES (UPV) es_ES
dc.title A nested-polymerase chain reaction protocol for the detection of Mycosphaerella nawae in persimmon es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1007/s10658-013-0237-0
dc.rights.accessRights Cerrado es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Ecosistemas Agroforestales - Departament d'Ecosistemes Agroforestals es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Agroforestal Mediterráneo - Institut Agroforestal Mediterrani es_ES
dc.description.bibliographicCitation Berbegal Martinez, M.; Mora Sala, B.; García Jiménez, J. (2013). A nested-polymerase chain reaction protocol for the detection of Mycosphaerella nawae in persimmon. European Journal of Plant Pathology. 137(2):273-281. doi:10.1007/s10658-013-0237-0 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion http://dx.doi.org/10.1007/s10658-013-0237-0 es_ES
dc.description.upvformatpinicio 273 es_ES
dc.description.upvformatpfin 281 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 137 es_ES
dc.description.issue 2 es_ES
dc.relation.senia 251647 es_ES
dc.contributor.funder Fundación de la Comunitat Valenciana para la Investigación Agroalimentaria, Agroalimed es_ES
dc.contributor.funder Universitat Politècnica de València es_ES
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