Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.)
Abstract Ficus carica L. is considered one of the oldest fruit trees in the Mediterranean basin and is widely grown and harvested for the consumption of its fruits dry and fresh. This species is affected by different virus diseases, especially by Fig mosaic disease (FMD), for which Fig leaf mottle-a...
| Autor: | |
|---|---|
| Tipo de recurso: | tesis doctoral |
| Fecha de publicación: | 2017 |
| País: | España |
| Institución: | Universitat Politècnica de València (UPV) |
| Repositorio: | RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia |
| Idioma: | inglés |
| OAI Identifier: | oai:riunet.upv.es:10251/79876 |
| Acceso en línea: | https://riunet.upv.es/handle/10251/79876 |
| Access Level: | acceso abierto |
| Palabra clave: | Fig Mosaic RT-PCR Virus distribution Cytokinins Encapsulation Micropropagation Synthetic seed Sanitation |
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| dc.title.none.fl_str_mv |
Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.) |
| title |
Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.) |
| spellingShingle |
Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.) Yahyaoui, Emna Fig Mosaic RT-PCR Virus distribution Cytokinins Encapsulation Micropropagation Synthetic seed Sanitation |
| title_short |
Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.) |
| title_full |
Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.) |
| title_fullStr |
Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.) |
| title_full_unstemmed |
Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.) |
| title_sort |
Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.) |
| dc.creator.none.fl_str_mv |
Yahyaoui, Emna |
| author |
Yahyaoui, Emna |
| author_facet |
Yahyaoui, Emna |
| author_role |
author |
| dc.contributor.none.fl_str_mv |
Alfaro Fernández, Ana Olvido Burruano, Santella D'Onghia, Anna Maria Germana, Maria Antonietta Departamento de Ecosistemas Agroforestales Instituto Agroforestal Mediterráneo Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural Repositorio Institucional de la Universitat Politècnica de València Riunet |
| dc.subject.none.fl_str_mv |
Fig Mosaic RT-PCR Virus distribution Cytokinins Encapsulation Micropropagation Synthetic seed Sanitation |
| topic |
Fig Mosaic RT-PCR Virus distribution Cytokinins Encapsulation Micropropagation Synthetic seed Sanitation |
| description |
Abstract Ficus carica L. is considered one of the oldest fruit trees in the Mediterranean basin and is widely grown and harvested for the consumption of its fruits dry and fresh. This species is affected by different virus diseases, especially by Fig mosaic disease (FMD), for which Fig leaf mottle-associated virus 1 (FLMaV-1), Fig leaf mottle-associated virus 2 (FLMaV-2), Fig mild mottling-associated virus (FMMaV), Fig mosaic virus (FMV), Fig latent virus 1 (FLV-1), Fig badnavirus 1 (FBV-1) and Fig fleck-associated virus (FFkaV) are associated. FMD is the most widespread disorder of this species, which represents a threat and a constraint for healthy fig production and germplasm exchange. Thus, the objective of the present doctoral research was the establishment of an efficient and rapid in vitro F. carica propagation, sanitation and conservation of free-FMD plant material for future large-scale commercialization. Initially, FMD-related viruses distribution was screened within the different fig plant organs (buds, leaves, syconia and seeds) of 14 Mediterranean genotypes (Palazzo, Severoni precoce, Bianca, Pilusedda, Dottato bianco, Bifera, Zidi, Baiyadi, Biancu, Brogiotto nero, Catalanisca, Houmairi, Triboiti and Turca 'Serilop') which were utilized afterward as in vitro plant source material. RT-PCR assays revealed that all the aforementioned viruses were present without any exception in seeds, whereas only 4 viruses (FBV, FFkaV, FLMaV-1 and FMV) were detected in buds, leaves and syconia with highly variable infection rates. Moreover, encapsulation technology proved to be a powerful multiplication technique to sustain standard fig tissue culture protocol for three cultivars (Catalanisca, Palazzo and Bifera) and it gave high, almost similar, viability, regrowth and conversion rates. Microcutting rooting in one-step was achieved and conversion rate was comparable for the three cultivars. Furthermore, in order to eliminate FMD associated viruses, with the exception of FBV-1 which resisted to all the sanitation attempts, Caulogenesis and Meristem Tip Culture Protected by the Synthetic Seeds technique (MTC-SS) gave the best sanitation rates. Finally, F. carica (cv. Houmairi) artificial seeds conservation, for final delivery, was achieved. A high viability and moderate regrowth rates were registered with a lesser conversion rate strictly related to the plant growth regulators (PGRs) used. Keywords: Fig, mosaic, RT-PCR, virus distribution, cytokinins, encapsulation, micropropagation, synthetic seed. |
| publishDate |
2017 |
| dc.date.none.fl_str_mv |
2017 2017-04-21 2017 2017-03-14 |
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doctoral thesis http://purl.org/coar/resource_type/c_db06 AM http://purl.org/coar/version/c_ab4af688f83e57aa |
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info:eu-repo/semantics/doctoralThesis |
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doctoralThesis |
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https://riunet.upv.es/handle/10251/79876 |
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https://riunet.upv.es/handle/10251/79876 |
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Inglés eng |
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Inglés |
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eng |
| dc.rights.none.fl_str_mv |
open access http://purl.org/coar/access_right/c_abf2 Reserva de todos los derechos http://rightsstatements.org/vocab/InC/1.0/ |
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info:eu-repo/semantics/openAccess |
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open access http://purl.org/coar/access_right/c_abf2 Reserva de todos los derechos http://rightsstatements.org/vocab/InC/1.0/ |
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openAccess |
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application/pdf application/vnd.openxmlformats-officedocument.wordprocessingml.document application/pdf |
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Universitat Politècnica de València |
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Universitat Politècnica de València |
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reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia instname:Universitat Politècnica de València (UPV) |
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Use of standard and setup of non conventional techniques for the elimination of viruses associated with Fig Mosaic Disease (FMD) in fig germplasm (Ficus carica L.)Yahyaoui, EmnaFigMosaicRT-PCRVirus distributionCytokininsEncapsulationMicropropagationSynthetic seedSanitationAbstract Ficus carica L. is considered one of the oldest fruit trees in the Mediterranean basin and is widely grown and harvested for the consumption of its fruits dry and fresh. This species is affected by different virus diseases, especially by Fig mosaic disease (FMD), for which Fig leaf mottle-associated virus 1 (FLMaV-1), Fig leaf mottle-associated virus 2 (FLMaV-2), Fig mild mottling-associated virus (FMMaV), Fig mosaic virus (FMV), Fig latent virus 1 (FLV-1), Fig badnavirus 1 (FBV-1) and Fig fleck-associated virus (FFkaV) are associated. FMD is the most widespread disorder of this species, which represents a threat and a constraint for healthy fig production and germplasm exchange. Thus, the objective of the present doctoral research was the establishment of an efficient and rapid in vitro F. carica propagation, sanitation and conservation of free-FMD plant material for future large-scale commercialization. Initially, FMD-related viruses distribution was screened within the different fig plant organs (buds, leaves, syconia and seeds) of 14 Mediterranean genotypes (Palazzo, Severoni precoce, Bianca, Pilusedda, Dottato bianco, Bifera, Zidi, Baiyadi, Biancu, Brogiotto nero, Catalanisca, Houmairi, Triboiti and Turca 'Serilop') which were utilized afterward as in vitro plant source material. RT-PCR assays revealed that all the aforementioned viruses were present without any exception in seeds, whereas only 4 viruses (FBV, FFkaV, FLMaV-1 and FMV) were detected in buds, leaves and syconia with highly variable infection rates. Moreover, encapsulation technology proved to be a powerful multiplication technique to sustain standard fig tissue culture protocol for three cultivars (Catalanisca, Palazzo and Bifera) and it gave high, almost similar, viability, regrowth and conversion rates. Microcutting rooting in one-step was achieved and conversion rate was comparable for the three cultivars. Furthermore, in order to eliminate FMD associated viruses, with the exception of FBV-1 which resisted to all the sanitation attempts, Caulogenesis and Meristem Tip Culture Protected by the Synthetic Seeds technique (MTC-SS) gave the best sanitation rates. Finally, F. carica (cv. Houmairi) artificial seeds conservation, for final delivery, was achieved. A high viability and moderate regrowth rates were registered with a lesser conversion rate strictly related to the plant growth regulators (PGRs) used. Keywords: Fig, mosaic, RT-PCR, virus distribution, cytokinins, encapsulation, micropropagation, synthetic seed.Resumen La higuera (Ficus carica L.) es considerada como uno de de los árboles frutales más antiguos de la cuenca mediterránea y es ampliamente cultivado y cosechado para el consumo de sus frutos tanto secos como en fresco. Esta especie se ve afectada por diversas enfermedades virales, especialmente por la denominada "Fig mosaic disease" (FMD) asociada actualemnte a los virus: Fig leaf mottle-associated virus 1 (FLMaV-1), Fig leaf mottle-associated virus 2 (FLMaV-2), Fig mild mottling-associated virus (FMMaV), Fig mosaic virus (FMV), Fig latent virus 1 (FLV-1), Fig badnavirus 1 (FBV-1) y Fig fleck-associated virus (FFkaV). Esta enfermedad representa una amenaza y un obstáculo para la producción de higos y el intercambio de germoplasma. El principal objetivo del presente trabajo fue establecer un método de propagación de higuera in vitro para el saneamiento y la conservación de material vegetal libre de FMD para su posterior comercialización. Inicialmente, se estudió la distribución de los virus implicados en la enfermedad en diversos órganos de 14 genotipos de F. carica (Palazzo, Severoni precoce, Bianca, Pilusedda, Dottato bianco, Bifera, Zidi, Baiyadi, Biancu, Brogiotto nero, Catalanisca, Houmairi, Triboiti y Turca 'Serilop'), los cuales fueron utilizados posteriormente como fuente material vegetal in vitro. Los resultados obtenidos mediante RT-PCR revelaron que todos los virus mencionados estaban presentes sin excepción en las semillas, mientras que sólo cuatro de ellos (FBV, FFkaV, FLMaV-1 y FMV) fueron en brotes, hojas y siconios con tasas de infección variables. Además, la tecnología de encapsulación demostró ser una técnica de multiplicación eficaz para poder aplicar el protocolo estándar de cultivo de tejidos de higo para tres cultivares (Catalanisca, Palazzo y Bifera) dando altas tasas de viabilidad, rebrote y conversión. Se logró el enraizamiento de microcortes en un solo paso y el índice de conversión fue comparable para los tres cultivares. La callogénesis y el culñtivo de meristemos con la técnica de la semilla sintética (MTC-SS) fueron las técnicas que proporcionaron mayores tasas de desinfección para los virus estudiados a excepción de con FBV-1, entidad viral que no fue eliminada con ninguna de las técnicas ensayadas. Por último, se logró la conservación de las semillas artificiales de higuera (cv Houmairi), registrándose una alta viabilidad y tasas de rebrote moderadas con un menor grado de conversión estrictamente relacionado con hormonas utilizadas. Palabras clave: Higuera, mosaico, RT-PCR, la distribución de los virus, hormonas, encapsulación, micropropagación, y la semilla sintética.Resum La figuera (Ficus carica L.) és considerada un dels arbres fruiters més antics de la conca mediterrània i és àmpliament conreat i collit per al seu consum fresc i sec. Les malalties virals, especialment "Fig mosaic disease" (FMD), associada amb els viruses: Fig leaf mottle-associated virus 1 (FLMaV-1), Fig leaf mottle-associated virus 2 (FLMaV-2), Fig mild mottling-associated virus (FMMaV), Fig mosaic virus (FMV), Fig latent virus 1 (FLV-1), Fig badnavirus 1 (FBV-1) i Fig fleck-associated virus (FFkaV). Esta malaltia representa una amenaça per a la producció de figues i l'intercanvi de germoplasma. El principal objectiu d'aquest treball va ser estableixerun mètode de propagació de figuera in vitro per al sanejament i la conservació de material lliure de FMD per a su posterior commercialització. Inicialment, es va estudiar la distribució dels virus associats a FMD en diversos òrgans en 14 genotips de F. carica (Palazzo, Severoni Precoce, Bianca, Pilusedda, Dottato bianco, Bifera, Zidi, Baiyadi, Biancu, Brogiotto diners, Catalanisca, Houmairi, Triboiti i Turca 'Serilop'), els quals van ser utilitzats posteriorment com a font de material vegetal in vitro. Els resultats obtinguts del anàlisis realitzats per RT-PCR van revelar que tots els virus eren presents sense excepció en les llavors, mentre que només quatre virus (FBV, FFkaV, FLMaV-1 i FMV) van ser detectats en brots, fulles i siconis amb taxes d'infecció variables. A més, la tecnologia d'encapsulació va demostrar ser una tècnica de multiplicació eficaç per poder aplicar el protocol estàndard de cultiu de teixits de figa per a tres cultivars (Catalanisca, Palazzo i Bifera) donant taxesadequades de viabilitat, rebrot i conversió. Es va aconseguir l'arrelament de microtalls en un sol pas i l'índex de conversió va ser comparable per als tres cultivars. La calogènesi i el cultiu de meristems protegits per llavors sintètiques (MTC-SS)van ser les tècniques que proporcionarem millores tases de desinfecció per als virus estudiats amb l'excepció de FBV-1 que es va resistir a tots els mètodes de sanejament. Finalment, es va aconseguir la conservació de la llavors artificials de figuera (cv. Houmairi), registrant-ne una alta viabilitat i taxes de rebrot moderades amb un menor grau de conversió estrictament relacionat amb hormones utilitzades. Paraules clau: Figuera, mosaic, RT-PCR, la distribució dels virus, hormones, encapsulació, micropropagació, i la llavor sintètica.Universitat Politècnica de ValènciaAlfaro Fernández, Ana OlvidoBurruano, SantellaD'Onghia, Anna MariaGermana, Maria AntoniettaDepartamento de Ecosistemas AgroforestalesInstituto Agroforestal MediterráneoEscuela Técnica Superior de Ingeniería Agronómica y del Medio NaturalRepositorio Institucional de la Universitat Politècnica de València Riunet20172017-04-2120172017-03-14doctoral thesishttp://purl.org/coar/resource_type/c_db06AMhttp://purl.org/coar/version/c_ab4af688f83e57aainfo:eu-repo/semantics/doctoralThesisapplication/pdfapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/pdfhttps://riunet.upv.es/handle/10251/79876reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valénciainstname:Universitat Politècnica de València (UPV)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Reserva de todos los derechoshttp://rightsstatements.org/vocab/InC/1.0/info:eu-repo/semantics/openAccessoai:riunet.upv.es:10251/798762026-06-13T07:49:27Z |
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