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Detection of mutants with some alteration in leaf development is the best strategy to identify the genes that control these characters. Among the possible alternatives to generate mutants (e.g., radiations, chemical products such as EMS, etc.), insertional mutagenesis is the most appropriate to address this objective. With this strategy, after obtaining a large number of transgenic plants, their phenotype is evaluated in order to detect alterations caused by the integration of T-DNA. If the disruption of an endogenous gene occurs, a loss-of-function mutant is usually obtained, whereas the insertion of the T-DNA in a control region or in the vicinity thereof can cause an alteration of its level of expression. In any case, since the endogenous gene is labeled by T-DNA, it is possible to reach cloning of the gene by means of PCR-based methods (eg Anchor- or Tail-PCR) and obtain valuable information on the role of that gene in the evaluated character. The development of an efficient genetic transformation protocol in tomato and related wild species (Solanum pimpinellifolium, S. pennellii and S. galapagense) has allowed obtaining more than 7,000 T-DNA lines in the context of several projects. We screened these collections of T-DNA lines to identify mutants altered in characters of vegetative and reproductive development, as well as in the level of tolerance to abiotic stresses, mainly salinity and drought. The frequency of mutants (dominant or recessive) for the evaluated characters is around 10% and Southern analysis reveals that the average number of inserts per line is moderate (1.7), which facilitates the identification of the T-DNA insert that generates the mutant phenotype. Within the framework of these projects, the identification of several genes has been achieved from mutants selected for different characters. In this work we propose the characterization of several altered mutants in leaf development through the screening of T-DNA lines.

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RiuNet. Repositorio institucional de la Universitat de Valencia  

Autor(es)

ROMERO CORTÉS, PABLO YUSSEF - 

Id.: 70997403

Idioma: spa  - 

Versión: 1.0

Estado: Final

Palabras claveInsertional mutagenesis - 

Tipo de recurso: info:eu-repo/semantics/masterThesis  - 

Tipo de Interactividad: Expositivo

Nivel de Interactividad: muy bajo

Audiencia: Estudiante  -  Profesor  -  Autor  - 

Estructura: Atomic

Coste: no

Copyright: sí

: Reserva de todos los derechos

Requerimientos técnicos:  Browser: Any - 

Fecha de contribución: 16-feb-2018

Contacto:

Localización:

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