Q-PCR

Identification of novel genes involved in Petunia flower development using transcript profiling and reverse genetics

Nombre: 
Izaskun
Primer Apellido: 
Mallona
Segundo Apellido: 
González
Nombre completo: 
Mallona González, Izaskun
Foto del perfil: 
Foto Principal
0034-868071085
País: 
ESPAÑA
Departamento/Centro/Universidad: 

Ciencia y Tecnología Agraria. Instituto de Biotecnología Vegetal. Universidad Politécnica de Cartagena.

Programa de doctorado: 
Técnicas avanzadas en investigación y desarrollo agrario y alimentario
Año de programa: 
2008
Fecha de inicio: 
01/01/2009
Fecha de finalización: 
14/07/2012
Area de conocimiento: 
Biología vegetal
Estancias Anteriores: 

Ha realizado en 2011 una estancia externa en la Universidad de Verona (Italia) bajo la supervisión del Prof. Dr. Mario Pezzotti para realizar estudios transcriptómicos en petunia mediante micromatrices de ADN.

Resumen de tesis: 

Petals are a key element on plant life cycle as, in many species, they attract pollinators, thus aiding to reproduction. Furthermore, they have economic importance in ornamental crops. In the present study, petal transcriptional patterns were compared within the flower organs in Arabidopsis thaliana. It was found that catalytic molecular functions were overrepresented in
petals. A shortlist comprising the top ten differentially expressed genes in petals were mapped to the model species with industrial value Petunia hybrida, and further downregulated by RNAi. The silencing phenotypes found permitted to assign functions in petal
development to seven novel genes: when silenced, they triggered alterations on flower size and shape (PhCYP76, PhNPH3, PhFeSOD, PhXTH, PhCYP96 and PhWAK), petal smoothness (PhPRA), color (PhNPH3 and PhWAK ) and symmetry
(PhCYP76 ). Pleiotropic phenotypes were found, such as changes in root morphology and leaf color (PhCYP76 ), flower number, capsule and seed morphology (PhCYP96 ) and plant height (PhCYP76 and PhCYP96 ).

To accomplish the experimental design, three methods were developed. First, the “pESTle” management system that assembles, annotates, stores and serves expressed sequence tag data. Second, a reference gene selection for real time PCR experiments that includes a new method for stability estimation based on rank aggregation of published algorithms, and concludes that a normalization
factor with two members of EF1α, SAND, CYP or RAN1 is stable enough under most conditions. And third, a PCR efficiency estimator based on amplicon characteristics which allows efficiency-driven primer design in a Web tool.

Estado de tesis: 
Defendida
Fecha de lectura de tesis: 
23/07/2012
Genética
Director: 
Marcos Egea Gutiérrez-Cortines
Codirector: 
Julia Weiss
Fotos de galeria: 
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