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Projecte llegit

Títol: Molecular and physiological study of light regulation of plant development

Estudiant que ha llegit aquest projecte:

Tutor/a o Cotutor/a: ROIG VILLANOVA, IRMA

Departament: DEAB

Títol: Molecular and physiological study of light regulation of plant development

Data inici oferta: 24-01-2020      Data finalització oferta: 03-02-2020

Estudis d'assignació del projecte:
    GR ENG SIS BIOLÒGICS

Lloc de realització:


En empresa (cal signar un conveni de cooperació)

        Tutor/a Extern: Jaume Martínez
        Institució/Empresa: CRAG

Segon tutor/a extern: Pedro Pastor Andreu

Paraules clau:
Shade Avoidance Syndrome, Arabidopsis, hypocotyl elongation, HFR1, HY5, PHYA

Descripció del contingut i pla d'activitats:
El estudiante participará en el proyecto de investigación de
¿Regulación por luz del desarrollo vegetal" en el que
estudiamos a nivel genético, molecular y bioquímico la
respuesta de la planta a la proximidad de otros vegetales.
Las técnicas que se manejarán son las básicas en BIOLOGÍA
MOLECULAR Y GENÉTICA de plantas: transformación de E. coli,
Agrobacterium tumefaciens, Cardamine y Arabidopsis, extracción
de DNA plasmídico, obtención de plásmidos por ligación
tradicional y por el sistema gateway (análisis por PCR,
digestión con enzimas de restricción, etc.), extracción de DNA
de plantas, selección de plantas transgénicas, realización de
cruces entre plantas de Arabidopsis, etc.

Overview (resum en anglès): A plant can detect a possible situation of shade by perceiving changes in the ratio between Red light and Far Red light (R:FR ratio). When it detects it, a wide network of components is activated. If the plant is a shade-avoider, the action of these components triggers changes in its development including vertical growth promotion, branching inhibition and early flowering, among others. Altogether these responses are known as the Shade Avoidance Syndrome (SAS) and are detrimental for agriculture.
A research group of the Centre for Research in Agricultural Genomics (CRAG) found the temporal and spatial action of three important components of this network. LONGHYPOCOTYL IN FAR-RED1 (HFR1), ELONGATED HYPOCOTYL 5 (HY5) and Phytochrome A (phyA) proteins act in the hypocotyl of Arabidopsis thaliana, mainly during the first stages of seedling development. Once this was determined, it remained to be seen whether these three genes were expressed in the same location where the proteins they encode act.
This bachelor thesis is focused on the study of hypocotyl elongation as a SAS response. The ultimate objective is the analysis of the spatial distribution of the expression of HFR1, HY5 and PHYA in the different organs of the Arabidopsis seedling.
To perform these experiments, Arabidopsis thaliana mutant plants deficient in each of these components were transformed with constructs containing the promoter of the gene fused to the coding sequence of the gene, fused to a reporter gene. We employed three binary plasmids: pHFR1:HFR1-GFP-3xHA (named pSP125), pHY5:HY5-YFP (named pB71) and pPHYA:PHYA-GFP (named pAPAG).
After that, we followed a pipeline to identify homozygous lines expressing the transgenes. This pipeline involved selection of transformants by identifying resistant lines to selective antibiotics or herbicides, molecular analyses via PCR and complementation tests.
It was possible to obtain segregating transgenic lines that showed partial complementation phenotypes for the pSP125 and the pAPAG constructs. Due to the situation of COVID-19 and some failed results, it was not possible to analyse the expression of the three genes in the seedlings.

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