CBL - Campus del Baix Llobregat

Projecte llegit

Títol: Caracterització de dos gens involucrats en la biogènesi de la paret cel·lular de Quercus suber

Estudiant que ha llegit aquest projecte:

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

Departament: DEAB

Títol: Caracterització de dos gens involucrats en la biogènesi de la paret cel·lular de Quercus suber

Data inici oferta: 05-02-2024      Data finalització oferta: 05-10-2024

Estudis d'assignació del projecte:
    GR ENG SIS BIOLÒGICS
    GR ENG SIS BIOLÒG 23

Lloc de realització:


En empresa (cal signar un conveni de cooperació)

        Tutor/a Extern: David Caparrós
        Institució/Empresa: Centre de Recerca en Agrigenòmica

Paraules clau:
Quercus suber, suberina, pectines , gelp72 , pectinesterasa, clonació i Biologia molecular.

Descripció del contingut i pla d'activitats:
L'estudiant estarà implicat en la clonació d'un gen de blat de moro que participa en la biogènesi de la paret cel·lular amb la finalitat de poder sobre-expressar-ho en plantes i estudiar la seva funció. Les tasques en les que participarà seran; creixement de plantes, extracció de RNA, producció de cDNA, clonació en vectors d'expressió i transformació de plantes.

Overview (resum en anglès): The cork oak (Quercus suber) is exploited for its bark, cork, which generates wealth in rural areas of the western Mediterranean. Despite its economic value, the sector faces challenges such as the off-flavor in wine caused by 2,4,6-trichloroanisole (TCA), which originates from fungi. The Center for Research in Agricultural Genomics (CRAG) investigated this problem by studying two populations of Quercus suber: Cassà de la Selva (Spain) and Porto Torres (Italy), with differences in TCA production. The transcriptomes of these populations were sequenced to identify genes involved in the production of this compound, and differences were observed in the structure of the cork cell walls, suggesting variability in the mobility of the fungi responsible for TCA.

Within the cellular part, the study focuses on suberin and pectins. This Bachelor's Thesis aimed to study the enzymes involved in the synthesis and degradation of these compounds. These enzymes are pectinesterases, in the case of pectins, and gelp72, for suberin. In this work, the first step was to clone the promoter region of the gelp72 gene and the complementary DNA (cDNA) of the PE57 pectinesterase of Quercus suber. The goal was to study the sequence and functionality of the gelp72 gene promoter region with a special focus on the factors that regulate its expression in response to auxin treatments and its involvement in suberin degradation. Secondly, the pectinesterases present in the cork of these two populations were compared using transcriptomic and biochemical analysis techniques to identify differences in the expression and functionality of these enzymes and how they might influence the quality and durability of the cork.

The results of this work concluded that the methodology used was not effective in amplifying the promoter sequence of the gelp72 gene, prompting the planning of whole-genome DNA sequencing of the two populations. On the other hand, a pectinesterase related to the cell wall and TCA accumulation was successfully amplified, with successful transformation of Agrobacterium tumefaciens strains. These results will facilitate the subsequent transformation of Arabidopsis thaliana plants as a model to study the function of these proteins, with the ultimate goal of studying the role of these pectinesterases in the cell wall.

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