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Projecte llegit

Títol: Cell-type specificity of plant antiviral components (Especificitat de tipus cel·lulars dels components antivirals vegetals)

Estudiant que ha llegit aquest projecte:

Tutor/Cotutor: LOZANO LUIS, LIDIA

Departament: DEAB

Títol: Cell-type specificity of plant antiviral components (Especificitat de tipus cel·lulars dels components antivirals vegetals)

Data inici oferta: 14-10-2022      Data finalització oferta: 14-05-2023

Estudis d'assignació del projecte:
    GR ENG SIS BIOLÒGICS

Lloc de realització:
EEABB

Paraules clau:
Arabidopsis, especificitat de tipus cel·lular, components antivirals vegetals

Descripció del contingut i pla d'activitats:
El estudiante realizará la amplificación de regiones reguladoras
de la expresion (promotores) de genes involucrados en defensa
antiviral en Arabidopsis thaliana por PCR, su clonaje en vectores
binarios dirigiendo la expresión del gen reportero UiAd (GUS),
transformación de plantas de arabidopsis por el método de
inmersión de inflorescencias, selección de semillas portadoras de
los
transgenes y ensayos histologicos para determinar el patrón de
actividad de dichos promotores. El estudiante se familiarizará
con técnicas de uso cotidiano en laboratorios de biologia
molecular, uso de herramientas informaticas y pensamiento
critico. El estudiante participara en las reuniones de
laboratorio semanales en las que se discute tanto las estrategias
cientificas empleadas en los distintos proyectos en vgor del
laboratorio, como literatura relativa a los mismos. El estudiante
adquirirá formación en la escritura y presentación oral de
resultados de investigación.

Overview (resum en anglès): Gene silencing is a cellular mechanism by which gene expression is inhibited in eukaryotic organisms. It is present in both endogenous and exogenous control nucleic acids and is classified as transcriptional gene silencing (TGS) and posttranscriptional gene silencing (PTGS). In both cases, the main component responsible for giving the silencing machinery target specificity is the small RNA, and it can be classified into microRNA (miRNA) and small interfering RNA (siRNA). For plant viruses, the ability to access the phloem vascular system is essential in order to establish a successful infection, but this access is highly regulated. In this context, gene silencing is an important defense mechanism of plants against viruses.
Previously, in other projects at the laboratory, the specific cell types which the pathogens preferentially target for the suppression of silencing in the infection were identified. In this project, we wanted to complement the information obtained and it was planned with the objective of studying the expression of seven genes (DCL3, RDR1, RDR2, RDR6, SDN1, SGS3 and NRPD1A) of the gene silencing machinery in Arabidopsis thaliana, determining the specificity of cell types. It was done by obtaining constructions with the GreenGate method to subsequently obtain transgenic plants and thus be able to observe the activity of the promoter and the 3'UTR region of the chosen genes, regulating the expression of the GUS reporter gene.
Finally, with the study of gene expression at three timepoints (11, 13 and 31 days), positive results have been seen for 4 of the genes studied: RDR1, RDR6, NRPD1A and SDN1, where expression could be observed mainly in the vascular bundles, although expression was also seen in apical and lateral meristems, ovules and anthers. Therefore, it was confirmed that there is cell-type specificity for parts of the gene silencing machinery, mainly in the vasculature.

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