Cloning of Diplosporous Apomixis Candidate Genes for Its Functional Characterization in the Model Plant Arabidopsis thaliana

PASTEN MARÍA CIELO; BELLIDO ANDRÉS MARTÍN; GARBUS INGRID; ECHENIQUE VIVIANA

Workshop; VII Ciclo de Seminarios sobre avances en la Caracterización Genética y Molecular de la APOMIXIS; 2020

Eragrostis curvula is a perennial forage grass naturalized in semiarid regions of Argentina,with genotypes that can be either sexual diploids or polyploids (4×?8×) reproducingby diplosporous apomixis. Using different approaches, our research group obtaineda list of candidate genes related to this reproductive mode that remain to be functionallycharacterized. The aim of this work is to optimize the construction of vectors for the introduction of these Eragrostis candidate genes into the model plant Arabidopsis thaliana inorder to analyze their effects during reproductive development. Two strategies were used,Gateway and GoldenBraid, and vectors with single and multiple genes were introducedin A. thaliana plants using Agrobacterium and the floral dip method. In relation to GoldenBraid, focusing on the current knowledge about the complex dynamics that regulategermline specification in the ovule of A. thaliana, different promoters were selected, relatedto these key pathways, which will guide ectopic expression into the regions surroundingthe MMC. Currently, using these promoters in the GoldenBraid system, constructsfor each gene were generated, and vectors containing different combinations ofgenes were successfully created, using kanamycin or hygromycin for the selection of thetransgenic plants. The phenotypic effects of these genes will be analyzed in the near future,using whole mount clearing and DIC microscopy.