Título provisional de la tesis

Establishment of a DNA-free system for mutant induction in tomato

Resumen

Tomato is an important agricultural species worldwide. World production is estimated at approximately 159 million tons, while average annual consumption of fresh tomatoes is 18 kg per capita in Europe and 8 kg per capita in the USA (USDA ERS 2008). In the last few years, the consumption of tomatoes has increased considerably, as tomatoes are supplied to the market both fresh and in the form of processed products such as soups, juices, purees, and sauces. Consumption of fresh tomatoes reduces the risk of inflammation, cancer, and chronic noninfectious diseases, including cardiovascular diseases such as coronary heart disease, hypertension, diabetes, and obesity (Canene-Adams et al. 2005).

Considering the valuable biochemical composition of tomatoes, it makes sense to improve further these plants using biotechnological methods (e.g., CRISPR/Cas9-medianted genome editing approach).

CRISPR/Cas 9 genome editing is the newest approach that allows to knock down specific genes, deliver other genes or complexes into plants. Then it will be possible to obtain not only the valuable components of natural origin of these plants, but also to obtain additional recombinant proteins (for example), which also can be used for medical purposes, create resistant plants to biotic factors (viruses, bacterial and fungal pathogens, pests), to abiotic factors: salinity, drought, high/ low temperatures); to understand the functions of different genes in tomato plants.

As part of this dissertation research, a methodology will be developed thanks to which it will be possible in the future to change the genome of tomatoes directly. In the experiments will be used popular cultivars of Solanum lycopersicum and one wild species of tomato, Solanum pennellii. The target will be PHYTOENE DESATURASE (PDS) gene which is present in many organisms. PDS encodes a phytoene desaturase enzyme that converts 15-cis-phytoene into zeta-carotene. PDS-silenced plants show photobleaching phenotype in leaves and diminishing of pigment formation in fruits, so knock down of this gene will give opportunity easy visually detect the effectiveness of our procedure of genome editing. Other gene which is planned to be used is DsRed gene. DsRed is an interesting fluorescent marker protein which is easily detectable.

Director/a: JOSE MANUEL PEREZ PEREZ

codirector/a: EDUARDO LARRIBA TORNEL

Publicaciones derivadas de la tesis: Larriba, E.; Yaroshko, O.; Pérez-Pérez, J.M. Recent Advances in Tomato Gene Editing. Int. J. Mol. Sci. 2024, 25, 2606. https://doi.org/10.3390/ijms25052606

Yaroshko O, Pasternak T, Larriba E, Pérez-Pérez JM. Optimization of Callus Induction and Shoot Regeneration from Tomato Cotyledon Explants. Plants (Basel). 2023 Aug 14;12(16):2942. doi: 10.3390/plants12162942. PMID: 37631154; PMCID: PMC10459365.

Yaroshko, Olha. (2021). Achievements in Genetic Engineering of Amaranthus L. Representatives. International Journal of Secondary Metabolite. 8. 172-185. 10.21448/ijsm.925737.

Código ORCID: https://orcid.org/0000-0003-2517-4472