Aislamiento y caracterización funcional del antiporte NHX1 de Serendipita indica

Sanz García, Eugenio (2019). Aislamiento y caracterización funcional del antiporte NHX1 de Serendipita indica. Proyecto Fin de Carrera / Trabajo Fin de Grado, E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM), Madrid.


Title: Aislamiento y caracterización funcional del antiporte NHX1 de Serendipita indica
  • Sanz García, Eugenio
  • Haro Hidalgo, Rosario
Item Type: Final Project
Degree: Grado en Biotecnología
Date: June 2019
Faculty: E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM)
Department: Biotecnología - Biología Vegetal
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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The current human population is rising so that it is calculated to get to 9.5-10 billion people by year 2050, and therefore, demand for food and clean water can be expected to ascend a 50% in the next decades. In addition to the population increasing, we must regard the fact that every year lots of lands get useless for agricultural purposes due to contamination and salinization. To face this situation, actions must be taken to implement strategies that allow a higher crop production. Salinity affects negatively to plants and reduces their productivity. Saline soils alter seed germination and plant development, growth and reproduction. This is due to osmotic stress, saline stress, nutritional imbalances or a combination of those factors. The main responsible for these stresses is the cation Na+, which, at high concentration, is toxic for cells. K+ is one of the main nutrients that organisms need for correct growth and development. Physicochemical properties of Na+ are very similar to those of the K+ cation. For these reasons, cells expend energy trying to obtain a high intracellular concentration of K+ while keeping a low cytosolic Na+/K+ ratio. For that task, most living organisms use transporters to introduce K+ into cells, and other transporters to get the Na+ outward or into subcellular structures (endosomes, vacuoles). NHX is a type of Na+/H+ antiporter, usually located in the membrane of pre-vacuolar vesicles or endosomes. This work has focused in the NHX1 family, which is broadly present in many eukaryotic organisms, including fungi, plants and metazoans. Many soil microorganisms (fungi for instance) can associate with plant roots in symbiotic relationships that can enhance crops performance. The benefits of this mutualistic relationship include enhanced efficiency of nutrient uptake, drought tolerance, broader pH tolerance, and resistance to certain pathogens. Serendipita indica (formerly named Piriformospora indica) is a recently discovered endophytic fungus that has a great potential to improve crop performance when interacting with plant roots. Previous studies suggest that this mutualistic fungus could lead to growth enhancement, early flowering, and biotic and abiotic stress tolerance in its host plants. S. indica is known to be able to stablish symbiotic relations with some of the most important crops, such as rice or barley, as well as with the model plant, Arabidopsis thaliana, which facilitates the study of the symbiotic process. Arabidopsis plants inoculated with S. indica show salt tolerance but the molecular mechanisms behind it remain unknown. Recently the group in which this work was carried out, characterized two Na+ transporters of S. indica, which are candidates to mediate the low Na+ accumulation in the plant induced by the fungus. This project worked on the study and functional characterization of the Serendipita indica transporter SiNhx1. In order to accomplish such quest, the cDNA of the gene was cloned and sequenced. Then an in silico study was performed to determine the genomic structure of the gene, and the transmembrane domains and tridimensional structure of the protein. In addition, a phylogenetic analyse was made. To get to corroborate the function of SiNhx1, complementation essays were done in mutant yeasts and bacteria. Furthermore, the role of SiNhx1 in symbiosis was studied. S. indica and A. thaliana were co-cultivated in saline conditions, and in the roots inoculated the content of the cations and the expression of SiHNX1 were analyzed.

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Item ID: 57140
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Deposited by: Biblioteca ETSI Agrónomos
Deposited on: 30 Oct 2019 15:01
Last Modified: 30 Oct 2019 15:01
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