{"id":97412,"date":"2018-03-11T10:17:55","date_gmt":"2018-03-11T10:17:55","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/estudio-de-los-transportadores-de-sodio-y-potasio-implicados-en-la-tolerancia-a-la-salinidad-de-physcomitrella-patens\/"},"modified":"2018-03-11T10:17:55","modified_gmt":"2018-03-11T10:17:55","slug":"estudio-de-los-transportadores-de-sodio-y-potasio-implicados-en-la-tolerancia-a-la-salinidad-de-physcomitrella-patens","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/agronomia\/estudio-de-los-transportadores-de-sodio-y-potasio-implicados-en-la-tolerancia-a-la-salinidad-de-physcomitrella-patens\/","title":{"rendered":"Estudio de los transportadores de sodio y potasio implicados en la tolerancia a la salinidad de physcomitrella patens"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Ana Fraile Escanciano <\/strong><\/h2>\n<p>Resumen la mayor\u00eda de las plantas de cultivo son sensibles a la sal y se estima que en el a\u00f1o 2050 m\u00e1s del 50% de las tierras cultivables sufrir\u00e1n problemas de salinizaci\u00f3n. Con estas consideraciones, se hace indispensable incrementar la tolerancia a la salinidad de los distintos cultivos para poder satisfacer la necesidad de producci\u00f3n de alimentos para una poblaci\u00f3n mundial que alcanzar\u00e1 los 9.000 millones de habitantes.  en este trabajo nos hemos propuesto esclarecer algunos de los mecanismos implicados en la tolerancia a la salinidad de las plantas. Para ello hemos elegido como  planta modelo una briofita que ha resultado ser muy tolerante a la salinidad y se llama physcomitrella patens. Physcomitrella es capaz de soportar concentraciones de na+ que superan la concentraci\u00f3n del agua del mar. Es una planta de estructura sencilla, no vascular, con una fase dominante haploide, que se puede mantener en condiciones ax\u00e9nicas en el laboratorio. Adem\u00e1s, este musgo posee una alta frecuencia de recombinaci\u00f3n hom\u00f3loga lo que permite obtener mutantes por reemplazamiento g\u00e9nico con relativa facilidad. Todo ello, le convierte en un buen modelo para el estudio de la homeostasis i\u00f3nica a nivel de planta y a nivel celular. En esta tesis se han recogido y puesto a punto varios protocolos para la manipulaci\u00f3n in vitro, transformaci\u00f3n y estudio de los flujos de iones en physcomitrella.   en el presente trabajo nos hemos centrado en el estudio de los principales sistemas de salida de na+ de physcomitrella. La homeostasis del sodio resulta de especial inter\u00e9s en physcomitrella ya que es la \u00fanica planta conocida hasta el momento en la que coexisten antiportadores na+\/h+ tipo sos1 (ppsos1) y las bombas de na+ tipo ena (ppena1, ppena2), hasta ahora s\u00f3lo identificadas en hongos y no en plantas vasculares. Recientemente, con la secuenciaci\u00f3n del genoma completo de physcomitrella por el doe joint genome institute, hemos podido identificar otra nueva bomba tipo ena que hemos llamado ppena3 y un segundo gen sos1 que hemos llamado ppsos1b.  para el estudio y la caracterizaci\u00f3n de todos estos transportadores hemos utilizado distintos sistemas de expresi\u00f3n heter\u00f3logos que han sido utilizados tradicionalmente como son los mutantes de levaduras, de bacterias y de arabidopsis. Sin embargo lo m\u00e1s novedoso de este trabajo ha sido la caracterizaci\u00f3n de estos transportadores in planta mediante la generaci\u00f3n de mutantes de physcomitrella defectivos en sus bombas de na+ ppena1 y\/o en su antiportador na+\/h+ ppsos1. La generaci\u00f3n de los mutantes ppena1, ppena1ppena2, ppsos1 y ppena1ppsos1 y su caracterizaci\u00f3n mediante ensayos de flujos de entrada y salida de na+ nos ha permitido demostrar que ppena1 es el principal sistema de salida de na+ a ph alcalino y que ppsos1 es el principal sistema de salida de na+ a ph \u00e1cido. Por otro lado, es la primera vez que se demuestra en plantas que la eliminaci\u00f3n de un transportador sos1 conlleva la desaparici\u00f3n de la salida de na+ (como ocurre en los mutantes ppsos1 y ppena1ppsos1). Adem\u00e1s hemos podido observar que en estos mutantes la exposici\u00f3n al na+ induce muerte celular y que, independientemente de la presencia de na+, tienen afectado el transporte de k+ de alta afinidad. Pero sin duda, una de las mayores aportaciones de este trabajo ha sido la generaci\u00f3n de estos mutantes que podr\u00e1n convertirse en una nueva herramienta muy \u00fatil para el estudio y la caracterizaci\u00f3n de otros transportadores de na+ de plantas.  abstract the majority of crop plants are sensitive to salinity and it is expected that by year 2050 more than 50% of arable land will suffer from salinization problems. Considering this, it is essential to increase the salinity tolerance of different crops in order to satisfy the food production demands of a world-wide population that will reach 9,000 million people. in this work, we aim to clarify some of the mechanisms involved in salinity tolerance in plants. We have chosen as a model plant the bryophyte physcomitrella patens that was proven to be very tolerant to salinity. Physcomitrella is able to tolerate concentrations of na+ surpassing that of sea water. It is a nonvascular plant of simple structure with a dominant haploid phase that can be maintained under axenic conditions in the laboratory. In addition, this moss has a high frequency of homologous recombination thus allowing a rather simple generation of knock-out mutants. In this thesis, we have gathered and developed several protocols for the in vitro manipulation, transformation and study of the ion fluxes in physcomitrella. All this characteristics make from it a good model for the study of ionic homeostasis at both cellular and plant level. plant cells eliminate excess na+ by extruding it outside the cytosol or by sequestering it inside the vacuole. In the present work, we have focused in studying the main systems of na+ extrusion in physcomitrella. Sodium homeostasis in physcomitrella is of special interest since it is the only plant characterized until the moment in which na+\/h+ antiporters type sos1 (ppsos1) and na+ pumps type ena (ppena1, ppena2) coexist which, until the present time, were only identified in fungi but not in vascular plants. Recently, with the sequencing of the complete genome of physcomitrella by the doe joint genome institute, we were able to identify another new na+ pump type ena, that we called ppena3 and also a second sos1 gene, that we called ppsos1b.  for the study and characterization of these transporters we have used various routinely implemented systems of heterologous expression such as mutants of yeast, bacteria and arabidopsis. Nevertheless, the most novel part of this work is the characterization of these transporters in planta by generating physcomitrella mutants defective in na+ pump ppena1 and\/or na+\/h+ antiporter ppsos1. The generation and characterization of the mutants ppena1, ppena1ppena2, ppsos1 and ppena1ppsos1 by means of na+ efflux and influx assays allowed us to demonstrate that ppena1 is the main system of na+ efflux at alkaline ph while ppsos1 is the main system of na+ efflux at acidic ph. On the other hand, we have demonstrated for the first time in planta that the elimination of sos1 transporter leads to the impairment of na+ efflux (as in the case of the mutants ppsos1 and ppena1ppsos1). Moreover, we have observed that exposure of these mutants to na+ induces cell death as well as affecting their high affinity k+ transport independent of the presence or absence of na+. Indeed, one of the major contributions of this work has been the generation of these mutants that may become a new and very useful tool for the study and characterization of other na+ transporters in plants.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Estudio de los transportadores de sodio y potasio implicados en la tolerancia a la salinidad de physcomitrella patens<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Estudio de los transportadores de sodio y potasio implicados en la tolerancia a la salinidad de physcomitrella patens <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Ana Fraile Escanciano <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Polit\u00e9cnica de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 24\/11\/2009<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li>Bego\u00f1a Benito Casado<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: alonso Rodriguez navarro <\/li>\n<li>Francisco Rubio mu\u00f1oz (vocal)<\/li>\n<li>ram\u00f3n Serrano salom (vocal)<\/li>\n<li>Francisco Javier Quintero toscano (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Ana Fraile Escanciano Resumen la mayor\u00eda de las plantas de cultivo son sensibles a la sal y 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