Identificación y caracterización de genes inducidos por exceso de Boro en Chenopodium quinoa
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Archivos
Fecha
2015
Profesor/a Guía
Facultad/escuela
Idioma
es
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Universidad Andrés Bello
Nombre de Curso
Licencia CC
Licencia CC
Resumen
Quínoa o Chenopodium quina es una planta nativa del altiplano andino que se adapta a las
condiciones extremas de su entorno como a altos contenidos de boro en el suelo, de hecho
se determinó previamente que el ecotipo R-49 presenta tolerancia a altas concentraciones
de Boro. El boro es un elemento esencial para las plantas por formar parte de la pared de las
células vegetales, pero su exceso produce toxicidad. Existen mecanismos descritos por los
cuales algunas especies vegetales poseen tolerancia al exceso de boro en su medio de
crecimiento, entre los que destacan la variación en la expresión de algunos transportadores
de boro como NIP5;1, BOR1 y BOR4.
A través de técnicas de secuenciación masiva como RNA-Seq se han identificado
transcritos que caracterizan la respuesta a distintos estrés abióticos. Por esta razón, se
utilizó esta técnica para estudiar la respuesta de quínoa frente a una situación de exceso de
boro con el objetivo de identificar posibles transcritos asociados a la tolerancia de esta
planta. A partir del estudio de los transcritos que más variaron su expresión en el RNA-Seq,
se identificaron una serie de posibles proteínas asociadas a respuesta a estrés. Destacó el
componente c186654 gl, el cual codifica para una proteína putativa denominada CgREB.
A través de análisis in silico se identificaron entre 2 a 3 dominios transmembrana y posibles
modificaciones post-traduccionales dentro de las que destaca la miristoilación. Por lo que
se propone que CgREB podría formar parte de la cadena de transducción de señales en
respuesta a un exceso de boro. Por otro lado, a través del estudio de los transportadores de
boro en el transcriptoma de referencia construido a través del RNA-Seq, se identificó el
componente c171835_gl_i2 como un posible ortólogo de NIP5;1 en quínoa (CgNIP5;1), debido a que presenta las características aminoacídicas que posee este canal en otros
organismos vegetales. Por último, se confirmó por PCR en tiempo real el aumento de la
expresión relativa frente a un exceso de boro de los componentes que codifican para
CgREB y CgNIP5;1. Se sugiere que CgNIP5;1 participaría en la tolerancia de
Chenopodium quina a las altas concentraciones de ácido bórico.
Chenopodium quinoa or quinoa is a plant native from Andean highlands and is adapted to the extreme conditions of their environment; in fact previously in the laboratory it has been determinate that the R-49 ecotype has more tolerance to boron stress. Boron is essential for plants because is part of the plant cell wall, but high concentrations are toxic. Mechanisms described by some plant tolerance to treatments with excess of boron has been related to the boron transporters, such as NIP5;1, BOR and BOR4. Using massive sequencing techniques such as RNA-Seq has made the identification of transcripts that characterize the response to some abiotic stress, for this reason this technique was used to study the response of quinoa in excess of boron because we want to identify potential transcripts related in tolerance of this plant. From the study of the components that alter their expression in RNA-Seq, we identified different putative proteins related with stress response in plants. One of the highlights was the component c186654 g1 which encodes a putative protein called CgREB. In silico analysis have been proposed that this protein have between 2 to 3 transmembrane domains and could be post translationally modified for myristoylations. We propose that it may form part of a signal transduction pathway in response to an excess of boron. Furthermore, we identified B transporters, one of them encoded the component c171835~g1_i2 a possible ortholog of N1P5;1 in quinoa because it has all structural features presented in that channel contains in other plants. Finally, we confirmed by real-time PCR the increase of the relative expression predicted by the results of RNA-Seq for the component encoding CgREB and CgNIP5; 1 in high boron stress. The identified ortholog of NIP5;1, CgNIP5;1, would play a role in high B tolerance in Chenopodium quinoa.
Chenopodium quinoa or quinoa is a plant native from Andean highlands and is adapted to the extreme conditions of their environment; in fact previously in the laboratory it has been determinate that the R-49 ecotype has more tolerance to boron stress. Boron is essential for plants because is part of the plant cell wall, but high concentrations are toxic. Mechanisms described by some plant tolerance to treatments with excess of boron has been related to the boron transporters, such as NIP5;1, BOR and BOR4. Using massive sequencing techniques such as RNA-Seq has made the identification of transcripts that characterize the response to some abiotic stress, for this reason this technique was used to study the response of quinoa in excess of boron because we want to identify potential transcripts related in tolerance of this plant. From the study of the components that alter their expression in RNA-Seq, we identified different putative proteins related with stress response in plants. One of the highlights was the component c186654 g1 which encodes a putative protein called CgREB. In silico analysis have been proposed that this protein have between 2 to 3 transmembrane domains and could be post translationally modified for myristoylations. We propose that it may form part of a signal transduction pathway in response to an excess of boron. Furthermore, we identified B transporters, one of them encoded the component c171835~g1_i2 a possible ortholog of N1P5;1 in quinoa because it has all structural features presented in that channel contains in other plants. Finally, we confirmed by real-time PCR the increase of the relative expression predicted by the results of RNA-Seq for the component encoding CgREB and CgNIP5; 1 in high boron stress. The identified ortholog of NIP5;1, CgNIP5;1, would play a role in high B tolerance in Chenopodium quinoa.
Notas
Tesis (Bioquímica, Magíster en Bioquímica)
Esta tesis se realizó en el Laboratorio de Biotecnología Forestal del Centro de Biotecnología Vegetal de la Universidad Andrés Bello fmanciada por el proyecto Basal PFB-16.
Esta tesis se realizó en el Laboratorio de Biotecnología Forestal del Centro de Biotecnología Vegetal de la Universidad Andrés Bello fmanciada por el proyecto Basal PFB-16.
Palabras clave
Genes, Boro, Quinoa