Análisis transcriptómico dual, de la interacción entre la mutante Δbcfet1 de Botrytis cinerea durante el proceso infectivo de Arabidopsis thaliana, en condiciones contrastantes de hierro
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2022
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es
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Universidad Andrés Bello
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Licencia CC
Resumen
Botrytis cinerea es un reconocido hongo fitopatógeno caracterizado por
provocar grandes pérdidas en la agricultura a nivel mundial y por tener un gran
número de hospederos. Es un patógeno capaz de colonizar distintos órganos
vegetales, produciendo necrosis del tejido con el objetivo de obtener nutrientes a
partir de células muertas. En este contexto, los nutrientes disponibles en el encuentro
entre B. cinerea y su hospedero modulan el comportamiento de ambos organismos
durante la interacción. El hierro, un mineral fundamental para procesos metabólicos
y celulares, además de jugar un papel fundamental en la virulencia de los patógenos,
tiene también un rol importante en las respuestas de la planta frente a una infección.
Investigaciones recientes han demostrado que B. cinerea requiere de un sistema de
captación de hierro de alta afinidad denominado RIA (del inglés, Reductive Iron
Assimilation), el cual juega un papel en el proceso infectivo. Este sistema se compone
de una ferroxidasa de membrana (BcFET1) y una permeasa de hierro BcFTR1.
Mediante la generación de una mutante de deleción para la ferroxidasa (Δbcfet1), se
evidenció un fenotipo hipervirulento dependiente de la disponibilidad de hierro en la
planta, un resultado inesperado debido principalmente a que antecedentes
demostraban que la interrupción en componentes de RIA disminuye la virulencia en
otros patógenos. Para poder entender más a fondo el comportamiento de la mutante
anteriormente mencionada es que, en este trabajo, se realizó un análisis
transcripcional mediante RNA-seq dual para evaluar el comportamiento
transcripcional de la mutante durante la infección de A. thaliana en condiciones
contrastantes de disponibilidad de hierro. Los resultados evidencian que, a pesar de
la deleción del gen bcfet1, en ausencia de hierro, la mutante induce genes
relacionados al mantenimiento (bcfem1) y absorción transmembrana de este metal
(bcftr1), ambos regulados por un mismo factor de transcripción. Al mismo tiempo, A.
thaliana presenta un perfil de expresión diferencial asociado a una respuesta de
defensa hipersensible, lo que podría estar explicando el mayor daño que se provoca
en el tejido vegetal al ser infectado por la mutante de B. cinerea. Si bien este estudio
dilucida el comportamiento transcriptómico que ocurre en la interacción entre la
mutante Δbcfet1 y A. thaliana, los mecanismos regulatorios y cómo estos se ven
afectados por la disponibilidad de hierro, aún no están totalmente claros, por lo cual
es de gran interés seguir identificando y caracterizando la regulación de los
mecanismos de captación de hierro y cómo estos se relacionan a la virulencia de los
patógenos
Botrytis cinerea is a recognized phytopathogenic fungus characterized by causing great economic losses in agriculture worldwide and by having a large number of hosts. It is a pathogen capable of colonizing in different climatic conditions, producing necrosis of plant tissue, thus obtaining the nutrients required for its subsistence from dead cells. In this context, the nutrients available in the encounter between B. cinerea and its host modulate the behavior of both organisms during the interaction. Iron, a fundamental mineral for metabolic and cellular processes, in addition to playing a fundamental role in the virulence of pathogens, also has an important role in plant responses to infection. Recent research has shown that B. cinerea requires a high-affinity iron uptake system called RIA (Reductive Iron Assimilation), which also plays a role in its infective process. This system is composed of a membrane ferroxidase (bcfet1) and an iron permease bcftr1. Through the generation of a deletion mutant for the Δbcfet1 ferroxidase, a hypervirulent phenotype dependent on the availability of iron in the plant was evidenced, an unexpected result due to previous evidence that the interruption in RIA components decreased virulence in other pathogens. In order to fully understand the behavior of the aforementioned mutant, through this work a transcriptional analysis was performed using dual RNA seq to evaluate the molecular behavior of the mutant during A. thaliana infection under contrasting conditions. iron availability. The results show that, despite the deletion of the bcfet1 gene, in the absence of iron, the mutant induces genes related to the maintenance (bcfem1) and transmembrane absorption of this metal (bcftr1), both regulated by the same transcription factor. At the same time, A. thaliana presents a differential expression profile associated with a hypersensitive defense response, which could explain the greater damage caused to plant tissue when reduced by the B. cinerea mutant. Although this study elucidates the transcriptomic behavior that occurs in the interaction between the Δbcfet1 mutant and A. thaliana, the regulatory mechanisms and how they are affected by iron availability are still not completely clear, which is why it is of great interest. continue to identify and characterize the regulation of iron uptake mechanisms and how these are related to the virulence of pathogens
Botrytis cinerea is a recognized phytopathogenic fungus characterized by causing great economic losses in agriculture worldwide and by having a large number of hosts. It is a pathogen capable of colonizing in different climatic conditions, producing necrosis of plant tissue, thus obtaining the nutrients required for its subsistence from dead cells. In this context, the nutrients available in the encounter between B. cinerea and its host modulate the behavior of both organisms during the interaction. Iron, a fundamental mineral for metabolic and cellular processes, in addition to playing a fundamental role in the virulence of pathogens, also has an important role in plant responses to infection. Recent research has shown that B. cinerea requires a high-affinity iron uptake system called RIA (Reductive Iron Assimilation), which also plays a role in its infective process. This system is composed of a membrane ferroxidase (bcfet1) and an iron permease bcftr1. Through the generation of a deletion mutant for the Δbcfet1 ferroxidase, a hypervirulent phenotype dependent on the availability of iron in the plant was evidenced, an unexpected result due to previous evidence that the interruption in RIA components decreased virulence in other pathogens. In order to fully understand the behavior of the aforementioned mutant, through this work a transcriptional analysis was performed using dual RNA seq to evaluate the molecular behavior of the mutant during A. thaliana infection under contrasting conditions. iron availability. The results show that, despite the deletion of the bcfet1 gene, in the absence of iron, the mutant induces genes related to the maintenance (bcfem1) and transmembrane absorption of this metal (bcftr1), both regulated by the same transcription factor. At the same time, A. thaliana presents a differential expression profile associated with a hypersensitive defense response, which could explain the greater damage caused to plant tissue when reduced by the B. cinerea mutant. Although this study elucidates the transcriptomic behavior that occurs in the interaction between the Δbcfet1 mutant and A. thaliana, the regulatory mechanisms and how they are affected by iron availability are still not completely clear, which is why it is of great interest. continue to identify and characterize the regulation of iron uptake mechanisms and how these are related to the virulence of pathogens
Notas
Tesis (Ingeniería en Biotecnología)
Tesis financiada por el proyecto FONDECYT Regular N° 1190611.
Tesis financiada por el proyecto FONDECYT Regular N° 1190611.
Palabras clave
Botrytis Cinerea, Genética, Arabidopsis Thaliana, Efecto del Hierro, Transcripción Genética