Análisis de expresión de genes de uva de mesa (Vitis vinífera L.) relacionados con la expansión de la baya en crecimiento
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2013
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es
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Universidad Andrés Bello
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Licencia CC
Licencia CC
Resumen
La uva de mesa es el cultivo frutal de mayor importancia en Chile tanto en superficie
cultivada como en retornos por exportaciones, siendo nuestro país uno de los principales
productores y exportadores mundiales. Sin embargo, su competitividad en el mercado
mundial está en riesgo por diversos factores, como el pago de derechos de propiedad
intelectual por el uso de nuevas variedades desarrolladas en el extranjero. El tamaño de la
fruta es un valioso rasgo comercial en muchas especies, siendo determinante para la
preferencia de los consumidores de uva de mesa. Este hecho determina que éste sea un rasgo
clave durante la selección de nuevas variedades. El crecimiento de la baya está determinado
tanto por factores genéticos como ambientales, sobre los cuales el desarrollo y rearreglo de
la pared celular tienen un rol fundamental. Previo a esta tesis se determinaron mediante RNAseq
qué genes se expresan diferencialmente en etapas tempranas del desarrollo de las bayas,
en segregantes obtenidos de un cruzamiento de 'Ruby Seedless' x ' Sultanina' (Rx.S),
enfocándose particularmente en aquellos genes que participan en el rearreglo de la pared
celular, y que presentarían relación al tamaño de la baya. Diversos estudios han sugerido que
las expansinas (EXP) son reguladores endógenos del crecimiento celular catalizando la
expansión de la pared sin eventos hidrolíticos o transglicolíticos asociados. Por otro lado las
xiloglucanos endotransglicosilasa/hidrolasa (XTH) llevan a cabo la transglicosilación de los
xiloglucanos, permitiendo un deslizamiento transiente entre las microfibrillas de celulosa.
Ambas enzimas participarían en el rearreglo de la pared celular. Por otra parte, el inhibidor
de la pectinmetilesterasa (PMEI) es una proteína que actuaría bloqueando el sitio activo de
la proteína PME promoviendo la rigidez de la pared celular. El presente estudio determinó el
nivel de expresión de los genes VvEXPA8-like, VvEXPA15-like, VvXTH4-like, VvPMEI4
hipotéticamente relacionados al tamaño de la baya en uva de mesa en dos fenotipos
contrastantes (baya grande y baya pequeña) mediante PCR en tiempo real (qPCR) en
segregantes de la población RxS. Todos los genes mostraron diferentes niveles de expresión,
aunque VvEXP A8-like fue el que mostró mayores diferencias entre los fenotipos de baya
grande y baya pequeña, con un comportamiento en su expresión muy similar al descrito por
la literatura, convirtiéndose en el mejor candidato para la búsqueda de posibles marcadores
de selección para este fenotipo.
Table grape is one of the most important fruit crop in Chile, both in acreage and in export returns. However, its competitiveness in the world markets is at risk due to severa} factors, such as payments for intellectual property rights for the use of new varieties developed abroad. Fruit size is a valuable commercial trait for table grape consumers, so berries of larger sizes are preferred by producers. This fact determines that this is a key feature for the selection of new varieties. Berry's growth is determined by molecular and environmental factors, in which cell wall development has a central role. Previous to this thesis, it was determined by RNA-seq genes which are differentially expressed in early stages of berry development in siblings from a crossing between 'Ruby Seedless' x 'Sultanina' (RxS), focusing particularly on those genes involved in the rearrangement of the cell wall, and would present relation to the berry size. Studies suggest that expansines (EXP) are the key endogenous regulators of cell growth by catalyzing the expansion of the wall without hydrolitics or trans-glycolytics events. Otherwise, xyloglucan endotransglycosylase-hydrolase (XTH) would carry transglycosylation of xyloglucan (XyG) out, allowing a transient slip between cellulose microfibrils. Both enzymes would participate in cell wall rearrangement. Furthermore, the inhibitor of pectinmethylesterase (PMEI), is a protein that acts by blocking the active site of the pectinmethylesterase (PME) promoting the rigidity of the cell wall. In the present study, it was determined the expression level of the genes VvEXPA8-like, VvEXPA15-like, VvXTH4-like, VvPMEI4 hypothetically related to berry size in table grapes in two contrasting phenotypes (small and large berry size) by real time PCR (qPCR) in siblings of the crossing. All genes analyzed showed differing expression levels, although VvEXPA8-like was the one that showed major differences between the phenotypes of large and small berry size, with behavior expression very similar to that described in the literature becoming in the best candidate for the search selection markers for this phenotype.
Table grape is one of the most important fruit crop in Chile, both in acreage and in export returns. However, its competitiveness in the world markets is at risk due to severa} factors, such as payments for intellectual property rights for the use of new varieties developed abroad. Fruit size is a valuable commercial trait for table grape consumers, so berries of larger sizes are preferred by producers. This fact determines that this is a key feature for the selection of new varieties. Berry's growth is determined by molecular and environmental factors, in which cell wall development has a central role. Previous to this thesis, it was determined by RNA-seq genes which are differentially expressed in early stages of berry development in siblings from a crossing between 'Ruby Seedless' x 'Sultanina' (RxS), focusing particularly on those genes involved in the rearrangement of the cell wall, and would present relation to the berry size. Studies suggest that expansines (EXP) are the key endogenous regulators of cell growth by catalyzing the expansion of the wall without hydrolitics or trans-glycolytics events. Otherwise, xyloglucan endotransglycosylase-hydrolase (XTH) would carry transglycosylation of xyloglucan (XyG) out, allowing a transient slip between cellulose microfibrils. Both enzymes would participate in cell wall rearrangement. Furthermore, the inhibitor of pectinmethylesterase (PMEI), is a protein that acts by blocking the active site of the pectinmethylesterase (PME) promoting the rigidity of the cell wall. In the present study, it was determined the expression level of the genes VvEXPA8-like, VvEXPA15-like, VvXTH4-like, VvPMEI4 hypothetically related to berry size in table grapes in two contrasting phenotypes (small and large berry size) by real time PCR (qPCR) in siblings of the crossing. All genes analyzed showed differing expression levels, although VvEXPA8-like was the one that showed major differences between the phenotypes of large and small berry size, with behavior expression very similar to that described in the literature becoming in the best candidate for the search selection markers for this phenotype.
Notas
Tesis (Magíster en Biotecnología)
Palabras clave
Uva de Mesa, Factores genéticos, Crecimiento celular, Análisis de mejoramiento, Chile