Calle Calderón, Alejandro
Wunsch Blanco, Ana (dir.)
Universidad de Zaragoza,
2019
Abstract: La cereza (Prunus avium L.) es una fruta muy apreciada, con un alto valor nutricional y de interés económico, cuyo cultivo ha aumentado durante los últimos años. Para poder proporcionar nuevos cultivares adaptados a las demandas de los productores y consumidores, y que puedan hacer frente a desafíos como el calentamiento global, es necesario desarrollar herramientas que puedan ayudar a optimizar el proceso de mejora. Con este propósito, el objetivo de este trabajo es investigar la genética de algunos caracteres fenológicos y de calidad de fruto de interés en cerezo y avanzar en el conocimiento de los mecanismos biológicos que los regulan. Para alcanzar este objetivo se han utilizado siete familias intra-específicas de cerezo que derivan de polinizaciones cruzadas y autopolinizaciones de material vegetal local (‘Cristobalina’ y ‘Ambrunés’) que proporcionan variabilidad fenotípica adicional para éstos caracteres. Estas familias se genotiparon utilizando las plataformas genómicas ‘RosBREED cherry 6K y/o 15 SNP Illumina® Infinium’, lo que permitió desarrollar cinco mapas genéticos de alta densidad, que fueron utilizados para analizar la sintenia entre cerezo y melocotonero (P. persica), y para análisis de QTLs. Las siete familias fueron además fenotipadas para caracteres fenológicos (fecha de floración, período de desarrollo de fruto y fecha de maduración), y caracteres de calidad del fruto [firmeza, tamaño, color, contenido en sólidos solubles, acidez titulable y contenido en polifenoles (antocianinas y ácidos fenólicos)]. Estos datos se utilizaron para realizar análisis anuales y/o multianuales de QTLs utilizando diferentes estrategias de mapeo, el análisis de familias únicas se realizó utilizando MapQTL®, y el análisis de varias familias de manera combinada se realizó utilizando FlexQTL™. Estos análisis permitieron identificar QTLs principales y menores para todos los caracteres investigados, validar algunos QTLs previamente descritos para los mismos caracteres, identificar nuevos QTLs para caracteres estudiados por primera vez, e identificar nuevas variantes de QTLs de interés para la mejora. Se identificaron QTLs principales en los grupos de ligamientos (GL) 1 y 2 para fecha de floración, en el GL4 para el período de desarrollo de fruto y fecha de maduración, en los GL 1 y 2 para tamaño de fruto, en los GL 1 y 4 para firmeza, en el GL4 para el contenido en sólidos solubles, en GL6 para acidez titulable, en GL3 para color de fruto y contenido en antocianinas, y en GL1 para el contenido en ácidos fenólicos. Haplotipos de interés para la mejora en estos QTLs principales fueron identificados en algunos cultivares, como en los GL 1 y 2 de ‘Cristobalina’ para bajos requerimientos de frío y floración temprana, en el GL4 de ‘Cristobalina’ y ‘Burlat’ para período de desarrollo de fruto corto y fecha de maduración temprana, y en el GL1 de ‘Ambrunés’ para tamaño y firmeza. El período de desarrollo de fruto destacó como un carácter esencial para la fenología y la calidad del fruto, ya que la fecha de maduración, la firmeza y el contenido de sólidos solubles están correlacionados con el desarrollo de fruto y se asociaron a los mismos QTLs. La identificación de QTLs principales permitió proponer y confirmar genes candidatos para estos caracteres en estos QTL, y los genes candidatos para fecha de floración fueron investigados. Utilizando la secuencia del genoma de ‘Regina’, genes candidatos para requerimientos de frío y fecha de floración, PavDAM, ortólogos a los genes DAM de P. persica y P. mume, fueron identificados y caracterizados en el QTL principal de fecha de floración en GL1. La comparación de secuencias de los genes PavDAM de varios cultivares de cerezo con diferentes requerimientos térmicos y fechas de floración, permitió detectar polimorfismos que pueden estar asociados a las diferencias fenotipicas, y una deleción en el promotor de los PavDAM (DPD) en ‘Cristobalina’ que está asociado a floración temprana. También se desarrolló un marcador de ADN para esta mutación, que puede ser usado para la selección asistida por marcadores de floración temprana en ‘Cristobalina’.
Abstract (other lang.): Sweet cherry (Prunus avium L.) is a very appreciated fruit, with a high nutritional value and economic interest, which cultivation has increased during last years. In order to provide new cultivars adapted to producer and consumer demands, and that confront challenges like global warming, is necessary to develop tools that can help optimizing the breeding process. For this purpose, the objective of this work is to investigate the genetics of some relevant phenology and fruit quality traits of sweet cherry and to advance in the understanding of the biological mechanisms that regulate them. To achieve this goal, seven sweet cherry intraspecific populations that derive from cross- and self-pollinations of local plant material (‘Cristobalina’ and ‘Ambrunés’), and that provide additional phenotypic variation for these traits, were used. These populations were genotyped with the whole genome RosBREED cherry 6K and/or 15K Illumina® Infinium SNP arrays, which allowed developing five high-density genetic maps that were then used for analyzing synteny between sweet cherry and peach (P. persica) and for QTL analyses. The seven populations were also phenotyped for phenology traits (bloom time, fruit development period and maturity date), and for fruit quality traits [firmness, size, color, solid soluble content, titratable acidity and polyphenols (anthocyanins and phenolic acids) content]. These data were used to perform single and/or multi-year QTL analysis, using different mapping strategies, which included single bi-parental analysis with MapQTL®, or combined multi-parental populations using FlexQTL™. These analyses allowed identifying major and minor QTLs for all the traits investigated, validating some QTLs previously reported for the same traits, reporting new QTLs for newly investigated traits, and identifying new QTLs variants of breeding interest. Major QTLs were identified on linkage groups (LGs) 1 and 2 for bloom time, on LG4 for fruit development period and maturity date, on LGs 1 and 2 for fruit size, on LGs 1 and 4 for fruit firmness, on LG4 for solid soluble content, on LG6 for titratable acidity, on LG3 for fruit color and anthocyanins content, and on LG1 for phenolic acids content. Relevant QTL haplotypes for breeding purposes were identified in these major QTLs in some cultivars, like for low chilling and early blooming in ‘Cristobalina’ LGs 1 and 2, for short development period and early maturity date in ‘Cristobalina’ and ‘Burlat’ LG4, and in ‘Ambrunés’ LG1 for size and firmness. Fruit development period reveal itself as an essential trait for phenology and fruit quality, as maturity date, firmness and soluble solids contents were correlated with fruit development and were associated to the same QTLs. The identification of major QTLs allowed proposing and confirming candidate genes for these traits at these QTLs, and candidate genes for bloom time were investigated. Using the ‘Regina’ sweet cherry genome sequence, candidate genes for chilling requirements and bloom time, PavDAM genes, orthologous to P. persica and P. mume DAM genes, were identified and characterized in the major bloom time QTL on LG1. Sequence comparison of PavDAM genes of various sweet cherry cultivars with different chilling requirements and bloom times allowed detecting sequence polymorphisms that may be associated to their phenotypic differences, and a deletion in the ‘Cristobalina’ PavDAM promoter (DPD) that is associated to early blooming. A DNA marker for this mutation, that can be used for marker-assisted selection of early blooming from ‘Cristobalina’, was also developed.
Abstract (other lang.): Sweet cherry (Prunus avium L.) is a very appreciated fruit, with a high nutritional value and economic interest, which cultivation has increased during last years. In order to provide new cultivars adapted to producer and consumer demands, and that confront challenges like global warming, is necessary to develop tools that can help optimizing the breeding process. For this purpose, the objective of this work is to investigate the genetics of some relevant phenology and fruit quality traits of sweet cherry and to advance in the understanding of the biological mechanisms that regulate them. To achieve this goal, seven sweet cherry intraspecific populations that derive from cross- and self-pollinations of local plant material (‘Cristobalina’ and ‘Ambrunés’), and that provide additional phenotypic variation for these traits, were used. These populations were genotyped with the whole genome RosBREED cherry 6K and/or 15K Illumina® Infinium SNP arrays, which allowed developing five high-density genetic maps that were then used for analyzing synteny between sweet cherry and peach (P. persica) and for QTL analyses. The seven populations were also phenotyped for phenology traits (bloom time, fruit development period and maturity date), and for fruit quality traits [firmness, size, color, solid soluble content, titratable acidity and polyphenols (anthocyanins and phenolic acids) content]. These data were used to perform single and/or multi-year QTL analysis, using different mapping strategies, which included single bi-parental analysis with MapQTL®, or combined multi-parental populations using FlexQTL™. These analyses allowed identifying major and minor QTLs for all the traits investigated, validating some QTLs previously reported for the same traits, reporting new QTLs for newly investigated traits, and identifying new QTLs variants of breeding interest. Major QTLs were identified on linkage groups (LGs) 1 and 2 for bloom time, on LG4 for fruit development period and maturity date, on LGs 1 and 2 for fruit size, on LGs 1 and 4 for fruit firmness, on LG4 for solid soluble content, on LG6 for titratable acidity, on LG3 for fruit color and anthocyanins content, and on LG1 for phenolic acids content. Relevant QTL haplotypes for breeding purposes were identified in these major QTLs in some cultivars, like for low chilling and early blooming in ‘Cristobalina’ LGs 1 and 2, for short development period and early maturity date in ‘Cristobalina’ and ‘Burlat’ LG4, and in ‘Ambrunés’ LG1 for size and firmness. Fruit development period reveal itself as an essential trait for phenology and fruit quality, as maturity date, firmness and soluble solids contents were correlated with fruit development and were associated to the same QTLs. The identification of major QTLs allowed proposing and confirming candidate genes for these traits at these QTLs, and candidate genes for bloom time were investigated. Using the ‘Regina’ sweet cherry genome sequence, candidate genes for chilling requirements and bloom time, PavDAM genes, orthologous to P. persica and P. mume DAM genes, were identified and characterized in the major bloom time QTL on LG1. Sequence comparison of PavDAM genes of various sweet cherry cultivars with different chilling requirements and bloom times allowed detecting sequence polymorphisms that may be associated to their phenotypic differences, and a deletion in the ‘Cristobalina’ PavDAM promoter (DPD) that is associated to early blooming. A DNA marker for this mutation, that can be used for marker-assisted selection of early blooming from ‘Cristobalina’, was also developed.
Pal. clave: bioquimica
Titulación: Programa de Doctorado en Bioquímica y Biología Molecular
Plan(es): Plan 485
Department: Bioquímica y Biología Molecular y Celular
Nota: Presentado: 29 11 2019
Nota: Tesis-Univ. Zaragoza, Bioquímica y Biología Molecular y Celular, 2019
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