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Tesis Doctoral 2017 MEJORA GENÉTICA DE PATATA PARA COMPUESTOS BIOACTIVOS Y CAPACIDAD ANTIOXIDANTE Roberto Tierno Fernández Tesis doctoral 2017 MEJORA GENÉTICA DE PATATA PARA COMPUESTOS BIOACTIVOS Y CAPACIDAD ANTIOXIDANTE Roberto Tierno Fernández Tesis doctoral Director: Dr. D. José Ignacio Ruiz de Galarreta Tutora: Dra. Dª. Mª Teresa Lacuesta Calvo (c)2017 ROBERTO TIERNO FERNANDEZ AGRADECIMIENTOS Este trabajo no hubiera sido posible sin la participación de muchos compañeros que, en mayor o menor medida, han contribuido a la realización de esta tesis. En primer lugar, mi reconocimiento al director de tesis, José Ignacio Ruiz de Galarreta, que apostó por mí desde el primer momento y durante este tiempo, me ha dado toda la confianza, libertad y apoyo que he necesitado. A la dirección de Neiker, por haberme acogido durante estos cuatro años y al INIA, organismo que financió este proyecto y posibilitó mi formación. Al Departamento de Biología Vegetal y Ecología de la UPV-EHU, por permitirme presentar este trabajo y especialmente a la Dra. Mª Teresa Lacuesta Calvo por aceptar ser tutora de esta tesis. Quiero agradecer la paciencia y buena disposición de muchas personas sin cuyas enseñanzas y consejos este trabajo jamás hubiera podido realizarse. Entre ellas, quiero hacer una mención muy especial a Isi, Carlos Castaño, Carlos Herrán y Bego. Otras personas con las que he tenido la oportunidad de trabajar y formarme son Ainara, Carmen, Silvia, Patrick Riga, Leire, Berdaitz y Jon. Por supuesto, quiero y debo reconocer también el inestimable aporte de Néstor, Raquel López, Mikel González, Emma López de Armentia, Jon Lemos y otros camaradas becarios, cuya guía, consejo y conocimiento ha sido determinante. Tampoco me olvido de la gente de campo, particularmente Manolo y Pascual, pero también Mari Ángeles, Andoni y compañía, siempre dispuestos a echar una mano aportando una experiencia y conocimiento imprescindibles. Además, quiero expresar un agradecimiento muy especial a toda la gente que durante estos años ha pasado por el centro y con los que he tenido la oportunidad de trabajar, compartir muchos ratos y aprender: Sara, Irune, Yuri, Bea, etc. Sin embargo, siento una gratitud especial hacia mis colegas Álvaro e Iker, con los que he trabajado más estrechamente y durante más tiempo. Además de haber contribuido a este trabajo de forma decisiva, tanto directa como indirectamente, son maestros de muchas cosas y grandes personas. Quiero dar las gracias también a mis compañeros de pasillo, muchos de los cuales ya han sido mencionados, y hacer una mención muy especial a mi tocayo Roberto. A pesar de que nuestra coexistencia como vecinos de despacho ha sido relativamente breve, añoraré nuestras conversaciones en las que pude comprobar que, además de un pozo de sabiduría, es una gran persona. Por último, quiero dar las gracias al equipo de la oficina (Ana, Cristina, Deiene, Yolanda, Iratxe y Ernesto), por su paciencia y amabilidad. A Leire, Mónica, Ana, Iratxe, Sonia, Enrique y demás gente de biotecnología, por encontrar siempre un momento para echarme una mano y resolver mis dudas. A Raquel, María Eugenia, Ion, Imanol y demás miembros del departamento de sanidad vegetal, siempre dispuestos y con buenas palabras. Al personal de mantenimiento, José Ignacio, Carlos y Kike, por estar ahí siempre que tuve algún problema técnico. A Iranzu y Alex, integrantes del equipo de marketing y comunicación, con quienes he tenido la oportunidad de trabajar codo con codo en la organización del Congreso de Mejora de Plantas 2016, pasando muy buenos ratos y aprendiendo a valorar la enorme tarea que supone conseguir que todo salga bien. También a Ana Carrasco, que me desveló el mundo del cultivo sin suelo y, por supuesto, a Maite, siempre atenta y con una sonrisa. Por último, dar las gracias a mi familia y amigos. Sólo ellos son siempre realmente imprescindibles. ABREVIATURAS Y ACRÓNIMOS ADN: ácido desoxirribonucleico AFLP: Amplified Fragment Length Polymorphism AKT: Arabidopsis K+ transporter (Transportador de K+ de Arabidopsis) AMMI: Additive Main Effects and Multiplicative Interactions AMV: Alfalfa Mosaic Virus AN1: Anthocyanin 1 AOAC: Association of Analytical Comunities (Asociación de Comunidades Analíticas) APLV: Andean Potato Latent Virus APMV: Andean Potato Mottle Virus ANS: Anthocyanin synthase (Antocianina sintasa) bch: ß-carotene 3-hydroxylase (ß-carotene 3-hidroxilasa) BCTV: Beet Curly Top Virus bHLH: Basic helix-loop-helix BLUP: Best Linear Unbiased Predictor (Mejor Predictor Linear Insesgado) ºC: grados Celsius C3H: p-coumarate 3-hydroxylase (p-coumarato 3-hidroxilasa) C4H: Cinnamate 4-hydroxilase (Cinnamato 4-hidroxilasa) Ca: Calcio CCD: Carotenoid cleavage dioxygenases CCX: Cation exchanger (Intercambiador de cationes) CDF: Cation diffusion facilitator (Facilitador de la difusión de cationes) CG1: First Clonal Generation (Primera Generación Clonal) CG2: Second Clonal Generation (Segunda Generación Clonal) CG3: Third Clonal Generation (Tercera Generación Clonal) CHI: Chalcone isomerase (Chalcona isomerasa) CHS: Chalcone synthase (Chalcona sintasa) + + CHX: Cation/H exchanger (Intercambiador de cationes/H ) Chy: ß-carotene hydroxylase (ß-caroteno hidroxilasa) CIP: Centro Internacional de la Papa CL1: Clon de primer año CL2: Clon de segundo año CL3: Clon de tercer año cm: centímetro CMV: Cucumber Mosaic Virus CNGC: Cyclic-nucleotide gated channel CrtB: Phytoene synthase (Fitoeno sintasa) CrtI: Phytoene desaturase (Fitoeno desaturasa) CrtY: Lycopene β-ciclase (Licopeno β-ciclasa) CRTISO: Caroteno cis-trans isomerasa (Carotene cis-trans isomerase) Cu: Cobre cvs: cultivares DArT: Diversity Arrays Technology DFR: Dihydroflavonol 4-reductase (dihidroflavonol 4-reductasa) DXS: 1-deoxy-d-xylulose-5-phosphate synthase (1-deoxi-D-xilulosa 5-fosfato sintasa) EBN: Endosperm Balance Number (Número de Balance del Endospermo) EBV: Expected Breeding Value (Valor de Mejora Esperado) EDTA: Ácido etilendiaminotetraacético EFSA: European Food Safety Authority (Autoridad Europea de Seguridad Alimentaria) ELISA: Enzyme-Linked Immuno Sorbent Assay (Ensayo por Inmunoabsorción Ligado a Enzimas) F1: Primera generación filial de la descendencia de dos parentales diferentes F2: Segunda generación filial procedente de la F1 i Abreviaturas y acrónimos F3H: Flavanone 3-hydroxylase (Flavanona 3-hidroxilasa) FAO: Organización para la Agricultura y la Alimentación Fe: Hierro FPN: Ferroportin (Ferroportina) FRO: Ferric reductase-oxidase g: gramos GCA: General Combining Ability (Aptitud Combinatoria General) GEI: Genotype ∙ Environment Interactions (Interacciones Genotipo ∙ Ambiente) H: Hidrógeno H2: Broad- sense heritability (Heredabilidad en sentido amplio) h2: Narrow-sense heritability (Heredabilidad en sentido estricto) ha: hectárea HAK: High affinity K+ transporter (Transportador de K+ de alta afinidad) HKT: High-affinity K+ transporter (Transportador de K+ de alta afinidad) HPTLC: High Performance Thin Layer Chromatography (Cromatografía en Capa Fina de Alta Resolución) HCT: Hydroxycinnamoyl-CoA: shikimate hydroxycinnamoyl transferase (Hidroxicinamoil-CoA: shikimato hidroxicinamoil transferasa) HQT: Hydroxycinnamoyl-CoA: quinate hydroxycinnamoyl transferase (Hidroxicinamoil-CoA: quinato hidroxicinamoil transferasa) IREG: Iron-regulated protein (Proteína regulada por hierro) ISSR: Inter Simple Sequence Repeats ITP: Iron transporter protein (Proteína transportadora de hierro) K: Potasio KCO: K+ channel outward-rectifier kg: kilogramo KIR: K+ Inward Rectifier KORC: Outward Rectifying K Channel KT: K+ transporter (Transportador de K) KUP: K+ uptake permease l: litro LCYb: Lycopene ß-ciclase (Licopeno ß-ciclasa) LCYe: Lycopene ε-ciclase (Licopeno ε-ciclasa) LSD: Least Significant Difference (Mínima Diferencia Significativa) M: molar MAGRAMA: Ministerio de Agricultura, Alimentación y Medio Ambiente MAS: Marker Assisted Selection (Selección Asistida por Marcadores) MATE: Multi-drug and toxin efflux mg: miligramo Mg: Magnesio MHX: Mg2+/H+ antiporter ml: mililitro mM: milimolar Mpb: megapares de bases MRS2: Mitochondrial RNA splicing 2 MYB: Proteína proto-oncogen de la familia de factores de transcripción de la familia mieloblastosis N: Nitrógeno NAS: Nicotianamine synthase (Nicotianamina sintasa) ng: nanogramo NHX: Na+/H+ exchanger (Intercambiador de Na+/H+) NJ: Neighbor Joining nm: nanómetros NRAMP: Natural-resistance-associated macrophage protein OPT: Oligopeptide transporter (Transportador de oligopéptidos) P: Fósforo PAL: Phenylalanine ammonia-lyase (Fenilalanina amonio-liasa) PAMV: Potato Aucuba Mosaic Virus ii Abreviaturas y acrónimos pb: pares de bases PCA: Principal Component Analysis (Análisis de Componentes Principales) PCN: Potato Cyst Nematodes (Nematodos de Quiste de la Patata) PCR: Polymerase Chain Reaction (Reacción en Cadena de la Polimerasa) PF: Peso fresco PS: Peso seco pH: Logaritmo negativo de la concentración de iones hidrógeno (H+) PLRV: Potato Leafroll Virus PMTV: Potato Mop-Top Virus ppm: partes por millón PSTV: Potato Spindle Tuber Viroid PSY: Phytoene synthase (Fitoeno sintasa) PVA: Potato Virus A PVM: Potato Virus M PVS: Potato Virus S PVT: Potato Virus T PVX: Potato Virus X PVY: Potato Virus Y PYDV: Potato Yellow Dwarf Virus PYVV: Potato Yellow Vein Virus QRL: Quantitative Resistance Loci QTL: Quantitative Trait Loci RAPD: Random Amplified Polymorphic DNA REML: Restricted Maximum Likelihood RFLP: Restriction Fragment Length Polymorphism SCA: Specific Combining Ability (Aptitud Combinatoria Específica) SD: Standard Deviation (Desviación estándar) SE: Standard Error (Error Estándar)
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