Thesis (M.Sc) (Agriculture)--University of Limpopo,2005. / Host-status of wild cucumber (Cucumis myriocarpus) to the root-knot nematode (Meloidogyne incognita) was evaluated under microplot conditions. The 30-cm plastic diameter pots were placed into holes leaving 10-cm above the soil surface. Pots were filled to the 10-cm mark (soil surface) using 3:1 sand: Hygromix (v/v). Two-week old seedlings of C. myriocarpus were transplanted and irrigated with 2 L tapwater every other day. On the transplanting day, treatments were initiated by inoculating seedling with 0, 25, 125, 625 and 3 125 juveniles of M. incognita. The treatments were arranged in a randomised complete block design, with 10 replications. The experiment was terminated 56 days after initiating the study. At all levels of inoculation, the reproductive factor (Pf/Pi) was below unity, suggesting that nematodes failed to reproduce on this plant. Gall formation occurred, but the galls did not develop. Nematode had no effect on shoot and fruit weight, but significantly reduced stem diameter. Results of this study suggested that C. myriocarpus was a non-host to M. incognita. The failure of the galls to develop suggested that this plant is resistant to M. incognita. / sponsored by the South African National Research Foundation, through the Scarce Skills fellowship.

Thesis (Ph.D. (Plant Protection)) --University of Limpopo, 2012 / Global withdrawal of synthetic fumigant nematicides like methyl bromide due to their eco-unfriendliness resulted in serious consequences in production of crops which do not have genotypes that are resistant to plant-parasitic nematodes. Watermelon (Citrullus lanatus) is one such crop, where infection by highly aggressive root-knot nematodes (Meloidogyne species) invariably results into as high as 50% yield loss, with occasional total crop failures. Initial screening for nematode resistance in Cucumis species indigenous to South Africa suggested the possibility of the existence of nematode resistance, with the probability of these species being compatible with Citrullus species in inter-generic grafting technology. Uses of indigenous genera in Cucurbitaceae family as nematode-resistant seedling rootstocks in watermelon production could promote the South African watermelon industry as outlined in ISO 9001 certification guidelines to have competitive advantage in lucrative watermelon export markets. The objectives of this study were to determine the: (1) host-status and host-sensitivity of C. africanus and C. myriocarpus seedlings using a series of inoculation levels of M. incognita race 2 under various conditions, (2) host-status and host-sensitivity of C. africanus and C. myriocarpus seedlings using a series of inoculation levels of M. incognita race 4 and M. javanica, including the resistance form in these plant species, at least, under selected environmental conditions, (3) host-status and host-sensitivity of C. africanus and C. myriocarpus seedlings using a series of inoculation levels of M. incognita race 2 with multi-nematode xxviii infestations in order to establish whether the observed nematode resistance was sustainable when the plant was attacked by various pests at the root system level, (4) compatibility of inter-generic grafting of Citrullus and Cucumis seedlings in order to establish the potential uses of Cucumis species in olericulture, and (5) influence of the greenhouse whitefly (Trialeurode vaporariorum) infection on resistance of C. africanus to Meloidogyne species in order to establish whether the observed nematode resistance was sustainable when the plant was attacked by pests on complimentary organs. Reliability of measured variables was ensured by using statistical levels of significance (P ≤ 0.05) and coefficient of determination (R2), with validity being ensured by conducting experiments at the same location over two seasons or conducting one experiment during one season at two different locations, viz. the University of Limpopo and the Agricultural Research Council – Institute for Industrial Crops, and/or by setting up factorial treatments. Results consistently demonstrated that C. africanus and C. myriocarpus were non-hosts to M. incognita races 2 and 4 and M. javanica, without the test nematodes inflicting any damage to plants, which in plant-parasitic nematodes is described as nematode resistance. Quadratic relationships between RF values and log10(Pi + 1) transformations, in addition to confirming the density-dependent growth patterns of plant-parasitic nematodes, also suggested that chemical compounds responsible for suppression of nematodes in the two Cucumis species were different. The two Cucumis species were resistant to M. incognita races 2 and 4 and M. javanica, regardless of the environment under which the experiments were conducted. In field studies, the xxix two Cucumis species supported the ring nematodes (Criconema mutabile) and the spiral nematodes (Helicotylenchus dihystera), without these exo-parasitic nematodes inflicting any damage to plants, which in plant-parasitic nematodes is described as tolerance. Interactions among Meloidogyne species, C. mutabile and H. dihystera were either stimulatory or inhibitory, depending on whether Meloidogyne species were in the soil or inside the roots. Mechanisms of nematode resistance in the two Cucumis species were different, with C. africanus and C. myriocarpus depicting pre-infectional and post-infectional forms of resistance, respectively, without any sign of hypersensitivity in roots. When, seeds of Citrullus species were primed in water to hasten germination. Using the developed technology, survival of grafts improved from 36% to 100%, translating to relative improvement of 178%, with nematode-resistant rootstocks retaining their nematode resistant capabilities, while watermelon scions flowered earlier, with relatively higher fruit yield, without any deleterious effect on accumulation abilities of essential nutrient elements in leaves. Resistance of C. africanus to M. javanica was invariably broken by the greenhouse whitefly infection at high population levels, possibly through loss of non-structural carbohydrates, which are essential in synthetic pathways of secondary metabolites. Cucumis africanus and C. myriocarpus contain cucurbitacin B (C32H48O8) and cucurbitacin A [cucumin (C27H40O9), leptodermin (C27H38O8)], respectively, which have high demand for carbon and energy. Consequently, the efficacy of indigenous Cucumis species as nematode-resistant rootstocks in suppression of Meloidogyne species would be dependent upon the management of the xxx greenhouse whitefly population densities. In conclusion, C. africanus and C. myriocarpus have the potential for use as nematode-resistant rootstocks in the production of watermelon cultivars ‘Congo’ and ‘Charleston Gray’ in South Africa, where nematode population densities of M. incognita races 2 and 4 and M. javanica are widely distributed and are highly injurious to watermelons. Although nematode resistance in the two Cucumis species had attributes of sustainability, populations of the greenhouse whitefly broke the resistance. Proposed future research areas included influence of cucurbitacins in fruit quality of watermelons and protocols for mass culturing the nematode-resistant Cucumis rootstocks using tissue culture technology. / the National Research Foundation,the Agricultural Research Council (ARC) and the Landbank Chair of Agriculture-University of Limpopo

Thesis (M.Sc. (Plant Protection )) --University of Limpopo, 2011 / Crude extracts of wild watermelon (Cucumis africanus) and wild cucumber (C. myriocarpus) fruits are widely used for both medicinal and ritual purposes in South Africa. Fruits are collected fresh from the wild, but have high incidence of post-harvest decay. A study was conducted to isolate and identify the pathogen responsible for post-harvest fruit decay, followed by the pathogenicity tests. Decayed fruits were individually surface-sterilised using 0.5% NaOCl, incubated at 25ºC to allow for decay, small rotten pieces were severed and placed on solidified plates of potato dextrose agar and incubated. At harvest, seven days after incubation, isolated fungus was repeatedly cultured for 21 days for verification of diagnostic characteristics. Based on the morphological characteristics, the pathogen associated with fruit rot of both Cucumis species was identified through the assistance of an expert as Penicillium simplicissimum (Oudem) Thom. Pathogenicity results suggested that P. simplicissimum was responsible for the observed fruit decay in both species, with the higher incidence being in C. africanus, probably due to its low pH. Due to the antibiotics that P. simplicissimum releases and its reduction of medium pH, the culture retained its purity, without any contamination. In conclusion, the pathogen that induces post-harvest fruit decay in C. africanus and C. myriocarpus is P. simplicissimum, which has the ability to reduce the pH of the growing medium and also produce antibiotics. / National Research Foundation

Thesis (Ph.D. Agriculture (Horticulture)) -- University of Limpopo, 2012 / High yield losses in various crops due to plant-parasitic nematodes are associated with high initial nematode population densities (Pi). Uses of synthetic nematicides to reduce Pi were dependent on the physiological effect of materials on the protected crops, resulting into the coining of pre-emergent and post-emergent nematicides. Crude extracts of wild cucumber (Cucumis myriocarpus) fruit consistently reduced nematode population densities of the southern root-knot nematode (Meloidogyne incognita) when used as a post-emergent bio-nematicide. The purpose of this study was to investigate the compatibility of crude extracts of C. myriocarpus fruit when used as a pre-emergent bio-nematicide on germination and emergence of commercially important dicotyledonous and monocotyledonous crops using empirical tests and computer-generated models. Studies were conducted over a period of three years to assess the effects of this material on growth of various seedlings. Seven treatments comprising crude extracts of C. myriocarpus fruit (0, 2.5, 5, 7.5, 10, 12.5 and 15 g/pot) and test solutions (0, 25, 50, 75, 100, 125 and 150 g/ℓ distilled water) were used for emergence and germination in initial studies. Generally, 18 days after the treatments, variables measured and levels of crude extracts of C. myriocarpus fruit had negative quadratic relationships, which suggested that they had density-dependent growth responses. Subsequent studies were conducted using three selected crops each from the families Alliaceae, Gramineae and Solanaceae under greenhouse conditions, each with reduced concentration of 10 treatments (0, 0.25, 0.50, 0.75, 1.00, 1.25, 1.50, 1.75, 2.00 and 2.25 g material/pot). Using variables of various organs and crops, significant means were subjected to the Curve-fitting Allelochemical Dosage Response (CARD) computer model, which was characterised by six biological indices, viz. threshold stimulation (Dm), xli saturation level (Rh), 0% inhibition (D0), 50% inhibition (D50), 100% inhibition (D100) and transformation level (k). The model demonstrated that the responses of the three crops from each family when regressed to dosages of crude extracts of C. myriocarpus fruit exhibited the density-dependent growth patterns, characterised by responses that included stimulation, saturation and inhibition. The integrated sensitivities (Σk) of the tested crops to crude extracts of C. myriocarpus fruit ranged from Σk = 9 to Σk = 51, with eggplant (Solanum melongena) and sorghum (Sorghum bicolor) being the most sensitive, while tomato (Solanum lycopersicum) was the least sensitive. Using the data depicting the stimulation range from CARD model, viz. (Dm), which is a threshold stimulation dosage and (Rh), which is a saturation dosage, mean dosage stimulation response (MDSR) was determined for chive (Allium schoenoprasum), leek (Allium ampeloprasum), onion (Allium cepa), maize (Zea mays), millet (Panicum miliaceum), sorghum, eggplant, pepper (Capsicum annum) and tomato as being 1.19, 0.68, 0.45, 1.13, 0.86, 1.12, 0.74, 1.11, and 0.53 g, respectively. These MDSR values are dosages which when applied for respective crops at direct seeding would not affect germination or emergence. MDSR values were validated for onion, millet and tomato, resulting in approximately 100% suppression of nematodes in all three test crops. In contrast, 100% emergence occurred in millet and tomato, while the validated MDSR reduced emergence on onion by 15%, which confirmed the sensitivity of this crop to crude extracts of C. myriocarpus fruit. In conclusion, crude extracts of C. myriocarpus fruit have the potential for use as pre-emergent bio-nematicide in suppression of plant-parasitic nematodes in various crops.

Thesis (M.Sc. (Horticulture)) -- University of Limpopo, 2017. / Root-knot (Meloidogyne species) nematodes are economically destructive pathogens of over 3000 species, whereas others have resistance to Meloidogyne species. Wild watermelon (Cucumis africanus) and wild cucumber (Cucumis myriocarpus) are highly resistant to Meloidogyne species, particularly M. incognita and M. javanica. The two Cucumis species are used in inter-generic grafting with watermelon (Citrullus lanatus) as nematode resistant rootstocks. Also, the two Cucumis species are used in traditional medicine and in plant-parasitic nematode management as phytonematicides. The form of nematode resistance, which is essential in plant breeding, is not documented for the two Cucumis species. The objective of this study was to determine the form of nematode resistance in the two Cucumis species to M. incognita and M. javanica under greenhouse conditions. Four parallel experiments were each conducted under greenhouse conditions. Uniform six-week old Cucumis seedlings were transplanted into 250 ml polystyrene cups filled with 200 ml growing medium of steam-pasteurised fine sand. A week after transplanting, Cucumis seedlings were each infested by dispensing approximately 100 M. incognita second-stage juveniles (J2) or M. javanica J2 using a 20 ml plastic syringe by placing into 5-cm-deep furrow around the seedling stem and covered with growing medium. Treatments (periodic harvest intervals) were arranged in a randomised complete block design, replicated five times. Five seedlings from each experiment were harvested every second day, for 30 days, with stained roots being assessed for necrotic spot (suberised cells) number, giant cell number, proliferation of rootlet interference number and root gall number. Periodic harvest intervals were highly significant (P ≤ 0.01) on necrotic spot number, proliferation of rootlet interference number and root gall number in C. africanus-M. incognita relations, but were not significant for giant cell number. Treatments contributed 59, 64 and 50% in total treatment variation (TTV) of necrotic spot number, proliferation of rootlet interference number and root gall number, respectively. Harvest period had highly significant effects on necrotic spot number, giant cell number, proliferation of rootlet interference number and root gall number in C. africanus-M. javanica relations. Treatments contributed 55, 71, 63 and 59% in TTV of necrotic spot number, giant cell number, proliferation of rootlet interference number and root gall number, respectively. Periodic harvest intervals were significant (P ≤ 0.05) on giant cell number and highly significant on root gall number in C. myriocarpus-M. incognita relations. However, there were no significant treatment differences on necrotic spot number and proliferation of rootlet interference number. Treatments contributed 57 and 57% in TTV of root gall number and giant cell number, respectively. Harvest period had highly significant effects on giant cell number, proliferation of rootlet interference number and root gall number, but were not significant on necrotic spot number in C. myriocarpus-M. javanica relations. Treatments accounted for 67, 49 and 53% in TTV of giant cell number, proliferation of rootlet interference number and root gall number, respectively. In conclusion, the mechanism of resistance to M. incognita and M. javanica in both C. africanus and C. myriocarpus was post-infectional nematode resistance, which has attributes for introgression into commercial nematode-susceptible Cucumis cultivars. / Agricultural Research Council (ARC), National Research Foundation of South Africa; and the ARC-Universities Collaboration Centre for Smallholder Farmers

[ES] El melón (Cucumis melo L.) es una de las especies hortícolas más demandadas en el mundo, y como tal el mejoramiento vegetal tiene el desafío continuo de generar alternativas para el agricultor por medio del desarrollo de nuevas variedades que presenten, tolerancia/ resistencia a factores de estrés biótico (plagas y enfermedades) y abiótico (temperatura, humedad, suelo) y a su vez atributos de calidad apreciados por el consumidor. El mejoramiento convencional en melón por medio de cruzamientos entre líneas seleccionadas es una estrategia ampliamente utilizada, pero que presenta algunas limitaciones, como las barreras de cruzabilidad entre las variedades comerciales y el germoplasma silvestre, además de los períodos extensos de tiempo requeridos para evaluar las progenies. En este contexto, el injerto y el desarrollo de nuevos patrones están siendo adoptados a nivel mundial como estrategias alternativas de mejora en cultivos hortícolas, que permite aprovechar caracteres de interés presentes en especies no cultivadas o no comerciales sin necesidad de realizar largos y complejos planes de mejora (cruzamiento y retrocruzamiento). El uso de la diversidad intra e interespecífica como portainjertos para sandía ha dado muy buenos resultados, documentados en una amplia bibliografía científica. Sin embargo, las ventajas y beneficios del injerto presentan inconsistencias en los estudios realizados en melón. Esto se explica por los diferentes métodos de cultivos evaluados, así como por la amplia gama de variedades y tipos comerciales de melón existentes; esto exige a su vez, mayor inversión y fortalecimiento de la investigación en esta área del conocimiento. La presente Tesis Doctoral tiene por objetivo la generación de información relacionada al injerto en melón mediante la evaluación de portainjertos desarrollados en los grupos de mejora del Instituto de Conservación y Mejora de la Agrodiversidad Valenciana (COMAV), donde se ha llevado a cabo este trabajo, utilizando para ello diferentes condiciones de cultivo, y por lo tanto estrés, y utilizando dos tipos de melón muy consumidos en España y Europa como el ‘Piel de Sapo’ y el ‘Cantalupo’, además de una variedad tradicional muy apreciada por su calidad, el ‘Meló d’Or d’Ontinyent’. La recuperación de variedades tradicionales es un reto para la agricultura moderna y gracias al injerto validamos el potencial de esta técnica para producir ‘Meló d’Or d’Ontinyent’ bajo condiciones de estrés por la presencia de un hongo patógeno como Monosporascus cannonballus Pollack et Uecker en el suelo. El patrón experimental UPV-PRMc, desarrollado por el grupo de mejora de cucurbitáceas del COMAV de la Universidad Politécnica de Valencia, mediante la hibridación de un melón tipo ‘Piel de Sapo’ y una accesión de un melón silvestre del tipo agrestis, mostró su capacidad para proporcionar resistencia y vigor a la variedad tradicional, que sin el uso de un portainjerto resistente sería incapaz de culminar su ciclo de cultivo bajo estas condiciones. Ensayos de campo complementarios permitieron evaluar el comportamiento de diferentes portainjertos experimentales y comerciales de Cucurbita y Cucumis, en condiciones de estrés bajo sistemas de producción convencional en campo abierto. Con relación al vigor y al desarrollo de las plantas injertadas, los patrones de Cucurbita mostraron un retraso en el vigor temprano del cultivo y tanto Ma x Mo EF como Pe x Pe ZS tuvieron los más bajos rendimientos. Destaca el uso del híbrido Ma x Ec como una buena alternativa a los híbridos comerciales normalmente utilizados entre C. maxima x C. moschata. Por su parte los portainjertos de C. melo mostraron una mejor afinidad y mayor vigor consistentemente a lo largo de los 3 años de estudio, al igual que los híbridos interespecíficos Fi x My y Fi x An. Adicionalmente al estudio de la compatibilidad entre el melón Piel de Sapo y los diferentes patrones experimentales y comerciales, también se abordó el impacto sobre la calidad del fruto. Este aspecto es quizá el más controvertido en relación al injerto por el uso de patrones filogenéticamente distantes que pueden pertenecer a otra especie o género distinto al melón. Nuestros datos reflejaron que los patrones de Cucurbita tuvieron efecto sobre la forma del fruto y la cavidad seminal, mientras que los patrones de Cucumis melo presentaron un menor impacto sobre los parámetros de calidad del fruto. También es importante destacar que los patrones de especies silvestres o no cultivadas de Cucumis no afectaron significativamente la calidad, a pesar de que C. metuliferus E. Meyer ex Naudin mostró signos de colapso en el segundo año de evaluación y consistentemente produjo los frutos más pequeños en comparación a las plantas sin injertar. El estudio complementario sobre el perfil metabólico y aromático de los frutos ha permitido ampliar los criterios de selección de portainjertos de melón. Se observó la variación en el contenido de ácidos y azúcares principalmente debido al grado de maduración y en menor medida a una combinación de patrón-variedad específica. La modificación del perfil de compuestos volátiles mostró un efecto claro de los portainjertos de Cucurbita sobre la variedad de melón, con el incremento de compuestos aromáticos relacionados a la calabaza y la reducción de otros compuestos claves en el aroma del melón Piel de Sapo. Con la perspectiva de aprovechar el germoplasma silvestre como portainjerto, en el Capítulo 2 de la presente tesis doctoral, presentamos los estudios enfocados en la caracterización de especies silvestres o no cultivadas del género Cucumis, que son una fuente de resistencias a patógenos y enfermedades de importancia económica como nematodos, oído o Fusarium, y que están ausentes en las variedades comerciales de melón. La estandarización de metodologías para optimizar la germinación de accesiones de Cucumis silvestres ha sido un primer paso para maximizar el rendimiento de la semilla como material de partida, ya que la dormancia presente en el germoplasma no cultivado suele ser un factor limitante. Basados en los estudios preliminares de campo en los que se observó la diferencia de vigor entre una accesión de C. metuliferus y el melón ‘Piel de Sapo’, se estudió si los híbridos interespecíficos desarrollados por el grupo de mejora suponían una opción para el incremento del vigor y la combinación de características deseables en una misma planta capaz de soportar el injerto de una variedad comercial de melón. Las barreas de incompatibilidad, impidien los cruzamientos viables con C. metuliferus (filogenéticamente separado del resto de Cucumis africanos) pero los híbridos interespecíficos de los cruzamientos entre Cucumis ficifolius A. Rich. x Cucumis anguria L. y Cucumis ficifolius x Cucumis myriocarpus Naudin son viables. Estos materiales F1, mostraron un mayor vigor, un incremento en el diámetro de su hipocótilo y una buena compatibilidad con melones de tipo ‘Cantalupo’ y ‘Piel de Sapo’, bajo condiciones de invernadero y a campo abierto. Un alto grado de resistencia a Fusarium oxysporum f. sp. melonis como resultado de la combinación de sus genomas parentales, así como una buena tolerancia al estrés osmótico bajo condiciones in vitro, destacan como características deseables en estos dos híbridos interespecíficos. Utilizando especies silvestres e híbridos de Cucumis, se evaluaron distintas metodologías para la obtención de poliploides, con el objetivo de potenciar el vigor de las plántulas (tamaño y diámetro del hipocótilo) y estudiar los efectos de la poliploidización. La colchicina fue el compuesto antimitótico más efectivo aplicado sobre semillas pregerminadas de dos híbridos interespecíficos de Cucumis. En líneas homocigóticas de C. anguria, C. ficifolius, C. myriocarpus, o C. metuliferus, la duplicación del genoma no fue estable en ningún caso. La evaluación del nivel de ploidía confirmó que los alopoliploides obtenidos fueron hexaploides (6n=72). Los nuevos materiales obtenidos, son alopoliploides sintéticos que resultaron de la duplicación de un genoma híbrido. A diferencia de sus antecesores triploides, los alopoliploides mostraron la recuperación de su fertilidad, asociado posiblemente a la presencia de SNPs concretos relacionados a genes que codifican procesos reproductivos. La mejora de características de interés en especies silvestres (mayor diámetro de los hipocótilos) ha sido posible por medio de la poliploidización, lo que favorece el procedimiento de realización del injerto y la producción de la planta injertada. Los resultados presentados en esta tesis doctoral han sido posibles gracias a la colaboración de grupos de investigación interdisciplinarios de la Universidad Politécnica de Valencia, la Universidad Jaime I de Castellón y la Politécnica de Cataluña, en el marco del proyecto AGL2014-53398-C2-2-R APROXIMACIONES BIOTECNOLOGICAS Y CULTURALES PARA LA MEJORA DE LAS RESISTENCIAS Y EL CONTROL DE ENFERMEDADES EN MELON Y SANDIA y con apoyo de los proyectos AGL2017- 85563-C2-1-R de FEDER/Ministerio de Ciencia, Innovación y Universidades–Agencia Estatal de Investigación, y proyecto para Grupos de Excelencia PROMETEO 2017/078 además del apoyo de la SECRETARIA DE EDUCACIÓN SUPERIOR, CIENCIA, TECNOLOGÍA E INNOVACIÓN (SENECYT-ECUADOR) que por medio del Instituto de Fomento al Talento Humano (IFTH), proporcionó una beca de estudio al doctorando. / [EN] The melon (Cucumis melo L.) is one of the most demanded horticultural species in the world and as such, plant improvement has the constant challenge of thinking up alternatives for the farmer through the development of new varieties with tolerance / resistance to biotic (pests and diseases) and abiotic (temperature, humidity, soil) stress factors and quality attributes appreciated by the consumer. Conventional breeding in melon through crossings between selected lines is a widely used strategy, but it has some limitations, such as cross barriers between commercial varieties and wild germplasm, in addition to the extended periods of time required to evaluate the progeny. In this context, grafting and the development of new rootstocks are being adopted worldwide as alternative breeding strategies in horticultural crops, which allows to take advantage of relevant characteristics present in non-cultivated or non-commercial species without extensive and complex breeding programs (crossing and backcrossing). Development of rootstocks for watermelon crop derived from intra and interspecific diversity have been successful and well documented in a wide scientific bibliography. However, advantages and benefits from grafting show inconsistent data in melon assays. These results can be explained since different cultivation methods have been evaluated and because of the large number of commercial varieties of melon; this fact motivates a higher investment and strengthening of research in this area of knowledge. The present Doctoral Thesis has as objective the generation of information related with melon grafting, through the development, characterization and study under stress conditions of novel rootstocks used for two commercial cultivars consumed in Spain and Europe, like ‘Piel de Sapo’ and ‘Cantaloupe’, besides a traditional variety as the ‘Meló d’Or d’Ontinyent’. Recovery of traditional cultivars is a challenge for modern agriculture and thanks to grafting we have validated the potential of this technique to crop ‘Meló d’Or d’Ontinyent’ in soil stressed by Monosporascus cannonballus Pollack et Uecker. The experimental rootstock UPV-PRMc, developed by Institute for the Preservation and Improvement of Valencian Agro-diversity (COMAV) of the Polytechnic University of Valencia, from an hybridization between one ‘Piel de Sapo’ melon and one accession of wild agrestis melon, provided resistance and vigor to the traditional variety, which is not able to complete the crop cycle without be grafted onto a resistant rootstock. Complementary field trials allowed to test the behavior of experimental and commercial rootstocks of Cucurbita and Cucumis under stress conditions in conventional production systems. Regarding vigor and growth of grafted plants, Cucurbita rootstocks showed a delay in early vigor of plants and Ma x Mo EF and Pe x Pe ZS had the lowest yields. The Ma x Ec hybrid highlights as a good alternative to the C. maxima x C. moschata hybrids, commonly used in commercial grafting. On the other hand, C. melo rootstocks showed better affinity and higher vigor consistently along these 3 years, as well as the interspecific hybrids Fi x My and Fi x An. Additionally to the compatibility behavior between ‘Piel de Sapo’ melon and different experimental and commercial rootstocks, also the impact on fruit quality was studied. This issue is one of the most controversial about grafting because of the use of rootstocks phylogenetically distant that belong to other species or genera than melon. Our data showed an effect over shape and size of seed cavity when using Cucurbita rootstocks, while Cucumis melo ones showed a lower impact on fruit quality traits. It is important to note that rootstocks derived from wild or non-cultivated Cucumis species did not alter significantly the quality, despite of that C. metuliferus E. Meyer ex Naudin showed vine decline symptoms in the second year and consistently the smallest fruits compared to non-grafted plants. The complementary analysis of the aromatic and metabolic profile from fruits have allowed to increase the selection criteria for melon rootstocks. Variation in acids and sugars was observed mainly due to maturation degree and to a lesser extent to a specific rootstock-scion combination. The modification in volatile compounds levels displayed an effect of the Cucurbita rootstocks on melon cultivar, increasing aromatic compounds related to pumpkin and decreasing other key compounds in ‘Piel de Sapo’ aroma melon. With the prospect to take advantage from wild germplasm to develop new rootstocks, in the Chapter 2 in this Doctoral Thesis, we showed the studies focused on the characterization of wild or non-cultivated species in the Cucumis genus, which are a resistance resource against pathogens and diseases of economic importance like nematodes, powdery mildew or Fusarium, and that are absent in commercial melon. The standardization of methodologies to optimize the germination of wild Cucumis accessions has been the first step to maximize the seed yield as a started material, since the dormancy in non-cultivated germplasm is usually a limiting factor. Based on preliminary field studies, where the differences in vigor between C. metuliferus and ‘Piel de Sapo’ melon were observed, the development of interspecific hybrids was approached as an option to increase the vigor and combine characteristics of interest in one same plant able to bear the grafting with a commercial variety of melon. The incompatibility barriers prevented viable crosses with C. metuliferus (phylogenetically distant of the rest of African Cucumis), although two interspecific hybrids were viable from crosses between Cucumis ficifolius A. Rich. x Cucumis anguria L. and Cucumis ficifolius x Cucumis myriocarpus Naudin. These F1 plants had higher vigor, hypocotyl diameter and good compatibility with ‘Cantaloupe’ and ‘Piel de Sapo’ melons under field and greenhouse conditions. High level of resistance to Fusarium oxysporum f. sp. melonis as result of the combination of their parental genomes, as well as a good tolerance to osmotic stress under in vitro conditions, highlight as desirable characteristics in these two new interspecific hybrids. With the preliminary results on the improvement of traits in wild Cucumis germplasm from the hybridization, some methodologies were evaluated to obtain polyploids, in order to enhance the desirable features in these hybrids (vigor, hypocotyl size and diameter). Colchicine was the most effective antimitotic agent applied over pregerminated seeds of two Cucumis interspecific hybrids. In homozygous lines of C. anguria, C. ficifolius, C. myriocarpus, or C. metuliferus, genome duplication was not stable. The polyploidy level confirmed that allopolyploids obtained were hexaploids (6n=72). The novel materials obtained are synthetic allopolyploids that result from the duplication of a hybrid genome. Unlike their triploid ancestors, allopolyploids showed the recovery of their fertility, possibly associated with the presence of specific SNPs related to genes that encode reproductive processes. The improvement of characteristics of interest in wild species has been possible through polyploidization, which favors the development of new improved materials capable of adapting to the grafting process. The results presented in this Doctoral Thesis have been possible thanks to the collaboration of interdisciplinary research groups from Polytechnic University of Valencia, Jaime I University of Castellón and Polytechnic University of Catalonia, within the framework of the AGL2014-53398-C2-2 project. -R BIOTECHNOLOGICAL AND CULTURAL APPROACHES FOR THE IMPROVEMENT OF RESISTANCES AND THE CONTROL OF DISEASES IN MELON AND WATERMELON and with the support of the projects AGL2017-85563-C2-1-R of FEDER / Ministry of Science, Innovation and Universities-State Agency of Research and project for Groups of Excellence PROMETEO 2017/078, in addition to the support of the SECRETARIA DE EDUCACIÓN SUPERIOR, CIENCIA, TECNOLOGÍA E INNOVACIÓN (SENECYT-ECUADOR) which through the Institute for the Promotion of Human Talent (IFTH), provided a scholarship for the doctoral candidate. / [CA] El meló (Cucumis melo L.) és una de les especies hortícoles més demandades en el món, i consegüentment el millorament vegetal té el desafiament continu de generar alternatives per a l’agricultor per mitjà del desenvolupament de noves varietats que presenten, tolerància/resistència a factors d’estrès biòtic (plagues i malalties) i abiòtic (temperatura, humitat, sòl) i al seu torn atributs de qualitat apreciats pel consumidor. El millorament convencional en meló mitjançant creuaments entre línies seleccionades és una estratègia àmpliament utilitzada, però que presenta algunes limitacions, com la impossibilitat de creuar-se d’algunes varietats comercials i determinat germoplasma silvestre d’interès, a més dels períodes extensos de temps requerits per a avaluar les progènies. En aquest context, l’empelt i el desenvolupament de nous patrons estan sent adoptats a nivell mundial com a estratègies alternatives de millora en cultius hortícoles. L’empelt permet aprofitar caràcters d’interès presents en espècies no cultivades o no comercials sense necessitat de realitzar llargs i complexos plans de millora (creuament i retrocruzamiento). L’ús de la diversitat intra i interespecífica com a portaempelts de meló d’Alger ha donat molt bons resultats, documentats en una àmplia bibliografia científica. No obstant això, els avantatges i beneficis de l’empelt presenten inconsistències en els estudis realitzats en meló. Això s’explica pels diferents mètodes de cultius avaluats, així com per l’àmplia gamma de varietats i tipus comercials de meló existents; això exigeix al seu torn, major inversió i enfortiment de la investigació en aquesta àrea del coneixement. La present Tesi Doctoral té per objectiu la generació d’informació relacionada amb l’empelt en meló, mitjançant l’avaluació de nous portaempelts que s’han obtingut en els grups de millora del COMAV (Instituto de Conservación y Mejora de la Agrodiversidad Valenciana) on s’ha dut a terme aquest treballs fent servir diferents condicions de cultiu i per tant de estressos i dos tipus de meló molt consumits a Espanya i Europa com el ‘Piel de Sapo’ i el ‘Cantalupo’ a més d’una varietat tradicional reconeguda per la seua excel·lent qualitat, el ‘Meló d’Or d’Ontinyent’. La recuperació de varietats tradicionals és un repte per a l’agricultura moderna i mijançant l’empelt validem el potencial d’aquesta tècnica per a produir ‘Meló d’Or d’Ontinyent’ sota condicions d’estrès degudes a la presència en la terra d’un fong patogen com es el Monosporascus cannonballus Pollack et Uecker. El patró experimental UPV-PRMc, desenvolupat pel grup de millora de cucurbitàcies del COMAV, mitjançant la hibridació d’un meló tipus ‘Piel de Sapo’ i una accessió d’un meló silvestre del tipus agrestis, va mostrar la seua capacitat per a proporcionar resistència i vigor a la varietat tradicional, que sense l’ús d’un portaempelt resistent no completaria el cicle de cultiu sota aquestes condicions. Assajos de camp complementaris van permetre avaluar el comportament de diferents portaempelts experimentals i comercials de Cucurbita i Cucumis, en condicions d’estrès en sistemes de producció convencional a camp obert. En relació amb el vigor i al desenvolupament de les plantes empeltades, els patrons de Cucurbita van mostrar un retard en el vigor a l’inici del cultiu i tant Ma x Mo EF com Pe x Pe ZS van tindre els rendiments més baixos. Destaca l’ús de l’híbrid Ma x Ec com una bona alternativa als híbrids comercials normalment utilitzats que deriven de creuaments C. maxima x C. moschata. Per la seua part els portaempelts de C. melo van mostrar una millor afinitat i major vigor consistentment al llarg dels 3 anys d’estudi, igual que els híbrids interespecífics Fi x My i Fi x An. Addicionalment a l’estudi de la compatibilitat entre el meló ‘Piel de Sapo’ i els diferents patrons experimentals i comercials, també es va estudiar l’impacte sobre la qualitat del fruits. Aquest aspecte és potser el que més controvèrsia crea al voltant de l’ús de l’emplet on s’utilitzen portampelts filogenèticament distants que poden pertànyer a una altra espècie o gènere diferent al meló. Les nostres dades van reflectir que els patrons de Cucurbita van tindre efecte sobre la forma del fruit i la cavitat seminal, mentre que els patrons de Cucumis melo van presentar un menor impacte sobre els paràmetres de qualitat del fruit. També és important destacar que els patrons d’espècies silvestres o no cultivades de Cucumis no van afectar significativament la qualitat, a pesar que C. metuliferus E. Meyer ex Naudin va mostrar signes de col·lapse en el segon any d’avaluació i consistentment va produir els fruits més xicotets en comparació a les plantes sense empeltar. L’estudi complementari sobre el perfil metabòlic i aromàtic dels fruits ha permès ampliar els criteris de selecció de portaempelts de meló. Es va observar la variació en el contingut d’àcids i sucres principalment com a consequència del grau de maduració i en menor mesura a una combinació de patró-varietat específica. La modificació del perfil de compostos volàtils va mostrar un efecte clar dels portaempelts de Cucurbita sobre la varietat de meló, amb l’increment de compostos aromàtics relacionats amb la carabassa i la reducció d’altres compostos claus en l’aroma del meló tipus ‘Piel de Sapo’. Amb la perspectiva d’aprofitar el germoplasma silvestre per al desenvolupament de nous portaempelts, al Capítol 2 de la present tesi doctoral, presentem els estudis enfocats a la caracterització d’espècies silvestres o no cultivades del gènere Cucumis, que són una font de resistències a patògens i malalties d’importància econòmica com a nematodes, oïdi o Fusarium, i que no tenen en les varietats comercials de meló. L’estandardització de metodologies per a optimitzar la germinació d’accessions de Cucumis silvestres ha sigut un primer pas per a maximitzar el rendiment de la llavor com a material de partida, ja que la dormancia present en el germoplasma no cultivat sol ser un factor limitant. Basats en els estudis preliminars de camp en els quals es va observar la diferència de vigor entre una accessió de C. metuliferus i el meló ‘Piel de Sapo’, es van avaluar els híbrids interespecífics desenvolupats als grups de millora com una opció per incrementar el vigor i combinar característiques desitjables en una mateixa planta que a l’hora fora capaç de suportar l’empelt d’una varietat comercial de meló. Les barreas d’incompatibilitat, no fan posible la hibridació amb C. metuliferus (filogenèticament separat de la resta de Cucumis Africans) però els híbrids interespecífics viables dels creuaments entre Cucumis ficifolius A. Rich. x Cucumis anguria L. i Cucumis ficifolius x Cucumis myriocarpus Naudin són viables. Aquests materials F1, van mostrar un major vigor, un increment en el diàmetre del seu hipocòtil i una bona compatibilitat amb melons de tipus ‘Cantalupo’ i ‘Piel de Sapo’, en condicions d’hivernacle i a camp obert. També aquests materials han mostrat un alt grau de resistència a Fusarium oxysporum f. sp. melonis i una bona tolerància a l’estrès osmòtic (avaluada en condicions de cultiu in vitro), característiques desitjables en aquestos dos nous híbrids interespecífics. Partint del germoplasma de Cucumis silvestre i els híbrids obtinguts, es va avaluar diferents metodologies per a l’obtenció de poliploides, amb l’objectiu de potenciar el vigor de les plàntules (grandària i diàmetre dels hipocòtil) i estudiar els efectes de la poliploidització. La colxicina va ser el compost antimitòtic més efectiu al aplicar-lo a llavors pregerminadas dels dos híbrids interespecífics de Cucumis. En línies homozigòtiques de C. anguria, C. ficifolius, C. myriocarpus, o C. metuliferus, la duplicació del genoma no va ser estable en cap cas. L’avaluació del nivell de ploidía va confirmar que els al·lopoliploides obtinguts van ser hexaploids (6n=72). Els nous materials obtinguts, són al·lopoliploides sintètics que van resultar de la duplicació d’un genoma híbrid. A diferència dels seus antecessors triploides, els al·lopoliploides van mostrar la recuperació de la seua fertilitat, associat possiblement a la presència de SNPs concrets relacionats a gens que codifiquen processos reproductius. La millora de les característiques d’interès (major diàmetre dels hipocòtils) ha sigut possible mitjançant l’obtenció de poliploides, la qual cosa facilita el procediment per a dur a terme l’empelt i per tant la producció de planta empeltada. Els resultats presentats en aquesta Tesi Doctoral han sigut possibles gràcies a la col·laboració de grups d’investigació interdisciplinaris de la Universitat Politècnica de València, la Universitat Jaume I de Castelló i la Politècnica de Catalunya, en el marc del projecte AGL2014-53398-C2-2-R APROXIMACIONS BIOTECNOLOGICAS Y CULTURALES PARA LA MEJORA DE LAS RESISTENCIAS Y EL CONTROL DE ENFERMEDADES EN MELON Y SANDIA i amb suport dels projectes AGL2017- 85563-C2-1-R de FEDER/Ministeri de Ciència, Innovació i Universitats–Agència Estatal d’Investigació, i projecte per a Grups d’Excel·lència PROMETEO 2017/078 a més del suport de la SECRETARIA DE EDUCACIÓN SUPERIOR, CIENCIA, TECNOLOGIA E INNOVACIÓN (SENECYT-L’Equador) que per mitjà del ‘Instituto de Fomento del Talento Humano’ (IFTH), va proporcionar una beca d’estudi al doctorand. / Finalmente agradecer a mi país Ecuador y a la Secretaria De Educación Superior, Ciencia, Tecnología e Innovación (SENECYT) y al Instituto Nacional De Fomento Al Talento Humano (IFTH) que me apoyaron con una beca de estudios para realizar mi tesis Doctoral, con la que espero aportar además de conocimiento científico, alternativas y soluciones para la realidad agrícola del país. / Cáceres Burbano, AE. (2020). Desarrollo de nuevos portainjertos para melón mediante el uso de estrategias y herramientas biotecnológicas [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/147863 / TESIS