Anther-derived monoploids of Solanum phureja were utilized to investigate factors essential to an efficient method of regenerating doubled monoploids (DMs). The presence of silver thiosulfate (STS) in the MS basal medium did not affect the frequency of cells with 2x nuclei but increased the proportion of cells with 1x nuclei and decreased the proportion with 4x nuclei. Results indicated that STS lower the occurrence of endopolyploid nuclei. The production of DMs was not affected by the presence of STS in MS basal medium on which the source of leaf explants was maintained. The incubation of leaf cultures in the dark or light during the callus induction phase did not influence the subsequent regenerative ability of the monoploids. However, there was a significant genotype by incubation condition interaction. Overnight incubation on MS medium with benzyladenine (BA) pulse prior to transfer to regeneration medium did not affect regeneration. Field evaluation showed various responses of DMs in terms of growth and yield compared to their anther donors or corresponding F₁ progeny. Female fertility was observed in a majority of the DMs verifying their applicability as parental genotypes in practical breeding. Efforts to generate potato hybrids based on selection of genetically diverse parents using RAPD markers and to develop high yielding diploid potato germplasm that may be instrumental in new cultivar development were addressed. Genetic diversity among in vitro plantlets of S. phureja monoploids (which represent DM maternal genotypes) and diploid heterozygous pollinators (ID lines) was estimated using RAPD markers. Field evaluation revealed significant differences among fourteen F₁ hybrids of S. phureja DM x ID with respect to total tuber number, total tuber yield, average tuber weight and vigor. Using simple matching or Jaccard coefficients, the largest parental genetic distance was associated with the highest total tuber yield among the progenies of DM parents. Based on our results, RAPDs have the potential to facilitate the identification of diverse parents to maximize the expression of heterosis in S. phureja hybrids. SSR markers were used to analyze the genetic composition of anther-derived potato plants. Anther-derived monoploids exhibited a single allele as expected. Both homozygous and heterozygous diploids were identified. SSRs were also utilized to study allelic segregation in an F₁ population. Results of this experiment revealed Mendelian inheritance of SSR alleles. / Ph. D.

Tesis por compendio / [ES] En el primer capítulo, se caracterizó una colección de seis accesiones de berenjenas, 21 accesiones de 12 especies silvestres y 45 híbridos interespecíficos de berenjena con especies silvestres utilizando 27 descriptores morfológicos y 20 descriptores morfométricos de frutos basados en la herramienta fenómica Tomato Analyzer. Observamos diferencias significativas entre los tres grupos, con híbridos que muestran valores intermedios para la mayoría de los descriptores. Las especies silvestres mostraron una amplia diversidad para ampliar la base genética de la berenjena. En el segundo capítulo, el mismo material se caracterizó para el contenido en compuestos fenólicos del fruto, el color de la pulpa de la fruta y los caracteres relacionados con el pardeamiento. Mientras que el ácido fenólico predominante en la berenjena cultivada fue el ácido clorogénico (> 65.0%), en los parientes silvestres fue de menos del 50% del área del pico del cromatograma HPLC. Las variedades cultivadas presentaron un color de carne más claro y baja actividad de polifenol oxidasa (PPO). Los híbridos interespecíficos fueron intermedios para todos los caracteres estudiados. En el tercer capítulo, realizamos un estudio genético Linea por Probador (L × P) utilizando dos líneas cultivadas, una con citoplasma oriental y otra occidental con cuatro probadores que representan tres especies silvestres: S. insanum, S.anguivi y S . lichtensteinii. Además, evaluamos el material para 3 descriptores bioquímicos, 12 morfológicos y 8 de Tomato Analyzer. Encontramos diferencias significativas para los 23 caracteres estudiados. Los valores más altos para la componente SCA se determinaron en comparación con la componente GCA. En general, los probadores del genepool secundarios fueron mejores para la mejora de los caracteres bioquímicos, mientras que las especies de genepool primarias fueron mejores para los caracteres morfológicos. En el cuarto capítulo, para obtener información sobre la genética de caracteres importantes en la berenjena, realizamos el primer estudio genético detallado de la berenjena con 10 genotipos diversos de berenjena, entre ellos una accesión de S. insanum, mientras que el resto fueron variedades tradicionales con forma del fruto muy diversa. Cuando se cruzaron en un diseño de medio dialelo, los 10 padres produjeron un conjunto de 45 híbridos. Evaluamos padres e híbridos para 14 descriptores de características morfológicas y 14 características morfométricas del fruto. También determinamos las distancias genéticas utilizando 7,335 marcadores de SNP polimórficos. Del mismo modo, en el quinto capítulo, estudiamos los compuestos fenólicos del fruto, el color de la pulpa del mismo y los caracteres relacionados con el pardeamiento en el mismo material. Se determinó que la accesión de S. insanum accession INS2 presenta valores altamente significativos para los compuestos fenólicos totales y el contenido de ácido clorogénico. Se encontraron efectos significativos para la aptitud combinatoria específica y general para los caracteres bioquímicos estudiados. De manera similar a lo encontrado anteriormente, la distancia genética entre los padres no fue útil para predecir el comportamiento de los híbridos.Finalmente, en el sexto capítulo, hemos desarrollado un protocolo de agroinfiltración para la expresión transitoria de un gen en el fruto de la berenjena utilizando GUS; Vector pCAMBIA1304. Posteriormente, para probar la utilidad del protocolo, utilizamos la hidroxicinamoil CoA-quinato transferasa (SmHQT) de berenjena, que es la enzima central estudiada para aumentar el contenido de ácido clorogénico, en la construcción del gen con el promotor específico en un vector de transformación de plantas (pBIN19). Además, en nuestro casete, también coexpresamos la proteína P19 del Tomato bushy stunt virus para sobreexpresar la proteína. En esta tesis proporcionamos información sobre los parientes sil / [CA] En el primer capítol, es va caracteritzar una col·lecció de sis accessions d'albergínies, 21 accessions de 12 espècies silvestres i 45 híbrids interespecífics d'albergínia amb espècies silvestres utilitzant 27 descriptors morfològics i 20 descriptors morfomètrics de fruits basats en l'eina fenómica Tomato Analyzer. Observarem diferències significatives entre els tres grups, amb híbrids que mostren valors intermedis per a la majoria dels descriptors. Les espècies silvestres van mostrar una àmplia diversitat per a ampliar la base genètica de l'albergínia.En el segon capítol, el mateix material es va caracteritzar per al contingut en compostos fenòlics del fruit, el color de la polpa de la fruita i els caràcters relacionats amb el pardejament. Mentre que l'àcid fenòlic predominant en l'albergínia cultivada va ser l'àcid clorogènic (> 65.0%), en els parents silvestres va ser de menys del 50% de l'àrea del pic del cromatograma HPLC. Les varietats cultivades van presentar un color de carn més clar i baixa activitat de polifenol oxidasa (PPO). Els híbrids interespecífics van ser intermedis per a tots els caràcters estudiats. Curiosament, no trobem correlacions significatives entre el contingut de compostos fenòlics totals o àcid clorogènic i el pardejament de la polpa de la fruita. En el tercer capítol, realitzem un estudi genètic Línia per Emprovador (L × P) utilitzant dues línies cultivades, una amb citoplasma oriental i una altra occidental amb quatre emprovadors que representen tres espècies silvestres: S. insanum, S. anguivi i S. lichtensteinii. A més, avaluem el material per a 3 descriptors bioquímics, 12 morfològics i 8 de Tomato Analyzer. Trobem diferències significatives per als 23 caràcters estudiats. Els valors més alts per a la component SCA es van determinar en comparació amb la component GCA. En el quart capítol, per a obtindre informació sobre la genètica de caràcters importants en l'albergínia, realitzem el primer estudi genètic detallat de l'albergínia amb 10 genotips diversos d'albergínia, entre ells una accessió de S. insanum, mentre que la resta van ser varietats tradicionals amb forma del fruit molt diversa. Quan es van creuar en un disseny de mitjà dial·lel, els 10 pares van produir un conjunt de 45 híbrids. Avaluem pares i híbrids per a 14 descriptors de característiques morfològiques i 14 característiques morfomètriques del fruit. També determinem les distàncies genètiques utilitzant 7,335 marcadors de SNP polimòrfics. En l'estudi de l'herència dels caràcters morfològics importants per al desenvolupament d'híbrids en albergínia, es va trobar que la distància genètica entre els pares té un valor limitat per a predir el rendiment dels híbrids en aquest cultiu. Es va determinar que l'accessió de S. insanum accessió INS2 presenta valors altament significatius per als compostos fenòlics totals i el contingut d'àcid clorogènic. Es van trobar efectes significatius per a l'aptitud combinatòria específica i general per als caràcters bioquímics estudiats. De manera similar a l'oposat anteriorment, la distància genètica entre els pares no va ser útil per a predir el comportament dels híbrids. Finalment, en el sisé capítol, hem desenvolupat un protocol d'agroinfiltració per a l'expressió transitòria d'un gen en el fruit de l'albergínia utilitzant GUS; Vector pCAMBIA1304. Posteriorment, per a provar la utilitat del protocol, utilitzem la hidroxicinamoil CoA-quinat transferasa (SmHQT) d'albergínia, que és l'enzim central estudiat per a augmentar el contingut d'àcid clorogènic, en la construcció del gen amb el promotor específic en un vector de transformació de plantes (pBIN19). A més, en el nostre casset, també coexpresem la proteïna P19 del Tomato bushy stunt virus per a sobreexpresar la proteïna. En aquesta tesi proporcionem informació sobre els parents silvestres d'albergínies per a caràcters morfològics i bioq / [EN] In the first chapter, a collection of six eggplant accessions, 21 accessions of 12 wild species (the only primary genepool species S. insanum and 11 secondary genepool species) and 45 interspecific hybrids of eggplant with wild species were characterized using 27 morphological descriptors and 20 fruit morphometric descriptors based on the phenomics tool Tomato Analyzer. We observed significant differences among the three groups with hybrids showing intermediate values for most of the descriptors. The wild species showed an extensive diversity for broadening the genetic base of eggplant. In the second chapter, the same material was characterized for the fruit phenolics, fruit flesh colour and browning related traits. Wild relatives showed greater variation for the fruit phenolics than cultivated eggplant. While, the predominant phenolic acid in cultivated eggplant was chlorogenic acid (>65.0%), in the wild relatives it was less 50% of HPLC chromatogram peak area. Cultivated varieties had lighter flesh colour and low polyphenol oxidase (PPO) activity. The interspecific hybrids were found to be intermediate for all the characters studied. Interestingly, we found no significant correlations between total phenolics or chlorogenic acid contents and fruit flesh browning. In the third chapter, we performed a Line by Tester (L ×T) genetic study using two cultivated lines one with oriental and another with occidental cytoplasm along with four testers representing three wild species namely, S. insanum, S.anguivi, and S. lichtensteinii. Further, we evaluated the material for 3 biochemical, 12 morphological and 8 Tomato Analyzer based descriptors. We found a significant amount of variation for all the 23 traits studied. The higher values for the SCA component were determined as compared to the GCA component. Overall, the secondary genepool testers were better for the biochemical traits improvement whereas, the primary genepool species was better for the morphological traits. In the fourth chapter, in order to gain information regarding the genetics of important traits in eggplant, we performed the first detailed genetic study of eggplant with 10 diverse eggplant genotypes among them an S. insanum accession, while the remaining nine were highly diverse shaped fruits of popular eggplant cultivars. When crossed in a half-diallel matting design 10 parents produced a set of 45 hybrids. We evaluated parents and hybrids for 14 morphological and 14 fruit morphometric traits descriptors. We also determined the genetic distances using 7,335 polymorphic SNP markers. In the study of the genetics of important morphological traits for hybrid development in eggplant, we found that genetic distance among parents had limited value to predict hybrid performance in this crop. Likewise, in the fifth chapter, we studied the fruit phenolics, fruit flesh colour and browning related traits in the same material. We determined that S. insanum accession INS2 displayed values highly significant for the total phenolics and CGA content. Significant specific and general combining ability effects were found for the biochemical traits studied. Similarly, the genetic distance among parents was nonsignificant to predict hybrids performance. Finally, in the sixth chapter, we have developed an agroinfiltration protocol for the transient expression of a gene in the eggplant fruit using GUS bearing; pCAMBIA1304 vector. Thereafter, to prove the usefulness of the protocol, we have used the eggplant hydroxycinnamoyl CoA-quinate transferase (SmHQT), which is the central enzyme studied to increase the chlorogenic acid content, in a gene construct with the specific promoter in a plant transformation vector (pBIN19). Also, in our cassette, we also co-expressed the P19 protein of Tomato bushy stunt virus to overexpress the protein. Overall, in this thesis, we have provided information regarding the wild relatives of eggplants from a morphological and biochemical prospective. / Kaushik, P. (2019). Application of Conventional, Biotechnological and Genomics Approaches for Eggplant (Solanum melongena.L). Breeding with a Focus on Bioactive Phenolics [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/122295 / Compendio

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Potato cyst nematodes (PCN), Globodera rostochiensis and G. pallida, are economically important pests of potato (Solanum tuberosum) crops in potato growing regions worldwide. Integrated management is under threat, with effective nematicides increasingly being withdrawn on environmental and health grounds. Alternative strategies are urgently needed and trap cropping could be one of them. The non-tuber-bearing Solanum sisymbriifolium is regarded as an effective trap crop for PCN with strong hatching ability and immunity to PCN infection and has been used in the UK and The Netherlands. However, its mode of action is unknown. In order to shed light on the mode of action so that a novel control strategy could be identified, the interactions between G. pallida and S. sisymbriifolium were investigated using in vitro bioassays. In choice assays, G. pallida J2s were equally attracted to the roots of S. sisymbriifolium and to those of S. tuberosum. However, potato root diffusate (PRD), which is routinely used to induce PCN hatch, failed to attract G. pallida J2s in chemotaxis bioassays, indicating hatching factors (HFs) and soluble compounds present in PRD are not involved in attraction of G. pallida J2s to potato roots. The J2s invaded the roots of S. sisymbriifolium in large numbers but failed to develop further. To facilitate continuous observation of nematode development, a novel in vitro method was devised with the use of Pluronic F-127, which requires no sterilisation, and the life cycle of G. pallida was successfully observed in S. tuberosum roots. Quantitative real-time polymerase chain reaction analyses of defence related genes of S. tuberosum and S. sisymbriifolium infected with G. pallida revealed up-regulation of the chitinase gene (ChtC 2.1) at 3 days post inoculation in S. sisymbriifolium but not in S. tuberosum. Electrospray ionisation-mass spectrometry analyses of root exudate extracts of the two Solanum species and subsequent bioassay-guided fractionation showed that the HF of S. sisymbriifolium differs from that of S. tuberosum. Previously, attention had been solely paid to the hatching ability of the root exudate of S. sisymbriifolium, but this study revealed for the first time that the aerial part extract possesses a significant hatching ability.

La tomate (Solanum lycopersicum) est l’espèce modèle pour l’étude des fruits charnus. Le développement du fruit de tomate a été décrit comme une succession de phases de développement distinctes, dans lesquelles les régulations hormonales jouent un rôle crucial. Afin de mieux comprendre la régulation du développement du fruit de tomate, des approches ciblées sur des catégories particulières de protéines régulatrices (F-Box) ainsi que des approches non ciblées de génomique fonctionnelle ont été menées dans le laboratoire. L'objectif de cette thèse est de contribuer à la validation fonctionnelle de gènes candidats mis en évidence dans ces travaux: trois protéines à F-Box (SlFB2, SlFB11 et SlFB24) et deux facteurs de transcription (SlHAT22 et SlTGA2.1). Au cours de cette thèse, la caractérisation de lignées RNAi précédemment générés pour les protéines SlFB2, SlFB11 et SlFB24 a été poursuivie. Ce travail n’a pas permis pas de conclure sur le rôle de ces protéines à F-Box dans le développement du fruit de tomate. Mais elle a permis d'isoler un mutant d'insertion dans une lignée RNAi SlFB2, spécifiquement affecté au niveau des fruits. Ce mutant est caractérisé par l'absence de développement de tissu loculaire, ce qui entraine une altération de la forme des graines, ainsi qu'une augmentation de la fermeté du fruit. Le site d'insertion de l'ADN-T dans ce mutant n'est pas encore identifié. En outre, la caractérisation des lignées RNAi SlFB11, a permis de proposer l'implication de cette protéine dans la régulation des méristèmes apicaux et floraux. La caractérisation fonctionnelle de SlHAT22 et SlTGA2.1 a été initiée par la génération de lignées transgéniques présentant une altération du niveau d'expression de ces facteurs de transcription (sur-expression et RNAi) ou une altération de la fonction du facteur de transcription par utilisation de la technologie CRES-T. La caractérisation phénotypique des lignées transgéniques, ainsi que des analyses métaboliques préliminaires ont révélé que SlTGA2.1 et SlHAT22 sont impliqués dans des processus de régulation au cours du développement précoce du fruit de tomate. En outre, ils ont également suggéré que SlTGA2.1 est impliqué dans la régulation du murissement du fruit de tomate. / Tomato (Solanum lycopersicum) is a model species for the study of fleshy fruits. Tomato fruit development has been described as a sequence of distinct developmental phases where different hormones play crucial regulatory roles. To further gain insight into the regulation of tomato fruit development, targeted approaches focused on particular classes of regulatory proteins (F-Box) as well as global functional genomics approaches were undertaken in the laboratory. The aim of this thesis was to contribute to the functional validation of candidate genes isolated from such approaches: three F-Box proteins (SlFB2, SlFB11 and SlFB24) and two transcription factors (SlHAT22 and SlTGA2.1).During this PhD thesis, the characterization of SlFB2, SlFB11 and SlFB24 RNAi lines previously generated was pursued. This work does not allow to conclude about the role of these F-Box in tomato fruit development. But it allowed to isolate an insertion mutant in a SlFB2 RNAi line, specifically affected at the fruit level. This mutant is characterized by the absence of locular tissue development and subsequent alteration of seed shape, as well as by an increase in fruit firmness. The insertion site of the T-DNA in this mutant is not yet identified. In addition, characterization of SlFB11 RNAi lines, allowed to propose the implication of this protein in the regulation of the shoot apical and floral meristems.Functional characterization of SlHAT22 and SlTGA2.1 was initiated by the generation of transgenic lines carrying an alteration of the transcription factor (TF) expression level (OE and RNAi lines) or with an alteration of TFs function using the CRES-T technology. Phenotypical characterization of the transgenic lines, together with preliminary metabolic analyses revealed that SlTGA2.1 and SlHAT22 are implicated in regulatory processes during tomato early fruit development. In addition, they also suggested that SlTGA2.1 is involved in the regulation of tomato fruit ripening.

Le mildiou de la pomme de terre, causé par l'agent pathogène Phytophthora infestans est l'une des maladies les plus préjudiciables de la culture. Jusqu'à présent, la lutte chimique reste le moyen de contrôle le plus utilisé pour la maîtriser, classant la pomme de terre au premier rang en termes d'Indices de Fréquence de Traitement en grande culture. Par ailleurs, l'utilisation de variétés résistantes, comportant notamment des gènes de résistances spécifiques, a également démontré son efficacité pour limiter les dégâts engendrés par cette maladie. Mais leur efficacité est peu durable avec une durée moyenne de 4 ans avant l'apparition du phénomène de contournement par les isolats plus virulents. Il est donc nécessaire de développer des stratégies de contrôle de la maladie en combinant un ensemble d'approches génétiques, culturales, physiques, et chimiques afin de satisfaire au mieux les objectifs agronomiques, environnementaux et socio-économiques. Un modèle, nommé SIPPOM (Simulator for Integrated Pathogen POpulation Management), avait été développé dans le cas de la gestion durable du phoma du colza. Le présent travail a consisté à adapter la structure générique de SIPPOM au cas de la gestion intégrée du mildiou de la pomme de terre en développant des modules spécifiques à partir de modèles préexistants ou développés spécifiquement. Un modèle de culture (Spudgro), un modèle épidémiologique (Guntz-Divoux / Milsol), un modèle de nuisibilité (modèle de Shtienberg) et une fonction de dispersion (modèle de Scherm) ont été identifiés dans la littérature, adaptés et intégrés dans la structure générique de SIPPOM. Cet ensemble de modules a donné lieu à la réalisation du premier prototype opérationnel de la version informatisée de SIPPOM-de-terre sur la plate-forme de modélisation RECORD. Un nouveau modèle, appelé VOLPONE, a été développé pour représenter les dynamiques de repousses de pomme de terre dans une parcelle et sur un tas de déchets. Ce modèle permet de représenter les sources d'inoculum primaire à l'échelle du territoire sous l'influence du climat et des pratiques agricoles. La qualité prédictive du modèle de nuisibilité a été estimée à partir d'un jeu de données expérimentales générées au cours du travail de thèse et s'est montrée correcte. Des exemples de résultats issus de simulations réalisées à l'aide du premier prototype montrent la capacité du modèle, en fonction d'une combinaison de pratiques culturales et de conditions climatiques variables, à prédire la dynamique épidémique de la maladie, ainsi que les dégâts et dommages associés à l'échelle de la parcelle ou sur un parcellaire simple. Néanmoins, d'autres tâches restent à accomplir afin de disposer d'une version achevée de SIPPOM-de-terre sur la plate-forme RECORD. Le travail réalisé illustre la généricité du modèle d'origine SIPPOM et contribue à la production de connaissances et de références permettant l'optimisation d'outils existants tels Mileos®. Les avancées réalisées pourront contribuer à la conception de stratégies de gestion intégrée, collectives et durables pour cette maladie. / Potato late blight caused by the pathogen Phytophthora infestans is one of the most dreaded diseases to this culture. So far, chemical control is the most widespread method used against the pathogen. In turn, the amount of fungicides used to control this disease makes potato the crop with the highest Treatment Frequency Index of all arable crops. The use of resistant varieties, including specific resistance genes, has also demonstrated an effective limitation of injuries that this disease can cause. Still, specific resistances lack durability as there is an average span of 4 years before the emergence of a resistance breakdown phenomenon by more virulent strains. It is therefore necessary to develop control strategies that combine a set of methods (genetic, cultural, physical and chemical) to meet agronomic, environmental and socio-economic criteria. A model named SIPPOM (Simulator for Integrated Pathogen POpulation Management), was developed to address these issues for the control of phoma stem canker on oilseed rape. The present work consisted in adapting the generic structure of SIPPOM for the integrated control of potato late blight by designing specific sub-models using existing models or specifically developed models. A crop model (Spudgro), an epidemiologic model (Guntz-Divoux/Milsol), a damage model (Shtienberg's model) and a dispersal function (Scherm's model) were selected in the literature, adapted, and embedded in SIPPOM's generic structure. This set of sub-models led to an operational prototype of SIPPOM-de-terre under the RECORD modelling platform. A new model, named VOLPONE, was designed to simulate potato volunteer dynamics in a field or on a waste pile. This model permits to simulate inoculum sources at the territory scale under the influence of climate and cropping practices. The predictive quality of the damage model was assessed with data generated in specific field experimentations and proved to be good. Simulation examples produced with the first prototype of SIPPOM-de-terre illustrate the capacity of the model to represent epidemiological dynamics at the field scale or at a small regional scale under the influence of cropping practices and climate. However, further work is required before to totally achieve the design and the implementation of SIPPOM-de-terre under the modelling platform RECORD. The conducted study illustrates the genericity of the model SIPPOM and produced knowledge, references and tools for the integrated management of the disease. The outputs of this work will help design integrated, collective and durable control strategies of potato late blight.

La transformation de la paroi de l’ovaire en un péricarpe charnu implique une coordination entre les divisions cellulaires et l’expansion cellulaire. Des données considérables sur le développement et la maturation du fruit de tomate ont été établies, mais la coordination des divisions cellulaires, de l’expansion cellulaire et de l’endoréduplication durant la mise à fruit ainsi que durant la croissance du fruit de tomate reste grossièrement caractérisée au sein du péricarpe et de nombreuses questions ne sont pas résolues : comment ces deux processus sont-ils régulés et coordonnés durant le développement du fruit d’un point de vue cellulaire? Quand commence l’endoréduplication dans les tissus du fruit et quelle est sa fonction? La première partie de ce mémoire concerne la coordination des divisions cellulaires et de l’expansion cellulaire durant la fin du développement de l’ovaire et le début du développement du fruit. Une différenciation précoce des assises cellulaires composant la paroi de l’ovaire puis le péricarpe a été démontrée. Les divisions cellulaires se font principalement au sein de l’épiderme externe et montrent une synchronisation partielle tandis que l’expansion cellulaire se fait principalement dans le mésocarpe. L’endoréduplication semble être initiée avant l’anthèse. La deuxième partie est consacrée à l’analyse du RNA-seq nucléaire en fonction de quatre niveaux de ploïdie (4, 8, 16 et 32C). La majorité des gènes montrent une augmentation proportionnelle de leurs expressions en fonction des niveaux de ploïdie. Cependant, certains gènes révèlent une surexpression ou une sous-expression en fonction des niveaux de ploïdies. / The transformation of the ovary wall into a fleshy pericarp involves a coordinated pattern of cell division and cell expansion. Considerable data have been reported on tomato fruit development and ripening, but the pattern of cell division, cell expansion and endoreduplication at the tomato fruit set and during fruit growth remains grossly appreciated at the whole pericarp level and many questions are not yet resolved: How are cell division and cell expansion coordinated in tomato fruit a cellular level and according to developmental time? When does endoreduplication begin in fruit tissues and what is its function? The first part of this deals with the coordination of cell division and cell expansion during the end of tomato ovary development and the beginning of fruit growth. Evidence for early differentiation of cell layers in the ovary wall and then in fruit pericarp are presented. Cell division happens mainly in the external epidermis and shows partial synchronization, whereas cell expansion happens mostly in mesocarp cell layers. Endoreduplication is initiated as soon as before anthesis. The second part of this work is devoted to RNA-seq based transcriptome profiling of pericarp nuclei which have been sorted according to four ploidy levels (4, 8, 16 and 32C). We demonstrate that the expression of most of the pericarp-expressed genes shows a proportional increase according to ploidy level, on a nuclear basis. However, a significant amount of genes has been identified as over-expressed or under-expressed according to ploidy level.

O fungo Moniliophthora perniciosa, fitopatógeno responsável pela doença vassoura-de-bruxa no cacaueiro, é responsável por reduzir a produtividade de frutos de cacau nas Américas. Visto que os métodos atuais de controle desta doença não são eficientes, faz-se necessário a busca por compostos para seu controle químico. Os produtos naturais têm contribuído na síntese de novos pesticidas e a família Solanaceae é reconhecida como uma fonte promissora de compostos antifúngicos. Neste contexto, este trabalho teve por objetivo explorar o potencial biológico e químico de metabólitos secundários produzidos por plantas da família Solanaceae com atividade antifúngica à M. perniciosa. Para tanto, extratos aquosos de folhas de dez plantas foram avaliados quanto sua atividade inibitória a M. perniciosa e, observou-se maior atividade antifúngica pelos extratos de Cestrum nocturnum, Solanum seaforthianum e Brunfelsia uniflora. Destas, as plantas S. seaforthianum e B. uniflora mostram-se como promissoras e foram selecionadas para isolamento de seus compostos ativos. A partir do extrato aquoso de S. seaforthianum obteve-se duas frações constituídas por saponinas furostanas, a mistura (25R)-karatavioside C / (25R)-purpureagitoside, com CI50 de 40,9 &mu;g mL-1, e a mistura (25R)-timosaponin H1/ (25R)-uttroside B, com CI50 de 22,3 &mu;g mL-1 ao crescimento micelial. E, a partir do extrato aquoso de B. uniflora obteve-se três compostos da classe das saponinas espirostanas, o composto karatavioside A e os compostos parcialmente identificados BuM8i4, com aglicona identificada como (25R)-yucagenina, e BuM8i6, com aglicona identificada como (25R)-diosgenina, todos com CI50.< 15,63 &mu;g mL-1 ao crescimento micelial de M. perniciosa. Todos os compostos caracterizados neste trabalho estão sendo relatados pela primeira vez a partir das plantas estudas e também para atividade antifúngica. / The fungal phytopathogen Moniliophthora perniciosa is the causal agent of Witches\' Broom disease and the main responsible by limiting cacao production in Americas. The current disease control methods are not efficient and new antifungal compounds are necessary to the chemical management. Natural products has contributed to the development of natural product-based pesticides and the Solanaceae plants are known as a promising source of antifungal compounds. In this context, this work aimed to explore the biological and chemical potential of secondary metabolites produced by Solanaceae plants with antifungal activity against M. perniciosa. For this, antifungal activity of water extracts from leaves of ten plants was evaluated against M. perniciosa and the best results were observed by using extracts from Cestrum nocturnum, Solanum seaforthianum e Brunfelsia uniflora. Among these plants, S. seaforthianum and B. uniflora were selected for active compounds isolation due to their promising characteristics. From water extract of S. seaforthianum two furostan saponin fractions were obtained, the mixture (25R)-karatavioside C / (25R)- purpureagitoside, with IC50 of 40,9 &mu;g mL-1, and the mixture (25R)-timosaponin H1/ (25R)-uttroside B, with IC50 of 22,3 &mu;g mL-1 to the mycelial growth. From water extract of B. uniflora three spirostan saponin compounds were obtained, the compound karatavioside A and the compounds partially identified BuM8i4, with aglycone identified as (25R)-yucagenina, and BuM8i6, with aglycone identified as (25R)- diosgenina, all these compounds showed IC50< 15,63 &mu;g mL-1 to the mycelial growth of M. perniciosa. All compounds characterized in this study were obtained for the first time from these plants and also described about their antifungal activity.

Solanum lycocarpum A. St.-Hil. (Solanaceae Solanum), popular lobeira, é espécie arbustiva nativa e característica do Cerrado brasileiro. Seus frutos são empregados na medicina caseira como diurética, calmante, anti-espasmódica, antiofídica e antiepilética. As espécies do gênero Solanum são produtoras de heterosídeos alcaloídicos, os quais possuem atividade antitumoral, incluindo-se anticâncer de pele. O câncer de pele tem preocupado as autoridades no mundo com os crescentes índices atuais e, por esse motivo, a busca por novos ativos anticâncer é primordial. Dos frutos da lobeira, pode-se produzir extrato alcaloídico, o qual se constitui de uma mistura de glicoalcalóides composta majoritariamente de solasonina e solamargina. Tais compostos têm sido estudados quanto ao potencial anticancerígeno, são relativamente seletivos para células tumorais e, em consequência, apresentam baixa toxicidade às células sadias. Por estes motivos, o potencial antitumoral desta espécie deve ser investigado. Para isso, os frutos foram coletados, secos e triturados. A droga vegetal resultante foi submetida à extração tipo ácido-base para obtenção do extrato alcaloídico. O método analítico empregado para quantificação dos heterosídeos alcaloídicos foi desenvolvida e validada em CLAE-UV. A partir do extrato alcaloídico padronizado, foram desenvolvidas formulações para uso tópico. Estas formulações foram avaliadas por meio de ensaios de permeação e retenção dérmica in vitro, cujo método analítico para quantificação de solasonina e solamargina foi validado. Para escolha da formulação mais promissora para ensaio anticâncer in vivo, as formulações de melhor desempenho in vitro foram avaliadas quanto à retenção dérmica in vivo em camundongos hairless. A formulação B, contendo 1% de extrato alcaloídico, 5% de monoleína (monoleato de glicerol), 5% de propilenoglicol em base de gel de Natrosol (pH 6,5), apresentou retenção dérmica total adequada para futuros ensaios anticâncer de pele in vivo. / Solanum lycocarpum A. St.-Hil. (Solanaceae - Solanum), popular wolf-fruit is a native shrub species and characteristic of the Brazilian Cerrado vegetation. Its fruits are used in folk medicine as diuretic, sedative, anti-spasmodic, antiepileptic and antiophidic. This species belongs to the genus Solanum, known to produce alkaloid heterosides, which possess antitumor activity, including skin anticancer. Skin cancer has concerned the authorities in the world because of its growing rates. Therefore, the search for new active anticancer is paramount. The wolf-fruit furnish an alkaloidic extract, which is rich in a mixture of heteroside alkaloids and composed mostly of solasonine and solamargine. Such compounds have been studied as potential anticancer, because they are relatively selective for tumor cells and, consequently, have low toxicity to health cell. For these reasons, the antitumor potential of this species should be investigated. For that, the fruits were collected, dried and crushed. The dried plant biomass was submitted to acid-base extraction to obtain the alkaloidic extract. The analytical method for the quantitation of the alkaloids in both crude alkaloid extract and plant biomass was developed and validated by HPLC-UV. From the alkaloidic extract, formulations were developed for topical use. These formulations were evaluated by in vitro tests of skin permeation and retention in Franz diffusion cell, for which an analytical method for quantifying solasonine and solamargine was developed, as well. The selection of the most promising formulation for anticancer assay in vivo was based in both, the permeation performance of the formulation in vitro and its skin retention in vivo in hairless health mice. The formulation B, containing crude alkaloid extract 1%, Monoolein 5%, propyleneglycol 5% in Natrosol gel (pH 6.5), showed total dermal retention, which is suitable for future trials of skin anticancer in animal models.

O trabalho teve como objetivo avaliar a resposta à produção total, comercial e por classes de tamanho; ao teor de matéria seca e aos defeitos fisiológicos mancha-chocolate, coração-oco e coração-negro dos tubérculos do clone de batata IAC 2.5, submetido a três doses dos fertilizantes minerais 04-14-08 (N-P2O5-K2O) e sulfato de amônio, complementadas ou não com um composto orgânico de marca comercial Provaso. O experimento foi conduzido de outubro de 2012 a fevereiro de 2013, na Unidade de Pesquisa e Desenvolvimento de Itararé, da Agência Paulista de Tecnologia dos Agronegócios (APTA), em Itararé, SP, nas coordenadas geográficas 24º16\'06\"S e 49º12\'19\"W e altitude de 1200 m. Foi adotado o delineamento em blocos ao acaso em arranjo fatorial 3x2, com quatro repetições. Os tratamentos consistiram em três doses de 04-14-08 no plantio e de sulfato de amônio em cobertura, sendo respectivamente: 2150 kg ha-1 + 200 kg ha-1; 3225 kg ha-1 + 100 kg ha-1; e 4300 kg ha-1 + 0 kg ha-1, com (5000 kg ha-1) ou sem composto orgânico. As doses de 04-14-08 foram calculadas com base na análise de solo e recomendação de 100, 150 e 200% em P2O5 para a cultura da batata, e para a adubação em cobertura, adotou-se respectivamente as doses 40, 20 e 0 kg ha-1 de N e 44, 22 e 0 kg ha-1 de S. A parcela experimental foi composta por quatro fileiras de 3,5 m cada, totalizando 40 plantas. Como parcela útil, foram consideradas as 16 plantas das duas fileiras centrais, excetuando as plantas das extremidades, consideradas bordaduras de cabeceira. O espaçamento foi de 0,35 m entre plantas na fileira e de 0,8 m entre as fileiras. Os dados foram submetidos à análise da variância e as médias foram comparadas pelo teste de Tukey (p < 0,10), utilizando o programa estatístico Sisvar. Nas condições em que o experimento foi conduzido, a alta produtividade comercial de tubérculos (52,4 t ha-1) do clone IAC 2.5 apresentada pelo tratamento com 3225 kg ha-1 de 04-14-08 + 5000 kg ha-1 de composto orgânico no plantio e 100 kg ha-1 de sulfato de amônio em cobertura sugere a importância da complementação da matéria orgânica no aumento da eficiência da adubação mineral. / The study aimed to evaluate the response to total production, marketable production and production by tuber size classes; dry matter content and internal physiological disorders internal brown spot, hollow heart and black heart of potato tubers clone IAC 2.5 subjected to three rates of mineral fertilizers 04-14-08 (N-P2O5- K2O) and ammonium sulphate, supplemented or not with an organic compost trademark Provaso. The trial was conducted from October 2012 to February 2013, at the Research & Development Unity of Agência Paulista de Tecnologia dos Agronegócios (APTA), in Itararé, SP, Brazil (lat. 24º16\'06\'\'S; long. 49º12\'19\'\'W), and altitude of 1200 m. The treatments were arranged in a 3x2 factorial scheme in a completely randomized block design, with four replications. The treatments consisted of three rates of 04-14-08 at planting and ammonium sulphate in dressing fertilization, follow respectively: 2150 kg ha-1 + 200 kg ha-1; 3225 kg ha-1 + 100 kg ha-1; e 4300 kg ha-1 + 0 kg ha-1, with (5000 kg ha-1) or without organic compost. 04-14-08 rates were calculated based on the soil analysis and 100, 150 and 200% on P2O5 recommendation for the potato crop, and for dressing fertilization, was adopted quantities respectively 40, 20 and 0 kg ha-1 of N and 44, 22 and 0 kg ha-1 of S. The experimental plot consisted of four rows of 3.5 m each, for a total of 40 plants. Useful plot was the 16 plants from central rows. The spacing was 0.35 m between plants in the row and 0.8 m between rows. The data were submitted to analysis of variance and treatment means were compared by Tukey test (p < 0,10) using the statistical program Sisvar. Taking in account the conditions in wich the trial was conducted, a high yield of marketable tubers (52.4 t ha-1) for the clone IAC 2.5 by 3225 kg ha-1 of 04-14-08 + 5000 kg ha-1 of organic compost in planting and 100 kg ha-1 of ammonium sulphate in dressing fertilization suggests the importance of organic matter complementing in increasing the efficiency of mineral fertilization.