Nos frutos, a cor é fundamental para a aceitabilidade inicial do consumidor, especialmente para os chamados \"pequenos frutos\" como morangos e framboesas, sendo um critério tradicional para a apreciação e seleção do estado de maturação. Em framboesas (Rubus idaeus), as pesquisas têm focado a síntese de antocianinas, os pigmentos mais abundantes nesta espécie. Seus precursores provem da via de biossíntese dos fenilpropanóides, porém os fatores que regulam a atividade dessa via metabólica ainda são pouco conhecidos. Sabe-se que alguns hormônios como o etileno, o ácido indol-3-acético (AIA), o ácido abscísico (ABA) e o metil-jasmonato (MJ) estão relacionados à inibição da síntese ou aumento no acúmulo destes flavonóides. Neste contexto, o objetivo do trabalho é avaliar o efeito destes fatores hormonais sobre a biossíntese de antocianinas em framboesas da cultivar (cv.) Autumn Bliss. Foram avaliados os efeitos dos hormônios sobre as características físico-químicas e fisiológicas dos frutos (açúcares solúveis, ácidos orgânicos, respiração, síntese de etileno, cor), conteúdo de compostos fenólicos e flavonóides. Também foram quantificados os níveis de AIA livre, ABA e MJ e a expressão relativa das enzimas antocianidina sintase (ANS), dihidroflavonol redutase (DFR) e do fator de transcrição MYB10, regulador da transcrição de genes da via de biossíntese de antocianinas em framboesas. As alterações mais significativas foram observadas no tratamento com MJ, para o qual os frutos apresentaram acentuada produção de etileno (sem efeito sobre a respiração) e também na cor, mais acentuada nos frutos maduros deste grupo em comparação com os frutos do grupo controle. O MJ também induziu aumento nos níveis de expressão relativa de MYB, ANS e DFR, nos dois primeiros dias pós-colheita. Outro grupo que se destacou foi o tratado com AIA por apresentar inibição na síntese de antocianinas e compostos fenólicos, a qual foi correlacionada à inibição da transcrição do MYB e ANS. O ABA por sua vez, inibiu o acúmulo de transcritos da DFR, de forma correlata ao menor acúmulo de antocianinas. Exceto pelo ACC, que não produziu efeitos significativos em nenhum dos parâmetros avaliados, os resultados indicam que o metabolismo de antocianinas deve ser regulado por múltiplos sinais hormonais. / The color is fundamental for the fruits, for the initial acceptability of the consumers, is a factor that influence the appreciation and selection of the maturity stage, especially for berries, like strawberries and raspberries. In raspberries (Rubus idaeus), the research have focused on the anthocyanin biosynthesis, the pigments most frequent on this species. Their precursors came from the phenylpropanoid pathway but the factors that regulate this metabolic pathway remain unclear. There are evidences that some phytohormones as ethylene, indole-3-acetic acid (IAA) abscisic acid (ABA) and methyl jasmonate (MJ) are related to the inhibition or increase of these flavonoids on fruits. In this context, the aim of this study is to evaluate the effect of these phytohormones on the anthocyanin pathway in raspberries Autumn Bliss. It was evaluated the effects of these phytohormones in some physico-chemical and physiologic characteristics of these fruits (soluble sugars, organic acids, respiration, ethylene biosynthesis, color), phenolic compounds and flavonoid content. Also, it was quantified the auxin, ABA and MJ levels, and the relative transcript levels of anthocyanidin synthase (ANS), dihydroflavonol 4-reductase (DFR) and the transcriptional factor MYB10, a transcriptional regulator of the anthocyanin pathway in raspberries. The most significant alterations were observed on MJ treatment, in which the ethylene production in berries reach the highest levels (without effect on the respiration) and the most intense red color on ripe fruits in comparison with the control group. MJ also induced the increase on relative expression of MYB, ANS and DFR two days after harvest prior to the anthocyanin accumulation. Other noticeable effect was detected in the IAA group in which anthocyanin synthesis and phenolic compound content were inhibited and related to the inhibition of MYB and ANS transcription. ABA inhibited the DFR transcript accumulation, related to the delay in anthocyanins synthesis in relation to control group. Except by ACC, that did not have significant effects in any of the evaluated parameters, the results suggest that the anthocyanin metabolism might be regulate for multiple hormonal signs.

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Secondary metabolism in plants produces numerous compounds with wide-ranging activities, including the antineoplastic compound taxol and related taxanes. The biotechnological production of taxol has so far been based on empirical studies. The aim of the present work has been to study how the factors that enhance taxane production affect the metabolic profiles and gene expression in productive cells. As a consequence of this work, new potential candidates have been obtained for unknown taxane biosynthetic genes, some bottle-neck steps of taxane biosynthesis (in vitro and in silico) have been identified and a master regulator, not only for taxane biosynthesis, but also for other secondary metabolism routes, has been characterized. Coronatine, a powerful and less harmful elicitor than methyl jasmonate, has been successfully assayed and found to increase taxane production. In the different studies of this work, the expression level of genes that participate in taxol biosynthesis has been determined, clarifying their involvement in the production of this anti-cancer agent. / El metabolismo secundario de las plantas produce numerosos compuestos con un amplio rango de actividades, entre los que se encuentra el compuesto antineoplásico taxol y los taxanos relacionados, la producción biotecnológica del cual se basa en estudios empíricos. El objetivo de este trabajo ha sido estudiar como los factores que incrementan la producción de taxanos afectan los perfiles metabólicos y la expresión génica en los cultivos. De esta manera se han identificado nuevos genes candidatos que codifican para los genes desconocidos de la biosíntesis, algunos pasos limitantes de ésta y se ha caracterizado un regulador relacionado con el metabolismo secundario. Se ha ensayado la coronatina, un elicitor más eficiente para mejorar la producción de taxanos y menos dañino que el jasmonato de metilo. En los diferentes ensayos de este trabajo han sido determinados los niveles de expresión de genes que participan en la biosíntesis de taxol, ayudando a comprender su papel en la producción de este anticancerígeno.

The chemical elicitor methyl jasmonate (MeJA) could replace insecticides in Europe and Asia for protection of young conifers against the pine weevil (Hylobius abietis). This thesis mainly focuses on the effects of MeJA treatment on chemical defenses of conifers from seedling batches with documented field resistance. Tissues of three pine species and one spruce species, with various treatments, were here extracted in hexane, whereafter volatile contents of tissues were separated and analyzed by 2D GC-MS. Induced responses of seedlings of Maritime pine (Pinus pinaster) and Monterey pine (Pinus radiata) to the folivore pine processionary (Thaumetopoea pityocampa), and the phloem-feeder H. abietis, have been studied. Amounts of mono- and sesqui-terpenes (and also non-volatile resin) in conifer tissues (needles and phloem) were less induced by T. pityocampa than by H. abietis. The MeJA-treated seedlings of Scots pine (Pinus sylvestris) changed their composition of phloem monoterpenes (induction of (-)-β-pinene), and were better protected in field than the seedlings of Norway spruce (Picea abies), which increased their total amounts of monoterpenes. Orientation bioassays with H. abietis showed deterrent effects of (-)-β-pinene, (+)-3-carene, (-)-bornyl acetate and 1,8-cineole. Conversely, (-)-α-pinene (induced in P. abies but not in P. sylvestris) was non-deterrent. MeJA-treated seedlings fed on by H. abietis contained higher amounts of the H. abietis antifeedant 2-phenylethanol, in tissues and emissions. Phloem of control seedlings instead induced the (+)-α-pinene, which is one of the enantiomers of the H. abietis attractant α-pinene. In response to MeJA treatment, the relative amounts of the (+)-α-pinene increased in the phloem of  P. radiata, while it decreased in P. pinaster phloem. The preference of H. abietis in the field for P. radiata before P. pinaster may be explained by these changes in enantiomers of the H. abietis attractant α-pinene. / Barrträd är viktiga för skogsindustrin. I Sverige består 57% av ytan av produktiv skogsmark, och av dessa träd utgörs 80% av gran och tall. Som försvar mot insekter och sjukdomar, producerar barrträden kådämnen. För unga barrträd i Europa och Asien är snytbaggen (Hylobius abietis) en allvarlig skadegörare, som orsakar skogsnäringen stora kostnader. För att skydda små gran- och tallplantor före utplanteringen, skulle växthormonet metyljasmonat (MeJA) kunna användas. I denna avhandling har effekter av MeJA-behandling och insektsangrepp undersökts på de två viktigaste barrträden i Sverige, tall och gran (Pinus sylvestris och Picea abies) och på två arter i medelhavsområdet (Pinus radiata och Pinus pinaster). De kemiska försvarsämnena i barrträd har analyserats för att utvärdera effekterna av MeJA-behandling. Vävnader från barrträd med olika behandling har extraherats i organiska lösningsmedel och dofterna från plantorna har insamlats genom fastfas-mikroextraktion. De flyktiga ämnena har separerats och identifierats med hjälp av gaskromatografi och masspektrometri (GC-MS). Dessutom har optiskt aktiva doftämnen separerats med hjälp av en tvådimensionell GC-MS. Plantornas olika försvarsreaktioner har studerats vid angrepp av snytbaggen, som är en floemätare, och av tallfjärilslarv (Thaumetopoea pityocampa), som huvudsakligen äter barr. De lättflyktiga ämnena (mono- och seskviterpener) och de icke-flyktiga ämnena (kådsyror) ökade mer i stam och barr från angrepp av H. abietis än av T. pityocampa. Sammansättning av monoterpener ändrades i de MeJA-behandlade tallplantorna i Sverige, specifikt ökade (-)-β-pinene i stamfloemet. Tallplantorna var bättre skyddade i fält än granplantorna, där den totala mängden monoterpener ökade. (-)-β-Pinene, (+)-3-carene, (-)-bornyl acetate och 1,8-cineole visade sig ha en negativ effekt på snytbaggens orientering mot talldoft i orienteringstester. En av monoterpenerna, (-)-α-pinene, som av MeJA-behandlingen inducerats i granen, men inte i tallen, hade ingen negativ effekt på snytbaggen. MeJA-behandlade tallplantor som angripits av snytbaggen, innehöll större mängder av snytbagge-äthämmaren 2-fenyletanol, både i vävnader och i emissioner. Stam från kontrollplantor inducerade istället (+)-α-pinene, vilken är en av de optiska isomererna av snytbagge-attrahenten α-pinene. För de spanska arterna, som svar på MeJA-behandling, ökade de lättflyktigaste terpenerna i barren hos P. radiata men inte hos P. pinaster, vilken redan innehöll en hög terpenhalt. De relativa mängderna av (+)-α-pinene ökade också i stammen hos P. radiata, medan de minskade i P. pinaster. I fält väljer snytbaggen att gå till P. radiata- framför P. pinaster-plantor, vilket kan vara kopplat till de skillnader i α-pinene-isomerer, vilka här har observerats. / <p>QC 20160304</p>