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Tree peony is known worldwide for its excellent ornamental and medical values, but recent reports that their seeds contain over 40% α-linolenic acid (ALA), an essential fatty acid for humans drew additional interest of biochemists. To understand the key factors that contribute to this rich accumulation of ALA, we carried out a comprehensive study of oil accumulation in developing seeds of nine wild tree peony species. The fatty acid content and composition was highly variable among the nine species; however, we selected a high- (P. rockii) and low-oil (P. lutea) accumulating species for a comparative transcriptome analysis. Similar to other oilseed transcriptomic studies, upregulation of select genes involved in plastidial fatty acid synthesis, and acyl editing, desaturation and triacylglycerol assembly in the endoplasmic reticulum was noted in seeds of P. rockii relative to P. lutea. Also, in association with the ALA content, transcript levels for fatty acid desaturases (SAD, FAD2 and FAD3), which encode for enzymes necessary for polyunsaturated fatty acid synthesis were higher in P. rockii compared to P. lutea. We further showed that the overexpression of PrFAD2 and PrFAD3 in Arabidopsis increased linoleic and α-linolenic acid content, respectively and modulated their final ratio in the seed oil. In conclusion, we identified the key steps that contribute to efficient ALA synthesis and validated the necessary desaturases in P. rockii that are responsible for not only increasing oil content but also modulating 18:2/18:3 ratio in seeds. Together, these results will aid to improve essential fatty acid content in seeds of tree peonies and other crops of agronomic interest.

The increasing need for healthy edible oil has driven us to identify α-linolenic acid (ALA)-rich species and identify key biochemical steps in ALA synthesis. Seeds of tree peony species are rich in unsaturated fatty acid content with > 40% ALA in the total fatty acid. However, fatty acid content and composition is variable among the tree peony germplasm. To this extent, a comparative study was carried out to identify the key genes responsible for differential oil accumulation among nine wild tree peony species. Subsequent to analyzing fatty acid content and composition of the seeds from nine tree peony species, a high- (P. rockii) and low-oil (P. lutea) accumulating species were selected for transcriptome analysis. Gene expression analysis revealed upregulation of select genes involved in plastidial fatty acid synthesis, and acyl editing, desaturation and triacylglycerol assembly in the endoplasmic reticulum in seeds of P. rockii relative to P. lutea. Also, in association with ALA content in seeds, transcript levels for fatty acid desaturases (SAD, FAD2 and FAD3), which encode for enzymes necessary for polyunsaturated fatty acid synthesis were higher in P. rockii compared to P. lutea. Additionally, we showed that the overexpression of PrFAD2 and PrFAD3 in Arabidopsis increased linoleic and α-linolenic acid content, respectively and modulated their final ratio in the seed oil. In conclusion, we identified the key steps that contribute to efficient ALA synthesis and validated the necessary desaturases in P. rockii that are responsible for not only increasing oil content but also modulating 18:2/18:3 ratio in seeds. Together, these results will aid to improve essential fatty acid content in seeds of tree peonies and other crops of agronomic interest.

L‟acide docosahexaénoïque (DHA) s‟accumule principalement dans les membranes cérébrales humaines durant la période périnatale et les 2 premières années de la vie. Optimiser l‟incorporation des acides gras polyinsaturés à longue chaîne (AGPI-LC) n-3 dans le système nerveux central, notamment le DHA, constitue l‟un des objectifs majeurs de la nutrition néonatale et infantile. Ce projet étudie l‟effet de régimes à base de matière grasse laitière anhydre (MGLA) afin d‟améliorer la bioconversion des AGPI n-3 et l‟incorporation du DHA dans le cerveau. L‟autre objectif, au-delà des effets sur les acides gras, est d‟évaluer si l‟impact des apports destinés au jeune n‟exercent pas un effet plus large tels qu‟appréciés au niveau du lipidome du cerveau.Nous avons mis au point un modèle d‟expérimentation animale permettant d‟étudier les effets de mélanges de MGLA et d‟huiles végétales. Des mères sont nourries avec un régime déficient en acide α-linoléique (ALA) (0,4% des acides gras totaux (AGT)) pendant 6 semaines avant accouplement et pendant toute la gestation et la lactation. Après sevrage, quatre-vingts petits rats mâles et femelles ainsi carencés en AGPI n-3 sont séparés en 4 groupes et reçoivent des régimes à 10% de lipides pendant 6 semaines, soit à base d‟huile de palme mélangée avec des huiles vegétales pour un apport à 1,5%ALA (P1), ou 1,5% ALA supplementé avec 0,12%DHA et 0,4% acide arachidonique (ARA) (P2); soit à base de MGLA et d‟huiles végétales apportant 1,5%ALA (B1) ou 2,3%ALA (B2). Les acides gras cérébraux, plasmatiques et érythrocytaires ainsi que le profil lipidomique cérébral sont mesurés, et les résultats sont analysés en statistiques multivariées. Le régime B1 est supérieur aux deux régimes à base d‟huile de palme à 1,5 %ALA, pour restaurer la quantité de DHA du cerveau (augmentation de 14.38%, P < 0.05) ; le régime (B2) présente un bénéfice supplémentaire sur ce paramètre. Les concentrations cérébrales en DHA chez les rats mâles sont significativement plus faibles que chez les femelles en raison des interactions de l‟effet sexe sur l‟effet régime, mais cet effet s‟atténue avec les régimes MGLA ou l‟ajout de DHA préformés dans le régime palme (P2).Nous avons calculé un nouvel indice à l‟aide des profils en acides gras des globules rouges et du plasma pour prédire le contenu en DHA cérébral, et dont la performance est meilleure que celle des indices existants. Pour la première fois, nous avons pu mettre en évidence un effet majeur et inconnu jusqu‟alors, des régimes sur le lipidome du cerveau (analyse des espèces moléculaires de lipides), affectant près de 15% des espèces analysées. Ces changements semblent être liés, entre autres, au métabolisme du cholestérol, des acides gras et des messagers lipidiques.En conclusion, nos données sont susceptibles d„améliorer les formules infantiles. La première utilisation de l‟approche de lipidomique sans a priori que nous avons mise en oeuvre ouvre des perspectives nouvelles en nutrition infantile. / The accretion of docosahexaenoic acid (DHA) in brain membranes mainly occurs around delivery and during the first two years of life. One of the main goals of neonatal nutrition is to optimize the incorporation of n-3 long chain polyinsaturate fatty acids (LC-PUFA) into the central nervous system, including DHA. Our goal was to study the impact of several kinds of diets based on dairy-fat to improve the n-3 LC-PUFA bioconversion and DHA accretion into brain. The other endpoint was to assess if beyond the brain fatty acid profiles, the dietary intakes would bring about a wider effect such as the one that can be appreciated through a lipidomic approach.We compared the nutritional effect of dairy-fat based diets to that of palm-oil based diets in the rat reproductive model. Mother rats were made deficient in α-linoleic acid (ALA) (0.4% of fatty acids (FA) for 6 weeks prior to mating and throughout gestation and lactation. After weaning, the resulting deficient 40 rat pups of either gender were split into 4 groups and received 10% fat diets made with either 1.5%ALA palm oil blend (P1), same added with 0.12% DHA and 0.4% arachidonic acid (ARA) (P2) , 1.5% ALA dairy-fat blend (B1) or 2.5% ALA dairy-fat blend (B2). The brain, red blood cell (RBC) and plasma fatty acid profiles were analyzed and treated using multivariate statistics. B1 was superior to both palm-oil based diets to improve the brain DHA contents (14.4% increase, P < 0.05). B2 brought an additional benefit for this parameter. The brain DHA contents in males were significantly lower than for the female because of a diet x gender interaction. This effect was smoothed with the dairy-fat diets or the palm-oil based diet augmented with preformed DHA.We calculated a new fatty acid index to predict the brain DHA contents, based on the fatty acid profiles measured in RBC and plasma, and which demonstrated a better performance than the existing published index. For the first time, we showed a profound and yet unsuspected effect of diets until now on the brain lipidome (lipids molecular species), affecting about 15% of the features detected. These changes were ascribed to the cholesterol and fatty acid metabolism, and to the lipid messengers, among others.In conclusion, our data appear highly relevant to improve infant formulas. The first use of the lipidomic approach in neonatal nutrition open the paths of new researches in the area of infant nutrition

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-09-05T12:33:17Z No. of bitstreams: 1 arquivototal.pdf: 2400513 bytes, checksum: c311d90765b0ac86d9ce66ac470ae075 (MD5) / Made available in DSpace on 2017-09-05T12:33:17Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2400513 bytes, checksum: c311d90765b0ac86d9ce66ac470ae075 (MD5) Previous issue date: 2016-07-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Functional foods are highlighted for promoving beneficial effects on human health. Among them, chia seed (Salvia hispanica L.) is a natural source of polyunsaturated fatty acids, with high levels of α-linolenic acid, and other important nutritional components, such as vitamins, minerals and antioxidants. Considering the growing interest in the use of chia in human diet and its many benefits reported in the literature, this study determined the physicochemical characteristics and nutritional properties of chia seed grown in southern Brazil. It also investigated the phenolic compounds present in the seed and developed a microcapsule containing chia oil. A review was prepared addressing the main components: lipids, proteins, fibers and phenolic compounds, emphasizing the benefits of the functional properties of chia seed on human health and in the preparation of food products. The characterization of chia seeds grown in Brazil revealed considerable amounts of protein (25,03 g /100 g), lipids (34,41 g /100 g) and fibers (36,50 g/100 g), as well high amounts of essential amino acids and nutritionally important minerals, such as calcium, potassium, zinc and selenium. The fatty acid profile of chia oil revealed a high degree of unsaturation, consisting of oleic, linoleic and, predominantly, α-linolenic acids (65,61 g/100 g). The chia oil showed a high tocopherol content (586,78 mg/g), with γ-tocopherol isomer representing more than 90% of all isomers and phenolic compounds such as myricetin, quercetin, and kampferol synaptic acid. The efficiency of the extraction of phenolic compounds from seeds of chia was determined utilizing the statistical tool of mixture planning, simplex-lattice design. The solvents used were acetone, ethanol and water, and the responses analyzed were total phenolic content (TPC), antioxidant activity by the capture of the free radical DPPH· and ferric reduction ability (FRAP). The polarity of the solvents and their mixture strongly influenced the extraction, in which moderately polar mixtures such as acetone-water and ethanol-water were highly efficient to extract the antioxidant phenolic compounds. The phenolic profile of the extracts showed that synaptic, caffeic, salicylic, protocatequímico, hydroxybenzoic and gentisic acids, as well flavonoids, myricetin, quercetin and kaempferol are the major compounds responsible for high antioxidant activity. The addition of antioxidants naturally present in the seeds of chia and a combination with α-tocopherol and myricetingave oxidative stability to the chia oil when evaluated by the accelerated method. The spray-dry technique was evaluated as a way to protect the chia oil, as well the influence of the use of rotor-stator type homogenizersand ultrasound on the formation of emulsions and on the properties of the microcapsules. Application of the microencapsulation technology presented encapsulated oil results percentage (28,76%) and encapsulation efficiency (96%) similarly to commercial microparticles. Also, it was efficient in preserving the fatty acid profile of the oleaginous matrix and increased the time of the oxidative stability of the oil in natura (0,6 hours) to 8,2 hours of microencapsulated oil, especially when ultrasound was used. Nutritional information reported in this study illustrates the potential benefits that chia seed can promote on human health, and shows the potential use in industrial applications. / Os alimentos funcionais são destaque para a promoção de efeitos benéficos a saúde. Entre eles, a semente de chia (Salvia hispanica L.) é uma fonte natural de ácidos graxos poli-insaturados, com altos teores de ácido α-linolênico, além de outros componentes nutricionais importantes, tais como vitaminas, minerais e antioxidantes. Considerando o interesse crescente do uso de chia na dieta humana pelos benefícios a saúde relatados na literatura, o presente trabalho determinou as características físico-químicas do óleo e a composição de nutrientes de sementes de chia cultivadas no Brasil, investigou os componentes fenólicos presentes na semente, bem como desenvolveu uma microcápsula contendo óleo de chia. Uma revisão foi elaborada abordando os principais componentes: lipídios, proteínas, fibras e compostos fenólicos, com ênfase aos benefícios das propriedades funcionais da semente de chia sobre a saúde humana e na elaboração de produtos alimentares. A caracterização das sementes de chia cultivadas no Brasil revelou quantidades consideráveis de proteína (25,03 g/100g), lipídios (34,41 g/100g) e fibras (36,50 g/100g), além de quantidades elevadas de aminoácidos essenciais e minerais nutricionalmente importantes como cálcio, potássio, zinco e selênio. O perfil de ácidos graxos do óleo revelou um elevado grau de insaturação, composto pelos ácidos oleico, linoleico e, predominantemente, pelo ácido α-linolênico (65,61 g/100g). O óleo apresentou um elevado teor de tocoferois (586,78 μg/g), com o isômero γ-tocoferol representando mais de 90% de todos os isômeros e compostos fenólicos como miricetina, quercetina, campferol e ácido sináptico. O potencial antioxidante das sementes desengorduradas foi determinado a partir de um delineamento estatístico de mistura, modelo simplex lattice, de modo a produzir o extrato seco com maior conteúdo de fenólicos totais (CFT) e atividade antioxidante, aplicando os ensaios de captura do radical livre DPPH· e poder redutor do ferro (FRAP), assim como, identificando e quantificando por cromatografia líquida de alta eficiência (CLAE) os ácidos fenólicos e flavonoides. A polaridade dos solventes e suas misturas influenciaram fortemente a extração, onde misturas de solventes moderadamente polares como acetona-água e etanol-água foram altamente eficientes. O perfil fenólico dos extratos mostrou que os ácidos sinápico, cafeico, salicílico, protocatequímico, hidroxibenzoico, gentísico e os flavonoides, miricetina, quercetina e campferol são os compostos majoritários responsáveis pela elevada atividade antioxidante. A adição de compostos antioxidantes, presentes naturalmente nas sementes de chia, e a combinação destes como α-tocoferol e miricetina, proporcionou estabilidade oxidativa ao óleo de chia quando avaliado por método acelerado. Como forma de proteção para o óleo de chia foi avaliada a técnica de spray-dry, assim como a influência no emprego de homogeneizadores do tipo rotor-estator e ultrassom, na formação das emulsões e propriedades das microcápsulas. A utilização da tecnologia de microencapsulação apresentou resultados de porcentagem de óleo encapsulado (28,76%) e eficiência de encapsulação (96%), similarmente à micropartículas comerciais, e mostrou-se eficiente em conservar o perfil de ácidos graxos da matriz oleaginosa e elevou o tempo de estabilidade do óleo in natura de 0,6 horas para 8,2 horas o óleo microencapsulado, sobretudo quando utilizado o ultrassom. As informações nutricionais relatadas neste estudo ilustram os possíveis benefícios que a semente de chia pode promover à saúde humana, além de mostrar o potencial uso em aplicações industriais.