CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A Caatinga possui uma vegetaÃÃo heterogÃnea cuja biodiversidade taxonÃmica conta com mais de 2.000 espÃcies de plantas. Dentre essas, cerca de 220 pertencem à famÃlia das leguminosas com 80 espÃcies endÃmicas, Ãnicas desse bioma. Muitas sÃo usadas para diversas finalidades de forma indiscriminada, reduzindo consideravelmente a diversidade e o nÃmero de espÃcies antes mesmo do conhecimento de suas potencialidades. Estudos que possam agregar valor econÃmico e viabilizar o uso racional, sustentÃvel e a conservaÃÃo das mesmas, aliada à constante busca por novas fontes de proteÃnas vegetais para atender à demanda crescente da populaÃÃo, bem como a grande necessidade de descoberta de compostos naturais que auxiliem no combate aos patÃgenos humanos e de plantas, sÃo de extrema relevÃncia. Assim, o presente estudo objetivou avaliar o potencial nutricional e bioativo de sementes de dez espÃcies vegetais da Caatinga destacando a espÃcie mais promissora. Para tanto, dez espÃcies de leguminosas selvagens da Caatinga foram analisadas quanto a sua composiÃÃo nutricional, apresentando elevado percentual de proteÃna bruta (10,9  0,4 a 50,0  3,4 %), fibras (0,8  0,0 a 52,3 Â1,0 %) e energia (1.000 a 1.804 kJ/100g), com perfil de aminoÃcidos comparÃveis aos da soja, com maiores teores de lisina (1088 a 456 mg/gN) e histidina (199 a 918 mg/gN) e bom perfil de minerais por apreentar boas quantidades de (mg/100g de farinha) de todos eles, em especial, de ferro (3,8 a 20,2), cÃlcio (31 a 268), magnÃsio (102 a 244) e potÃssio (366 a 1.581). As sementes apresentaram baixas quantidades de lectinas (80 a 2.560 e 160 a 2.560 UH/gF, quando nÃo tratadas e tratadas com enzimas, respectivamente), inibidores de tripsina (4,1  0,4 a 27,4  0,2 gTI/mgF ), ureases (465  13 a 47.178  3.351 U/KgF) e atividade tÃxica, em apenas trÃs espÃcies, com DL50 variando de 0,72  0,03 a 1,12  0,04 g/Kg peso. Foi determinado um Ãndice de qualidade nutricional para todas as espÃcies, o qual apontou a espÃcie Piptadenia moniliformis Benth. (Catanduva) como detentora de melhor qualidade nutricional, sendo assim destacada e avaliada in vivo a qualidade das proteÃnas de suas sementes. Os processamentos tÃrmicos (fervura, cozimento em micro-ondas e autoclavagen) e o processo de extraÃÃo de -galactosÃdios nas sementes dessas espÃcies nÃo proporcionaram bom desempenho dos animais, tendo em vista a perda de peso apresentada. Melhoria nos parÃmetros nutricionais, como NPU e VB foi verificada apÃs a retirada de -galactosÃdios dessas sementes, sugerindo que a anÃlise de outros processamentos para o aproveitamento das proteÃnas de suas sementes, pode tornÃ-las uma fonte promissora. AlÃm do alto potencial nutricional, as dez espÃcies apresentam tambÃm potencial bioativo devido à presenÃa de metabÃlitos secundÃrios como alcalÃides, catequinas, calchonas, auronas, flavonÃis, fenÃis flavonas, xantonas, flavononÃis, saponinas e triterpenÃides. Possuem proteÃnas bioativas como proteases, quitinases (0,23  0,02 a 2,0  0,33 nKat/mgP), β-1,3-glucanases (0,01  0,0 a 0,8  0,01 nKat/mgP), alÃm de proteÃnas ativas contra microorganismos que tambÃm sÃo consideradas antinutricionais (lectinas, inibidores de tripsina, ureases e toxinas). A avaliaÃÃo dos extratos brutos (EB) das espÃcies mostrou que todas sÃo ativas contra larvas de Aedes aegipty com percentual de mortalidade variando de 13,33  0,54 a 100,00  0,00 %, exceto o EB de Caesalpinea bracteosa que foi ativo contra a cepa Bacillus subtilis e contra o fungo Fusarium oxysporum, juntamente como o EB de Dioclea megacarpa. A espÃcie Senna rugosa inibiu o crescimento das cepas Bacillus subtilis e Staphylococcus aureus. Os fungos fitopatogÃnicos Aspergilus niger e Colletotrichum truncatum foram inibidos pelos EBs de Piptadenia moniliformis e Enterolobium contortisiliquum, que alÃm destes, foi ativo frente a Neurospora sp. e Trichoderma viridae. A espÃcie P. moniliformis destacou-se por sua elevada atividade quitinÃsica (1,12  0,0 nKat/mgP) em adiÃÃo à atuaÃÃo contra modelos biolÃgicos susceptÃveis a essa enzima, tendo sido escolhida para sua purificaÃÃo. A fraÃÃo proteica purificada de P. moniliformis (PmFP) contÃm elevada atividade de quitinases e causou pequena reduÃÃo no crescimento das leveduras Saccharomyces cerevisiae e Candida tropicalis, bem como da bactÃria B. subtilis. Inibiu ainda, a eclosÃo de ovos de A. aegypti com CI50 de 204, 42  2,19 ÂgP/ml e alterou a estrutura dos ovos e morfologia das larvas de primeiro estÃdio. A investigaÃÃo do potencial nutricional e bioativo das espÃcies mostrou boa composiÃÃo de nutrientes, em especial de proteÃnas e confirmou a presenÃa de compostos bioativos de natureza proteica e de metabÃlitos secundÃrios, tornando-as promissoras fontes de nutrientes e compostos antimicrobianos e anti-inseticidas que podem ser utilizados biotecnologicamente para fins agrÃcolas e industriais / The Caatinga Biome shows an heterogeneous vegetation with a taxonomic biodiversity of over 2,000 species of plants. Among these, approximately 220 belong to the Leguminosae family with 80 endemic species, unique to that biome. Many are used for various purposes in an indiscriminate manner, greatly reducing the variety and number of species even before the knowledge of their potential uses. Studies that can add economic value and enable the rational, sustainable use of these species, coupled with the constant search for new sources of plant protein to meet the ever increasing demand of the population are extremely important. Similarly important is the search for natural compounds which may help to combat human and plant pathogens. Thus, this study aimed to assess the nutritional and bioactive value of the seeds of ten plant species from Caatinga, highlighting the most promising ones. For this, the seeds were analyzed for nutritional composition, showing a high percentage of crude protein (10.9  0.4 to 50.0  3.4%), dietary fiber (0.8  0 , 0 to 52.3  1.0%) and energy (1,000 kJ/100g a1.804), with amino acid profile similar to that of soybeans, with higher amounts of lysine (1088-456 mg/gN) and histidine (199-918 mg/gN) and good mineral profile, with good content (mg/100 g flour) for all of them, especially, iron (3.8 to 20.2), calcium (31 to 268), magnesium (102-244) and potassium ( 366-1581). The seeds showed low amounts of lectins (80-2560 and 160-2560 UH / gF, when untreated and treated with enzymes, respectively), trypsin inhibitor (4.1  0.4 to 27.4  0.2 GTI / mgF), urease (465  13 to 47,178  3,351 U / KGF) and toxic activity in only three species, with LD50 ranging from 0.72  0.03 to 1.12  0.04 g / kg body weight . Was given an index of nutritional quality for all species, which pointed to Piptadenia moniliformis Benth. species (Catanduva) as the most promising one and because of that the seeds of this species was had the quality of its proteins evaluated in vivo. The thermal processing (boiling, microwave cooking and autoclaving) as well as the removal of α-galactosides did not improve animals performance. The nutritional parameters NPU and BV were improved when the animals were fed the seeds diet after removal of the α-galactosides. This may indicate that the search for apropriate processing methods may turn these seeds a promising source of proteins. Besides the high nutritional potential, the seeds of the ten studied species also have bioactive potential due to the presence of secondary metabolites such as alkaloids, catechins, calchonas, Auron, flavonols, flavones phenols, xanthones, flavononols, saponins and triterpenoids. These seeds also have bioactive proteins such as proteases, chitinases (0.23  0.02 to 2.0  0.33 nkat / mg P), β-1,3-glucanase (0.01  0.0 to 0.8  0, 01 nkat / mg P), and proteins active against microorganisms that are also considered antinutritional factors (lectins, trypsin inhibitors, urease and toxins). The evaluation of the crude extracts (CE) of the seeds showed that all species are active against the larvae of Aedes aegipti with mortality rates ranging from 13.33  0.54 to 100.00  0.00%, except that of Caesalpinea bracteosa which similarly to the CE of Dioclea megacarpa, was active against the bacterium Bacillus subtilis and against the fungus Fusarium oxysporum. Seeds extract of Senna rugosa species was able to inhibit the growth of Bacillus subtilis and Staphylococcus aureus. The pathogenic fungi Aspergillus niger and Colletotrichum truncatum were inhibited by the CE of Piptadenia moniliformis and Enterolobium contortisiliquum. The latter was also active against Neurospora sp. and Trichoderma viridae. The species P. moniliformis was distinguished for its high chitinase activity (1.12  0.0 nkat / mg P) in addition to its activity against biological models susceptible to this enzyme. For these reasons attempts were made for its purification. The purified protein fraction of P. moniliformis (PmFP) contains high activity of chitinases and caused a small reduction in the growth of the yeasts Saccharomyces cerevisiae and Candida tropicalis, and of the the bacterium B. subtilis. This protein fraction also inhibits the hatching of A. aegypti eggs with IC50 of 204. 42  2.19 μgP / ml. It causes changes in the eggs structure and in the morphology of first stage larvae. Thus, investigation of bioactive and nutritional potential of the species showed good composition of nutrients, especially of proteins, and confirmed the presence of bioactive compounds from protein nature and secondary metabolites, making them promising sources of nutrients, antimicrobial and insecticides that can biotechnologically be used for agricultural and industrial purposes.

A família Fabaceae abundante nos trópicos, desperta grande interesse para o uso em recuperação de áreas degradas, já que em sua maioria, são lenhosas e perenes, se adaptam aos mais diversos ecossistemas brasileiros e são capazes de fazer associações simbiônticas com bactérias pertencentes ao gênero Rhizobium, que fixam nitrogênio atmosférico fornecendo-o para a planta em uma forma reativa. Dentro deste contexto, o presente estudo avaliou em um experimento em casa de vegetação como a fixação biológica de nitrogênio em três espécies pertencentes a família Fabaceae variou em função da adição de nitrogênio mineral em diferentes doses e através de tratamentos incluindo-se a inoculação com bactérias do gênero Rhizobium e fungos micorrízicos arbusculares (FMAs). Os resultados obtidos através da atividade da redução de acetileno indicaram que os nódulos se encontravam ativos no último mês de experimento nas três espécies fixadoras de N2. Houve uma grande variação na porcentagem de N na planta proveniente da fixação (NPPfix). Quando as plantas cresceram em meio onde a disponibilidade de N era baixa, sem a aplicação deste nutriente no solo, a FBN foi favorecida, variando de 34 a 84%. Pelo contrário, foi possível observar que a fixação foi inibida em vasos onde a adição de nitrogênio foi elevada. Devido ao uso do solo não esterilizado, os inoculantes aplicados (bactérias e fungos) parecem ter competido com a microbiota nativa do solo, assim sendo, diferenças entre os tratamentos inoculados e o controle (sem inoculação) não foram encontradas neste estudo / Legumes (Fabaceae) are abundant in the tropics and considered as of great interest as restorer of degraded areas, since most of them are woody and perennials, adapted to several Brazilian ecosystems and able to make associations with bacteria belonging to Rhizobium genus, which fix atmospheric nitrogen, and giving it to the plants in a reactive form. In this context, this study evaluated in a greenhouse experiment how nitrogen biological fixation (FBN) varied in three species of Fabaceae family according to the N-mineral addition in different doses and according to treatments including inoculation with bacteria of Rhizobium genus and arbuscular mycorrhizal fungi (AMF). The results obtained from the acetylene reduction activity indicate that the nodules were active in the last month of experiment in the three N2-fixing species. There was a wide variation in the plant N percentage derived from fixation (NPPfix). When plants grew in a soil where N availability was low, without application of this nutrient in the soil, the BNF was favored, ranging the NPPfix from 34 to 84%. On the contrary, it was possible to observe that FBN was inhibited in the pots where N additions were higher. Due to the use of unsterilized soil, the applied inoculants (bacteria and fungi) appear to have competed with the native soil microbiota, thus differences between inoculated and control (without inoculation) could not be established in this study

As hortaliças são alimentos amplamente consumidos na dieta dos brasileiros. Há alguns anos, estudos sobre compostos presentes em vegetais, como os carotenóides, vitaminas, fibras e compostos fenólicos vem sendo bastante explorados devido aos seus possíveis efeitos biológicos e benefícios à saúde humana. Os compostos fenólicos são produtos do metabolismo secundário das plantas e se destacam principalmente por seu poder antioxidante. O objetivo deste trabalho foi avaliar os compostos fenólicos e a atividade antioxidante em algumas hortaliças consumidas no Brasil. Os métodos utilizados foram o do radical livre DPPH e ABTS, auto-oxidação do sistema -caroteno/ácido linoléico, redução do ferro, estabilidade oxidativa em Rancimat e identificação química por meio da técnica de cromatografia gasosa acoplada a espectrômetro de massas. Extratos etanólicos foram utilizados em todas as análises. O teor de compostos fenólicos variou entre 1,2 a 16,9 mg/g, sendo que a cenoura apresentou a menor quantidade e a alface a maior. As hortaliças que apresentaram a maior atividade antioxidante foram a alface (54,9% e 0,45 µmol Fe2+/mg), nos métodos do DPPH e FRAP , respectivamente, o açafrão (111,8µM trolox/g e 92,8%), nos métodos ABTS+ e -caroteno, o agrião e o brócolis no método do Rancimat (fator de proteção 1,29). Além dessas, a alcachofra, espinafre, e aspargo também apresentaram atividade antioxidante considerável. Os compostos fenólicos mais frequentemente encontrados nas amostras pela técnica de CG-EM foram os ácidos ferúlico, caféico e p-cumárico. O ácido ascórbico presente também contribuiu para a atividade antioxidante de alguns vegetais. Desta forma, pode-se verificar boa atividade antioxidante de várias hortaliças, ressaltando assim a importância em uma dieta equilibrada destes vegetais. / Vegetables are widely consumed foods in the diet of Brazilians. A few years ago, studies on compounds present in vegetables such as carotenoids, vitamins, fibers and phenolic compounds have been conducted due to their biological effects and benefits to human health. Phenolic compounds are distinguished mainly by their antioxidant power. The aim of this study was to evaluate the phenolic compounds and antioxidant activity in some vegetables consumed in Brazil. The methods for evaluating the antioxidant activity used were the DPPH free radical and ABTS, -carotene bleaching, reduction of Fe3+, oxidative stability in Rancimat, while the chemical composition used the gas chromatography - mass spectrometry technique (GC-MS). Ethanol extracts were used in all analysis. The content of phenolic compounds ranged from 1.2 to 16.9 mg / g, with carrots presenting the lowest amount and lettuce the highest. The vegetables with the highest antioxidant activity were lettuce (54.9% and 0.45 mmol Fe2+/ mg), through DPPH and FRAP methods, respectively, saffron (111.8 mM trolox / g and 92.8%) through the ABTS+ and -carotene methods, watercress and broccoli through the Rancimat method (protective factor 1.29). Besides these, the artichoke, spinach, and asparagus also showed considerable antioxidant activity. Phenolic compounds most frequently found through the GCMS technique were ferulic acid, caffeic acid and p-coumaric acid. Ascorbic acid also contributed to the antioxidant activity of some vegetables. Generally, it is confirmed the good antioxidant activity of some vegetables, emphasizing its importance in a balanced diet.

Orientador: Marlene Aparecida Schiavinato / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-21T08:53:28Z (GMT). No. of bitstreams: 1 Longatti_ClaudiaAparecida_M.pdf: 1449126 bytes, checksum: be3aefaf910ddbe46531e367581164f3 (MD5) Previous issue date: 2012 / Resumo: O metabolismo geral de uma planta pode ser afetado pelo excesso de sais no solo, visto que a salinidade pode causar reduções no crescimento e acelerar a senescência. Os brassinosteróides são hormônios vegetais envolvidos no aumento da tolerância a vários tipos de estresses, inclusive o estresse salino. Com o objetivo de avaliar a influência da aplicação de brassinosteróide no crescimento e metabolismo do nitrogênio em plantas de Cajanus cajan sob diferentes condições de estresse salino, plantas noduladas de C. cajan foram cultivadas em casa de vegetação na ausência ou presença de brassinosteróide (24-epibrassinolídeo), sob diferentes concentrações de NaCl (0 mM, 100 mM e 200 mM). A salinidade afetou o crescimento, reduzindo a área, o comprimento do caule, a longevidade foliar, o teor de umidade de folhas, caules e raízes, bem como a atividade da nitrogenase, os teores de pigmentos fotossintetizantes, de malondialdeído e de nitrogênio total em folhas verde maduras, e a proficiência e eficiência do uso de nitrogênio. A aplicação de brassinosteróide amenizou o efeito do NaCl nas reduções da área foliar e teores de clorofila a e total. Também atenuou o efeito do estresse salino na atividade da nitrogenase. Esse efeito positivo do brassinosteróide sobre a atividade da enzima refletiu-se no aumento da concentração de compostos nitrogenados nas folhas verde-maduras e reduziu, ainda, a perda de N nas folhas senescentes. A maior concentração de N total nas folhas verde-maduras de plantas tratadas com o 24-epibrassinolídeo levou à redução da eficiência de utilização desse elemento / Abstract: The general metabolism of a plant can be affected by salinity excess in soil, due to the fact that salinity can cause growth reduction and accelerate senescence. The brassinosteroids are plant hormones involved in increasing tolerance in several types of stress, including salt stress. In order to verify the influence of the application brassinosteroid on development and nitrogen metabolism in Cajanus cajan plants under different levels of salt stress, nodulated plants of C. cajan were cultivated in the greenhouse in the absence or presence of 24-epibrassinolide, under different concentrations of NaCl (0 mM, 100 mM and 200 mM). Salinity affected growth reducing leaf area, branch length, leaf longevity, moisture content of leaves, stems and roots, as well as activity of nitrogenase, amounts of photosynthetic pigments, malondialdehyde and total nitrogen in mature green leaves; nitrogen use proficiency and efficiency. The application of brassinosteroid attenuated the effect of NaCl on the leaf area and on the concentrations of chlorophyll a and total chlorophyll. The hormone also attenuated the effect of salt stress on the activity of nitrogenase. The positive effect of the hormone on the activity of the enzyme led to an increase in the concentration of nitrogenous compounds in the mature green leaves and reduced the lost of nitrogen in senescent leaves. The higher concentration of total nitrogen in mature green leaves of the plants treated with 24-epibrassinolide led to a reduction in nitrogen use efficiency / Mestrado / Biologia Vegetal / Mestre em Biologia Vegetal

Made available in DSpace on 2014-08-20T14:38:51Z (GMT). No. of bitstreams: 1 Dissertacao_Caroline_Moreira_ Rodrigues.pdf: 942864 bytes, checksum: 8281e84c8318c352dab069ed9fc6abee (MD5) Previous issue date: 2010-05-31 / The aim of this study was to evaluate morphogenesis, structural and forage production of persian clover (Trifolium resupinatum L.) intercropped with ryegrass (Lolium multiflorum Lam) in the field, subjected to different heights and cutting intervals. The experiment was conducted during 2009 in the area ceded by Embrapa Clima Temperado in Capão do Leão, RS. Sowing was done by throwing in plots of 8 m2, with a density of 10 kg ha-1 of persian clover and 15 kg ha-1 of ryegrass. The design was randomized complete block design in a 3x3 factorial design with five replicates, totaling 45 experimental units. We used three cutting height, 4 cm, 8 cm and 12 cm and three cutting intervals, corresponding to the accumulation of heat 320, 420 and 520 GD. Technique was used in the branches / tillers marked and made ratings weekly in clover on count the number of leaves (open living, senescent, dead, emerged), number of nodes, plant height and length of branching, and in ryegrass on count the number of leaves (fully expanded, growing, senescent fully expanded, growing senescent dead), plant height and length of the tiller. On the occasion of the cuts was made that production of dry matter and botanical separation. The rate of leaf appearance (TAF) of Persian clover is modified by the interval (GD) and cutting height. Intervals of 320 and 420 GD have higher rates of leaf appearance (TAF) of Persian clover. The cutting height does not change the TAF of ryegrass. The range of GD 320 has a better TAF of ryegrass. In the cutting of 420 GD-Persian clover has a higher number of green leaves open fewer dead leaves, branches of greater length, greater number of nodes and greater forage production, and the recommended range for management by producers. The highest yield of intercropping forage ryegrass and Persian clover is obtained with GD 520 but with a high ratio stem / leaf. / O objetivo do presente trabalho foi avaliar características morfogênicas, estruturais e produção de forragem do trevo-persa (Trifolium resupinatum L.) consorciado com azevém (Lolium multiflorum Lam.) a campo, submetidos a diferentes alturas e intervalos de cortes. O experimento foi conduzido no ano de 2009, em área cedida da EMBRAPA Clima Temperado Estação Terras Baixas, no município de Capão do Leão, RS. A semeadura foi feita a lanço, em parcelas de 8 m2, com densidade de 10 kg.ha-1 de trevo-persa e 15 kg.ha-1 de azevém. O delineamento utilizado foi blocos completos ao acaso, em esquema fatorial 3x3, com cinco repetições, totalizando 45 unidades experimentais. Foram utilizadas três alturas de corte, 4 cm, 8 cm e 12 cm e três intervalos de corte, correspondentes ao acúmulo térmico de 320, 420 e 520 GD. Foi utilizada a técnica das ramificações/perfilhos marcados e feitas avaliações semanais no trevo quanto ao número de folhas (vivas abertas, senescentes, mortas, surgidas), número de nós, altura de planta e comprimento de ramificação; e no azevém quanto ao número de folhas (completamente expandidas, em expansão, completamente expandidas senescentes, em expansão senescente, mortas), altura de planta e comprimento de perfilho. Na ocasião dos cortes, foi feita verificação da produção de matéria seca e separação botânica. A taxa de aparecimento de folhas (TAF) do trevo-persa é modificada pelo intervalo (GD) e altura de corte. Intervalos de 320 e 420 GD apresentam melhor taxa de aparecimento de folhas (TAF) do trevo-persa. A altura de corte não altera a TAF do azevém. O intervalo de 320 GD apresenta melhor TAF do azevém. No intervalo de corte de 420 GD o trevo-persa apresenta maior número de folhas vivas abertas, menor número de folhas mortas, maior comprimento de ramos, maior número de nós e maior produção de forragem, sendo o intervalo recomendado para o manejo pelos produtores. A maior produção de forragem da consorciação azevém e trevo-persa é obtida com 520 GD mas com alta relação caule/folha.

La productivité et la qualité des céréales biologiques sont soumises à deux principaux facteurs limitants dans les systèmes sans élevage : des déficits chroniques en azote (N) du sol et des infestations par les adventices. Des légumineuses telles que les trèfles ou les luzernes peuvent servir à la fois de plantes de couverture et d'engrais verts grâce à leur fixation symbiotique d'N atmosphérique. Cependant, leur substitution aux céréales présente un moindre intérêt économique dans les systèmes de grandes cultures en l'absence d'animaux pour les valoriser. L'association relais de couverts de légumineuses dans un blé d'hiver nous a semblé être une option intéressante pour à la fois enrichir le système sol-plante en N, couvrir le sol dès la récolte du blé associé et limiter le risque de compétition avec le blé en décalant au printemps la date de semis des légumineuses sous couvert de blé. Pour évaluer l'efficacité de ces associations, quatre espèces de légumineuses (Medicago lupulina L., M. sativa L., Trifolium pratense L. et T. repens L.) ont été semées au tallage du blé d'hiver sur huit parcelles réparties dans la région Rhône-Alpes. Leurs effets sur la maîtrise des adventices, l'enrichissement, la préservation et la restitution d'N au système sol-plante et les performances des cultures ont été observés, durant une succession blé d'hiver-culture de printemps. Les résidus des couverts ont été enfouis à la fin de l'hiver, 9 à 12 semaines avant le semis d'une culture de printemps. Nos travaux ont montré l'absence d'effet des couverts associés sur le rendement en grains du blé d'hiver. Mais des diminutions du taux protéique des grains sont apparues dans un tiers des situations d'association notamment avec M. lupulina et T. pratense, les espèces les plus développées à la récolte du blé. Notre suivi de la disponibilité des ressources trophiques principales (eau, N, lumière) nous a permis d'identifier une compétition pour l'eau et l'N du sol. Nous avons également noté l'efficacité des couverts de légumineuses dans le contrôle de la densité des adventices dès le stade de floraison du blé et de leur biomasse durant l'interculture. Le meilleur contrôle des adventices a été permis par M. lupulina et T. pratense, à la récolte du blé, et T. pratense et T. repens, à la fin de l'automne, associé aux biomasses aériennes observées les plus importantes. Enfin, nous avons observé une forte proportion d'N issu de la fixation symbiotique dans la biomasse aérienne des légumineuses à la fin de l'automne (80 à 94%), représentant un apport d'N exogène au système sol-plante évalué entre 37 et 77 kg N ha-1. Cet enrichissement en N n'a pas entrainé d'aggravation de la lixiviation d'N durant l'hiver. Les couverts de légumineuses n'ont pas non plus diminué la lixiviation comparativement à l'absence de couvert. Après leur destruction, les résidus des couverts ont restitué une partie de l'N accumulé (+28 à +42 kg ha-1 d'N minéral sur les 90 premiers centimètres de sol par rapport au témoin à l'émergence de la culture de printemps, 12 semaines après leur destruction). Cette restitution a permis un enrichissement en N des pailles et grains de la culture de printemps et une augmentation de 30% du rendement lorsqu'il s'agissait de maïs. Finalement nous concluons sur l'intérêt des couverts de légumineuses associés en relais dans un blé d'hiver pour apporter une réponse positive aux problèmes principaux des rotations de grandes cultures biologiques (adventices, déficits d'N et diminutions des performances des cultures). Nous terminons en proposant des voies d'évolution des associations testées, notamment pour limiter les risques de compétition durant l'association. Nous évoquons également les implications scientifiques et pratiques de ce travail pour de futures études sur ce sujet. / Cereal productivity and quality are subject to two main problems in organic stockless systems: chronic soil nitrogen (N) deficiencies and weed infestation. Legume species as clovers or alfalfas can be used as cover crops and green manures due to their natural ability to fix atmospheric N. Nevertheless, their substitution to cereals in crop rotations is less economically viable without animals to use it. Relay intercropping of legume cover crops (RIL) in winter wheat was expected to simultaneously enrich the soil-plant system in N, cover the soil from the wheat harvest onwards and limit the risk of competition with wheat by delaying the undersowing of legumes in spring. To evaluate the efficiency of RIL, four legume species (Medicago lupulina L., M. sativa L., Trifolium pratense L. and T. repens L.) were undersown at wheat tillering stage, in eight fields organically managed in the Rhône-Alpes region, France. Their effects on weed control, on N enrichment, preservation and restitution in the soil-plant system and on crop performance were observed during the succession of the winter wheat and a spring crop. RIL residues were incorporated in soil in late winter, 9 to 12 weeks before the sowing of spring crops. Our work illustrated the absence of detrimental effect of RIL on wheat grain yield despite the decrease of the grain protein content in one third of the situations. This decrease was mainly observed with M. lupulina and T. pratense as they were the most developed legume species at wheat harvest. Our monitoring of trophic resources (water, N and light) enlightened the competition for soil water and N during the intercropping period. We also showed the efficiency of RIL in the control of weed density from wheat flowering stage onwards and of weed aerial biomass in late autumn. The best weed control was observed with M. lupulina and T. pratense, at wheat harvest, and with T. pratense and T. repens in late autumn, in relation to the highest aerial biomasses observed with these species. Finally, we noted an important proportion of N derived from atmosphere (Ndfa) in legumes' shoots in late autumn (80 to 94%), representing an input of exogenous N in the soil-plant system ranging from 37 to 77 kg Ndfa ha-1. The N enrichment of the system did not increase mineral N lixiviation during winter. However, legume cover crops did not significantly prevent any lixiviation of mineral N relative to the control without cover crop. After their ploughing in, legumes residues returned a part of the accumulated N (+28 to +42 kg ha-1 of mineral N in the first 90 cm of soil compared to the control at the emergence of the spring crop, 12 weeks after ploughing). This restitution of mineral N allowed a significant N enrichment of straw and grains of the spring crop with all previous legumes cover crop species. With maize as spring crop, the grain yield increased by 30%, on average, compared to the control treatment. We finally conclude on the interest of RIL in winter wheat to bring a positive response to the problems of weed control, N deficiency and crop performances in organic crop rotations. We then suggest possible improvements of the RIL system, especially against the competition for trophic resources during the relay intercropping period. We also mention scientific and practical implications of this work for future studies on this subject.

Les légumineuses permettent de réduire les intrants en élevage (engrais, concentrés) en raison notamment de leurs niveaux élevés en protéines. Cependant, à la fois pendant le processus d'ensilage et celui de fermentation dans le rumen, les protéines peuvent subir une importante dégradation, ce qui affecte la valeur nutritive des fourrages et induit des rejets d'azote (N) importants, notamment dans l'urine. Certaines légumineuses peuvent alors être d'un intérêt particulier car elles produisent des composés secondaires qui peuvent modifier positivement les processus fermentaires et digestifs. Ainsi, les tannins condensés (CT) présents dans le sainfoin (SF, Onobrychis viciifolia) sont capables de se lier aux protéines, réduisant leur dégradation dans le silo et le rumen et se traduisant par un transfert de l'excrétion d'azote de l'urine vers les fèces. Le trèfle violet (RC, Trifolium pratense) contient la polyphénoloxydase (PPO), une enzyme qui catalyse l'oxydation de différents composés phénoliques en quinones. Comme les CTs, les quinones sont capables de former des complexes avec les protéines permettant de réduire leur dégradation dans le silo et le rumen. L'objectif de cette thèse était alors d'étudier et de quantifier les bénéfices potentiels de l'utilisation de ces deux espèces de légumineuses bioactives sur i) la qualité et la conservation des ensilages, ii) la fermentation ruminale, l'efficacité digestive et les performances des ovins, et iii) l'empreinte environnementale (excrétion d'N et de CH4). Nous avons effectué deux essais in vitro et deux essais in vivo, basés sur des ensilages composés de ces deux légumineuses, seules ou en mélange avec une graminée (la fléole- T, Phleum pratense L.) qui nous a servie de contrôle. Les essais in vitro nous ont permis de nous focaliser sur la qualité et la conservation des ensilages ainsi que sur la fermentation ruminale, tandis que les essais in vivo se sont concentrés sur la performance et l'efficacité digestive des agneaux, ainsi que sur leur bilan azoté et leurs émissions de CH4. L'inclusion de légumineuses bioactives dans les ensilages d'herbe a amélioré la qualité du fourrage, la fermentation pendant le processus d'ensilage ainsi que la protection des protéines contre une dégradation au sein du silo et du rumen. Globalement, l'alimentation des agneaux avec des mélanges comportant ces légumineuses s'est traduite par une augmentation de l'ingestion de matière sèche, en comparaison des agneaux alimentés avec la graminée pure. Néanmoins, en raison de la digestibilité nettement plus faible de T-SF, probablement due à une composition et une nature des fibres différentes ainsi qu'à la présence de CT, les agneaux ayant reçu T-SF ont montré une ingestion et des performances plus faibles que ceux ayant reçu les ensilages contenant RC. Dans le rumen, il semble que les protéines de RC aient été plus protégées de la dégradation que celles de SF, tandis que dans la suite du tractus digestif, les complexes formés entre protéines et CT (avec SF) se seraient moins dissociés que ceux formés entre protéines et quinones (avec RC), ce qui pourrait en partie expliquer le transfert d'excrétion de l'N de l'urine vers les fèces, observé chez les agneaux alimentés avec T-SF et bénéfique pour l'environnement. SF a également permis de réduire légèrement les émissions de CH4. Ainsi, utiliser des légumineuses bioactives dans les pratiques d'alimentation des ruminants apparaît une stratégie prometteuse pour fournir des produits animaux de façon plus durable. Nos résultats montrent que chaque espèce apporte des avantages différents, plutôt orientés vers la qualité de l'aliment et les performances animales pour RC mais plutôt orientés vers la réduction des rejets pour SF. Des recherches complémentaires sont donc nécessaires pour mieux caractériser ces avantages et élargir les investigations à d'autres espèces, d'autres mélanges et d'autres bénéfices potentiels. (...) / Fodder legume species allow to reduce inputs in livestock breeding systems (fertilizer, concentrates) notably because they contain high levels of crude proteins which are of primary importance in ruminant nutrition. However, during both silage and rumen fermentation processes, proteins are submitted to degradation which affects forage nutritive value and leads to nitrogen (N) losses notably via urine. Some specific legumes can then be of particular interest as they produce plant secondary compounds that can positively affect silage and digestive processes. Condensed tannins (CTs) present in sainfoin (SF, Onobrychis viciifolia) are able to bind with proteins thereby reducing their degradation in the silo and the rumen, resulting in a shift in N excretion from urine to faeces. Red clover (RC, Trifolium pratense) contains polyphenol oxidase (PPO), an enzyme that catalyses the oxidation of different phenolics into quinones. As CTs, quinones are able to form complexes with proteins that will similarly reduce their degradation in the silo and the rumen. The aim of this thesis was to investigate and quantify the potential benefits of using these two bioactive legume species on i) quality and conservation of silages, ii) rumen fermentation, digestive efficiency and sheep performance, and iii) environmental footprint (N excretion and CH4 emissions). We conducted two in vitro and two in vivo trials which were based on silages of pure legumes or of different mixtures with the grass species (timothy T, Phleum pratense L.), which served as control. In the in vitro trials, we focussed on silage quality, silage conservation and rumen fermentation, while in the in vivo trials, we focussed on lambs' performance, digestion efficiency, N balance and CH4 emissions. Including bioactive legumes in mixtures with grass improved, compared to pure grass, forage quality and fermentation during the silage making process, as well as proteins' protection from degradation within both the silos and the rumen. Lambs fed with the mixtures involving legumes responded with an increase in DM intake compared to their counterparts fed with T. Nevertheless, due to a possibly different fibre composition and to the presence of CT which impaired SF digestibility, lambs that consumed T-SF showed lower intake and performance than those that received RC-containing silages. In the rumen, RC proteins appeared more protected from degradation than SF ones, while in the subsequent parts of the digestive tract, the proteins-CT complexes (from SF) might less dissociate than the proteins-quinones ones (from RC). This could partly explain the environment-friendly shift in N excretion from urine to faeces when animals are fed with T-SF. SF also allowed to slightly reduce CH4 emissions. Thus, utilizing bioactive legumes in livestock feeding practices is a promising strategy to produce animal products more sustainably. Our results show different benefits relative to the bioactive legume species involved, directed towards boosted forage quality and animals' performance for RC but towards lowered wastes for SF. Further research is thus needed to better characterize these benefits and enlarging investigations to other plant species, mixtures and potential benefits (e.g. health). This will help to determine the appropriate choice of plant species according to the objectives.

Foi estudada a fração não-protéica de sementes de feijão (Phaseolus vulgaris L. ), tendo por objetivo identificar a presença e conhecer o teor de (γ-glutamil peptídeos sulfurados. Estes peptídeos (γ-glutamil-S-metilcisteína e (γ-glutamil metionina) podem interferir com o doseamento da metionina pelo método da clivagem com brometo de cianogênio e análise do metiltiocianato por cromatografia em fase gasosa. O extrato etanólico contém 21 % do nitrogênio total do feijão. A lavagem do extrato com clorofórmio reduziu o teor de nitrogênio para 5,5%, pela remoção de proteínas co-extraídas. O extrato não-proteico assim obtido foi purificado sucessivamente por resinas de troca catiônica e aniônica, ocorrendo a separação dos constituintes pelos seus valores de pK. A purificação foi acompanhada por testes químicos, cromatografia em camada delgada e análise de aminoácidos. Foi identificada com base nas suas características químicas, a presença de S-metilcisteína, principalmente na forma de dipeptídeo com ácido glutâmico. A quantificação do peptídeo após hidrólise ácida (3h / 110° C), resultou num teor de S-metilcisteína total de 0,67g /100g proteína, que corresponde a aproximadamente 50% do teor de metionina no feijão, determinado por cromatografia de troca iônica. Estes resultados podem indicar que o teor de metiltiocianato total, liberado pela metionina e pela S-metilcisteína do feijão (1 ,6%) (determinado em trabalho anterior), descontado do teor de S-metilcisteína (0 ,7%), equivale à metionina quimicamente disponível, correspondendo a aproximadamente 64% da metionina total presente no feijão. No ensaio de germinação das sementes do feijão foi observado um decréscimo da (γ-glutamil-Smetilcisteína, indicando uma mobilização dos aminoácidos constitutintes. / The non-protein fraction of Phaseolus vulgrias L. seeds has been analysed to obtain information on the identity and level of (γ-glutamyl peptides containing a sulfur amino acid. These peptides (γ-glutamyl-S-methylcysteine and (γ-glutamyl methionine) could interfere with the determination of methionine measured by cleavage with cyanogen bromide and subsequent gas-chromatographic identification of methylthiocyanate. The ethanolic extract of (γ-glutamyl peptides contains 21% of the total seed nitrogen, which decrease to 5,5%, by removing the coextracted bean proteins with chloroform. This non-protein extract was then purified by ion-exchange chromatography on basic and acid resins, taking advantage of the different pK values of the amino acids. The isolation procedure was followed up by chemical analysis, thin layer chromatography and amino acid analysis. Based on their chemical properties we were able to identify the presence of S-methylcysteine, mainly in the form of a dipeptide with glutamic acid. Quantitative analysis of the peptide after acid hydrolysis (3h /110° C) showed a level of 0,67g /total S-methylcysteine /100gbean protein. This content accounted for approximately 50% of the bean methionine content, determined by ion-exchange chromatography. These results might indicate that total methylthiocyanate released from both, methionine and S-methylcysteine of beans, (1,6%) (from a previous paper) subtracted from S-methylcysteine (0,7%) could be recognized as chemically available methionine, corresponding to approximately 64% of total methionine. During seed germination the (γ-glutamyl-S-methylcysteine undergo a degradative process resulting in loss of the dipeptide.

MSCAGR (Animal Science) / Department of Animal Science

MSCAGR (Plant Production) / Department of Plant Production / Bambara groundnut (Vigna subterranea) is one of the most important legumes cultivated primarily for food by smallholder farmers in Africa. It is an affordable source of protein and contributes to income generation as well as soil fertility. Despite its importance, it is cultivated largely for subsistence purposes in South Africa. Growers use landraces. The agronomic performance of the traditional varieties depends on environmental factors prevailing in a particular area. In Limpopo and Mpumalanga Provinces, there is no adequate information regarding the performance of bambara groundnut germplasm. The objectives of the study were to (i) determine the agronomic performance of Bambara groundnut across three contrasting locations in Limpopo and Mpumalanga provinces over two cropping seasons (ii) determine the genotypic variation in the seed protein level among 42 bambara groundnut genotypes. Forty-two bambara groundnut genotypes were evaluated under three different environmental conditions (Syferkuil, Thohoyandou and Nelspruit) over two (2013/2014, 2014/2015) seasons in a 7 × 6 rectangular lattice design replicated three times. Eight agronomic traits including dry shoot weight (DSW), number of pods per plant (NPP), pod length (PL), number of seed per pod (NSP), pod weight per plant (PWT), seed weight per plant (SWT), 100 seed weight (100-SWT) and seed yield (SYLD) were measured. The results showed that there were significant genotype x location interactions which demonstrated that the prevailing agro-ecological conditions at the test locations were distinct from each other. Five genotypes (‘BGN-19‘, ‘BGN-11‘, ‘BGN-12‘, ‘BGN-4‘and ‘BGN-34‘) attained >25.0% seed yield advantage over the local check ‘BGN-39‘. The results also showed that light brown coloured genotypes attained relatively higher seed yield compared to the other seed colours types. The cultivar superiority index (CSI) showed that three genotypes (‘BGN-12‘, ‘BGN-19’ and ‘BGN-34’) were the most stable across the test locations and attained >900.0 kg/ha on average. There were significantly high positive correlations between PWT and each of the three other attributes (SWT, 100 SWT and SYLD). In terms of seed protein, the results showed a poor relationship between seed yield and protein levels. ‘BGN-12’ which produced the highest seed yield, attained the lowest percent seed protein while genotype. On average, the genotypes contained 21.72% protein. The highest and lowest seed protein quantities were attained by the genotypes ‘BGN-42’ (25.17%) and ‘BGN-12’ (19.89%) respectively. / NRF