Zuzana Jindřišková The Study of Secondary Metabolites in Explant Culture of Trifolium pratense L. The basic prerequisite for a successful elicitation that is used to increase the production of secondary metabolites is, among others, finding a suitable elicitor, its concentration and optimal duration of effect of the elicitor on the plant in vitro culture, which was the main subject of this diploma thesis. The focus of our observations was the influence of 6-, 24- , 48- and 168-hour effect of nickel chloride solution (in the concentrations of 0.1 mmol, 1 mmol, 10 mmol and 100 mmol) and zinc sulphate (in the concentrations of 0.1 µmol, 1 µmol, 10 µmol a 100 µmol) on the production of flavonoids and isoflavonoids in the suspension culture of Trifolium pratense L. variety Tempus. The culture was cultivated on the Gamborg nutrien medium with the addition of 2 mg.l-1 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 6- benzylaminopurine at 25řC and the light period of 16 hours light/ 8 hours dark. The maximum content of flavonoids, which was found out by the photometric determination of the Czech Pharmacopoeia 2009, was proved in the suspension culture of Trifolium pratense L. variety Tempus (0.406%) after 48-hour elicitation of nickel chloride solution in the concentration of 0.1 mmol, when there was a...

1 ABSTRACT Kateřina Muráriková Influencing of production of plant tissue culture of Trifolium pratense L. I The aim of this thesis was to check the influence of cinnamic acid, as a precursor of the phenylpropanoids biosynthesis, on the flavonoids production of Trifolium pratense L. suspension culture, to compare two varieties of this plant, the DO-8 one and the Tempus one, in terms of the effect of this precursor, and to help achieve the successful increase of the production of secondary metabolites by the Trifolium pratense L. explant cultures. Concerning the DO-8 variety, the changes of the isoflavonoids production as a result of the cinnamic acid influence have been also tested. The suspension cultures were being cultivated on the Gamborg's nutrient medium with the addition of 2 mg.l-1 2,4-dichlorphenoxyacetic acid and 2 mg.l-1 6-benzylaminopurine. The cultivation was being realised on a slow moving roller at temperature of 25 řC and in the period of 16 hours of light and 8 hours of dark. The cinnamic acid was added in the four concentrations and its influence was observed after 6, 24, 48 and 168 hours. The flavonoids determination was being applied on the check samples and the samples with the addition of cinnamic acid by the spectrophotometric method in agreement with the Český lékopis 2009 and the...

1 Abstract Veronika Machová Effect of 2-benzylthiopyridine-4-carbothioamide derivatives on isoflavonoids and flavonoids accumulation in suspension culture of Trifolium pratense L. The paper examinate the effect of three concentration of 2-(2-fluoro-6- nitrobenzylsulfanyl) pyridine-4-carbothioamide on the production of flavonoids and isoflavonoids by the Trifolium patense L. (variety DO-8, variety Tempus). The culture were cultivated in the Gamborg nutrient media with addition of 2 mg·l-1 2,4- dichlorophenoxyacetic acid a 2 mg·l-1 6-benzylaminopurine, at the temperature of 25řC, 16-hr light/8-hr dark period. The elicited and the inspection samples underwent the photometric determination of flavonoids in accordance with the Czech Pharmacopoeia 2009 ant the determination of isoflavonoids via the HPLC method. The results show that the maximum content of flavonoids in suspension cultures of Trifolium pratense L. (variety DO-8) was caused by 48-hour elicitation by 100 µmol·l-1 solution of 2-(2-fluoro-6-nitrobenzylsulfanyl) pyridine-4-carbothioamide, when the a statistically significant increase in production was 93% compared to control cultures. The maximum content of flavonoids in variety Tempus was caused by 48-hour elicitation by 10 µmol·l-1 solution of 2-(2-fluoro-6-nitrobenzylsulfanyl) pyridine-4-...

Lucie Částková Influencing of production of plant tissue culture of Trifolium pratense L. II The aim of this thesis was to observe the effect of the abiotic elicitor vanadyl sulfate on the production of flavonoids in the suspension culture of Trifolium pratense L., variety DO-8 and variety Sprint. The effects on the production of isoflavonoids caused by the elicitor have been also studied. The cultures were cultivated at the temperature of 25 žC, 16 hour light/8 hour dark period, in the Gamborg's nutriet medium with the addition of 2 mg.l-1 of 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 of 6-benzylaminopurine. Vanadyl sulfate was added in four concentrations and its influence was observed after 6, 24, 48 and 168 hours. The quantity of flavonoids was determined spectrophotometrically according to the Pharmacopoeia Bohemica 2009. The quantity of isoflavonoids was determined using HPLC. The maximal content of flavonoids, 0,459 %, was measured in the suspension culture of Trifolium pratense L., variety DO-8, taken after 48 hours of action of elicitor vanadyl sulfate, which concentration was 1 µmol.l-1 . The statistically significant increase in production was 106 % compared to the control. The production of flavonoids in variety Sprint was most influenced by the action of vanadyl sulfate, which...

The experiment objectified the termination of lambs to the foot of the mother in pasture cultivated of ryegrass and red clover with and without creep-feeding to the lamb. Evaluations in the performance, characteristics of carcass, production of commercial cuts and production of the components had been carried through not-carcass in the lambs kept to the foot of the mother and submitted to the alimentary treatments: PAST = pasture cultivated of ryegrass and red clover; APT = pasture cultivated of ryegrass and red clover + creep-feeding to the lamb during all the period of use of the grass; AP42 = pasture cultivated of ryegrass and red clover + creep-feeding to the lamb from 42 days after beginning of the use of the grass. For all the evaluated parameters of the grass, did not have interaction (P>0.05) between treatment and period, did not have difference between treatments (P>0.05) for the parameters evaluated in the grass. Animals that had received creep-feeding had presented greater (P<0.05) profit of daily average weight and lesser (P<0.05) number of days to reach the weight established of slaughter in relation to that they had only remained in pasture. The components carcass had not differed (P>0.05) in the alimentary treatments. The weight of slaughter of 28 kg was only reached in 43, 71 and 86% of the animals of treatments PAST, AP42 and APT, respectively. The job of creep-feeding better does not provide to quality and nor carcass yield, for the lambs slaughtered with 28kg. In handling pastures of similar form, the creep-feeding promotes additive effect on the efficiency of termination of the animals. / O experimento objetivou a terminação de cordeiros ao pé da mãe em pastagem cultivada de azevém e trevo vermelho com e sem a alimentação privativa ao cordeiro. Foram realizadas avaliações no desempenho, características de carcaça, produção de cortes comerciais e produção dos componentes não-carcaça nos cordeiros mantidos ao pé da mãe e submetidos aos tratamentos alimentares: PAST = pastagem consorciada de azevém e trevo vermelho.; APT = pastagem consorciada de azevém e trevo vermelho + alimentação privativa ao cordeiro durante todo o ciclo de uso do pasto; AP42 = pastagem consorciada de azevém e trevo vermelho + alimentação privativa ao cordeiro a partir de 42 dias após inicio da utilização do pasto. Para todos os parâmetros avaliados do pasto, não houve interação (P>0,05) entre tratamento e período, não houve diferença entre tratamentos (P>0,05) para os parâmetros avaliados no pasto. Animais que receberam alimentação privativa apresentaram maior (P<0,05) ganho de peso médio diário e menor (P<0,05) número de dias para atingir o peso estabelecido de abate em relação a aqueles que somente permaneceram em pastagem. Os componentes não carcaça não diferiram (P>0,05) nos tratamentos alimentares. O peso de abate de 28 kg foi atingido somente em 43, 71 e 86 % dos animais dos tratamentos PAST, AP42 e APT, respectivamente. O emprego da alimentação privativa não proporciona melhor qualidade e nem rendimento na carcaça, para cordeiros abatidos aos 28kg. Em pastagens manejadas de forma similar, a alimentação privativa promove efeito aditivo sobre a eficiência de terminação dos animais.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2018-06-14T13:19:23Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) resumo_tese_natalia_silveira_correa.pdf: 16569 bytes, checksum: ed2405236282a6dfb013ce0c61b1b3aa (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-06-14T20:52:30Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) resumo_tese_natalia_silveira_correa.pdf: 16569 bytes, checksum: ed2405236282a6dfb013ce0c61b1b3aa (MD5) / Made available in DSpace on 2018-06-14T20:52:30Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) resumo_tese_natalia_silveira_correa.pdf: 16569 bytes, checksum: ed2405236282a6dfb013ce0c61b1b3aa (MD5) Previous issue date: 2017-06-12 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O objetivo deste trabalho foi avaliar a viabilidade, o vigor da semente e o crescimento de plantas de cornichão e trevo vermelho, provenientes de áreas da Embrapa Pecuária Sul, submetidas a diferentes doses de herbicidas pré e pós emergentes (diclosulam, bentazona e iodosulfurom-metílico), com a finalidade de identificar os herbicidas seletivos à estas forrageiras. Na primeira etapa do experimento foi avaliado o efeito do herbicida pré-emergente diclosulam no potencial fisiológico das sementes e crescimento inicial das plântulas. As sementes das forrageiras, foram semeadas em substrato previamente umedecido com solução de diclosulam nas concentrações de 0; 0,5; 1,0 e 1,5 vezes a dose registrada do produto pelo fabricante. Na segunda etapa, após o cultivo em casa de vegetação por 107 dias, foi realizada a aplicação dos pósemergentes, por meio de pulverizador costal de pressão constante em condições atmosféricas de 23º C de temperatura e 71% de umidade relativa do ar (UR). Foram utilizadas concentrações do herbicida referentes a zero; 0,5; 1,0 e 1,5 vezes a dose registrada pelo fabricante. Os resultados da primeira etapa, demostraram que as variáveis comprimento da parte aérea, comprimento de raiz e massa seca total foram alteradas negativamente pela exposição às diferentes doses de diclosulam, enquanto o potencial germinativo e a capacidade de emergir não foram prejudicadas, em ambas as espécies estudadas. Na segunda etapa os resultados obtidos mostraram maior sensibilidade das plantas de cornichão ao herbicida bentazona e das plantas de trevo vermelho ao herbicida iodosulfurom-metílico. Portanto é possível concluir que, as diferentes doses de diclosulam não comprometem o potencial germinativo, nem a capacidade de emergir das sementes de cornichão e trevo vermelho, porém, interferem negativamente no seu desenvolvimento. Já com a aplicação dos pósemergentes, o cornichão apresentou sensibilidade ao herbicida bentazona e características de tolerância ao herbicida iodosulfurom-metílico, enquanto o trevo vermelho respondeu de forma contrária, sendo mais sensível a aplicação do iodosulfurom-metílico e tolerante ao bentazona, ativando mecanismos de adaptação e permitindo o desenvolvimento da planta. / The objective of this research was to evaluate viability, seed vigor and growth of birdsfoot trefoil and red clover, plants from Embrapa Pecuária Sul, was submitted to different doses of pre and post emergent herbicides (diclosulam, bentazon and iodosulfuron-methyl), with purpose to identify herbicides selective to these forages. In the first stage of the experiment the effect of the pre-emergent herbicide diclosulam in physiological potential of seeds and initial growth of seedlings was evaluated. The seeds of this forages were sown on a substrate previously moistened with diclosulam solution at concentrations 0; 0,5; 1,0 and 1,5 times the dose registred by manufacturer. In second stage, after the cultivation in the greenhouse for 107 days, the postemergence application was carried out by a costal sprayer with constant pressure at atmospheric conditions of 23 ºC and 71% relative humidity). Herbicide concentrations used were zero; 0,5; 1,0 and 1,5 times the dose registred by manufacturer. The results of the first stage showed that the variables shoot length, root length and total dry mass were negatively altered by exposure to different doses of diclosulam, while germination potential and emergence capacity were not impaired in both species studied. In the second stage, the results obtained showed greater sensitivity of birdsfoot trefoil plants to herbicide bentazon and the red clover plants to iodosulfuron-methyl herbicide. Therefore, it is possible to conclude that, different doses of diclosulam do not compromise germination potential, neither ability to emerge from birdsfoot trefoil and red clover seeds, but negatively interfere in its development. With the post-emergence application, the birdsfoot trefoil showed sensitivity to herbicide bentazon and characteristics of tolerance to herbicide iodosulfuron-methyl, while the red clover responded in a contrary way, being more sensitive the application of the herbicide iodosulfuron-methyl and tolerant to bentazon, activating mechanisms of adaptation and allowing plant development.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-06-06T14:03:30Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tese_fernanda_reolon_tonel.pdf: 1524534 bytes, checksum: 4326518bd7bd55acb90e0f2630de6ac4 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-06-06T14:30:22Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tese_fernanda_reolon_tonel.pdf: 1524534 bytes, checksum: 4326518bd7bd55acb90e0f2630de6ac4 (MD5) / Made available in DSpace on 2017-06-06T14:30:22Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tese_fernanda_reolon_tonel.pdf: 1524534 bytes, checksum: 4326518bd7bd55acb90e0f2630de6ac4 (MD5) Previous issue date: 2014-12-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O estudo objetivou avaliar a capacidade fitorremediadora do trevo vermelho (Trifolium pratense L.) e do feijão miúdo (Vigna unguiculata L.) a compostos derivados de petróleo. Foram realizados experimentos em laboratório e em casa de vegetação, onde as sementes das duas espécies foram expostas as concentrações de 0,0; 0,1; 0,2 e 0,3 % (v/v) de óleo diesel e em seguida submetidas às análises de viabilidade, vigor e atividade de enzimas hidrolíticas. Em uma segunda etapa, as duas espécies, foram submetidas à contaminação por 0,0; 0,5; 1,0 e 1,5 % (v/v) de óleo diesel. O trevo vermelho foi cultivado durante 112 dias e a partir de coletas periódicas foram realizadas análises de parâmetros de crescimento, teor de pigmentos fotossintéticos, anatomia foliar e radicular e análise de enzimas antioxidantes. O feijão miúdo permaneceu em casa de vegetação durante 60 dias, para as análises periódicas dos parâmetros de crescimento, teor de pigmentos fotossintéticos, fotossíntese e enzimas antioxidantes. Para ambas, no início e no final do experimento foi realizada análise de hidrocarbonetos de petróleo no substrato de cultivo. Os resultados obtidos mostraram maior sensibilidade das duas espécies submetidas ao contaminante em condições controladas, no entanto, a influência negativa foi observada apenas nas maiores concentrações do composto tóxico. Os demais parâmetros foram diferentemente influenciados nas duas espécies, destacando-se para ambas, os efeitos negativos ocorridos na maior concentração de óleo diesel (1,5 % (v/v)) e benefícios ocasionados na menor concentração (0,5 % (v/v)). Portanto, os efeitos fisiológicos ocasionados permitiram a determinação do limite de tolerância das plantas de trevo vermelho e feijão miúdo ao óleo diesel. A eficiência na remoção dos compostos tóxicos, verificada para as duas espécies permitiu a confirmação de ambas como fitorremediadoras de compostos derivados de petróleo. / The study aimed to evaluate the phytoremediation capacity of red clover (Trifolium pratense L.) and cowpea (Vigna unguiculata L.) derived from petroleum compounds. Experiments were conducted in laboratory and greenhouse, where the seeds of both species concentrations of 0.0 were exposed; 0.1; 0.2 and 0.3% (v/v) of diesel and then subject to feasibility analysis, strength and activity of hydrolytic enzymes. In a second step, the two species were submitted to contamination by 0.0; 0.5; 1.0 and 1.5 % (v/v) of diesel oil. Red clover was grown for 112 days and from periodic samples were taken analyzes of growth parameters, photosynthetic pigments content, leaf and root anatomy and analysis of antioxidant enzymes. Cowpea remained in a greenhouse for 60 days for regular reviews of the growth parameters, photosynthetic pigments content, photosynthesis and antioxidant enzymes. For both the beginning and end of the experiment was performed petroleum hydrocarbon analyzer in the cultivation substrate. The results showed higher sensitivity of both the dopant species under controlled conditions, however, the negative effect was observed only at higher concentrations of the toxic compound. The other parameters were influenced differently in the two species, standing out for both the negative effects occurring in the highest concentration of diesel (1.5 % (v/v)) and benefits incurred in lower concentration (0.5 % (v/v)). Therefore, the physiological effects caused allowed the determination of the tolerance limit for red clover plants and cowpea to diesel oil. The efficiency of removal of toxic compounds found in these species permitted confirmation both as phytoremediator of petroleum compounds.

<p>Hemijska karakterizacija ekstrakata&nbsp; <em>T. pratense&nbsp;</em>L.&nbsp; određena je LC-MS-MS, a etarskih ulja GC-MS analizom, pri čemu je&nbsp; identifikovano vi&scaron;e&nbsp;bioaktivnih jedinjenja.&nbsp; Spektrofotometrijskim&nbsp;metodama ispitan je antioksidantni potencijal&nbsp;ekstrakata i etarskih ulja. Hepatoprotektivni&nbsp;efekat određen je&nbsp; na&nbsp; homogenatu i hemolizatu&nbsp;jetre laboratorijskih mi&scaron;eva.&nbsp; Budući da&nbsp; ova&nbsp;biljka ima visok sadržaj izoflavona, određena je&nbsp;<br />estrogena, antiestrogena aktivnost, kao i&nbsp;citotoksičnost ekstrakata. Antimikrobna&nbsp;<br />aktivnosta ekstrakata i etarskih ulja ispitana je&nbsp;na&nbsp; &scaron;est bakterijskih sojeva.&nbsp; Ekstrakti i etarska&nbsp;ulja su bili umereno aktivni u pogledu biolo&scaron;kih&nbsp;aktivnosti, osim u slučaju antimikrobne&nbsp;aktivnosti koju nije pokazao nijedan ispitivani&nbsp;uzorak.</p> / <p>The chemical characterization of&nbsp; <em>T.&nbsp; pratense</em> extracts was determined by LC-MS-MS, and the essential oils of the GC-MS analysis, wherein the&nbsp; various&nbsp; bioactive compounds were identified.&nbsp; Antioxidant potential of extracts and essential oils was tested&nbsp; by spectrophotometric methods.&nbsp; Hepatoprotective&nbsp; effect&nbsp; was determined&nbsp; in&nbsp; haemolysate and liver homogenate of&nbsp; laboratory&nbsp; mice.&nbsp; Estrogen, antiestrogen activity, as well&nbsp; as the cytotoxicity of the extracts were&nbsp; determined&nbsp; due to the high&nbsp;content of isoflavones.&nbsp; Antimicrobial activity of extracts and essential oils was&nbsp; investigatied&nbsp; on six bacterial strains. Essential oils and extracts were moderately active in terms of biological activity, except&nbsp; in the case of&nbsp;&nbsp; antimicrobial activity&nbsp; where none of the extracts were active against tested bacterial strains.</p>

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The ingestive behavior and the ingestion of forage by beef heifers were evaluated in the vegetative and flowering stages of ryegrass (Lolium multiflorum L.). The treatments were: grazing exclusively on ryegrass, ryegrass intercropping with vetch (Vicia sativa L.) or red clover (Trifolium pratense L.). The grazing method was continuous with variable stocking. The experimental design was the completely randomized with repeated measures arrangement, three forage systems and three repetitions of area. For evaluations of forage intake, chromic oxide was used as an indicator of fecal production and heifers were considered as experimental units. The forage mass, allowance of forage and allowance of leaf blades were, 1596.2 kg / ha DM, 12 kg DM / 100 kg BW and 4.5 kg DM / 100 kg BW respectively. The participation of vetch and red clover in the forage mass corresponded to 13.2% and 4.5% respectively in the vegetative stage. In the flowering stage, the participation of legumes corresponded to 0.6%. In forage as grazed, crude protein (17.8%), neutral detergent fiber (55.8%) and in situ dry matter digestibility (76.7%) were similar among forage systems. These variables differed between phenological stages of ryegrass. The grazing time differed between the systems being superior in the system with vetch. The forage intake rate in the clover system was higher than the other systems. The dry matter intake was similar among forage systems and was higher in the vegetative stage compared to the flowering stage. The average daily gain was similar among forage systems and phenological stages. With little participation of red clover or vetch in the herbage mass intercropped with ryegrass, it does not modify the intake of dry matter, crude protein and neutral detergent fiber, resulting in the same performance of beef heifers. / Foram avaliados o comportamento ingestivo e a ingestão de forragem por bezerras de corte, nos estádios vegetativo e de florescimento do azevém (Lolium multiflorum L.). Os tratamentos foram: pastagem exclusiva de azevém, azevém consorciado com ervilhaca (Vícia sativa L.) ou com trevo vermelho (Trifolium pratense L.). O método de pastejo foi o contínuo com lotação variável. O delineamento experimental foi o inteiramente casualizado com medidas repetidas no tempo, três sistemas forrageiros e três repetições de área. Para as avaliações da estimativa de ingestão de forragem, foi utilizado o óxido de cromo como indicador da produção fecal e as bezerras foram consideradas as unidades experimentais. A massa de forragem, oferta de forragem e oferta de lâminas foliares foram: 1596,2 kg/ha de MS, 12 kg de MS/100 kg PC e 4,5 kg de MS/ 100 kg PC, respectivamente. A participação de “ervilhaca” e “trevo vermelho” na massa de forragem correspondeu a 13,2% e 4,5% respectivamente no estádio vegetativo. No estádio de florescimento o percentual das leguminosas correspondeu a 0,6%. Na forragem da simulação do pastejo, a proteína bruta (17,8%), fibra em detergente neutro (55,8%) e digestibilidade in situ da matéria seca (76,7%), foram similares entre os sistemas forrageiros. Essas variáveis diferiram entre estádios fenológicos do azevém. O tempo de pastejo diferiu entre os sistemas sendo superior no sistema com ervilhaca. A taxa de ingestão de forragem no sistema com trevo foi superior aos demais sistemas. A ingestão de matéria seca foi similar entre os sistemas forrageiros e foi maior no estádio vegetativo em relação ao estádio de florescimento. O ganho médio diário foi semelhante entre sistemas forrageiros e estádios fenológicos. Com pequena participação de “trevo vermelho” ou “ervilhaca” na massa de forragem consorciada com azevém, a ingestão de matéria seca, de proteína bruta e de fibra em detergente neutro não são modificadas resultando em mesmo desempenho de bezerras de corte.