Thèse de doctorat--Sciences naturelles--Faculté des sciences de Paris, 1910. N°: 61. / Notes bibliogr.

10 ABSTRACT The subject of this study was to evaluate the efect of abiotc elicitor on rutn producton in callus and suspension cultures of buckwheat. The cultvar of buckwheat used for this research was Fagopyrum esculentum Moench var. Bambi, cultvated in Murashige and Skoog nutrient medium with the additon of growth regulator 2,4-dichlorfenoxyacetc acid (2,4-D) in concentraton of 1 mg/l. The elicitor used in this study was a soluton of methylviologen, 1 ml of it was added to the cultures in three diferent concentratons: c1 = 100.0 mg/100 ml, c2 = 10.0 mg/100 ml and c3 = 1.0 mg/100 ml. The elicitor was afectng the cultures for 6, 12, 24, 48, 72 or 168 hours. Afer the defned period of tme, cultures were collected, dried out and stored for further analysis of rutn content. To control samples (without elicitor treatment) 1 ml of ethanol 96% was added and they were collected afer 6, 24, 72 or 168 hours. Releasing of rutn into the nutrient medium was also investgated. Rutn content in each sample of cultures and in each sample of nutrient medium was later determined by HPLC. Any signifcant increase in the producton of rutn was not observed in this study. The maximum amount of rutn detected was 0.1 mg/g DW, thus the lowest quantty detectable, and was found in suspension cultures in three cases, afer the additon of...

Elicitation is one of the methods used for increasing the production of secondary metabolites in vitro cultures. The aim of this work is to evaluate the effect of abiotic elicitor methylviologen (paraquat) on the production of flavonoids in callus and suspension cultures of Fagopyrum esculentum Moench., variety Pyra. The cultures were cultivated in Murashige and Skoog nutrient medium with addition of 2,4-D in concentration of 1 mg/l as a growth regulator. Elicitor was added as a solution in three different concentrations (c1 = 2.1929 · 10-4 mol/l, c2 = 2.1929 · 10-3 mol/l and c3 = 2.1929 · 10-2 mol/l). The effect of elicitation on rutin production was monitored in six time intervals: 6, 12, 24, 48, 72 and 168 hours. The rutin content was determined by HPLC analysis. No rutin was produced in callus and suspension cultures without the presence of elicitor. Even after the elicitation, there was no statistically significant increase in the production of rutin. The maximum rutin content was detected in the suspension culture after 12 hours of elicitor treatment in c2 concentration, the content was 0.1 mg/g DW. The release of rutin into the nutrient medium was also not observed.

Plants are an important source of secondary metabolites, which are a valuable natural substance used in many fields. One way to increase their production is by the elicitation method. In this paper the effect of abiotic elicitor methylviologene in three different concentrations was studied on the rutin production in callus and suspension culture of Fagopyrum esculentum Moench variety Spačinki. The cultivation was carried out on Murashige and Skoog nutrient medium with the addition of a 1 mg/l 2,4- dichlorophenoxyacetic acid as growth regulator. Samples were taken at regular time intervals after 6, 12, 24, 48, 72 and 168 hours of elicitation. The rutin content was analyzed by HPLC. The maximum rutin production (0.3 mg/g DW) was recorded in the callus culture after 48 hours of methylviologene treatment at a concentration of 2.1929.10-4 mol/l. No increase in rutin content after methylviologene elicitation was observed in the suspension culture. The study also included monitoring of the rutin release into nutrient medium, but this was not demonstrated.

62 10 ABSTRACT The object of this study was the influence of abiotic elicitor on the production of rutin in suspension culture of Fagopyrum esculentum Moench. The culture was cultivated in Murashigeho and Skoog nutritive medium with growth regulator: 2,4−dichlorfenoxyacetic acid (1 ml/l). The ultrasound was used as abiotic elicitor (0,1 W/cm3 , 35 kHz) for time period of 1, 2, 3, 4 and 5 min. The samples were taken 0, 6, 12, 24, 48, 72 and 168 hours after elicitation. The kontrol samples (without the influence of ultrasound) were taken 24 and 168 hours after elicitation. The amount of rutin was analyzed by HPLC. Suspension culture of Fagopyrum esculentum Moench. didn't produce any rutin under the influence of ultrasound. No release of rutin into the nutritive medium was observed during this study.

The aim of this work was to investigate the effect of pyrazine derivative, 1-octyl-3-(pyrazin-2- yl)urea, as an abiotic elicitor on the production of flavonoid rutin in in vitro cultures of Fagopyrum esculentum Moench., Cultivar Bamby. Suspension and callus cultures were cultivated on Murashige and Skoog nutrient medium (MS) with the addition of 2,4-dichlorophenoxyacetic acid (2,4-D) as growth regulator at a concentration of 1 mg/l. The elicitor solution was added to the cultures at three concentrations: c1 (100,0 mg/100 ml), c2 (10,0 mg/100 ml) and c3 (1,0 mg/100 ml). The elicitor was monitored at six time intervals: 6, 12, 24, 48, 72 and 168 hours. To control samples 1 ml of ethanol 96% was added instead of elicitor solution and samples were collected after 24 and 168 hours. Samples were taken at given time intervals and dried. Subsequently, the rutin content was monitored by HPLC. The rutin release into the nutrient medium was also tested. During the experiment on the callus cultures no statistically significant increase in rutin production after elicitor treatment was observed. But elicitor increased rutin production in suspension cultures after treatment in all tested concentrations. The calluses always released rutin into the nutrient medium. The rutin content in the media ranged from 53,4 to...

Yang, Nan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (p. 94-117). / Abstracts in English and Chinese. / ACKNOWLEDGMENTS --- p.I / ABSTRACT --- p.II / LIST OF ABBREVIATIONS --- p.VII / TABLE OF CONTENTS --- p.IX / Chapter Chapter 1 --- general introduction / Chapter 1.1 --- Cholesterol and cardiovascular disease --- p.1 / Chapter 1.2 --- Functions of cholesterol and lipoprotein --- p.2 / Chapter 1.3 --- Cholesterol metabolism and regulation in the body --- p.5 / Chapter 1.3.1 --- General process of cholesterol metabolism --- p.5 / Chapter 1.3.2 --- Cholesterol metabolism in liver --- p.6 / Chapter 1.3.2.1 --- The uptake of LDL cholesterol into the liver --- p.6 / Chapter 1.3.2.2 --- Cholesterol synthesis --- p.7 / Chapter 1.3.2.3 --- Synthesis of bile acids --- p.8 / Chapter 1.3.2.4 --- RCT pathway --- p.9 / Chapter 1.3.3 --- Lipids absorption in the intestine lumen --- p.9 / Chapter 1.3.3.1 --- Niemann-Pick Cl like 1(NPC1L1) --- p.10 / Chapter 1.3.3.2 --- ABCG5/8 --- p.10 / Chapter 1.3.3.3 --- Acyl-CoA:cholesterol acyltransferase (ACAT) 2 --- p.11 / Chapter 1.3.4 --- Cholesterol homeostasis --- p.11 / Chapter 1.3.5 --- The regulation of the cholesterol metabolism --- p.11 / Chapter 1.3.5.1 --- The role of SREBP-2 --- p.11 / Chapter 1.3.5.2 --- The role of LXR --- p.13 / Chapter 1.3.5.3 --- Feedback regulation of cholesterol --- p.13 / Chapter 1.4 --- Bile acid metabolism --- p.13 / Chapter 1.4.1 --- The function of bile acid --- p.13 / Chapter 1.4.2 --- Bile acid synthesis --- p.14 / Chapter 1.4.3 --- Enterohepatic circulation of bile --- p.14 / Chapter 1.5 --- Effect of Dietary composition on the blood cholesterol --- p.15 / Chapter 1.5.1 --- Dietary cholesterol --- p.15 / Chapter 1.5.2 --- Dietary protein --- p.15 / Chapter 1.5.2.1 --- Research history of dietary protein on the cholesterol --- p.15 / Chapter 1.5.2.2 --- Dietary casein --- p.17 / Chapter 1.5.2.3 --- Soy protein --- p.18 / Chapter 1.5.2.4 --- Buckwheat protein --- p.18 / Chapter 1.5.2.5 --- Mechanism of dietary protein on the cholesterol --- p.18 / Chapter 1.5.3 --- Dietary fiber --- p.18 / Chapter 1.5.4 --- Other functional components in the diet --- p.19 / Chapter 1.5.4.1 --- Phytosterol --- p.19 / Chapter 1.5.4.2 --- Dietary flavonoids --- p.21 / Chapter 1.6 --- Chemical composition of Tartary buckwheat --- p.22 / Chapter 1.6.1 --- Buckwheat protein --- p.22 / Chapter 1.6.2 --- Dietary fiber --- p.23 / Chapter 1.6.3 --- Phytosterols --- p.23 / Chapter 1.6.4 --- Flavonoids --- p.23 / Chapter Chapter 2 --- Effect of Tartary Buckwheat Flour on Blood Cholesterol Level in Male Hamsters / Chapter 2.1 --- Introduction --- p.25 / Chapter 2.2 --- Objective --- p.27 / Chapter 2.3 --- Materials and methods --- p.27 / Chapter 2.3.1 --- Hamsters --- p.27 / Chapter 2.3.2 --- Diets --- p.28 / Chapter 2.3.3 --- "Determination of plasma lipid, lipoproteins" --- p.30 / Chapter 2.3.4 --- Determination of cholesterol concentration in organs --- p.31 / Chapter 2.3.5 --- Determination of fecal neutral and acidic sterols output --- p.31 / Chapter 2.3.6 --- "Western blotting of liver SREBP-2, LDLR, HMGR, LXR and CYP7A1 proteins" --- p.36 / Chapter 2.3.7 --- "Real-Time PCR Analysis of mRNA or Liver SREBP-2, LDLR, HMGR, and CYP7A1 and Small Intestine NPC1L1, ABCG5, ABCG8, ACAT2, and MTP" --- p.37 / Chapter 2.3.8 --- Intestinal ACAT2 activity measurement --- p.37 / Chapter 2.3.9 --- Statistics --- p.39 / Chapter 2.4 --- Results --- p.40 / Chapter 2.4.1 --- Nutritional composition of different flours --- p.40 / Chapter 2.4.2 --- "Growth, food intake and relative organ weights" --- p.44 / Chapter 2.4.3 --- Effect of different flour diets on the plasma lipid profile --- p.44 / Chapter 2.4.4 --- Effect of different flour diets on organ cholesterol of hamsters --- p.44 / Chapter 2.4.5 --- Cholesterol balance and excretion of fecal neutral and acidic Sterols --- p.44 / Chapter 2.4.6 --- "Effect of different flour diets on hepatic SREBP-2, HMGR, LDLR and CYP7A1 immunoreactive mass" --- p.51 / Chapter 2.4.7 --- "Effect of different flour diets on intestinal ABCG5, ABCG8, NPC1L1, MTP, and ACAT2 immunoreactive mass" --- p.54 / Chapter 2.4.8 --- Effect of different diet group on intestinal ACAT activity --- p.54 / Chapter 2.5 --- Discussion --- p.57 / Chapter 2.6 --- Summary --- p.61 / Chapter Chapter 3 --- Effect of DefattedTartary Buckwheat Protein Extract on Blood Cholesterol Level in Male Hamsters / Chapter 3.1 --- Introduction --- p.62 / Chapter 3.2 --- Objective --- p.63 / Chapter 3.3 --- Materials and methods --- p.63 / Chapter 3.3.1 --- Hamsters --- p.63 / Chapter 3.3.2 --- Diets --- p.63 / Chapter 3.3.3 --- "Determination of plasma lipid, lipoproteins" --- p.66 / Chapter 3.3.4 --- Determination of cholesterol concentration in organs and fecal neutral and acidic sterols output --- p.66 / Chapter 3.3.5 --- "Western blotting of liver SREBP-2, LDLR, HMGR and CYP7A1 proteins" --- p.66 / Chapter 3.3.6 --- "Real-Time PCR Analysis of mRNA or Liver SREBP-2, LDLR, HMGR, and CYP7A1 and Small Intestine NPC1L1, ABCG5, ABCG8, ACAT2, and MTP" --- p.66 / Chapter 3.3.7 --- Intestinal ACAT2 activity measurement --- p.67 / Chapter 3.3.8 --- Protein digestibility determination --- p.67 / Chapter 3.3.9 --- Statistics --- p.67 / Chapter 3.4 --- Results --- p.68 / Chapter 3.4.1 --- Diet composition --- p.68 / Chapter 3.4.2 --- "Growth, food intake, fecal excretion" --- p.72 / Chapter 3.4.3 --- Relative organ weights and organ cholesterol concentration --- p.72 / Chapter 3.4.4 --- Effect of different defatted protein extracts on the plasma lipid profile --- p.76 / Chapter 3.4.5 --- Cholesterol Balance and Excretion of Fecal Neutral and Acidic Sterols --- p.76 / Chapter 3.4.6 --- "Apparent protein digestibility in casein, TBP, WP and RP diet groups" --- p.77 / Chapter 3.4.7 --- "Effect of different defatted protein extracts on hepatic SREBP-2, HMGR, LDLR and CYP7A1 immunoreactive mass" --- p.83 / Chapter 3.4.8 --- "Effect of different defatted protein extracts on intestinal ABCG5, ABCG8, NPC1L1, MTP, and ACAT2 immunoreactive mass" --- p.83 / Chapter 3.5 --- Discussion --- p.87 / Chapter 3.6 --- Summary --- p.91 / Chapter Chapter 4 --- Conclusion --- p.92 / References --- p.94

Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Tereza Ošťádalová Supervisor: Doc. PharmDr. Lenka Tůmová, CSc. Title of diploma thesis: The selenium effect on secondary metabolites production in in vitro cultures of medicinal plants - II. Key words: callus, suspension culture, abiotic elicitor, selenium, Fagopyrum In vitro plant cultures usually produce only a small amount of secondary metabolites. The method of elicitation is one of the options how to increase the production of these substances. The effect of selenium as abiotic elicitor on rutin production in callus and suspension culture of Fagopyrum esculentum Moench., variety Pyra was observed in this study. The release of rutin into the nutrient medium was studied as well. The cultivation was performed in Murashige and Skoog (MS) nutrient medium which was enriched with 1 mg/l of 2,4-dichlorophenoxyacetic acid. The solution of selenium in three different concentrations (c1 = 9.012.10-3 mol/l, c2 = 9.012.10-4 mol/l, c3 = 9.012.10-5 mol/l) was used. The samples were taken after 6, 12, 24, 48, 72 and 168 hours of elicitor treatment. The rutin content was determined by high-performance liquid chromatography (HPLC). The elicitation led to increasing of rutin amount in callus and also in suspension...

Higher plants represent an important source of valuable substances, so called secondary metabolites, which can be obtained through explant cultures of plants. Elicitation is a method of increasing the secondary metabolites production. This study aims to evaluate the secondary metabolites production in Fagopyrum esculentum variety Spacinska cultures in vitro after abiotic elicitor treatment. The experiment was focused on alteration of rutin production in callus and suspension cultures of F. esculentum var. Spacinska after selenium application. Murashige and Skoog nutrient medium supplemented with 1 mg l-1 2,4-dichlorophenoxyacetic acid was used for the cultivation. Selenium solutions of various concentrations (c1 = 9.012×10-3 mol l-1 , c2 = 9.012×10-4 mol l-1 , c3 = 9.012×10-5 mol l-1 ) were affecting the cultures for 6, 12, 24, 48, 72 and 168 hours. The content of rutin was determined by HPLC. The release of secondary metabolites into the nutrient medium was studied as well. After elicitor application, the rutin production increased in both callus and suspension cultures. Higher levels of rutin content were detected in callus culture. The maximum rutin content (0.6 mg g-1 DW) was reached after 12 h of elicitor treatment of c2 concentration in callus culture. Concerning suspension culture, the...

De nos jours, le sarrasin ou « blé noir », est toujours associé aux fameuses galettes bretonnes, cependant, son histoire demeure méconnue. Introduit comme culture vivrière dans l’ouest de la France (Bretagne et Normandie occidentale) au cours des XVe et XVIe siècles, le sarrasin y devient la principale culture et denrée alimentaire à partir du XVIIe siècle. Cette étude s’intéresse au rôle du sarrasin dans les différents pans des sociétés de l’Ouest du XVIe au XXe siècle, autour de quatre grandes parties : l’introduction et la diffusion du sarrasin en Europe, sa place dans les systèmes agraires du Massif armoricain, son incidence dans l’alimentation des populations de cette région, et son utilisation dans l’atténuation des crises de subsistances. Au fil des chapitres, les pratiques bretonnes et normandes font l’objet d’une comparaison, afin d’identifier les éléments constitutifs d’une identité commune aux populations de l’Ouest et ceux qui relèvent de particularismes territoriaux. Cette approche comparative est reproduite au sein même de ces deux espaces, pour appréhender leurs spécificités locales. / Today, buckwheat is unavoidably associated with the famous Breton galettes (buckwheat pancakes), yet we know little of its history. Introduced as a subsistence crop in western France (Brittany and west Normandy) during the 15th and 16th centuries, buckwheat became the predominant crop and foodstuff in the region from the 17th century. This study examines the role of buckwheat in various aspects of societies in western France from the 16th century to the 20th century, via four main sections: the introduction and spread of buckwheat in Europe, its place in the agrarian systems of the Armorican Massif, its impact on the diets of populations in this region and its use in relieving subsistence crises. The chapters compare practices in Brittany and Normandy, to identify elements of a common identity among populations in western France, and elements that are specific to territories. This comparative approach is repeated within these two spaces, to identify local particularities.