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Optimization and application of in vitro techniques in selected members of the family Brassicaceae
Hilgert-Delgado, Alois Antonín ; Fernández Cusimamani, Eloy (advisor) ; Jana, Jana (referee)
This thesis is focused on the application and optimization of biotechnological methods within the family Brassicaceae. Most of this thesis is focused specifically the genus Brassica. The main objective of this dissertation thesis was to optimize and apply selected biotechnological methods in the resynthesis of oilseed rape (B. napus) from its baseline species (B. oleracea, CC and B. rapa, AA) and subsequently create valuable genetic plant materials.
The optimized procedure implemented in my work have exhibited better results of hybrid production (resynthesis) than in similar published experiments and provides a simplified and less laborious method via simple ovule culture and early hybrid verification.
The next accomplished step was to work with a wider range of materials using a broader source of different and contrasting genotypes from Brassica rapa (spring and winter oilseed and vegetable turnips) and B. oleracea (green and purple curly kales and cabbages) for new combinations and wider genetic diversity.
It was concluded that the optimized ovule culture protocol with early verification, developed in the framework of this thesis, is satisfactorily sufficient enough to be applicable in breeding programmes, aimed at diversity expansion of winter oilseed rape gene pool, as the resynthesized embryos were derived in most combinations.
New resynthesized verified lines were colchicine treated in order to obtain diploid regenerants and the fertile plants were self-pollinized and crossed with elite oilseed rape lines for further research and breeding.
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Traditional Use of Coca (Erythroxylum sp.)
Jelínek, Petr ; Fernández Cusimamani, Eloy (advisor) ; Pavla, Pavla (referee)
Coca (Erythroxylum sp.) is a genus of plants in the family Erythroxylaceae, whose most important species come from the Andean region of South America. Coca leaves are traditionally used as a medicament, a stimulant or for the production of food products. In some areas, coca is an important part of the culture. On the other hand, coca is notoriously known as a raw material for the production of cocaine which is then illegally distributed throughout the world. The aim of this thesis was to objectively summarize and evaluate information about coca shrub, its expansion, growing, and especially about the use and importance of the coca leafs. This information was obtained from specialist literature, scientific articles and works of national and international organizations. Coca leaves contain high levels of lipids, protein, carbohydrates, fibre, vitamins A, E, B2, B5, B6 and B12, calcium, phosphorus, iron, magnesium, potassium and zinc in comparison with selected food (corn, beans, bananas, cow milk, chicken meat), they also have a high calorific value. Therefore, coca is an important nutritional additive. The leaves are chewed or used to manufacture traditional medicines, alcoholic drinks, mate de coca tea, Coca-Cola, cosmetic products, cigarettes, and concentrated cocaine, which occurs in coca leaves as an alkaloid to a concentration of 1,5%. Coca is part of traditional festivals and celebrations (e.g. baptisms, engagements, weddings and funerals), serves as a sacrifice and an idol, is offered at social gatherings and appears in folk tales. Coca is chewed mainly for its stimulant effects, it also suppresses hunger and fatigue and generally improves mood. In terms of pharmaceutical effects, risks and other factors, coca cannot be regarded as comparable to cocaine. Thanks to its positive effects and profound cultural importance, coca should remain part of the Andean tradition. In addition, it could also find use in modern medicine. Coca would not have to be considered merely as a dangerous drug.
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Micropropagation of Satureja Montana L.
Zunová, Tereza ; Fernández Cusimamani, Eloy (advisor) ; Michaela, Michaela (referee)
Winter savory (Satureja montana L.) is a plant occurring in the Mediterranean and the Black Sea region. For its high content of essential oils is widely used in culinary and traditional medicine.
Savory is propagated generative and vegetative. Seeds germinate slowly, and therefore, the vegetative propagation is faster, especially in in vitro conditions may be propagated a large amount of plant material in a short time.
The aim of this thesis was to investigate the growth effect of cytokinins on regeneration and growth plants from nodal segments of S. montana L.
Nodal segments were cultured on six variants of medium supplemented with 0.5; 1 and 1.5 mg/l 6-benzylaminopurine (BAP) and 0.5; 1 and 1.5 mg/l kinetin (KIN). Variant without growth regulators served as a control. Culture conditions were 25/20 °C day/night, light intensity 3000 lx and daily period of 16/8 hours. After 6 weeks of measurements were evaluated parameters such as: number of shoots, plant height, the number of newly created nodes and the number and length of roots.
In respect of the coefficient of micropropagation (i.e. the number of newly created nodes on the plant) was statistically significantly best variant medium supplemented with 1.5 mg/l KIN where on average create 8.88 +/- 2.87 nodes. At the height of the plant which was highest in the variant with a concentration 1.5 mg/l KIN (2.27 +/- 1.33) and the number of shoots with highest average value in a concentration 1 mg/l BAP (2.87 +/- 1.19) encore of growth regulators were not statistically proven. High concentrations of BAP and KIN inhibited the induction of roots and from variants transferred to ex vitro conditions were regenerated 46 % plants.
Adding BAP and KIN into the medium has an influence on plant regeneration of Satureja montana L.
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Micropropagation of Lippia dulcis Trev.
Zubíková, Alžběta ; Fernández Cusimamani, Eloy (advisor) ; Michaela, Michaela (referee)
Lippia dulcis Trev. (Verbenacea) is a perennial herb with a creeping stem from Central America. It is known for its healing properties for problems with the respiratory or digestive system, but also its sweet taste, which causes terpene hernandulcin because of which the leaves of Lippia dulcis Trev. are 1,000 times sweeter than sugar. Generatively, the plant reproduces by seeds and vegetatively by cuttings.
The main objective of this study was to investigate the influence of cytokinins kinetin (KIN) and 6-benzylaminopurine (BAP) with the addition of auxin indole-3-acetic acid (IAA) on the growth and regeneration of plants from nodal segments Lippia dulcis Trev. in vitro conditions.
Nodal segments were cultured on 6-variants of MS medium supplemented with growth regulators at concentrations of 0.5; 0.75 and 1.0 mg / l BAP and KIN with combination 0.5 mg / l IAA and one variant of pure MS medium as a control. Nodal explants were cultured for 35 days (the time of subculturing) and measured every 7 days. The following parameters were evaluated: regeneration, number of shoots, length of shoots, number of nodes, number and length of roots.
The measurement results show that a combination of growth regulators auxin-cytokinin have no positive influence on the growth and regeneration Lippia dulcis Trev. in vitro conditions. The best regeneration (100%), growth (on an average length of the shoots 3.05 +- 1.42) and the highest number of newly created points (5.79 +- 2.4) occurred at the explants cultured on a clean MS medium without addition of growth regulators. Among the variant with pure MS and media variants with the addition of growth regulators are statistically significat defferences, but without a favorable influence on the development of nodal explants.
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The importance and use of sea buckthorn (Hippophae rhamnoides L.) in veterinary medicine
Lešnerová, Olga ; Lukešová, Daniela (advisor) ; Fernández Cusimamani, Eloy (referee)
Bachelors thesis mainly discussed the findings which are related to significance and potential utilization of the sea buckthorn (Hippophae rhamnoides L.) in veterinary medicine. We could appoint an antioxidant and immunomodulatory effects, positive effects on the cardiovascular system or treatment of skin diseases. Sea buckthorn could be also used in animal nutrition. For feeding purposes were mainly used leaves and residues after pressing juice and oils. The work also included informations about plant morphology, cultural management, harvesting, yield and processing. In a separate chapter were mentioned substances which were contained in various parts of the plant. Thanks to them, seabuckthorn is so familiar recently. Its potential is primarily in the use of bioactive substances in the field of medicine. In the last part of the thesis were created diagrams that showed quantity of selected substances contained in sea buckthorn fruits/juice and oils. For comparison were chosen vitamin C for fruits, vitamin E and carotenoids for oils within two subspecies H. rhamnoides L. subsp. rhamnoides and H. rhamnoides L. subsp. sinensis. The amount of extracted substances were often influenced by the used technology. In a detailed study and comparison of literature sources, it was found that for the subspecies H. rhamnoides L. subsp. rhamnoides literature indicates the quantity of vitamin C with a median of 229,5 mg/100 g, the content of vitamin E in the seed oil with a median of 351,8 mg/100 g and for pulp oil 147,5 mg/100 g. The carotenoid content in seed oil came with median 16,25 mg/100 g, for pulp oil it was 265 mg/100 g. Subspecies H. rhamnoides L. subsp. sinensis had the median value of vitamin C 1038 mg/100 g. For vitamin E in seed oil was the median value 119,3 mg/100 g, in pulp oil it was 171 mg/100 g. The median for carotenoids content in seed oil was 32,5 mg/100 g, the value for pulp oil was 282 mg/100 g. It will be necessary to ensure the subsequent research analysis about substances, especially in the field of veterinary medicine.
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Milk thistle (Silybum marianum) utilization in animal feeding
Klímová, Stefanie ; Lukešová, Daniela (advisor) ; Fernández Cusimamani, Eloy (referee)
Milk thistle (Silybum marianum (L.) Gaertn.) is a medicinal herb known for its hepatoprotective and antioxidant properties which not only have positive results in human medicine but also in veterinary medicine. Active substances of milk thistle are called silymarin complex and they are obtained by pressing the seeds of the plant. The bachelor thesis has a literature review of summarized findings of the use of substances contained from milk thistle in human medicine and further in animal nutrition. In the next section were selected case studies dealing with the influence of the addition of milk thistle to complete feed mixtures in laboratory animals. Milk thistle in experimental works was administered to rabbits as an antioxidant AV3 (0.2 ml.kg-1) and was followed by evaluation of metabolic parameters in comparison with the control group that was fed complete feed mixture without the addition of milk thistle. The investigated parameters were growth, biochemical and reproductive parameters and quality of rabbit meat. Addition of milk thistle did not affect the growth parameters, the quality of the flesh and blood biochemistry, compared to the control groups of animals, with the exception of the level of cholesterol, which increased by 38.6%. Reproduction parameters of milk thistle supplement increased the number of born and weaned kids and decreased offspring mortality compared to the control group. Milk thistle can therefore be used as a dietary supplement for improving reproductive parameters. Use of substance contents of milk thistle in human medicine has been evaluated in the survey from where 55 % of the 120 respondents use silymarin in tablets in the acute or chronic liver diseases for its hepatoprotective effect.
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