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In vitro cultures of medicinal plants - XIII.
Kubeš, Jan ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
Jan Kubeš Genista tinctoria in vitro - abiotická elicitace Cultures of medicinal plants in vitro - XIII. The plants cultures in vitro contain lesser amounts of secondary metabolites in compare with intact plants. The elicitors can affect these metabolites production. The effect of electric current (50mA) of different voltage (5, 7, 8, 10, 15, 20 and 24 V) and different time exposition (10, 30, and 60 minutes) on content of isoflavones was studied on (Genista tinctoria) suspension culture. The highest concentration of genistin (0.17 %) was measured in suspension culture after 30 min of elicitation by 10 V after 6 hours of cultivation. The highest concentration of daidzein (0.35 %) was found in suspension culture after 60 min of elicitation by 5 V after 24 hours of cultivation. The highest concentration of genistin (1.6 mg/100ml) was measured in medium after 30 min of elicitation by 5 V after 24 hours of cultivation. The highest concentration of daidzein (1.77 mg/100ml) was found in medium after 10 min of elicitation by 24 V after 6 hours of cultivation.
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In vitro cultures of medicinal plants - XII.
Janoutová, Martina ; Tůmová, Lenka (advisor) ; Dušková, Jiřina (referee)
Martina Janoutová In vitro cultures of medicinal plants - XII. Abstract The effect of ultrasound (US) as abiotic elicitor on the flavonolignans production in Silybum marianum L suspension culture was investigated. The culture was cultivated in Murashige and Skoog nutritive medium with ( - NAO) (g/l) as growth regulator at 25o C and luminous period 16 h of light and 8 h of darkness. The elicitor - ultrasound by frequency 35kHz and intensity 0,1Wcm-3 for a period 1, 2, 3, 4 and 5 min has been used. The samples were taken in 0, 6, 12, 24, 48, 72 and 168 h after US exposition. The control samples were taken in 0 and 48 h. The quantity of flavonolignans was determined by High performance liquid chromatography (HPLC). The highest increase of taxifolin content was apparent after 5 min of US elicitation and 48 h sampling (0,04%) - 400%, other increase was apparent after 5 min of elicitation and 72 h sampling. The higher content of silychristin was found after 1 min of US elicitation and 72 h sampling, the same level was observed after 2 min of elicitation and 24 h sampling. The higher level of silydianin was detected after 2 min of US elicitation and 6 h sampling and the silybin B after 2 min of elicitation and 12 h sampling after exposition. Taxifolin and flavonolignans release to the nutrient medium was...
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In vitro cultures of medicinal plants - XI.
Sojková, Kristýna ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
In vitro cultures of medicinal plants - XI. Elicitation is one of the few strategies that can be used in enhancement of secondary metabolites production from explant cultures. The effect of abiotic elicitor (silver nitrate) on flavonolignan and flavonoid taxifolin production in suspension culture of Silybum marianum L. (Gaertn.) and on isoflavones production in suspension culture of Genista tinctoria L. was tested. Silver nitrate in various concentrations (5.887.10-3 mol/l; 5.887.10-4 mol/l; 5.887.10-5 mol/l) was used as elicitor. Content of secondary metabolites in suspension cultures was determinated by high performance liquid chromatography (HPLC). The samples were taken after 6, 12, 24, 48, 72 and 168 hours after elicitor treatment. The highest content of taxifolin production (0.02 %) in suspension culture of Silybum marianum L. (Gaertn.) after silver nitrate (5.887.10-4 mol/l) treatment and 72 h sampling was detected. The highest content of genistin (0.05 %) in suspension culture of Genista tinctoria L. was found after silver nitrate (5.887.10-4 mol/l) treatment and 48 h sampling. The highest content of daidzein (0.09 %) was detected after elicitor (5.887.10-4 mol/l) treatment and 168 h sampling.
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Properties and in vitro Degradation of Metalic Biodegradable Materials
Ročňáková, Ivana ; Trojanová, Zuzanka (referee) ; Vojtěch,, Dalibor (referee) ; Podrábský, Tomáš (advisor)
Biodegradable metallic materials for medical applications have received considerable attention in recent years. The main reason is that they provide high potential for fabrication of temporal orthopedic implants such as bone fixation devices. Magnesium is an excellent candidate for fabrication of biodegradable implants due to its biocompatibility, mechanical properties similar to human bone and relevance for biological body functions. The fast degradation rate of magnesium and its biodegradable alloys in physiological environment limits its clinical application. Another attractive material in the field of biodegradable materials is zinc, which is among the essential elements in human body. Zinc exhibits an excellent corrosion resistance, and inferior biocompatibility compared to with magnesium. Hence, surface modification to form a hard, dense/porous, biocompatible and corrosion resistant modified layer has become an interesting topic in magnesium base biomaterials. Since hydroxyapatite is well tolerated by living organisms and in addition, improves the bone growth, it appears to be excellent candidate for such coatings on surface of biodegradable materials (e.g. Mg, Zn). This thesis is focused on comparison of corrosion behavior of pure non ferrous metals (Mg, Zn) and metals coated with hydroxyapatite, in simulated body fluids. The present approach is the use of modified atmospheric plasma spray technology to produce the hydroxyapatite coatings – suspension spraying. Composition and structure of the coatings and corrosion products were studied by light microscopy, scanning electron microscopy equipped with energy dispersive microanalyzer and X-ray diffraction. Corrosion of Mg and Zn samples was monitored by weight loss and determined by X ray and micro-tomography. The application of the HA coating resulted in decrease of corrosion rate of pure Mg. The corrosion rate of coated Mg samples was lower by 27,3 % in comparison with the corrosion rate of pure non coated Mg. Corrosion degradation of uncoated and coated Zn samples was minimal. The aplication of HA on the non ferrous surface appears to be a very promising method to improve corrosion and biological properties of these biodegradable materials.
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Glycaemic index of food and its determination
Špiláčková, Aneta ; Kouřimská, Lenka (advisor) ; Monika, Monika (referee)
The purpose of this thesis was to elaborate a literature recherche on a given theme with focus on analytical methods to estimate the glycaemic index of food.
This bachelor thesis is divided into three main chapters. First chapter describes general characteristics of glycaemic index of foods and influencing factors. In this chapter states the influence of high and low glycaemic index foods on health, wellbeing and glycaemia in humans. Subsequent part of this chapter is dealing with many other factors that influence the absolute amount of glucose in blood in 2 hour interval after meal.
Chapter two initiates into conventional and continual methods of glycaemia measurement.It highlights the advantages and limitations of each of these methods.
Third, the most extensive chapter is dealing with methods of glycaemic index assessment. This chapter is broken down into two parts. First part specifies clinical methods that are also called in vivo, and the second part is focusing on in vitro methods. Clinical methods require blood glycaemia measurement in healthy volunteers after consuming a test meal. With this method we are able to reliably state the glycaemic index of certain foods.
The limiting factor of this method is that it is rather time-consuming and it requires higher financial means. These measurements take place under a strict control, according to guidelines and only with the approval of ethical committee.
Analytical methods are based on simulation of human digestion. In vitro methods use hydrolysis index to obtain the glycaemic index. The final values are only an assessment of glycaemic index of given food. Other method that is described is measurement of fast and slowly available glucose and starch in given test foods. It is based on measurement of released glucose from the test food with HPLC method, time incubation and standardized conditions. Last method includes partitioning of a test food, and subsequent HPLC analysis of carbohydrates and sugar alcohols. The HPLC results are processed in artificial neural network to reveal glycaemic index values.
The advantages and disadvantages of each mentioned method are stated in conclusion of this work. I compared and contrasted reliability, elaborateness, budget demands
of analytical methods with clinical assessment methods.
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Healing of cultural varieties paniculate phlox (Phlox paniculata) from nematodes using in-vitro cultures
Nogly, David ; Matiska, Pavel (advisor) ; Augustinová, Ludmila (referee)
Phlox paniculata belongs to the family Poleminiaceae. Phlox paniculata is very important perenial which is used in commons gardens, park landscaping and also to product cut flowers. Although garden phlox are classified as resistant plants to pests and pathogens, they suffer from leaf and stem nematodes of genus Ditylenchus spp. Method eliminating these nematodes from plants for 100 % is still unknown. This thesis investigate the possibility of eliminating nematodes by using explant cultures and creating healthy plants for future mother plants of Phlox paniculata. For this experiment was chosen 10 varieties, 'Aňa Gaganova', 'Blue Paradise', 'Errötense Mädchen', 'Molodost', Igor Talkov', 'Jubilee', 'Laura', 'Mies Copijn', 'Sekret' and 'Winsdor'. These plants were also researched for regeneration of calluses and growing ratio of shoots according to their varietes.
Plants were tested for presence of nematodes Ditylenchus dipsaci by using modified Baermann Technique. All varietes of Phlox paniculata were grown in greenhouse and all of them were tested. For experiment were chosen leaf segments measuring 0,5 - 1 cm2 and the cultivation took place on MS medium (Murashige and Skoog 1962) + 30 g sucrose, pyridoxine 0,5 mg. l-1, thiamine 0,5 mg. l-1, glycine 2 mg. l-1 , nicotinic acid 0,05 mg. l-1. Leaf segments were placed to MS medium enriched by 1,5 mg . l-1 TDZ (Thidiazuron) and 0,5 mg . l-1 IAA (Indole-3-acetic acid). Callus regeneration at the level of plant growth regulators was almost fully vertified during this experiment. Best results were obtained from follow varieties 'Aňa Gaganova', 'Blue Paradise', 'Errötense Mädchen' and 'Molodost'. These results were also obtained from repeated experiment. Positive influence of controlled conditions to growing shoots and callus regeneration was also proved. Explants which were grown in controlled conditions reached higher efficiency on average 0,33 shoots per callus.
By successful recovery of the plants from pests Ditylenchus dipsaci using explantate cultures, it was verifed the initial hypothesis, where resulting explants are transferred to ex-vitro conditions were 100 % pest free. Although, callus cultures grown on MS medium of 1,5 mg . l-1 TDZ and 0,5 mg . l-1 IAA showed some degree of nematode occurrence during testing, consequent passage eliminated the remaining population of nematodes.
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In vitro embryo production in horses
Babická, Dominika ; Šichtař, Jiří (advisor) ; Šimoník, Ondřej (referee)
The aim of this work is to write a literature review covering the latest findings in the field of in vitro embryo production in horses. In order to perform in vitro embryo production, you first need to know the anatomy of a mare and management of sexual activity. Reproductive systém of mares consists of two ovaries, two oviducts, uterus, vagina and vulva. Unlike other livestock, the development of oocytes is in the cortex of the ovary. Sexual activity is driven neurohumoral and production of hormone is ensured by the central nervous system and endocrine glands.
For in vitro embryo production must be obtained first oocytes. These are collected from either the living or the dead mares. For live mares are used ovum pick up (OPU, transvaginal ultrasound guided aspiration), standing flank laparotomy or transcutaneous flank puncture. OPU is a noninvasive method, where each follicle is flushed several times, and in the obtained solution is looked for oocytes. Invasive, but a very successful method for obtaining oocytes is transcutaneous flank puncture. There are three ways for collecting oocytes from abbatoir ovaries - either by slicing ovaries or curettage or aspiration. Very good results in amounts of obtained oocytes was proved by slicing ovaries. First, after collecting oocytes must be sorted according to cumulus-oocyte complex (COC) on compact, expanded or degenerating oocytes. Compact and expanded oocytes are distinguished under a microscope based on the morphology of the edges of this COC. Compact oocytes have a clear line margins, while margins of expanded oocytes are deformed and have heterogeneous cytoplasm. Best results for the production of embryos exhibit expanded oocytes. First, it has to be matured in the medium to the stage of MII. The culture system for oocyte maturation consists of the M199 with Earl's salts, 10% fetal bovine serum and 5 mU - ml of bovine follicle stimulating hormone in a humidified atmosphere of 5% CO2 in air at 38.2 ° C. It was found out that regardless of the duration of cultivation, results of development of blastocysts after 24 or 48 hours were the same.
Standard IVF in horses can not be performed due to the zona pellucida and sperm capacitation. Therefore it is used intracytoplasmic sperm injection (ICSI), when one immobilized sperm is injected directly into the oocyte. Before ICSI sperm capacitation must be done by using the calcium ionophore. Oocytes were further cultured in vitro to the blastocyst stage. Another possibility is to transfer the fertilized oocyte directly to the uterine horn of the mare, or it is used cryopreservation for future.
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Studium možnosti využití termoterapie při eliminaci spály růžovitých u kdouloně
Nečasová, Aneta
The annual shoots of quinces were taken in the spring and summer of 2014 with vegetative sprouts. In the spring they were winter sprouts and in the summer the sprouts were ready for another season. The sources of the annual shoots were quinces grown in Školní zemědělský podnik Žabčice. 14 other varieties of quinces were chosen for testing. The originator of fire blight the Erwinia amylovora was located on these varieties. Different temperature intervals of cultivation were tested where the presence of the patogen could have been eliminated. Moreover the test of pathogenicity in cooperation with Fytopatologie rostlin, which have not proved the presence of that pathogen, while orchard in Žabčice is attacked by it. So there is no certainty in saying that thermotherapy is presenting itself as efficient way to treat this sickness.
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Studium konverze somatických embryí smrku v rostliny
Psotová, Martina
This work deals with somatic embryogenesis of Picea abies. Part of this work was to compare the viability of plant growth regulator when used before maturation and without them. The main observations of the effects of regulators focused on the final phase of maturation and phase conversion in the plants. The research was conducted in laboratories Mendel University at the Department of Plant Biology. For conditions in the early stages was artificially maintained darknes phase stable air temperature of 23+-2°C together with artificial air circulation. In the following phases photoperiod was controlled, stable air temperature at 23+-2°C with artificial air circulation in the cultivation room. Effect of growth regulators used before maturation was studiedon plants at the final stage of maturation and during conversion to plants. Slower development of roots and aerial parts of plants in the variant with added growth regulators was observed.
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