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Influence of total leaf nitrogen and non-structural carbohydrates on Rubisco content and its activity under ambient and elevated concentrations of carbon dioxide
Vičíková, Magda ; Babák, Libor (referee) ; Hrstka, Miroslav (advisor)
This diploma thesis examines influence of content of leaf nitrogen and non-structural carbohydrates on Rubisco activity and its content under impact of ambient (A = 385 µmolCO2 mol-1) and elevated (E = 700 µmolCO2 mol-1) concentrations of carbon dioxide in needles of Norway spruce (Picea abies L.). The Rubisco activity was determined spectrophotometrically, the Rubisco content by using SDS-PAGE, the content of non-structural carbohydrates by using the anthron method and total leaf nitrogen was determined by using the Kjeldahl method. The Rubisco activity in needles of Norway spruce did not show acclimation on elevated concentration of carbon dioxide, but the activity was elevated concentration of carbon dioxide stimulated during the growing season. The acclimation was proved by Rubisco content, its content was significantly lower in needles cultivated under elevated carbon dioxide. Content of total leaf nitrogen was statistically significantly lower in needles of Norway spruce cultivated under elevated carbon dioxide then in needles cultivated under ambient concentration of carbon dioxide. On the contrary, the content of non-structural carbohydrates was significantly higher in needles cultivated under elevated concentration of carbon dioxide then in variant A. Seasonal changes of Rubisco content depended on content of non-structural carbohydrates, the Rubisco content decreased with increasing content of non-structural carbohydrates. No dependence was detected between seasonal changes of the total leaf content and the Rubisco content.
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The dynamics of non-structural saccharides accumulation and Rubisco activity under the elevated carbon dioxide concentration and sink manipulation at beech
Uhrová, Lucie ; Vítová, Eva (referee) ; Hrstka, Miroslav (advisor)
This diploma thesis deals with dynamic of accumulation of non-structural carbohydrates and activity of Rubisco enzyme at elevated concentration of CO2 on beech (Fagus sylvatica L.). Three years old seedlings of beech were cultivated in minisphere with ambient (385 µmol•mol-1, variant A), and with elevated concentration CO2 (700 µmol•mol-1, variant E) for four months. In every variant the first half of plants was fertilized by nitrogen (variant N+) and the second half was control (variant N-). Plants used for experiment were at first adapted for darkness for 12 hours. Subsequently tested leaves were cut off, leafstalk including short segment of branch (approximately 1 cm) was inserted into 0.7 M solution of sucrose (variant S) or water (variant V) and exposed to radiation 200 mol•m-2•s-1 for 0, 30, 60, 120, 180 minutes. Then leaf area and fresh mass of leaf blade were established, samples were fixed in liquid nitrogen and stored in deep freezer to analysis in –70 °C. Rubisco content was determined by SDS-PAGE method, Rubisco activity spectrofotometrically and content of non-structural carbohydrates by anthrone method and HPLC method. Rubisco content was significantly lower in the N- variant than in N+ variant. Rubisco content was also significantly lower in E than in A variant, which is an evidence of down-regulation. Rubisco activity is moderately stimulated at E variant with time, but differences between variant A and E are not statistically significant. Influence of sucrose feeding to Rubisco activity was not proved. Significant differences were detected by anthrone method in non-structural carbohydrates content between variants S and V, but not between variants A and E. Statistically significant increase of sucrose content with time was detected by HPLC method at variant AS, but not at variant ES.
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Alpine treelines in the High Asia and their relation to temperatures
Yakubouskaya, Alina ; Treml, Václav (advisor) ; Romportl, Dušan (referee)
Treeline is a gradual transition from closed forest to treeless areas at a certain elevation above the sea. According to its forms, we distinguish diffuse, abrupt, island treelines and krummholz. Depending on the form we can assume the main factors affecting its occurrence and possible response to climatical changes. Generally, the position of the treeline in various latitudes relates to the thermal characteristics, mainly to the temperatures of the growing season, limiting tree growth, the availability of necessary nutrients and seedling survival. Limitation by an insufficient carbon balance as a cause of the ecotone, is not considered as a major or decisive factor by many authors. In most cases they proved a sufficient carbon balance at the treelines and fixed an incresing amount of non-structural carbohydrates, needed for trees to activate their growth after a period of dormancy. This work was focused primarily on identifying the positions of the treeline in the mountains of High Asia, and determining the basic thermal characteristics relating to these sites. The best indicator, connected to the position of the treeline in this region was founded. The results were compared with global research. Key words: treeline; diffuse, abrupt and island treelines, krummholz, sink limitation hypothesis,...
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Alpine treelines in the High Asia and their relation to temperatures
Yakubouskaya, Alina ; Treml, Václav (advisor) ; Romportl, Dušan (referee)
Treeline is a gradual transition from closed forest to treeless areas at a certain elevation above the sea. According to its forms, we distinguish diffuse, abrupt, island treelines and krummholz. Depending on the form we can assume the main factors affecting its occurrence and possible response to climatical changes. Generally, the position of the treeline in various latitudes relates to the thermal characteristics, mainly to the temperatures of the growing season, limiting tree growth, the availability of necessary nutrients and seedling survival. Limitation by an insufficient carbon balance as a cause of the ecotone, is not considered as a major or decisive factor by many authors. In most cases they proved a sufficient carbon balance at the treelines and fixed an incresing amount of non-structural carbohydrates, needed for trees to activate their growth after a period of dormancy. This work was focused primarily on identifying the positions of the treeline in the mountains of High Asia, and determining the basic thermal characteristics relating to these sites. The best indicator, connected to the position of the treeline in this region was founded. The results were compared with global research. Key words: treeline; diffuse, abrupt and island treelines, krummholz, sink limitation hypothesis,...
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The dynamics of non-structural saccharides accumulation and Rubisco activity under the elevated carbon dioxide concentration and sink manipulation at beech
Uhrová, Lucie ; Vítová, Eva (referee) ; Hrstka, Miroslav (advisor)
This diploma thesis deals with dynamic of accumulation of non-structural carbohydrates and activity of Rubisco enzyme at elevated concentration of CO2 on beech (Fagus sylvatica L.). Three years old seedlings of beech were cultivated in minisphere with ambient (385 µmol•mol-1, variant A), and with elevated concentration CO2 (700 µmol•mol-1, variant E) for four months. In every variant the first half of plants was fertilized by nitrogen (variant N+) and the second half was control (variant N-). Plants used for experiment were at first adapted for darkness for 12 hours. Subsequently tested leaves were cut off, leafstalk including short segment of branch (approximately 1 cm) was inserted into 0.7 M solution of sucrose (variant S) or water (variant V) and exposed to radiation 200 mol•m-2•s-1 for 0, 30, 60, 120, 180 minutes. Then leaf area and fresh mass of leaf blade were established, samples were fixed in liquid nitrogen and stored in deep freezer to analysis in –70 °C. Rubisco content was determined by SDS-PAGE method, Rubisco activity spectrofotometrically and content of non-structural carbohydrates by anthrone method and HPLC method. Rubisco content was significantly lower in the N- variant than in N+ variant. Rubisco content was also significantly lower in E than in A variant, which is an evidence of down-regulation. Rubisco activity is moderately stimulated at E variant with time, but differences between variant A and E are not statistically significant. Influence of sucrose feeding to Rubisco activity was not proved. Significant differences were detected by anthrone method in non-structural carbohydrates content between variants S and V, but not between variants A and E. Statistically significant increase of sucrose content with time was detected by HPLC method at variant AS, but not at variant ES.
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Influence of total leaf nitrogen and non-structural carbohydrates on Rubisco content and its activity under ambient and elevated concentrations of carbon dioxide
Vičíková, Magda ; Babák, Libor (referee) ; Hrstka, Miroslav (advisor)
This diploma thesis examines influence of content of leaf nitrogen and non-structural carbohydrates on Rubisco activity and its content under impact of ambient (A = 385 µmolCO2 mol-1) and elevated (E = 700 µmolCO2 mol-1) concentrations of carbon dioxide in needles of Norway spruce (Picea abies L.). The Rubisco activity was determined spectrophotometrically, the Rubisco content by using SDS-PAGE, the content of non-structural carbohydrates by using the anthron method and total leaf nitrogen was determined by using the Kjeldahl method. The Rubisco activity in needles of Norway spruce did not show acclimation on elevated concentration of carbon dioxide, but the activity was elevated concentration of carbon dioxide stimulated during the growing season. The acclimation was proved by Rubisco content, its content was significantly lower in needles cultivated under elevated carbon dioxide. Content of total leaf nitrogen was statistically significantly lower in needles of Norway spruce cultivated under elevated carbon dioxide then in needles cultivated under ambient concentration of carbon dioxide. On the contrary, the content of non-structural carbohydrates was significantly higher in needles cultivated under elevated concentration of carbon dioxide then in variant A. Seasonal changes of Rubisco content depended on content of non-structural carbohydrates, the Rubisco content decreased with increasing content of non-structural carbohydrates. No dependence was detected between seasonal changes of the total leaf content and the Rubisco content.
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