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Diurnal changes of Rubisco activity under ambient and elevated concentrations of carbon dioxide
Uhrová, Lucie ; Vítová, Eva (referee) ; Hrstka, Miroslav (advisor)
Diurnal changes of Rubisco activity under the impact of ambient (380 µmol mol-1) and elevated (700 µmol mol-1) concentrations of carbon dioxide were studied in beech (Fagus sylvatica). Samples from leaves were taken in the two-hour intervals from 4:00 to 21:30 on the 22. 7. 2009 and activity of Rubisco was determined spectrophotometrically. No statistically significant differences were found between Rubisco activity of leaves growing in ambient and elevated CO2 concentrations. No increase of Rubisco total activity after daylight shows that nocturnal inhibitor CA1P is not present or plays only negligible role in beech. During the day Rubisco activity showed only statistically insignificant deviations. A slight decrease of Rubisco activation state in the afternoon indicates the afternoon depression of Rubisco initial activity.
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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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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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Diurnal changes of Rubisco activity under ambient and elevated concentrations of carbon dioxide
Uhrová, Lucie ; Vítová, Eva (referee) ; Hrstka, Miroslav (advisor)
Diurnal changes of Rubisco activity under the impact of ambient (380 µmol mol-1) and elevated (700 µmol mol-1) concentrations of carbon dioxide were studied in beech (Fagus sylvatica). Samples from leaves were taken in the two-hour intervals from 4:00 to 21:30 on the 22. 7. 2009 and activity of Rubisco was determined spectrophotometrically. No statistically significant differences were found between Rubisco activity of leaves growing in ambient and elevated CO2 concentrations. No increase of Rubisco total activity after daylight shows that nocturnal inhibitor CA1P is not present or plays only negligible role in beech. During the day Rubisco activity showed only statistically insignificant deviations. A slight decrease of Rubisco activation state in the afternoon indicates the afternoon depression of Rubisco initial activity.
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