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Impact of elevated carbon dioxide concentration on Rubisco content in Fagus sylvatica and Picea abies
Hamříková, Dominika ; Vítová, Eva (referee) ; Hrstka, Miroslav (advisor)
In this bachelor’s thesis content of Rubisco in beech (Fagus sylvatica) and Norway spruce (Picea abies) was studied. The plants were cultivated in conditions with ambient carbon dioxide concentration (350 mol·mol-1) and elevated carbon dioxide concentration (700 mol·mol-1). The content of Rubisco in exposed and shaded leaves or needles was compared. The content of Rubisco was determined by SDS-PAGE method. A significant decrease in Rubisco content was demonstrated in conditions E versus A in shaded beech leaves. In other cases, the decrease was not so significant. A comparison of exposed and shaded needles also did not show a statistically significant difference, but in shaded beech leaves against exposed leaves Rubisco content was significantly lower, especially in conditions E.
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Influence of needle age at Rubisco activity and content in Norway spruce under the impact of ambient and elevated CO2 concentrations
Bošková, Martina ; Vítová, Eva (referee) ; Hrstka, Miroslav (advisor)
In this diploma work influence of needle age at Rubisco activity and content in Norway spruce (Picea abies) was studied. The plants were cultivated in conditions with ambient (A) CO2 concentration (350 µmol CO2/mol) and elevated (E) CO2 concentration (700 µmol CO2/mol). Sampling was done two times during the growing season (in the middle of June and in the end of September) were taken. Initial and total Rubisco activities were measured spectrophotometrically. Rubisco content was determined by SDS–PAGE method. Rubisco activity in 18-months-old needles was in E higher than in A. Rubisco contents in current-year needles and one-year-old needles were in A higher than in E in September. These differences were statistically significant that demonstrates the down-regulation of Rubisco content in conditions of elevated CO2 concentration. It seems the course of activities and content depending on age of the needles are antiparallel, that means that decrease of content is followed by increase of activity.
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The impact of UV radiation on activity and content of Rubisco enzyme in beech
Cigánková, Michaela ; Urban,, Otmar (referee) ; Hrstka, Miroslav (advisor)
This bachelor thesis deals with the influence of UV radiation on content and activity of Rubisco enzyme in leaves of European beech (Fagus sylvatica). Plants were cultivated in ambient (UV), lower (UV-) and higher (UV+) UV-B radiation both at ambient (AC) and elevated (EC = 700 micromol mol-1) CO2 concentrations. Leaves sampling was done four times during the growth season, in the second part of July, at the beginning and in the end of August and in the middle of September. Rubisco activity was assayed spectrophotometrically, the content of Rubisco was determined by SDS-PAGE method and quantified by computer densitometry. The results confirmed a reduction of Rubisco content and down-regulation on Rubisco level at elevated CO2 concentration. With the exception of September sampling, the content of Rubisco was lower in UV+ at ambient concentration of CO2. The same was not demonstrated at elevated CO2 conditions. Hypothesis that UV-B radiation changes a ratio of large to small subunit of Rubisco was confirmed. The influence of UV-B radiation on Rubisco activity was not proved.
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Diurnal changes of Rubisco content and its activity under ambient and elevated concentrations of carbon dioxide
Matulková, Zuzana ; Zemanová, Jana (referee) ; Hrstka, Miroslav (advisor)
In this diploma thesis, the diurnal changes of initial and total Rubisco activity and Rubisco enzyme content in beech (Fagus sylvatica) were studied under conditions of ambient (A) CO2 concentration (350 µmol.mol-1) and elevated (E) CO2 concentration (700 µmol.mol-1) during the day. Samples were taken on July 8th (from 10:00 to 21:30), on July 9th (from 04:00 to 12:00) and then on July 22th (from 04:00 to 21:30). The initial and total activity were measured spectrophotometrically and the activation level was calculated from the ratio of initial and total activities. Rubisco enzyme content was determined by SDS-PAGE method and the initial and total specific activity were calculated from the ratio of initial or total activity and Rubisco enzyme content. In our experiment no statistically significant difference was found between Rubisco activities in beeches cultivated under conditions of ambient CO2 concentration and elevated CO2 concentration, so any down-regulation of Rubisco activity did not appear under the influence of ambient CO2 concentration. Diurnal changes of Rubisco activities showed only statistically nonsignificant fluctuation. After daylight no significant increase of total Rubisco activity was observed, which demonstrates the absence or the immaterial effect of CA1P night inhibitor in beech. Intensive fluctuation of the activation level (40–90 %) proves the regulation of photosynthesis during the day via Rubisco enzyme carbamylation. Rubisco enzyme content in beech cultivated under conditions of ambient CO2 concentration was lower than that one in beech cultivated under conditions of elevated CO2 concentration. Then down-regulation of Rubisco enzyme content is presented under conditions of ambient CO2 concentration. Before daylight Rubisco enzyme content showed an indistinctive minimum on the level of statistical significance, during the day only nonrelevant fluctuation was noticed.
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Impact of elevated concentration of carbon dioxide on diurnal changes of Rubisco activity and its content
Zítková, Jana ; Vítová, Eva (referee) ; Hrstka, Miroslav (advisor)
Diurnal changes of initial and total activity and content of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) under conditions of ambient (350 µmol mol-1) and elevated (E = 700 µmol mol 1) concentration CO2 were measured in needles of Norway spruce. Needles of Norway spruce were taken on 22th July in two-hours from 3:30 a. m. till 10:30 p. m. The Rubisco activity was determined by spectrophotometry, the Rubisco content was detected by SDS-PAGE. The Rubisco activity was in most samples statistically significantly lower in needles cutlivated under ambient carbon dioxide then in needles cultivated under elevated carbon dioxide. Diagrams of the Rubisco activity shows absence of night inhibitor CA1P. The Rubisco activity was directly proportional to the intensity of photosynthetically active radiation in time from 7:00 a. m. till 7:30 p. m. The Rubisco content was statistically significantly higher in needles of Norway spruce cultivated under ambient carbon dioxide than in needles cultivated under elevated CO2, which shows acclimation of the Rubisco content during long-term effect of elevated CO2 concentration.
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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 impact of UV radiation on activity and content of Rubisco enzyme in beech
Cigánková, Michaela ; Urban,, Otmar (referee) ; Hrstka, Miroslav (advisor)
This bachelor thesis deals with the influence of UV radiation on content and activity of Rubisco enzyme in leaves of European beech (Fagus sylvatica). Plants were cultivated in ambient (UV), lower (UV-) and higher (UV+) UV-B radiation both at ambient (AC) and elevated (EC = 700 micromol mol-1) CO2 concentrations. Leaves sampling was done four times during the growth season, in the second part of July, at the beginning and in the end of August and in the middle of September. Rubisco activity was assayed spectrophotometrically, the content of Rubisco was determined by SDS-PAGE method and quantified by computer densitometry. The results confirmed a reduction of Rubisco content and down-regulation on Rubisco level at elevated CO2 concentration. With the exception of September sampling, the content of Rubisco was lower in UV+ at ambient concentration of CO2. The same was not demonstrated at elevated CO2 conditions. Hypothesis that UV-B radiation changes a ratio of large to small subunit of Rubisco was confirmed. The influence of UV-B radiation on Rubisco activity was not proved.
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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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Impact of elevated concentration of carbon dioxide on diurnal changes of Rubisco activity and its content
Zítková, Jana ; Vítová, Eva (referee) ; Hrstka, Miroslav (advisor)
Diurnal changes of initial and total activity and content of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) under conditions of ambient (350 µmol mol-1) and elevated (E = 700 µmol mol 1) concentration CO2 were measured in needles of Norway spruce. Needles of Norway spruce were taken on 22th July in two-hours from 3:30 a. m. till 10:30 p. m. The Rubisco activity was determined by spectrophotometry, the Rubisco content was detected by SDS-PAGE. The Rubisco activity was in most samples statistically significantly lower in needles cutlivated under ambient carbon dioxide then in needles cultivated under elevated carbon dioxide. Diagrams of the Rubisco activity shows absence of night inhibitor CA1P. The Rubisco activity was directly proportional to the intensity of photosynthetically active radiation in time from 7:00 a. m. till 7:30 p. m. The Rubisco content was statistically significantly higher in needles of Norway spruce cultivated under ambient carbon dioxide than in needles cultivated under elevated CO2, which shows acclimation of the Rubisco content during long-term effect of elevated CO2 concentration.
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Diurnal changes of Rubisco content and its activity under ambient and elevated concentrations of carbon dioxide
Matulková, Zuzana ; Zemanová, Jana (referee) ; Hrstka, Miroslav (advisor)
In this diploma thesis, the diurnal changes of initial and total Rubisco activity and Rubisco enzyme content in beech (Fagus sylvatica) were studied under conditions of ambient (A) CO2 concentration (350 µmol.mol-1) and elevated (E) CO2 concentration (700 µmol.mol-1) during the day. Samples were taken on July 8th (from 10:00 to 21:30), on July 9th (from 04:00 to 12:00) and then on July 22th (from 04:00 to 21:30). The initial and total activity were measured spectrophotometrically and the activation level was calculated from the ratio of initial and total activities. Rubisco enzyme content was determined by SDS-PAGE method and the initial and total specific activity were calculated from the ratio of initial or total activity and Rubisco enzyme content. In our experiment no statistically significant difference was found between Rubisco activities in beeches cultivated under conditions of ambient CO2 concentration and elevated CO2 concentration, so any down-regulation of Rubisco activity did not appear under the influence of ambient CO2 concentration. Diurnal changes of Rubisco activities showed only statistically nonsignificant fluctuation. After daylight no significant increase of total Rubisco activity was observed, which demonstrates the absence or the immaterial effect of CA1P night inhibitor in beech. Intensive fluctuation of the activation level (40–90 %) proves the regulation of photosynthesis during the day via Rubisco enzyme carbamylation. Rubisco enzyme content in beech cultivated under conditions of ambient CO2 concentration was lower than that one in beech cultivated under conditions of elevated CO2 concentration. Then down-regulation of Rubisco enzyme content is presented under conditions of ambient CO2 concentration. Before daylight Rubisco enzyme content showed an indistinctive minimum on the level of statistical significance, during the day only nonrelevant fluctuation was noticed.
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