Maciej Kocurek

Physiological and biochemical characterisation of watered and drought-stressed barley mutants in the HvDWARF gene encoding C6-oxidase involved in brassinosteroid biosynthesis

Brassinosteroids (BR) are plant steroid hormones that were discovered more than thirty years ago, but their physiological function has yet to be fully explained. The aim of the study was to answer the question of whether/how disturbances in the production of BR in barley affects the plants metabolism and development under conditions of optimal watering and drought. Mutants with an impaired production of BR are one of the best tools in research aimed at understanding the mechanisms of action of these hormones. The study used barley cultivars with a normal BR synthesis (wild type) and semi-dwarf allelic mutants with an impaired activity of C6-oxidase (mutation in HvDWARF), which resulted in a decreased BR synthesis. Half of the plants were subjected to drought stress in the seedling stage and the other half were watered optimally. Plants with impaired BR production were characterised by a lower height and developmental retardation. Under both optimal watering and drought, BR synthesis disorders caused the reduced production of ABA and cytokinins, but not auxins. The BR mutants also produced less osmoprotectant (proline). The optimally watered and drought-stressed mutants accumulated less sucrose, which was accompanied by changes in the production of other soluble sugars. The increased content of fructooligosaccharide (kestose) in optimally watered mutants would suggest that BR is a negative regulator of kestose production. The decreased level of nystose in the drought-stressed mutants also suggests BR involvement in the regulation of the production of this fructooligosaccharide. The accumulation of the transcripts of genes associated with stress response (hsp90) was lower in the watered and drought-stressed BR-deficient mutants. In turn, the lower efficiency of photosystem II and the net photosynthetic rate in mutants was revealed only under drought conditions. The presented research allows for the physiological and biochemical traits of two BR-barley mutants to be characterised, which helps BR function to be understood. The knowledge can also be a good starting point for some breeding companies that are interested in introducing new semi-dwarf barley cultivars.

Endogenous progesterone and its cellular binding sites in wheat exposed to drought stress.

Progesterone is a basic hormone that regulates the metabolism in mammals. The presence of this compound has also been found in certain plants. It is believed that progesterone can regulate growth processes and resistance to stress, however, its precise role in plants remains unknown. The research conducted in this study was aimed at analyzing the content of endogenous progesterone and its cellular binding sites in the leaves of spring wheat exposed to drought. Changes were studied in two cultivars of wheat - a cultivar sensitive to drought (Katoda) and tolerant cultivar (Monsun). Plants had undergone periodic droughts during the seedling stage or in the phase of heading. The occurrence of free progesterone as well as its conjugated forms was observed in wheat studied. The amount of progesterone ranged from 0.2 to 5.8pmolgFW(-1) and was dependent on the cultivar, age of the plants, stage of development and fluctuated as a result of the exposure to drought. Cv. Katoda responded to a water deficit by lowering the amount of progesterone and cv. Monsun by increasing its level. Progesterone in plants grown in limited water conditions occurred primarily in a free form. While in the optimal watering conditions, some of its pool was found in the form of conjugates. In the spring wheat the occurrence of binding sites for progesterone was detected in cell membranes, cytoplasm and nuclei in the range of 10-36fmol/mg of protein. The wheat cultivars tested, Monsun and Katoda, differ in their concentration of cellular binding sites for progesterone. This number varied in the individual fractions during different stages of plant development and due to the effect of drought stress. The number of binding sites for progesterone located in the membrane fraction of seedlings and flag leaves increased significantly under drought in the cv. Katoda (35-46%), but did not change in the cv. Monsun. Whereas the number of cytoplasmic progesterone binding sites increased during the drought in the cv. Monsun (about 50%), they did not change in the cv. Katoda. Changes in the amount of progesterone and its binding sites in the cell under the influence of drought were then different depending on whether the cultivar was tolerant or sensitive to drought. The possibility of utilizing these changes as markers of drought resistance is discussed. The results obtained suggest that progesterone is a part of wheat response to stress factors (drought).

Photosynthesis-related characteristics of the midrib and the interveinal lamina in leaves of the C3–CAM intermediate plant Mesembryanthemum crystallinum

BACKGROUND AND AIMS Leaf veins are usually encircled by specialized bundle sheath cells. In C4 plants, they play an important role in CO2 assimilation, and the photosynthetic activity is compartmentalized between the mesophyll and the bundle sheath. In C3 and CAM (Crassulacean acid metabolism) plants, the photosynthetic activity is generally attributed to the leaf mesophyll cells, and the vascular parenchymal cells are rarely considered for their role in photosynthesis. Recent studies demonstrate that enzymes required for C4 photosynthesis are also active in the veins of C3 plants, and their vascular system contains photosynthetically competent parenchyma cells. However, our understanding of photosynthesis in veins of C3 and CAM plants still remains insufficient. Here spatial analysis of photosynthesis-related properties were applied to the midrib and the interveinal lamina cells in leaves of Mesembryanthemum crystallinum, a C3-CAM intermediate plant. METHODS The midrib anatomy as well as chloroplast structure and chlorophyll fluorescence, diurnal gas exchange profiles, the immunoblot patterns of PEPC (phosphoenolpyruvate carboxylase) and RubisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase), H2O2 localization and antioxidant enzyme activities were compared in the midrib and in the interveinal mesophyll cells in leaves of C3 and CAM plants. KEY RESULTS Leaf midribs were structurally competent to perform photosynthesis in C3 and CAM plants. The midrib chloroplasts resembled those in the bundle sheath cells of C4 plants and were characterized by limited photosynthetic activity. CONCLUSIONS The metabolic roles of midrib chloroplasts differ in C3 and CAM plants. It is suggested that in leaves of C3 plants the midrib chloroplasts could be involved in the supply of CO2 for carboxylation, and in CAM plants they could provide malate to different metabolic processes and mediate H2O2 signalling.

Mammalian androgen stimulates photosynthesis in drought-stressed soybean

The aim of the present studies was to assess the possibility of compensating the negative effects of drought stress on gaseous exchange and efficiency of photosystem II in soybean seedlings by application of the androgen — androstenedione. Androstenedione (0.25 mg dm−3) was applied via presowing seed soaking (12 h). Control seeds were untreated with steroid. Plants were cultured in pots. On the 12th day of growth, the plants were watered for the last time. Drought symptoms occurred during the next 10 days. On the 22nd day of growth, leaf gaseous exchange and PSII measurements were taken. Afterwards the plants were watered. Two days later measurements were taken again. Androstenedione improved the intensity of leaf net photosynthesis. The effect of androstenedione was manifested during the rehydration of plants that have undergone a period of drought. An increase in net photosynthesis intensity was accompanied by higher transpiration. Possible mechanisms of androstenedione action — effect on aquaporin functionality and membrane stability — are discussed. The significance of ethanol and DMSO (solvents of steroid) in experiments on the physiological activity of androstenedione is also considered.

The content of photosynthetic pigments and the light conditions in the fruits and leaves of sweet pepper

Investigations were carried out on the fruits of sweet pepper at its two development stages: on green fruits, on mature red and yellow fruits and on leaves. The content of the photosynthetic pigments and the optical properties were examined.In the green fruits when compared with leaves the content of the photosynthetic pigments is smaller by 40 to 50 % and the value of the ratio: chlorophyll a/b is lower. Chlorophyll is absent in mature fruits, while the content of carotenoids is a few times higher.The optical properties of green fruits and of the leaves in the PAR (photosynthetically active radiation) range are the same. In the range 700–1100 nm the green fruits show smaller reflectance and transmittance and a few times greater absorptance of irradiation which contributes to the warming up of the seed bag, while small absorptance of leaves in this range protects them against overheating.In mature fruits, in the PAR range, the reflectance and transmittance are higher, while the absorptance of irradiation in comparison with that of green fruits is smaller. In the range 700 – 1100 nm the changes are rather small and refer to the increase of reflectance and reduction of absorptance, while the transmittance of irradiation remains unchanged.

Effects of exogenously applied hydrogen peroxide on antioxidant and osmoprotectant profiles and the C3-CAM shift in the halophyte Mesembryanthemum crystallinum L.

Exogenously applied H2O2 (50, 100 and 200mM) to Mesembryanthemum crystallinum root medium induced a transition from C3 to Crassulacean Acid Metabolism (CAM), as evaluated by diurnal malate (Δmal) fluctuations. A very high concentration of H2O2 (400mM) reduced Δmal below the value measured in control plants. An increase of malate content during the night in 400mM H2O2-treated plants might suggest that malate decarboxylation is crucial for CAM functioning. We conclude that malate plays a dual role: i) a protective and signaling function before CAM expression, and ii) a storage form of CO2 in plants performing CAM. A slight stimulation of photosystem II (PSII) photochemistry and net photosynthesis observed during the C3-CAM shift indicated that neither photoinhibition nor reduction of the photosynthetic rate were prerequisites for CAM. Moreover, CAM induction corresponded to a decrease of catalase activity. In CAM-performing plants, α-tocopherol, polyamines (putrescine and spermidine) and proline showed daily alterations and the content of α-tocopherol and polyamines was lower at the end of the day. In contrast, the proline concentration correlated with the applied H2O2 concentration and was higher at the end of the day in treated plants. The dynamic changes of antioxidant and osmolyte levels suggest their active role in preventing oxidative damage, stress acclimation mechanisms and involvement in metabolic regulation and/or signal transduction cascades.

The Photosynthetic Surface Area of Apple Trees

The development of stem and leaf surface areas were studied comparatively throughout a year’s cycle, and the proportions of these areas within the total area of trees were calculated. Depending on the variety, the total surface areas of trees: all branches, boughs and tree trunks after winter pruning, ranged from 27 dm2 to 118 dm2. At the beginning of the vegetation season, the proportion of leaves in newly developed stems amounted to 90%–92%, and the proportion of these new stems was 8%–10%. Later in the vegetation season, the leaf area amounted to 83%–89%, and that of new stems 11%–17%, and there were no differences between the varieties. Towards the end of the vegetation season, total leaf area on a tree ranged from 110–120 dm2 to 296 dm2, whereas the area of newly grown stems ranged from 42 dm2 to 168 dm2.