A protein, identifiable as calmodulin (CaM), has been isolated from the seedling tissue of Pharbitis nil. The method has been developed to isolate a high quality protein from plant tissue containing the high content of polyphenols. This protein was relatively heat-stable and bound to hydrophobic resin in calcium-dependent manner. It was recognized by the antibody against pea and carrot, but did not bind to antibody against Dictyostelium discoideum. This protein had M_r of 15 kDa and 18.5 kDa in the presence and absence of Ca₂₊, respectively, and was able to stimulate calmodulin-deficient cAMP phosphodiesterase. Based on its migration on SDS-PAGE gels, M_r and binding to anti-CaM antibodies it was deduced that calmodulin from P. nil is essentially identical to calmodulin isolated from other plants.

Relative water content (RWC), leaf water potential (Ψ_w) and osmotic potential (Ψ_s), contents of chlorophyll (Chl) a, Chl b, soluble sugars, and seed quality (gum content) were used to evaluate the role of phosphorus in alleviation of the deleterious effect of water deficit in clusterbean (Cyamopsis tetragonoloba L. Taub). Under water stress, Ψ_w, Ψ_s, and Chl and gum contents decreased and soluble sugar contents increased. Phosphorus application increased Chl and sugar contents in control plants and ameliorated negative effects of water stress.

Corchorus olitorius plants treated by 5 μg cm^-3 of Cd, Pb, Al or Cu in hydroponic culture accumulated in leaves 190, 150, 350 and 325 μg g^-1(d.m.) of these metals, respectively, after 6 d of exposure. Exposure of Corchorus plants to tested metals resulted in a sharp rise in content of amino acids in leaf tissues, however the magnitude of accumulation was different from one metal to another. Presence of sulphur in the growth medium significantly increased uptake of Cd and Pb and cysteine (cyst) was more effective than K₂SO₄. Similarly, addition of salicylic acid (SA) in the growth medium significantly enhanced the ability of Corchorus plants to accumulate all these metals. Growth of Corchorus plants was significantly reduced by treatment with any of the four metals except Cu and added cyst, K₂SO₄ or SA alleviated the growth retarding effect of metals.

Net photosynthetic rate (P_N), transpiration rate (E), and stomatal conductance (g_s) during water stress and after rehydration were measured in Phaseolus vulgaris, Beta vulgaris, and Zea mays. Immediately before imposition of water stress by cessation of watering, plants were irrigated with water (control), 100 μM abscisic acid (ABA), and/or 10 μM N⁶-benzyladenine (BA). In all three species, application of ABA decreased g_s, E, and P_N already 1 h after application. However, during water stress g_s, E, and P_N in plants pre-treated with ABA remained higher than in plants pre-treated with water. Positive effects of ABA application were observed also after rehydration. In contrast, the effects of pre-treatment with BA were species-specific. While in bean plants BA application ameliorated negative effect of water stress, only very slight effects were observed in maize, and in sugar beet BA even aggravated the effects of water stress.

A new protocol for micropropagation of endangered Daphne cneorum through multiple shoot formation has been developed. Two different types of explants (dormant apical buds and in vitro seed-derived young seedlings) from plants in two different localities were used for the initiation of multiple shoots on agar woody plant medium (WPM) with 0.2 mg dm^-3 benzylaminopurine (BAP), 0.1 mg dm^-3β-indolebutyric acid (IBA), 200 mg dm^-3 glutamine, and 200 mg dm^-3 casein hydrolysate. From 10 seeds only one germinated and the multi-apex culture bearing 12 shoots sprouted out from in vitro seed-derived young seedling. After 6-month cultivation 35 multi-apex cultures were achieved from in vitro seed-derived young seedling. On 1/3 strength WPM medium supplemented with 2.83 mg dm^-3 IBA 50 % of cultures (clusters of 3 - 5 shoots) rooted but no rooting occurred in the presence of α-naphthaleneacetic acid (NAA). The rooted plantlets were acclimatized for 4 weeks in the greenhouse and then transferred into natural conditions. The plants successfully survived the winter and flowered.

Elevated UV-B radiation (12.2 kJ m^-2 d^-1) as against the ambient level of 10 kJ m^-2 d^-1 affected flowering, productivity and biomass partitioning of green gram [Vigna radiata (L.) Wilczek cv. KM-2]. UV-B stress delayed flowering initiation and achievement of 50 % flowering, reduced flower retention by 25 %, potential yield by 18 % and all yield attributes such as pod number (25 %), pod mass (41 %), seed number (32 %) and seed mass (45 %). Harvest index and shelling percentage were also reduced by 31 and 7 %, respectively. Application of triadimefon (20 mg dm^-3) to unstressed plants accelerated flowering and enhanced flower retention (21 %), potential yield (15 %) and yield attributes (7 to 44 %). The partitioning of biomass between plant parts also showed improvement over the control plants. In UV-B-stressed plants, triadimefon treatment compensated the inhibitions to varying extents.

During ageing of bean (Phaseolus vulgaris L.) cotyledons in plants with modified life span the time-course of four cytokinins, ethylene, and the end products of free radical attack, lipofuscin-like pigments (LFP), were studied. UV irradiation shortened cotyledon life span, while epicotyl decapitation prolonged it. In controls, LFP increased at the senescence onset but at the end of life span it returned to the initial level. Ethylene increased more than 3-fold at the time of abscission. The content of individual cytokinins (zeatin, zeatin riboside, isopentenyl adenine, isopentenyl adenine riboside) varied differently during ageing but they did not decreased in any case under level observed in young cotyledons at the time of abscission. UV irradiation resulted in 14-fold increase in LFP concentration at the end. Ethylene increased 8-fold 2 h after irradiation. Individual cytokinins increased after UV irradiation to a different extent and time-course, nevertheless cotyledon life span was shortened. Decapitation induced LFP decrease. On day 13, LFP abruptly increased and than decreased and stayed lowered until abscission. Ethylene was maximum on day 24, at the time of abscission, it was above 200 % of control. Decapitation produced transient decrease in some cytokinins namely zeatin and isopentenyl adenine riboside.

Effects of phosphorus and thiourea application (either alone or in combination) were studied on clusterbean (Cyamopsis tetragonoloba Taub.) plants subjected to water stress by withholding irrigation at pre- and post-flowering stages in pot culture trial. Water stress significantly decreased shoot water potential, relative water content of leaves, net photosynthetic rate, contents of total chlorophyll, starch and soluble proteins as well as nitrate reductase activity at both the growth stages. Application of phosphorus and thiourea or combined application increased most of these parameters. Results revealed synergistic effects of P and thiourea in enhancing net photosynthesis, leaf area, chlorophyll content and nitrogen metabolism leading to significant improvement in plant growth and seed yield under water stress condition.

The work is concerned with the effect of low concentrations (10^-7 mol dm^-3) of sodium naphthenate on total content of Cd and its particular forms in the intercellular space and inside cells, as well as on some physiological and biochemical parameters of young soybean plants grown in the presence of 1 mmol dm^-3 solution of cadmium chloride. Presence of naphthenate reduced in average by 40 % content total and intracellular Cd in root, stem and leaves and alleviated the harmful effect of Cd on activity of nitrate reductase and content of photosynthetic pigments.

The effects of 1 mM cadmium, lead and nickel on dry mass, Cd, Pb, and Ni contents, and changes in leaf structure in young wheat plants were studied. In leaves, Cd content was highest, followed by Pb and Ni, in roots Cd content was also highest, but followed by Ni and Pb. Roots accumulated considerably larger amounts of the three heavy metals than leaves. Largest reductions of leaf and root mass were obtained with Cd. Pb and Ni effects were almost equal. Ni excess had a strong negative effect on mesophyll thickness, while Cd mostly reduced the number and size of vascular bundles and vessel diameter. High Pb reduced the diameter of vessels causing their different deformations.

The aim of the present work was to evaluate the feasibility of mannose as a selection system for the future genetic transformation of annatto (Bixa orellana L.). Hypocotyl segments, inverted hypocotyls and immature zygotic embryos were inoculated onto a Murashige and Skoog's medium supplemented with B5 vitamins, 87.6 mM sucrose and mannose in different combinations, 2.8 g dm^-3 Phytagel®, and 4.56 μM zeatin (organogenesis) or 2.26 μM 2,4-dichlorophenoxyacetic acid and 4.52 μM kinetin (somatic embryogenesis). Annatto explants did not regenerate on medium with mannose as the only carbon source when inverted hypocotyls and immature zygotic embryos were used. However, organogenesis (5 % frequency) occurred exclusively in hypocotyl-derived explants nearest to the crown (collar) of the seedlings. No further shoot development was achieved. Therefore the substitution of sucrose by mannose inhibited both organogenesis and embryogenesis, and thus the employment of mannose could constitute an alternative selective agent in protocols for genetic transformation of this species.

Multiple shoots were induced from the cotyledonary nodes derived from seedling of Sesbania rostrata on Nitsch (1969; N) medium supplemented with various concentrations of benzyladenine (BA). 1 mg dm^-3 BA proved to be the best, eliciting 5.8 ± 1.0 shoots per explant in 100 % cultures. The elongation of shoots was best at 2.0 mg dm^-3 BA. The shoot proliferation capacity increased to 7.5 shoots per explant following transfer of explants to the fresh shoot multiplication medium (MS + 1.0 mg dm^-3 BA), after an initial incubation of 30 d. To further enhance number of shoots per explant an alternative strategy of cultivation of mother explant on fresh shoot multiplication medium after excision of shoots was adopted. Following the repeated harvesting of shoots an average of 33 shoots per explant could be obtained. The in vitro regenerated shoots produced roots when transferred to half-strength MS medium supplemented with 3 % sucrose and 1 mg dm^-3 IBA. The developed plantlets were planted in the soil and transferred to the field after an acclimatization period of 3 - 4 months. These plants produced flowers and fruits in the field and exhibited the development of prominent and more organized stem nodules as compared to the in vivo raised plants of the same age.

Effect of phosphorus deficiency on photosynthetic and respiratory CO₂ exchanges were analysed in primary leaves of 2-week-old bean (Phaseolus vulgaris L. cv. Golden Saxa) plants under non-photorespiratory (2 % O₂) and photorespiratory (21 % O₂) conditions. Low P decreased maximum net photosynthetic rate (P_Nmax) and increased the time necessary to reach it. In the leaves of P-deficient plants the relative decrease of P_Nmax at 2 % O₂ was larger than at 21 % O₂. The results suggested the influence of photorespiration in the cellular turnover of phosphates.

The growth and the content of endogenous cytokinins (CKs) of current-year-old shoots from juniper plants (Juniperus communis L.) growing over and off ore site were compared. The juniper shoots from ore site (M plants) had higher metal content and exhibited delayed growth. Less bases and nucleosides of Z- and iP- type CK and more iP-conjugates were present in the M shoots. These changes were probably due to inhibited CK export from the roots and/or altered CK metabolism forming less biologically active CKs.

The effect of radiation quality (350 - 740 nm) and darkness (D) on in vitro rooting, and chemical composition of the peach rootstock GF 677 was studied. Shoot explants were exposed for four weeks to cool white (control) (W), red (R), blue (B), green (G) or yellow (Y) radiation from fluorescent tubes. Some of the explants were kept in D during the rooting stage and others were maintained only for the first 2- or 4-d under R, B, G, Y or D, and subsequently were transferred to W. W was the most effective radiation source for adventitious root formation of GF 677 explants. Rooting was inhibited in those plants that remained in continuous D, and R reduced root growth in all treatments. The 2- or 4-d exposure to D, Y or B followed by W helped adventitious root development similarly as did W. G significantly increased Fe concentration in roots.

Exogenously applied methyl jasmonate (MeJA) might induce the formation of necrotic lesions that closely resemble hypersensitive response lesions. Cellular damage, restricted to the infiltrated zone, was accompanied with the production of H₂O₂ from the oxidative burst. H₂O₂ generated in response to MeJA can be histochemically detected in cells surrounding the necrotic lesions as well as in the vascular tissues. The response is systemic and maximizes with time. Among 12 plant species from different families that were assayed for both hypersensitive reaction (HR)-like response and H₂O₂ generation, only woody species exhibited both MeJA-inducible HR cell death and the generation of H₂O₂. To assess the role of H₂O₂ in MeJA-induced HR-like cell death, a gain and loss of function strategy was employed. The cumulative results indicate that H₂O₂ is neither necessary nor sufficient for MeJA-inducible cell death and that O₂^- rather than H₂O₂ might be responsible.

The cloning of a 465 bp fragment from the 5'-flanking region of the gene encoding a cytosolic cyclophilin from periwinkle was achieved through inverse polymerase chain reaction. The DNA fragment was fused to a gusA-intron marker then introduced into tobacco by Agrobacterium tumefaciens-mediated transformation. Histochemical analysis of the transgenic shoot cultures demonstrated that the construct was able to drive GUS expression in stomata guard cells, but not in mesophyll cells when shoots were still attached to the callus from which they were initiated. In separated transgenic shoots and in seedlings, GUS was expressed in external and internal phloem and root hairs, respectively. GUS activity in transgenic tobacco seedlings was also investigated by fluorimetric assays. Treatments with NaCl or ABA decreased promoter activity whereas treatment with yeast extracts increased it.

Image analysis was used in studying stomatal morphology during acclimatization of tobacco plantlets to ex vitro conditions, 45 d after transfer leaf area was 15 times, and total number of stomata per leaf four times increased. During acclimatization stomatal density was decreased considerably on both leaf sides, and was compensated by an increase in stomatal sizes, e.g., in stomatal length and in stomatal area (both guard cells and pore). Elongation of stomata was increased indicating that the originally circular stomata of in vitro plantlets were changed into elliptical ones in ex vitro acclimatized plants.

The in vitro regeneration of three flax (Linum usitatissimum L.) breeding lines (cv. Jitka, cv. Areco and NLN 245) and selection of transgenic plants were studied. A. tumefaciens derived binary vector GV3101 (pPM90RK)(pPCVRN4) bearing tetramer of 35S promoter enhancer was used in transformation experiments. Following 3 weeks of cultivation on shoot inducing Murashige and Skoog agar medium containing BAP (0.1 µM) and NAA (0.005 µM) from 82.6 % to 98 % of hypocotyl segments formed shoots. While ticarcillin (500 mg dm^-3) used to eliminate Agrobacterium following the transformation decreased the organogenic response by about 10 % only, the addition of 20 mg dm^-3 hygromycin to ticarcillin efficiently suppressed the regeneration of untransformed control plants. To look up for genomic mutations caused by T-DNA insertion from Agrobacterium transformation or originated from somaclonal variation over 500 regenerated plants have been cloned, transferred into soil and evaluated especially for their morphological characteristics. Up to now among plants of cv. Areco-background at least 8 genotypes showed changes either in flower or filament and stigma colour and one clone of plants with pollen sterility was identified. Among fifty four plant clones evaluated in 7 clones the presence of transgene specific sequence hpt was detected and simultaneously Agrobacterium contamination of tissues was firmly excluded.

Potato (Solanum tuberosum L. cv. Bintje) was transformed with a cDNA clone encoding an osmotin-like protein. Transgenic and non-transgenic in vitro plants were subjected to NaCl for 3 weeks. The shoot and root development was slightly affected by salinity indicating that the salt condition used was a mild stress. The endogenous proline content of the osmotin-like transformed clone only raised slightly as compared to the non-transformed genotype, where a marked increase in proline content could be observed as a result to salt stress. These data provide evidence for the involvement of osmotin-like proteins in the mechanisms of salt tolerance in potato plants.

Various ecophysiological investigations are presented in Aldrovanda vesiculosa, a rootless aquatic carnivorous plant. A distinct polarity of N, P, and Ca tissue content per dry mass (DM) unit was found along Aldrovanda shoots. Due to effective re-utilization, relatively small proportions of N (10 - 13 %) and P (33 - 43 %) are probably lost with senescent leaf whorls, while there is complete loss of all Ca, K, and Mg. The total content of starch and free sugars was 26 - 47 % DM along adult shoots, with the maximum in the 7^th - 10^th whorls. About 30 % of the total maximum sugar content was probably lost with dead whorls. The plant was found to take up 5 - 7 times more NH₄⁺ to NO₃^- from a mineral medium. Under nearly-natural conditions in an outdoor cultivation container, catching of prey led to significantly more rapid growth than in unfed plants. DM of the fed controls was 48 % higher than in the unfed plants. The controls produced 0.69 branches per plant, while the unfed plants did not produced any. However, the N and P content per DM unit increased by 6 - 25 % in the apices and the first 6 whorls in the unfed variant, as compared to the fed controls. It may be suggested that carnivory is very important for Aldrovanda.

The seeds of two barley (Hordeum vulgare L.) cultivars (a drought resistant cv. Tokak-137/57 and a drought sensitive cv. Erginel-90) were imbibed either in distilled water (control) or in a solution containing 40 mg dm^-3 paclobutrazol (PBZ) and air dried. Seeds were germinated and grown in a glasshouse for 21 d and seedlings were subjected to salt stress by treating them with 100 and 200 mM NaCl for 12 d. The height of shoots was significantly decreased and root length was increased in PBZ-treated plants prior and after NaCl stress for 12 d leading to an increase in root to shoot ratio. Leaf chlorophyll and carotenoid contents in PBZ treated plants were increased in controls and especially in plants subjected to salt stress. PBZ induced increase in superoxide dismutase (SOD) activities was higher in cv. Tokak-157/37, than in cv. Erginel-90. However, an increase in SOD activity was not accompanied by an increase in peroxidase activity.

Contents of soluble proteins, proline and chlorophyll in winter wheat (Triticum aestivum cv. Doğu-88) and cabbage leaves (Brassica oleracea convar. acephala) during acclimation to low temperature were investigated. When both of the plants species were cold acclimated, soluble proteins, proline and chlorophyll contents were higher than in the controls (non-acclimated). Also protein patterns differed between the plants at control and cold conditions.

This study was conducted to determine effects of nitrogen supply (75 and 150 kg(N) ha^-1) and CO₂ enrichment on partitioning of macro and micro nutrients in wheat (Triticum aestivum L. cv. HD-2285). Plants were grown from seedling emergence to maturity inside open top chambers under ambient CO₂ (CA, 350 ± 50 μmol mol^-1) and elevated CO₂ (CE, 600 ± 50 μmol mol^-1). Leaves, stems and roots of the same physiological age were analyzed for carbon, nitrogen, calcium, copper, iron, zinc and manganese content at 40, 60 and 90 d after germination. C, Cu, Mn and Zn content was higher in the stem, leaves and roots on dry mass basis under CE than CA. However, N and Fe contents decreased in CE grown plants. Ca content was unaffected due to CE and variable N supplies.

Barley (Hordeum vulgare L. cv. Alfa) seedlings were treated for 4 d before UV-B irradiation with 0.05 mM proline or 150 mM NaCl. UV-B exposure induced synthesis of yellow coloured compounds with maximum absorbance at 438 nm. The content of these compounds was increased in proline-treated and decreased in NaCl-treated plants. UV-B radiation reduced chlorophyll/carotenoids ratio, oxygen evolution rate and photochemical efficiency of PS 2 as estimated by chlorophyll fluorescence and increased proline accumulation, H₂O₂ generation and lipid peroxidation. Exogenous proline had no effect on the parameters studied and did not change the response of plants to UV-B radiation. NaCl inhibited photochemical efficiency of PS 2, reduced oxygen evolution and increased H₂O₂ concentration and lipid peroxidation. The combination of NaCl and proline treatment led to lowering the inhibitory effect of NaCl in non UV-B irradiated seedlings. There was not relationship between the level of UV-B-induced compounds and UV-B tolerance of barley seedlings.

The activities of nitrate reductase and glutamine synthetase in leaves of greenhouse grown rose plants (Rosa hybrida cvs. Ilseta and Mercedes) grafted on various rootstocks were compared with those in leaves of non-grafted, own-root plants of these cultivars. The results obtained showed that the enzymatic activities as well as nitrate content in the leaves were altered by the grafting and by type of the rootstock used. These rootstock-imposed alterations differed between the two cultivars used in the study.

Two important enzyme in organic acid metabolism, phosphoenolpyruvate carboxylase (PEPC) and NADP-malic enzyme (NADP-ME), show marked diurnal rhythms in their activities during the establishment of the soybean - B. japonicum symbiosis. The pH of the nutrient solution changes in parallel with NADP-ME activity, being maximal during the night periods, whereas activity of PEPC was highest during the day periods. The results from the experiments with stem girdled plants indicated that the activity of root PEPC is modulated to a great extent by the supply of photosynthates from the shoots. It was also established that succinate application in the nutrient solution during inoculation altered significantly the pattern of assayed enzyme activities. Although our experiments did not reveal the precise mechanism of the involvement of root PEPC and NADP-ME in soybean response to inoculation with B. japonicum, they indicated the pattern of their activity during the first 72 h postinoculation which are critical for establishment and functioning of the symbiosis.