A full length cDNA (designated CrETR1) was isolated by polymerase chain reaction amplification of a cDNA library from periwinkle (Catharanthus roseus) cell cultures. CrETR1 cDNA encodes a polypeptide of 740 amino acids with a predicted molecular mass of 82 kDa. The deduced protein contains a hydrophobic ethylene-binding transmembrane region, a GAF domain, a third domain homologous to the histidine protein kinase domain of the prokaryotic two-component systems, and a fourth carboxyl-terminal domain homologous to the receiver domain of the response regulators, as found in the A. thaliana ethylene receptor ETR1. CrETR1 transcripts are strongly accumulated in petals and ovaries of C. roseus young plants whereas no significant changes are detected in cell cultures submitted to various stress or hormonal (including ethylene) treatments. The amount of the monoterpene indole alkaloid ajmalicine in the cells treated by ethylene is reduced after addition of inhibitors of histidine kinases showing a possible involvement of CrETR1 protein in the ethylene-related signalling pathway leading to alkaloid biosynthesis enhancement in C. roseus cell cultures.

Electrophoretic analyses of non-reduced and reduced seed storage proteins from Solanaceae and Cucurbitaceae species and cultivars were performed. High molecular disulfide bonded complexes between intermediary subunits of 11S globulins previously detected in Capsicum annuum cultivars, were found in Solanum melongena cultivars as well. The data obtained might be used for further elucidation of peculiarities of the 11S globulins in dicotyledonous plants.

Tortula ruralis is an important experimental system for the study of plant desiccation tolerance. EST gene discovery efforts utilizing desiccated gametophytes have identified a cDNA TrRpt2 encoding a predicted polypeptide with significant similarity to the 26S proteasome regulatory subunit IV. TrRPT2, the 446 amino acid deduced polypeptide, has a predicted molecular mass of 49.6 kDa, and a predicted pI of 8.15. Phylogenetic analysis demonstrated that previously characterized RPT2 polypeptide sequences could be reproducibly grouped into 3 major clades and that TrRPT2 forms a discrete evolutionary group. RNA blot hybridizations were used to analyze TrRpt2 expression in response to: 1) desiccation and rehydration, 2) abscisic acid-treatment, 3) increased NaCl concentration, and 4) NaCl-shock. TrRpt2 steady-state mRNA transcript levels are unchanged in response to all treatments and the gene is constitutively expressed.

In vitro protocols for plantlet regeneration and medium-term genotype conservation of eight wild species of Curcuma have been optimized. Both the phenomena were genotype-dependent and influenced significantly by type and concentration of cytokinins used. In general, benzyladenine (BA) was found superior to other cytokinins tested for plantlet regeneration and γ,γ-dimethylallylaminopurine (2iP) for conservation. Number of shoots per culture ranged from 1.3 to 7.2 and conservation period from 264 to 379 d. In 30-d-old cultures, highest frequency of shoot regeneration could be obtained in C. malabarica (7.2 shoots per culture) on MS + 11.4 μM zeatin. Curcuma sp. (unidentified wild species) could be conserved for maximum period (379 d) on MS + 24.6 μM 2iP followed by C. aromatica (363 d) on MS + 22.8 μM zeatin. The tissue culture-raised plantlets were morphologically similar to their parents. The in vitro-conserved plants multiplied rapidly in tissue cultures and produced normal rhizomes upon transfer to soil in net house.

We have assayed different combinations of nutrient media and growth regulators to induce callus and plant regeneration from explants of root, shoot and leaf, complete seed, and isolated mature embryo of barley (Hordeum vulgare L. cv. Hassan). The best results were obtained with mature embryo in J25-8 medium supplemented with 2.0 mg dm^-3 2,4-dichlorophenoxyacetic acid where about 75 % developed friable calli. Some 80 - 85 % of these calli regenerated barley plants in the same J25-8 medium supplemented with 1.0 mg dm^-3 indole-3-butyric acid and 0.1 mg dm^-3 kinetin.

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.

Stable transformation and expression of transgenes was achieved in Centaurium erythraea Gillib. using Agrobacterium rhizogenes-mediated system. Five hairy root clones exhibited the transformed phenotype. Shoot regeneration, with green organized structures, was apparent in 4 clones, after the first subculture on Murashige and Skoog (MS) half strength medium. The integration of Ri-plasmid T-DNA was confirmed by polymerase chain reaction (PCR) analyses.

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.

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.

In vitro regeneration of Melia azedarach L. was studied. Shoots were regenerated from calli initiated from leaflets of in vitro growing plants. The best medium for establishment of cultures was Murashige and Skoog (MS) medium with 4.44 μM benzylaminopurine (BAP) + 0.46 μM kinetin (KIN) + 16.29 μM adenine sulphate (ADE). Regenerated shoots were multiplied in MS + 0.44 μM BAP + 0.37 μM KIN + 3.26 μM ADE. Maximal rooting of 89 % was achieved by culture of regenerated shoots in MS + 12.26 μM indole-3-butyric acid for 3 d and subsequently in MS lacking growth regulators for 27 d. Rooted shoots were acclimatized and successfully transferred to soil.

This study deals with the effects of two cytokinins [kinetin (Kin) and N-(2-chloro-4-pyridyl)-N-phenylurea (CPPU)] and cytokinin antagonists [2-chloro-4-cyclobutyl-amino-6-ethylamino-1,3,5-triazine (ACK1) and N-(4-pyridyl)-O-(4-chlorophenyl)carbamate (ACK2)] in concentration of 1 μM on in vitro cultured Gypsophila. The application of Kin and CPPU stimulated bud opening and increased fresh and dry masses. Cytokinin antagonists reduced the number of sprouted buds and bud fresh and dry masses. In plants treated with CPPU the chloroplasts possessed well developed membrane system, which covered almost the entire chloroplasts volume. In ACK2 treated plants, the plastid apparatus in each cell was represented by two types of chloroplast in which the inner membrane system was differently organized. Cell wall adjacent chloroplasts possessed structure similar to the controls. In inner located chloroplasts part of thylakoids were semi-concentrically arranged and partially destructed.

Capsaicin, a possible allelochemical, caused growth inhibition of roots and shoots of alfalfa (Medicago sativa), cress (Lepidium sativum), lettuce (Lactuca sativa), crabgrass (Digitaria sanguinalis), timothy (Phleum pratense) and ryegrass (Lolium multiflorum), and suppressed their germination. Increasing the dose of capsaicin increased the inhibition. The concentrations for 50 % inhibition of the root growth were 2.7, 0.32, 2.1, 0.27, 0.29 and 0.57 mM for alfalfa, cress, lettuce, crabgrass, timothy and ryegrass, respectively, and the concentrations for 50 % inhibition of the shoot growth were 17, 0.87, 6.7, 2.3, 1.4 and 6.2 mM for alfalfa, cress, lettuce, crabgrass, timothy and ryegrass, respectively. Germination percentage was inhibited 50 % at the concentrations 82, 88, 68, 4.8, 22 and 11 mM for alfalfa, cress, lettuce, crabgrass, timothy and ryegrass, respectively. Thus, effectiveness of capsaicin on the plant growth differed with species and targets, and suggests that capsaicin may act as an allelochemical to other plants.

The effects of 50 mM NaCl and 5, 10, 15 and 20 mM boron on the rate of germination, growth rate, contents of boron, sodium, potassium and chloride, and membrane permeability in maize (Zea mays L.) and sorghum (Sorghum bicolor L.) were studied. Germination rate, lengths of roots and shoots as well as dry matter production in the two tested plants, decreased with the increasing B concentration in nonsaline conditions, and markedly under salinity. Membrane permeability increased by increasing B concentration only under salinity. Increase in B concentration of sorghum was lower under salinity when compared to nonsaline conditions. Contrary to this, increase in B concentration of maize was higher under salinity. Under salinity Na and Cl concentrations increased and K concentration decreased in the both tested plants. Potassium concentration was decreased by B treatments under salinity.

The effect of exogenous brassinosteroids (BR) on the flowering induction of Pharbitis nil was examined. Generally plants treated with brassinolide and castasterone form less number of flowers than control plants, but degree of flowering inhibition was depended on the concentration and the method of BR application as well as the length of the inductive dark period. In plants regenerated from sub-induced apices treated with brassinolide at concentration of 1 and 10 μM the flower formation was inhibited completely.

The greatest growth of wheat and rape plants in vitro was reached on media with 5 or 9 % sucrose, respectively. The highest efficiency for transfer of these plants to ex vitro conditions was found at the same sucrose concentrations. The content of endogenous non-structural saccharides (glucose, fructose, sucrose, starch and fructans) increased with increasing sucrose concentration in the medium up to 10 %.

Wild type of Arabidopsis thaliana plants were cultivated hydroponically in Hoagland and Arnon nutrient solution and treated with copper (5 - 100 µM) for 2, 4, 7 and 14 d. A progressive decrease of the root length and biomass was observed at increasing Cu concentration in the nutrient solution. Roots accumulated higher amounts of Cu than shoots at all Cu treatments. Changes of cell and chloroplast ultrastructure of Cu-treated plants were also observed. Cu application did not induce formation of Cu-phytochelatin complexes. Changes in glutathione and glutathione disulfide content observed in roots and shoots of Cu-treated plants suggest their participation in amelioration of metal-induced oxidative stress.

Multiple shoots were initiated from nodal and shoot tip explants collected from mature trees of Morus alba L. cultivars Chinese White, Kokuso-27 and Ichinose, and M. multicaulis Perr. cultivars Goshoerami and Rokokuyaso after 2 weeks of culture. Nodal explants were more responsive than shoot tip explants. Murashige and Skoog basal medium was found to be most suitable medium and 6-benzylaminopurine was the most effective cytokinin for shoot induction. Explants collected between April and September evoked better response than the explants collected between October and March. Shoots were multiplied by transferring nodal explants excised from in vitro raised shoots onto a medium containing cytokinins. Sucrose was the most suitable carbon source examined for shoot multiplication. An increase in shoot multiplication rate was noticed upto 4 - 5 subcultures. Nodal explants rooted on an auxin-supplemented medium. The acclimatized plants were successfully transplanted in the field.

A greenhouse pot experiment with different phosphorus supply was conducted to study growth, photosynthesis and free polyamine (PA) content in Plantago lanceolata L. plants in relation to arbuscular mycorrhizal (AM) colonization. Inoculum of Glomus fasciculatum (BEG 53) was used. Inoculated plants had high colonization intensities which were related to the P supply. Non-mycorrhizal (NM) plants showed a typical yield response curve for P availability. Dry masses of mycorrhizal (M) plants were higher at the lowest soil P content than those of NM plants, but the opposite was found at the highest P supply. P contents in M plants were always higher. There were no differences in chlorophyll (Chl) concentrations (except the lowest soil P content) and ratios of variable to maximum Chl fluorescence (Fv/Fm) values between M and NM plants, whereas M plants had higher ratios of leaf area to fresh mass (A/f.m.) at low soil P contents and they had significantly higher CO₂ fixation capacities per unit leaf area. Free putrescine (Put), spermidine (Spd) and spermine (Spm) contents in NM plants were usually highest at the lowest P supply. The ratios of Put/(Spd+Spm) were identical in M and NM leaves. They were significantly higher, however, in NM roots at the two low P doses. It is concluded, that a P nutritional status might exist, below which PA concentrations and ratio are increased drastically, possibly indicating P deficiency or a certain state of plant development with a higher demand for AM symbiosis.

γ-Aminobutyrate (GABA) was the only amino acid out of three amino acid intermediates of GABA shunt that increased significantly after 28 h from the beginning of osmotic stress induced by 20 % polyethylene glycol 6000 in wheat seedlings. At the same time specific activities of glutamate decarboxylase (GAD) and GABA aminotransferase (GABA-T) two enzymes of GABA pathway did not change as compared with the control plants. The response of two GABA-T activities (with pyruvate or 2-oxoglutarate as amino acid acceptor) to aminooxyacetate, 3-chloro-L-alanine and p-hydroxymercuribenzoate prompted us to suggest that at least two isoforms of GABA-T showing different substrate specificity do exist in wheat leaves.

Two wheat (Triticum aestivum L.) genotypes differing in their sensitivity to water deficit (stress tolerant - C306 and stress susceptible - HD2329) were subjected to osmotic stress for 7 d using polyethylene glycol (PEG-6000; osmotic potential -1.0 MPa), at initial vegetative growth. The plants were either supplemented with 1 mM CaCl₂ (Ca²⁺) alone or along with verapamil (VP; calcium channel blocker) to investigate the involvement of calcium in governing osmoregulation. Relative elongation rate (RER), dry matter (DM) production, water potential (Ψ_w), electrolyte leakage (EL), contents of proline (Pro) and glycine betaine (GB) and activities of γ-glutamyl kinase (GK) and proline oxidase (PO) in shoots and roots were examined during stress period. C306 showed relatively higher accumulation of Pro while HD2329 accumulated more GB under stress. RER, DM and Ψ_w were relatively higher in C306 than HD2329. Roots compared to shoots showed lower content of osmolytes but had faster rate of their accumulation. Presence of Ca²⁺ in the medium increased the activity of GK and decreased that of PO while in the presence of its inhibitor, decrease in activity of both the enzymes was observed. Ca²⁺ appeared to reduce the damaging effect of stress by elevating the content of Pro and GB, improving the water status and growth of seedlings and minimizing the injury to membranes. The protective effect of Ca²⁺ was observed to be more in HD2329 than C306.

Plants of maize (Zea mays L.) were waterlogged for 7 d and irrigated (root application) or sprayed (foliar application) with 0, 1, 3 and 6 mg dm-3 of boron. The stability of leaf membranes was assessed by determining leakage of electrolytes from leaf discs exposed to heat (51 °C) and dehydration (40 % PEG). Leaf membranes were more stable to heat than to dehydration. The membranes of waterlogged plants were more injured than those of control plants. Waterlogging reduced contents of dry mass, chlorophyll, soluble proteins, total free amino acids and soluble sugars and leaf relative water content (RWC). Application of boron increased the stability of leaf membranes, chlorophyll, soluble sugars, soluble proteins, amino acids contents, leaf RWC and dry mass accumulation. Foliar boron application was more effective. Application of boron alleviated the deleterious effect of waterlogging.

Distribution of cadmium between roots and shoots of barley was manipulated by the cadmium concentration (0.01 and 0.005 mM Cd²⁺), pH (4.6 and 5.9) as well as treatment duration. The prolongation of treatment increased dry mass and content of cadmium in plants. The cadmium is accumulated mainly in roots. Presence of both, 0.005 mM Cd²⁺ and 100 mM NaCl in medium at pH 5.9 (Cd-NaCl plants) resulted in the most severe growth inhibition of plants, but about one half accumulation of cadmium in roots then in a case of only Cd-treated plants. In the Cd-NaCl plants, the net photosynthetic and transpiration rates were less reduced then in a case of only NaCl-treated plants. The treatments also influenced uptake of Ca, Cd, Cu, K, Mg, Na and Zn predominantly in roots.