Radish (Raphanus sativus L. cv. Redondo Vermelho) seedlings were analysed by light and scanning electron microscopy to characterize the structural changes caused by the exposure to 0.5 or 1.0 mM cadmium chloride for 24, 48 and 72 h. The analyses showed changes in the anatomical and morphological characteristics of roots, stems and leaves of two-week-old seedlings. In all tissues, pressure potential was decreased. Premature death with the disintegration of the epidermis and an increase in the number of root hairs was observed in roots exposed to Cd. The stem of seedlings exposed to Cd exhibited more cells layers in the cambial region. The main effects observed in leaves in response to Cd were stomatal closure, lack of cell wall thickening and alterations in the shape of the chloroplasts. It is suggested that the structural changes observed in seedlings treated with Cd were mainly caused by a Cd-induced decrease in water uptake.

A study of the genetic variability of a population of Hypericum brasiliense was made using several isozyme systems as well as an investigation of the morphogenic potential of apical buds from plants at different development stages (juvenile and adult) using in vitro culture. The results from nine isozymes systems showed low polymorphism in the alleles. Apical buds from juvenile plants originated plantlets with apical dominance and fast growth while those from adults led to the development of flower buds.

The effect of an extended drought (from 1992 to 1995) on water relations was assessed on evergreen oak (Quercus ilex L.) in a dehesa ecosystem (Seville, Southern Spain). Diurnal and seasonal transpiration patterns were analysed at leaf (porometry) and whole-tree level (sap flow), focusing on the relationship between tree transpiration rates (E_t) and potential evapotranspiration rates (PET). Daily maximum E_t varied over the year, becoming higher between May and August, and lower between November and April. Annual E_t (169 - 205 mm y^-1) accounted for less than 40 % of annual rainfall. The prolonged drought did not affect the water relations of the Q. ilex, mainly due to strong stomatal regulation avoiding the loss of water. Stomatal control was found in all seasons, although it was stronger in summer. This behaviour leads to low water consumption and low E_t/PET ratios throughout the year (0.05 to 0.27).

Growth parameters of six fast growing trees showed that the roots responded to Cd treatment more sensitively than the shoots. Cd-treatment suppressed rooting and root growth (length and biomass production) as well as its development in all tested species. Root systems of Salix cinerea, Salix alba, and Populus cv. Robusta were more tolerant to Cd stress than the root system of the other studied species. Shoot growth parameters of Salix species were significantly reduced unlike Populus species, which were not affected by Cd treatment.

In vitro bulblet formation was studied using solid, liquid and bioreactor culture (immersion and periodic immersion in liquid media using ebb and flood) in order to develop a cost effective method for the mass propagation of Lilium oriental hybrid 'Casablanca'. Although the percent of bulblet formation was higher in solid culture, the increased growth rate and production of large number of bulblets in bioreactor makes it suitable for mass propagation. Four times per day and 15 min of medium supply was optimal for bulblet formation in ebb and flood bioreactor. Bulblet formation was also found to be effective in 16-h photoperiod. It was also observed that bulblet formation in the medium with 1.0 mg dm^-3 BA and 0.3 mg dm^-3 NAA was higher than in the medium without growth regulators, but formation of abnormal bulblets was higher in medium with BA and NAA.

The pattern of NADH- and NAD(P)H-specific nitrate reductase (NRs) activities in Indian mustard (Brassica juncea L. Czern. and Coss.) was monitored throughout growth stages. NAD(P)H:NR (EC 1.6.6.2) activity was maximum at early stages of growth (30 days after sowing, DAS), then declined gradually reaching to almost zero at 90 DAS. Contrary to this, NADH:NR (EC 1.6.6.1) activity was low at 30 DAS, then gradually increased till 90 DAS and thereafter, it became constant. The decrease in NAD(P)H:NR activity and increase in the NADH:NR activity were associated with the seasonal decrease in nitrate content in the soil.

Phenylmethylsulphonyl fluoride (PMSF), a well known inhibitor of both thiol- and serine-type proteases, in aqueous solutions either alone or with the plant growth regulators, methyl ester of jasmonic acid (MeJA) and N⁶-benzyl-aminopurine (BAP), significantly inhibited the growth of excised Cucurbita pepo L. (zucchini) cotyledons. SDS-PAGE analysis of the protein profiles showed that PMSF suppressed the gradual decline of the main 20 - 25 kDa polypeptide group and the low molecular mass polypeptides (below 15 kDa) while leupeptine was not able to affect the electrophoretic pattern of cotyledon proteins. On the other hand, in the presence of PMSF, the content of the polypeptides with higher molecular mass including the 97.4 kDa polypeptide and the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (55 kDa) decreased. Besides, when applied together with MeJA, PMSF prevented the appearance of the jasmonate-induced polypeptides (JIPs; 69, 60 and 43 kDa) thus suggesting that JIPs are synthesized from aminoacids released during the breakdown of cotyledon storage proteins.

Seed storage proteins of Ebenus cretica were fractionated to albumins, globulins, prolamins and glutelins according to their solubility in water, 0.5 M NaCl solution, 55 % propanol-2 and 0.125 M sodium borate (pH 9.0) containing 0.5 % SDS (sodium dodecyl sulfate) solution, respectively. Glutelins consist of the major (about 81 %) fraction of the total extracted proteins. Analysis by SDS-PAGE revealed that the total extracted protein patterns from different racemes of the same plant were similar, while those from seeds of different plants were different. In addition, distinct differences were observed within protein patterns of alkaline extractable glutelin fractions and salt soluble globulin fractions. In E. cretica four ecotypes (A - D) were distinguished by SDS-PAGE of total extracted seed proteins. The last method was more simple and rapid than others and was suggested for screening analysis.

Effect of free radical scavengers and metal chelators on polyethylene glycol (PEG, osmotic potential -1.5 MPa) induced oxidative damage in detached rice leaves was investigated. PEG treatment resulted in a decrease in relative water content and an increase in proline content, and lipid peroxidation. PEG treatment also decreased chlorophyll and protein contents. Free radical scavengers (ascorbate, sodium benzoate, reduced glutathione, and thiourea) retarded and metal chelators [2,2'-bipyridine (BP), 8-hydroxyquinoline, and 1,10-phenanthroline] prevented PEG-induced oxidative damage. Furthermore, the protective effect of BP was reversed by adding Fe²⁺ and Cu²⁺, but not by Mn²⁺ or Zn²⁺. The protective effect of BP is most likely mediated through chelation of iron. It seems that oxidative damage induced by PEG may require the participation of iron.

The relationship between nutrient composition, crop biomass, and glutamate dehydrogenase (GDH) isoenzyme pattern was investigated in soybean (Glycine max) and maize (Zea mays) by monitoring the nutrient induced isomerization of the enzyme from the seedling stage to the mature crop. GDH was extracted from the leaves of the plants, and the isoenzymes were fractionated by isoelectric focusing followed by native polyacrylamide gel electrophoresis. The isomerization V_max values for soybean GDH, similar to maize GDH increased curvilinearly from 200 - 400 μmol mg^-1 min^-1 as the inorganic phosphate nutrient applied to the soil decreased from 50 - 0 mM. In soybean, combinations of N and K, P, or S nutrients induced the acidic and neutral isoenzymes, and gave biomass increases 25 - 50 % higher than the control plant. GDH isoenzymes were suppressed in soybean that received nutrients without N, K, or P and accordingly the biomass was about 30 % lower than the control. Treatment of maize with NPK nutrients increased the GDH V_max values from 138.9 at the vegetative to 256.4 μmol mg^-1 min^-1 at the reproductive phase, and suppressed the basic isoenzymes, but induced both the acidic and neutral isoenzymes thereby inducing seed production (27.0 ± 1.4 g per plant); whereas both the acidic and basic isoenzymes were suppressed in the control maize, and seeds did not develop. Simultaneous induction of the acidic, neutral, and basic isoenzymes of GDH indicated the occurrence of senescence. Therefore in maize and soybean, the induction of the acidic and basic isoenzymes of GDH led to the enhancement of biomass.

The effect of organic (Fe-EDTA and Fe-EDDHA) and inorganic (FeCl₃) iron substances on rooting of the rootstock GF-677 (Prunus amygdalus × Prunus persica) in vitro was studied. Full rooting (100 %) was observed in explants nourished with Fe-EDDHA, while less rooting was found in the absence of iron or in the presence of FeCl₃. On the contrary, no root formation was observed in explants nourished with Fe-EDTA, which showed extremely lower chlorophyll and high iron contents at the end of the experiment.

The plant growth retardant, paclobutrazol at 8.5 or 17.0 μM concentrations effectively inhibited the stem elongation and primary leaf expansion of bean seedlings. Although the retardant reduced the relative water content in well-watered plants, the water and pressure potentials remained high in the primary leaves. K⁺, Na⁺, Mg²⁺ and Ca²⁺ contents in the primary leaves of the paclobutrazol-treated plants were not significantly different from those in the control. The stomatal density increased on both surfaces but the length of guard cells was not reduced significantly on the adaxial epidermes of the paclobutrazol-treated primary leaves. The inhibitory effect of paclobutrazol on the abaxial stomatal conductances became more pronounced with time during the light period but the adaxial surfaces displayed similar or slightly higher conductances than those of the control. The transpiration rate on a unit area basis did not change significantly or increased in the treated leaves thus the reduced water loss of paclobutrazol-treated plants was due to the reduced leaf area. Stomatal conductances of the adaxial surfaces responded more intensively to exogenous abscisic acid and the total leaf conductance decreased faster with increasing ABA concentration in the control than in the paclobutrazol-treated leaves. Paclobutrazol, an effective inhibitor of phytosterol biosynthesis, not only amplified the stomatal differentiation but increased the differences between the adaxial and abaxial stomatal conductances of the primary leaves.

The effect of maize root exudates on the toxicity of lead and manganese to arbuscular mycorrhizal fungus (AMF) Glomus intraradices was studied in vitro by observation of intraradical hyphae regrowth from colonised root segments. Higher heavy metal (HM) concentrations strongly reduced the hyphal growth, however, the inhibitory effect was to a large extent eliminated by the addition of maize root exudates to the media. However, the capacity of exudates to ameliorate HM toxicity was limited and did not operate when a threshold HM concentration was reached.

The ascorbate system, one of the major antioxidant systems, has been studied in two bryophytes; a moss, Brachythecium velutinum (Hedw.) B., S. & G., and a liverwort, Marchantia polymorpha L. The moss and liverwort gametophytes contain ascorbate both in the reduced and oxidized form; utilize ascorbate in removing hydrogen peroxide by means of ascorbate peroxidase and reconvert to ascorbate its oxidation products by means of dehydroascorbate reductase and monodehydroascorbate reductase. Ascorbate oxidase activity was measured in the cytosolic fraction suggesting a localization of the enzyme different from more evolved organisms. The ascorbate content was maintained in the moss after drought stress while it declines in the liverwort, which seems more sensitive to water stress. Since ascorbate recycling is more efficient in the moss than in the liverwort, this seems to suggest a correlation between efficiency of ascorbate recycling and water stress tolerance.

The histological events during adventitious bud and somatic embryo development in explants from Picea omorika seedlings cultivated in vitro have been examined. Meristematic activity in the superficial layers of cotyledons led to bud primordia formation. Bud primordia had characteristic zonal organization with needle primordia arising on the flanks of the meristem. Somatic embryo initial was formed on the cotyledon surface after an unequal transversal division of a cell. The smaller, apical cell gave rise to apical dome and later on secondary, filamentous suspensor while the larger vacuolated cell presented primary suspensor.

The effects of ethephon, an ethylene generating compound, and 2,5-norbornadiene (NBD), an inhibitor of ethylene action, on peroxidase (POD; EC 1.11.1.7), catalase (CAT; EC 1.11.1.6), polyphenol oxidase (PPO; EC 1.14.18.1) activities and proline content in salt-stressed spinach leaves were investigated. POD and PPO activities were increased by NaCl + ethephon + NBD combination and reduced by NBD. Also, ethephon increased the CAT activity while ethephon + NBD reduced CAT activity. NaCl + ethephon increased proline content. The antagonistic effect of ethephon and NBD was seen on POD and PPO activity and proline accumulation, but was not on CAT activity.

The effect of Pseudomonas fluorescens treatment and Fusarium oxysporum f. sp. cubense inoculation on induction of phenylalanine ammonia-lyase (PAL), peroxidase (POX), chitinase, β-1,3-glucanase and accumulation of phenolics in banana (Musa sp.) was studied. When banana roots were treated with P. fluorescens strain Pf10, a two-fold increase in phenolic content in leaf tissues was recorded 3 - 6 d after treatment. Challenge inoculation with F. oxysporum, the wilt pathogen, steeply increased the phenolic content in P. fluorescens-treated banana plants. Significant increase in POX activity was detected 6 - 9 d after P. fluorescens treatment. PAL, chitinase and β-1,3-glucanase activities increased significantly from 3 d after P. fluorescens treatment and reached the maximum 6 d after treatment. Challenge inoculation with F. oxysporum further increased the enzyme activities. These results suggest that the enhanced activities of defense enzymes and elevated content of phenolics may contribute to bioprotection of banana plants against F. oxysporum.

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.

Trigonelline (TRG) is a conjugate of nicotinic acid, and is postulated to function as a compatible solute in response to salinity- and water deficit-stresses. TRG concentrations and several agronomic characteristics were measured under irrigated field and non-irrigated field conditions within 18 soybean (Glycine max) genotypes using leaves taken from different growth stages (vegetative, flowering and pod development). Under irrigation, relative water content (RWC) ranged from 90.0 to 99.6 %. Under non-irrigation, RWC ranged from 86.3 to 97.5 %. TRG concentration ranged from 364 to 555 μg g^-1(d.m.) under irrigation, and from 404 to 570 μg g^-1(d.m.) under non-irrigation. TRG concentrations increased in the majority of genotypes (15 of 18) under non-irrigation even though RWC did not significantly differ in many genotypes between treatments. TRG decreased as plants progressed to pod development and seed filling. Mean seed yield under non-irrigated conditions declined 55 % relative to the irrigated controls. TRG concentrations among all genotypes were significantly correlated with seed yield.

Almost all processes in the life of a plant are directly or indirectly affected by both stresses and phytohormones. Nevertheless, apart from abscisic acid, the role of phytohormones in plant response to water stress is far from being fully elucidated. This review tries to answer the question whether interactions between abscisic acid and some other phytohormones might be important in the regulation of stomatal opening during water stress and subsequent rehydration. Firstly, it describes the changes in the contents of individual endogenous phytohormones during water stress. Then, it deals with the effects of applied phytohormones on stomatal opening, and on transpiration and photosynthetic rates in different plants species. Finally, it focuses on the alleviation or stimulation of absicic acid-induced stomatal closure by application of other phytohormones.

Efficiency of pretreatment as foliar spray of indole-3-acetic acid, gibberellic acid and kinetin, each ranging from 0.1 to 10.0 μM concentration, in restoring the metabolic alterations imposed by NaCl salinity was investigated in Vigna radiata (L.) Wilczek. Glycolate oxidase, superoxide dismutase, catalase and peroxidase activities increased under stress in leaves and roots also. Malondialdehyde content and total peroxide content also increased under stress. All the three hormones used were able to overcome to variable extents the adverse effects of stress imposed by NaCl to these parameters.

The nodal and internodal explants excised from the orthotropic shoots of Sesbania sesban var. bicolor elicited the development of shoots directly from the explants as well as via an intervening callus phase on Nitsch (N) medium. On benzyladenine (BA) supplemented media, the adventitious shoot buds developed involving a callus phase. An average of 8.9 ± 4.1 shoots developed per nodal explant on N medium containing 0.5 mg dm^-3 BA in 95 % cultures, whereas 65 % cultures of internodal explants developed shoots with an average of 5.9 ± 3.6 shoots per explant on N medium supplemented with 1.0 mg dm^-3 BA. On kinetin (Kn) supplemented medium shoots developed directly from the surface of both the explants at all the concentrations tried. Nodal explants on N medium supplemented with 1.5 mg dm^-3 Kn developed an average of 12.5 ± 7.9 shoots per explant in 100 % cultures, while internodal explants induced an average of 11.6 ± 7.4 shoots per explant in 75 % explants at 0.5 mg dm^-3 Kn. The in vitro regenerated shoots developed roots when implanted on N medium supplemented with 2 mg dm^-3 indole-3-butyric acid (IBA), after 30 d of inoculation. The in vitro developed plantlets were initially acclimatized under controlled conditions for four months, prior to their transfer to the field.

For most crops growing in polluted areas or treated with agricultural chemicals, no genotoxicity assays are available. We have studied the possibility of using the alkaline protocol of the plant-based molecular assay - the Single Cell Gel Electrophoresis (SCGE) assay (also called Comet assay) as a method for detecting induced DNA damage in 8 agronomic important plants (ordered according to the diameter of the nuclei): sugar beet, alfalfa, tobacco, lentil, maize, potato, hard wheat, and bread wheat. The monofunctional alkylating agent ethyl methanesulphonate (EMS) was applied as a model genotoxic agent on young excised leaves of the tested crops for 18 h at 26 °C in the dark. With increasing concentrations of 2 to 10 mM EMS, the DNA damage, expressed by the averaged median tail moment values, significantly increased in nuclei of all crops studied. No correlation between the diameter of nuclei and sensitivity to EMS treatment was observed. The data obtained demonstrate the feasibility of using the Comet assay for detecting induced DNA damage in crops.

Auxins were analysed in roots, stems with apical shoots, and leaves of beans (Phaseolus vulgaris L. cv. Zorin) cultured for three weeks on media supplied with different concentrations of jasmonic acid (JA). Morphology changes induced by applied JA were reflected in tissue-specific changes of auxins.

Rooting of Eucalyptus globulus shoots was influenced by the concentration of the indole butyric acid (IBA) and NH₄ ⁺ in the root-induction medium. Optimum plantlet vigor and survival were achieved using low concentrations (1 - 2.5 μM) of IBA and when NH₄NO₃ was removed. Removal of NH₄ ⁺ also had a significant effect on medium pH, its presence caused a decrease in pH as the culture period proceeded. When different nitrate compounds (excluding NH₄NO₃) were used as the nitrogen source, the medium pH was more stable and this was associated with higher root production. The higher root production, in association with appropriate IBA concentrations, produced plantlets with higher survival and better growth on transfer to soil.