The ultrastructure of chloroplasts in the primary leaf of 10-d-old bean plants (Phaseolus vulgaris L., cv. Cheren Starozagorski) was studied 3, 5, 24, 48, 72 and 168 h after a single treatment with simulated acid rain (pH 2.4, 2.2, 2.0 and 1.8). Different changes in chloroplast structure till full destruction of organelles were traced. A determining factor for these changes was the histological localization of chloroplasts. In the chloroplasts of palisade parenchyma different degrees of expansion of thylakoids (3, 5, and 24 h after the single treatment), and conformational changes of the inner membrane system (48, 72 and 168 h) were observed. The chloroplasts of spongy parenchyma showed a significantly higher degree of structure resistance. The expansion of thylakoids was weak and did not depend on the duration of treatment. The ultrastructural changes of chloroplasts confirmed relative resistance of this species till pH 2.0.

The growth retardant paclobutrazol (PBZ) inhibited stem internode growth of in vitro cloned potato plants. The extent of growth inhibition caused by10-9 M PBZ in Murashige and Skoog medium was genotype-specific, varying between 10 - 60% of the stem growth of untreated controls in ten cultivars examined. An increase in percentage of de novo bud regenerating stem internode segments (SIS) as well as in the total number of buds per explant was observed in SIS taken from PBZ pretreated plants. PBZ applied directly into the regenerative media had no stimulatory effect on there generation process. We assume that the enhancing effect of PBZ on regeneration may be attributed to its interaction with cytokinin metabolism.

A continuous propagation of juvenile larch in vitro was based on adventitious bud formation and different cytokinin combinations were tested concerning their effectiveness to induce elongation of adventitious buds. Zeatin (1.5 mg dm-3) combined with kinetin (0.15 mg dm-3) was found to be the best. Development and elongation of buds was achieved on a modified LP-medium. Using this system it was possible to propagate different larchspecies (Larix decidua, L. gmelinii and L. sukaczewii f. multiramosus) continuously. Shoots were successfully rooted and transferred to the soil.

Shoot and root fresh and dry matters and their Cu content were determined in 7-d-old seedlings of Triticum aestivum L. cv. Alkora treated with Cu (10,20, 40 µg cm-3) and Si (500 µg cm-3). Si significantly reduced the toxic effect of Cu on fresh and dry matter production of wheat seedlings. Moreover, plants treated with Cu and Si absorbed less Cu from the solution and had higher water content in shoots and roots than that treated with Cu only.

An efficient protocol was developed for in vitro plant regeneration via somatic embryogenesis from cell suspension cultures of metal tolerant grass Echinochloa colona (L.) Link. Callus was obtained by culturing leaf base on MS medium supplemented with 0.5 mg dm^-3 of 6-benzylaminopurine (BAP) and 2.0 mg dm^-3 of 1-naphthaleneacetic acid (NAA). Cell suspensions were initiated and established in MS liquid medium containing 0.5 mg dm^-3 BAP, 1.0 mg dm^-3 NAA and 2.0 mg dm^-3 2,4-dichlorophenoxyacetic acid (2,4-D). A reduction in the concentration of 2,4-D to 0.5 mg dm^-3 induced formation of somatic embryos. The embryos developed and grew into normal plants in the presence of half strength MS medium without growth regulators. The regenerated plants were hardened in the greenhouse and subsequently grown in the open. This system may be also used for isolation and culture of protoplasts as a first step in somatic hybridization.

We measured PAR-saturated CO₂ exchange rate (CER), and leaf N, P and chlorophyll (Chl) concentrations in 21 plant species, selected to encompass as broad a range in specific leaf area (SLA) as possible, and encompassing non-succulent C3 as well as succulent CAM plants. We worked with plants growing under uniform conditions in the facilities of a biological research station to ensure that any correlations found were due to inherent, genetically controlled, relationships between the measured parameters and not due to variations in resource availability in different habitats. We found CER to be strongly correlated to SLA, leaf N concentration and Chl concentration. CER increased much faster with increasing leaf N concentration (CER ≈ N^3.1) than with increasing SLA (CER ≈ SLA^1.2). CER also increased much faster with leaf N concentration than with increasing Chl concentration (CER ≈ Chl^1.3), indicating the photosynthetic N-use efficiency (NUE) to be higher for plants with high N concentration than for plants with low N concentration (NUE ≈ N^2.1). Analysis of covariance showed that these relationships exist even when comparing plants of widely different growth forms - succulent or non-succulent, and of different photosynthetic pathways, as the C3 and CAM plants compared here. Testing against scaling coefficients calculated using dimensional analysis, showed that the scaling of N, Chl and CER against SLA was not merely a result of diluting N and Chl with carbon in thicker leaves but that SLA, probably through influencing light absorptio and/or CO2 diffusion pathway, played an independent role in controlling CER.

Comparative analysis of the electrolyte efflux, as a screening test of the membrane tolerance to water stress, was carried out in poikilohydric plants Ramonda serbica Panč. and Ramonda nathaliae Panč. & Petrov. and homoiohydric plant Saintpaulia ionantha Wendl. from the same family Gesneriaceae. Water stress was induced by PEG 600. The high degree of solute leakage in the East-African drought-intolerant Saintpaulia ionantha points to the loss of membrane integrity. In contrast, Balkan endemites Ramonda serbica and R. nathaliae show high resistance to water stress due to the specific constitutional drought tolerance mechanisms.

Maize (Zea mays) was cultivated on lead-adultrated soil up to 600 mg(Pb) kg-1. At maturity, the maize seeds were harvested. The glutamate dehydrogenase (GDH) was fractionated to its isoenzyme population by Rotofor isoelectric focusing (IEF). The increasing Pb concentration progressively enhanced the more acidic isoenzymes (pI 6.3 - 6.5), and at the same time suppressed the less acidic isoenzymes (pI 7.3 - 7.8) and at the 600 mg(Pb) kg-1(soil) only the most acidic couple of isoenzymes (pI 6.3, and 6.5) were detectable. The NH4+ Km values of the GDH increased progressively from 6.2 in the control to 100 mM and the total glutathione content of maize seeds from 60 to 240 nmol g-1 in the 600 mg(Pb) kg-1(soil) treated maize. The orderly, and sequential isomerization of GDH in response to Pb suggests that the enzyme functions as a sensor in the monitoring of environmentally induced stress.

A simple procedure for the detection of extracellular plant proteolytic enzymes using insoluble dye stained gelatin substrates incorporated into an appropriate culture medium is described. Extracellular proteinases produced by the tested plant cells (callus culture and cell suspension) hydrolyzed the substrates and dyed peptide fragments were released. Dyed zones around and under the proteinase-producing callus cultures were formed on the agar medium. Similarly, coloration of the culture media using proteinase-producing cell suspensions was observed.

Genetic tumour tissues of Nicotiana glauca (Grah.) × N. langsdorffii (Weinm.), which grow on auxin and cytokinin-free medium, were incubated with [14C]-/[3H]-adenine or [3H]-hypoxanthine to investigate cytokinin biosynthesis. Adenine was supplied to tissues of two different ages (2- and 3.5-week-old) for 8, 24 or 30 h. The uptake was over 91.0 % (of "supplied radioactivity") by 2-week-old tissues as compared to around 50.0 % uptake by 3.5-week-old tissues. Incorporation into cytokinins could not be detected. While unmetabolized adenine accounted for only about 24.0 and 13.4 % of "extracted radioactivity" (following 8 and 30 h incubation, respectively) in 2-week-old tissues, relatively higher levels, i.e. 36.0 and 34.5 % (following 8 and 24 h incubation, respectively) were present in 3.5-week-old tissues. The metabolites formed were adenosine and its nucleotides (4.5 - 16.5 % and 37.4 - 60.2 % of the extracted radioactivity, respectively). Hypoxanthine was supplied to 3.5-week-old tissues for 8 and 24 h. While the uptake was low (<28.0 % of supplied radioactivity), the major proportion of extracted radioactivity was due to unmetabolized hypoxanthine (79.8 % and 85.9 % after 8 and 24 h incubation periods, respectively); the minor metabolites were inosine and adenosine (both <0.5 %) and their nucleotides (< 3.5 %). Radioactivity incorporation into cytokinins from hypoxanthine was not detected. Thus in the present investigations precursor incorporation from either adenine or hypoxanthine into cytokinins could not be demonstrated. It is possible that this may be due to slow rate of cytokinin turnover in these tissues.

Several pieces of research suggest that leaf liquid-path conductance (K1) may be smaller in evergreen species (EG) than in deciduous species (DC), coexisting in tropical dry forests. This was investigated in four DC and two EG in a tropical dry forest in Venezuela. The K1 (kg m-2 MPa-1 s-1) were consistently lower in EG (8.4 × 10-5) than in DC (12 to 20 × 10-5). Differences in water potential (ΔΨW) inside the leaves were calculated using K1 values in connection with maximum transpiration rates measured in the field. It was found that ΔΨW was about 1.9 MPa in EG, intermediate for one DC (Pithecellobium dulce, 1.7 MPa) and minimum for other DC (1.0 to 1.3 MPa).

The effect of cytokinins (CKs) and K+ on stomatal behaviour in darkness were studied in the root hemiparasite Melampyrum arvense before (the preparasitic stage) and after attachment to the host (Capsella bursa pastoris L. Med.). The solutes were applied with xylem stream. The stomatal apparatus of the attached hemiparasite was insensitive to externally supplied CKs and K+. Contrary to this finding, the stomatal aperture of hemiparasite in the preparasitic stage increased to about 25, 40 and 69% of the value obtained in light, respectively, after treatment with 200 mM KCl and 10-5 M zeatin riboside ([9R]Z), applied separately or together. CKs influenced K+ transport. The treatment with KCl and [9R]Z,separately or together, increased the content of K+ in guard cell pairs to about 32, 46 and 79 % of the value obtained in light, respectively. Other CKs had a smaller effect (45 - 16 %) in comparison with that of [9R]Z; isopentyladenine was nearly inactive.

We have examined the toxicity of ethanol in tissue culture of the apple rootstock 'Jork 9'. During proliferation through axillary branching, 0.2% (v/v) ethanol slightly stimulated proliferation whereas significant inhibition occurred at concentrations of 0.4 % or higher. In adventitious root formation, significant inhibition occurred at concentrations of 0.1 % or higher. The effect of ethanol was stage-dependent: during the induction period (i.e. from 24 to 72 h after the start of the rooting treatment), there was little or no inhibition. During autoclaving, ethanol evaporated to ca. 50 %.

Standard methods of isolation of chromatin histones from Arabidopsis thaliana yield strongly degraded proteins when applied to plants grown from seeds in axenic liquid media. For isolation of undegraded histones from Arabidopsis grown in liquid media we used extraction with guanidine hydrochloride followed by selective binding of histones on BioRex 70 resin in the batch system. The quality of obtained proteins is confirmed by SDS polyacrylamide gel electrophoresis.

The mechanisms of saccharose and sorbitol transport in Prunus persica leaves were investigated in plasma membrane vesicles purified by aqueous 2-phase partitioning and equilibrated in pH 7.5 buffer containing K+. The imposition of an artificial proton motive force energized an active uptake of both saccharose and sorbitol. The maximum uptake rate of saccharose was 2.5 times higher than that of sorbitol. Saccharose and sorbitol uptake exhibited saturation kinetics suggesting they were carrier-mediated. Apparent Km for the saccharose and the sorbitol uptake were 0.36 and 0.67 mM, respectively. Active absorption of saccharose was completely inhibited by a non-permeant thiol reagent, PCMBS, contrary to sorbitol absorption. These results suggested that saccharose and sorbitol were transported at least by two different carriers.

Suspension-cultured rice cells growth was markedly inhibited and ammonium content increased when rice cells were deprived of phosphate. When rice cells were cultured at increasing concentrations of ammonium chloride, ammonium content increased, however, no significant inhibition of cell growth was observed. Addition of D-arginine, an inhibitor of putrescine biosynthesis, resulted in a complete recovery of growth in rice cells under phosphate deprivation, but did not decrease the content of ammonium. Our results indicate that the growth inhibition induced by phosphate deprivation is not associated with ammonium accumulation.

Triazine (0.6 mM) quenched the chlorophyll fluorescence emission at 686nm, selectively reduced the biosynthesis of linolenic acid (C18:3), lowered the ratio of unsaturated/saturated fatty acids in thylakoid membranes and percentage of total glycerides in the extracted oils, and extremely enhanced ethylene production by seedlings. These effects were partly reversed by the addition of 100 g m-3 of vitamins (thiamine > nicotinic acid >pyridoxine).

In order to clarify the role of endogenous growth inhibitors A-2α and A-2β in a dwarf pea plant, red light (emission peak 657 nm) treated, 9-d-old seedlings of dwarf pea (Pisum sativum L. cv. Progress No. 9) were transferred to darkness, and the resulting changes in growth rate and concentrations of A-2α and A-2β were monitored. The growth rate of the epicotyls increased, and the concentration of the inhibitors in the epicotyls decreased, according to sigmoidal time courses. The relationship between the logarithms of the concentration of the inhibitors and the corresponding growth rate was linear. These results suggest that A-2α and A-2β, may play an important role in the growth recovery process of the dwarf pea cultivar after termination of red light irradiation.

The response of Brassica carinata hybrids and their parents to moisture stress at different growth stages was studied. B. carinata 226 was found to be susceptible to stress at pre-flowering and post-flowering stages while B. carinata 241 at flowering stage. Neither the changes in stomatal conductance nor in chlorophyll content could fully explain the reduction in net photosynthetic rate (P_N) induced by stress. B. carinata 241 had higher leaf water potential (ψ_w) although, it had lower P_N compared to B. carinata 226. Both the parents had lower P_N as well as leaf ψ_w. The stress response of PN in hybrids followed that in their respective female parents.

Seedlings of the salt sensitive wheat cultivar C-306 evolved more ethylene than the salt tolerant cultivar Kharchia-65 under different levels of both chloride- and sulphate-dominated types of salinity. Pre-sowing seed soaking treatments with kinetin, gibberellic acid and to a lesser extent indole-3-acetic acid alleviated salt stress effects as apparent from seedling dry mass. Treated seedlings also evolved more ethylene both under saline and non-saline conditions. Ethrel did not affect seedling growth as well as ethylene production. Abscisic acid inhibited seedling growth and ethylene production under both types of salinity.

The role of plant antioxidant system in water stress tolerance was studied in three contrasting wheat genotypes. Water stress imposed at different stages after anthesis resulted in a general increase in lipid peroxidation (LPO) and decrease in membrane stability index (MSI), and contents of chlorophylls (Chl) and carotenoids (Car). Antioxidant enzymes like glutathione reductase and ascorbate peroxidase significantly increased under water stress. Genotype C 306, which had highest glutathione reductase and ascorbate peroxidase activity, also showed lowest LPO and highest MSI, and Chl and Car contents under water stress in comparison to susceptible genotype HD 2329, which showed lowest antioxidant enzyme activity as well as MSI, Chl and Car contents and highest LPO. HD 2285 which is tolerant to high temperature during grain filling period showed intermediate behaviour. Thus, the relative tolerance of a genotype to water stress as reflected by its comparatively lower LPO and higher MSI, Chl and Car contents is closely associated with its antioxidant enzyme system.

Activity of Cu2+-diamine oxidase (DAO; E.C.1.4.3.6.) was measured in Vigna radiata (L.) Wilczek cultivars K851, MH8320 and Pusa Baisakhi in light and in dark during ontogeny of seedlings. DAO activity was always the highest in cv. K851. In both light and dark grown seedlings maximum DAO activity was detected on day 2 after germination. Thereafter, in light grown seedlings it declined consistently upto non-detectable levels. In dark, DAO activity was higher than in light and it had the second maximum on day 7 following a similar declining pattern as observed for the light grown seedlings. The DAO activity was higher in a shoot apex alongwith leaves than in roots and shoot axis.

Maturation and germination of somatic embryos of hybrids A. alba × A. alba and A. alba × A. nordmanniana were followed by protein analysis of single embryogenic -suspensor masses (ESM) and analysis of storage protein accumulation during somatic embryo development. Very important step was one week pre-cultivation of ESM on medium with polyethylene glycol (PEG) and abscisic acid (ABA). Low osmotic potential of maturation medium and addition of ABA supported development of somatic embryo. Also partial drying of somatic embryo during following three weeks was needed for its normal development. In spite of morphologically fully developed, the somatic embryos were not physiologically ready for germination at least in terms of storage protein accumulation.