The excised shoot tips of Beta vulgaris L. cvs. Ras poly, Gala and Lola were inoculated aseptically on Murashige and Skoog's (MS) medium with 0.25 mg dm-3 α-naphthaleneacetic acid (NAA) and 1.00 mg dm-3 6-benzylaminopurine (BAP). They produced multiple shoots at relatively high frequency. For shoot multiplication, a concentration of 1.00 mg dm-3 NAA and 0.25 mg dm-3 BA in the culture medium was found to be optimum for all the cultivars. MS medium supplemented with 3.0 mg dm-3 3-indolebutyric acid (IBA) and 0.02 mg dm-3 N-isopentenylaminopurine (2ip) was found most effective for rooting of micropropagated shoots. In vitro produced plantlets were successfully transferred to the soil. The percentage of survival was more than 95 % and micropropagated plants showed no morphological differences from those grown naturally.

The effect of irradiance on the expression of the Cucumis sativus pathogenesis-related (srPRX) anionic peroxidase was studied in germinating seeds at the period when seedlings start to be photosynthetically active. The diversity in the expression patterns of srPRX was observed in both dark- and light-grown seedlings using activity staining and immunoblotting: beside the three srPRX isoenzymes also other three, serologically unrelated, peroxidase isoforms were accumulated in dark-grown seedlings and one in light-grown seedlings. Furthermore, in light-grown seedlings, it was observed a marked difference in the expression of particular srPRX isoenzymes at higher irradiance (up to 260 W m-2, 400 - 700 nm) in comparison to low irradiance. By tissue printing on nitrocellulose paper it was demonstrated that irradiance during germination induced changes in the temporal and spatial distribution of the srPRX.

Shoots of Malus domestica Borkh. cv. Compact Spartan raised in vitro do not root in a single auxin medium. Components of the rooting medium were tested not only for root initiation but also for root elongation. Root emergence and further growth were inhibited by a too prolonged auxin treatment, the presence of NH4NO3 and the lack of substrate aeration. Saccharose was essential to achieve highly reproducible root growth on agar but was not necessary on watered vermiculite. Ca(NO3)2 stimulated root initiation, emergence and growth and improved their viability.

In vitro cultures of Calathea ornata Koern. cv. Sanderiana shoots were maintained on modified medium of Murashige and Skoog. With the aim to find effective micropropagation method the effects of cytokinins, paclobutrazol, saccharose, mineral salts and shoot-tip damage on the shoot growth and quality were studied. The highest number of shoots was obtained on a medium containing 2.5 mg dm-3 benzylaminopurine, 2.5 mg dm-3 kinetin, 4.5 % saccharose and raised concentrations of Ca, Mg, Fe and Mn. Shoot branching was markedly stimulated by the shoot-tip damage.

Differences in anatomy and morphology of the kiwifruit leaves and leaf petioles might play a considerable role in the sex-determination. Three months after bud break (June), the kiwifruit leaves of both male and female plants, grown on the vegetative and generative shoots showed different leaf area (128.6 ± 13.45 cm2 in male and 104.5 ± 4.02 cm2 in female plants) and shape. The most frequently leaf shape was determined as "folium cordatum" and "folium rotundato-cordatum". Higher values of total leaf thickness of the female leaves (190 ± 3.84 µm) in comparison to male leaves (174 ± 3.52 µm) were estimated, resulting in the thicker adaxial leaf epidermis and especially in thicker palisade parenchyma in female leaves (136 ± 2.76 µm in comparison to 104 ± 1.61 µm in male leaves). Typically bifacial leaves were observed in both male and female leaves. Anomocytic stomata in hypostomatic leaves were found. The reticulate venation appears to be the main type of leaf venation. Stalked stellate multicellular trichomes on the abaxial leaf side were frequently observed in the leaves of both sexes. No important differences between male and female plants were found in the structures of vascular system in leaves and leaf petioles. Thus leaf thickness and surface morphology of adaxial leaf epidermis can be considered as important structural parameters in the sex determination.

Both short-term and continuous red radiation stimulated while far red radiation inhibited growth and ethylene production in Chlorella pyrenoidosa. Kinetin and linuron also affected culture density and ethylene production, depending on their concentration. Phytochrome might participated in the regulation of growth and ethylene production.

The involvement of ethylene in shoot formation in vitro was studied in one year old lavandin (Lavandula officinalis Chaix x Lavandula latifoliaVillars) callus. A peak in ethylene evolution characterized thenon-regenerating leaf callus line, as compared to the shoot-forming calyxcallus line, on the growth medium provided with 2,4-dichlorophenoxyacetic acid (1 mg dm-3) and kinetin (0.5 mg dm-3). After one year in culture, calyxcallus attained the capacity to grow on auxin-reduced media, showing decreased ethylene production and faster shoot bud emergence, when transferred onto the regeneration medium, supplemented with 10 mg dm^-3 benzyladenine. Shoot formation was also inhibited by addition of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid, indicating an involvement of ethylene in the failure of regeneration.

Supply of 1, 2, 5, 10 or 20 mM nitrate to detached roots, scutella or shoots from 5- to 6-d-old Zea mays L. seedlings increased in vitro nitrate reductase (NR) activity in all the organs and NADPH specific NR (NADPH:NR) activity in roots and scutella but not in the shoots. Usually 2 to 5 mM nitrate supported maximum enzyme activity, the higher concentration did not increase it further. The protein content in the roots, scutella and shoots increased up to 5, 2 and 20 mM medium nitrate, respectively. Nitrate uptake also increased with increasing nitrate concentration in roots and shoots, but it increased only slightly in the scutella. In both roots and scutella, methionine sulfoximine had no effect, while cycloheximide and tungstate abolished nitrate induced NADH:NR activity completely and NADPH:NR partially. Methionine sulfoximine increased nitrate uptake by roots and scutella slightly, but other inhibitors had no effect. The depletion of dissolved oxygen from the medium was lower in the presence of nitrate than in its absence or in the presence of ammonium, especially in the scutella.

Five sets of synthetic oligonucleotide (20-to 24-mers containing no intenal repeats) primers of known gene sequences [yellow lupin nodule specific (hydroxyl) proline-rich protein, pearl millet alcohol dehydrogenase, Pisum sativum heat shock proteins, Drosophila homeobox, and tRNA] were used to differentiate 73 soybean accessions, including 56 Glycine max (L.) Merr. and 17 G. soja Zucc. & Sieb. The amplified genetic markers revealed polymorphic bands for most genotypes studied. The χ²-analyses of the results showed that several fragments produced with these gene primers were associated non-randomly with resistance to Phytophothora, maturity, seed size, flower colour, seed coat colour, seed hilum colour, growth type, and leaf shape. These markers appear to be valuable for differentiation of G. max and G. soja species and genotypes within these species.

Membrane lipid peroxidation, activity of free radical scavangers and ethylene evolution of Amaranthus lividus seedlings were used to determine the lead and cadmium (1, 10, 100 and 1000 µM) induced phytotoxicity. Malondialdehyde (MDA) accumulation and higher lipoxygenase activity (LOX) was found in the 7-d-old treated seedlings. The activities of free radical scavangers like peroxidase, catalase and superoxide dismutase declined considerably with the concomitant rise in hydrogen peroxide level. Heavy metal treatment also caused decline in ethylene evolution in germinating seedlings.

A floating green alga Ulva pertusa Kjallman was grown in the laboratory under various irradiations: "white light" (as reference, broad spectral band, WLC), red radiation (600-700 nm, RRC) and blue radiation (400-500 nm, BRC). During 15 d of culture, the specific growth rate of WLC varied highly when compared to BRC and RRC. The contents of chlorophyll (Chl) and proteins, and the nitrate reductase (NR) activity were significantly higher in BRC than in RRC while the content of saccharides was slightly higher in RRC than BRC. U. pertusa in WLC had the highest contents of saccharides, proteins, and Chl, and the highest NR activity. In the WLC, closely arranged well organized thylakoids were seen whereas in the BRC, although the number of thylakoid layers was similar to WLC, they were widely separated from each other. In contrast to this, in the RRC, the thylakoids were less prominent and were also densely covered with ribosomes.

Cytokinin-like effects of pure caffeine were tested in bioassays specific for this hormonal activity [radish cotyledon growth and chlorophyll (Chl) biosynthesis in cucumber cotyledon and tobacco cell suspension] and in cell elongation bioassays [elongation of segments from soybean internode and internode elongation in dwarf cultivars of guandu (Cajanus cajan) and mucuna (Mucuna deeringiana)]. 6-Benzyl-aminopurine and kinetin (KIN) were used for comparison with caffeine. Although weaker than those given by cytokinins, positive responses were observed in all specific bioassays and in elongation of soybean internodes. A remarkable synergistic effect between caffeine and KIN was observed for the synthesis of Chl in the tobacco cell suspension bioassay, in which different concentrations of the alkaloid were combined with a single concentration of KIN. The hormone-like effect of caffeine might be related to the resemblance between caffeine and adenine derivatives.

The effect of concentration of zinc ions on chlorophyll (Chl) and dry matter contents in Senna occidentalis (L.) Link plants was studied under various availability of soil water. Diurnally, the internal Zn2+ content was predominantly affected by supplied Zn2+ and by daytime (Tm). Chl content, Chl a/b ratio, and Chl stability to heat were mainly affected by soil water content, and the dry matter content depended mostly on Tm.

5-Aminolevulinic acid synthase (ALAS) has been detected in a normal (auxin- and cytokinin-dependent) green sugar beet callus under light and under darkness. ALAS activity was lower when the callus was grown under light. The supply of precursors of the Shemin pathway (glycine and succinate) to dark-grown callus enhanced considerably the capacity of the 5-aminolevulinic acid (ALA) formation. Glutamate, γ-aminobutyrate or α-ketoglutarate also increased ALA accumulation. Such an accumulation was also obtained after inhibition of polyamine synthesis. The results show that glutamate or its derivatives might feed the Shemin pathway in conditions preventing glutamate to be used through the Beale pathway.

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.

Difluoromethylornithine (DFMO) counteracted several processes that are promoted or inhibited by auxins or inhibitors of polar auxin transport: inhibition of asymmetric gene expression in carrot, stimulation of gametic embryogenesis inBrassica, inhibition of root elongation in tobacco, inhibition of the development of lateral roots in pea and adventitious roots in apple, and inhibition of floral bud formation inArabidopsis

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.