Leaf tissues of 38 genotypes, derived from four accessions, of the hexaploid species Helianthus tuberosus (2n=6x=102) responded to growth regulators (BA, NAA) chiefly by forming callus, while aventitious organogenesis or somatic embryogenesis were induced occasionally. A remarkable regeneration frequency (about 30 %) was achieved only from leaves of genotype HTPI-15. Explants of many regenerated plants of HTPI-15 subjected to a second culture cycle in vitro displayed a high morphogenetic potential (regeneration frequency > 90 %). White globular structures were initiated on the adaxial surface of these leaves without a callus phase. Somatic embryogenesis was asynchronous and embryoids, of different developmental stage, were simultaneously detected on each explant. Although many embryos developed single or malformed cotyledons or germinated precocciously, without the differentiation of a complete root system, phenotypically normal plants were regenerated after rooting on regulator-free half-strength MS medium.

Heat shock proteins (HSPs) ranging in molecular masses from 14 to 110 kDa were induced in embryonic axes of germinating Cajanus cajan (L.) Millspaugh seeds after exposure to 40 °C for 1 or 2 h. At 45 °C, there was a marked decline in synthesis of HSPs. A close relationship was observed between HSPs induced and the growth of the germinating seeds. Pretreatment of germinating seeds at 40 °C for 1 h or 45 °C for 10 min followed by incubation at 28 °C for 3 h led to considerable thermotolerance (45 °C, 2 h) and the recovery of protein synthesis.

In the leafstem moss Mnium affine two superoxide dismutase (SOD) isoforms were found in chloroplasts and two in mitochondria. Four other isozymes were probably cytosolic and two of them had high activity and thermostability and were very sensitive to H₂O₂. On the other hand, one of the mitochondrial isoenzymes was very sensitive to high temperature. The activity and thermosensitivity of SOD was considerably dependent on calcium and zinc ions. The effect was different for the individual isoforms and related to their subcellular distribution. Calcium ions predominantly activated and stabilized one cytosolic and the mitochondrial SODs, while zinc ions influenced one chloroplastic and two cytosolic SODs.

Water relations of two rapidly expanding species in deforested localities, Calamagrostis epigeios and Calamagrostis arundinacea, were compared. After short- or long-term water stress, water content and specific leaf area were more reduced in C. epigeios than in C. arundinacea. After short-term stress, osmotic potential was similar in both species, but after long-term stress, it was much lower in C. epigeios plants. Abscisic acid and proline contents were higher in stressed plants of C. epigeios than in those of C. arundinacea.

A transgenic expression system of Chlorella kessleri using the gene for β-glucuronidase (GUS) was developed. Cells of this unicellular green alga were bombarded with the plasmid pBI 121, which bears β-glucuronidase under the control of CaMV 35S promoter and the kanamycin resistant gene. Maximum GUS activity was obtained after 48 h of bombardment using a helium pressure of 900 kPa; GUS activity was then assayed for many generations. The stable transformants were able to grow on kanamycin containing medium after repeated passages between selective and nonselective medium and exhibited GUS activity comparable to that of control cells. Stable transformed cells were confirmed by polymerase chain reaction (PCR) and Southern hybridization of GUS probe with the genomic DNA of C. kessleri.

The short-term relationships of monoterpene emission with temperature and relative humidity were studied in Pinus halepensis L. and Quercus ilex L. seedlings grown in air-conditioned chamber. In P. halepensis terpene emission rate increased with temperature (from 15 to 35 °C) and relative humidity (from 40 - 60 to 65 - 95 %). In Q. ilex, a terpene non-storing species, it increased with temperature only at high relative humidities but not at relative humidities lower than 60 %.

After inoculation of Brassica juncea leaves with Alternaria brassicae, activities of the cell wall degrading enzymes, polygalacturonase (EC 3.2.1.15) and cellulase (EC 3.2.1.4) decreased in leaf blight resistant cultivar RC-781 and increased in the susceptible cultivar Varuna upto 3 d. In the leaves of both the cultivars 11 poly-peptides were observed in the absence of A. brassicae inoculation. After inoculation in the resistant cultivar RC-781 there was no change in the polypeptide pattern, while in the susceptible cultivar Varuna, four polypeptides (43.7 to 58.8 kDa) disappeared only at 3^rd day after inoculation.

The potassium uptake by potato tuber discs tissues freshly cut and after 24 h of ageing in the presence or not of abscisic acid was investigated. Uptake kinetics revealed a biphasic dependence on external K⁺ concentrations. At concentration less than 10 mM, uptake was mediated by a saturable component and a linear component became apparent at higher concentrations. At low K⁺ concentrations (lmM), the capacity of K⁺ uptake diminished by 2 times after ageing. Treatment of tissues with ABA increased the rate of K⁺ uptake. In both fresh and aged tissues the uptake was strongly enhanced by fusicoccin and decreased by several metabolic inhibitors and ATPase inhibitors, underlying the active nature of uptake and suggesting the involvement of a plasmalemma H⁺-ATPase in K⁺ transport system.

Two newly developed salt-tolerant genotypes of spring wheat, S24 and S36 and their salt-tolerant parents, LU26S (from Pakistan) and Kharchia (from India) along with a salt-sensitive cv. Potohar were grown in full strength Hoagland's nutrient solution with 0 or 125 mM NaCl. At the onset of the booting stage third leaf from top was sampled for protein analysis. Total soluble protein content increased due to salt treatment in all cultivars/lines but this increase was more marked in salt-sensitive cv. Potohar and low in salt-tolerant S24 as compared with the other lines. Patterns of labelled polypeptides in all cultivars/lines were identical; the differences were only quantitative (for instance, 29 kD and 48 kD polypeptides were reduced significantly due to NaCl treatment only in the cv. Potohar).

This work describes in vitro morphogenesis and plantlet regeneration from seeds of the naturally polyembryonic tree Syzgium alternifolium. The basal medium (BM) comprised half strength Murashige and Skoog's (MS) salts, B₅ vitamins, 2 mg dm^-3 glycine, 250 mg dm^-3 ascorbic acid and 20 g dm^-3 sucrose. Addition of auxins like indole-3-acetic acid (IAA), 1-naphthalene acetic acid or 2,4-dichlorophenoxyacetic acid to the basal medium induced formation of roots or callus from the dicotyledonous as well as tricotyledonous seeds. In contrast, cytokinins like N⁶-benzyladenine (BA), kinetin, 2-isopentenyl adenine, thidiazuron alone or a combination of BA and auxins induced development of adventitious shoots. The medium containing 3.0 mg dm^-3 BA and 0.5 mg dm^-3 IAA induced the highest number of adventitious shoots (32 - 33) from dicotyledonous seed with an average length of 4.1 cm within 6 weeks of incubation. Rooting of 80 % adventitious shoots was achieved by dipping the shoots in 100 mg dm^-3 IAA for 15 min. About 70 % of the rooted shoots were successfully established in pots after hardening.

Simulated acid rain (SAR) combined with higher concentration of aluminium (SAR+Al) influenced the ecophysiology of three arbuscular mycorrhizal fungi (AMF) in both the germination and symbiotic phases of their life cycle. Acaulospora tuberculata, an isolate from the soil with low pH, exhibited a higher tolerance to environmental stress as compared to Glomus mosseae and G. fistulosum. This higher tolerance may be related to the edaphic conditions of soil of the isolate origin. The histochemical staining of the alkaline phosphatase and NADH-diaphorase activities in the extraradical mycelium (ERM) of the AMF proved to be more sensitive indication of negative effects of the SAR or SAR+Al stress compared to commonly measured parameters of the AMF such as mycorrhizal colonisation or growth of the ERM.

21-d-old seedlings of the soybean (Glycine max) cvs. Essex and Forrest were treated with NaCl in a step-wise manner over 9 d (3 d 30 mM, 3 d 70 mM, and 3 d 100 mM) and maintained under 100 mM NaCl for an additional 14 d. During salt treatment, osmotic potential decreased more in cv. Forrest relative to cv. Essex. In non-stressed leaf tissue, cv. Forrest contained more trigonelline (TRG) relative to cv. Essex. During salt treatment, TRG amounts increased in cv. Forrest but were unchanged in cv. Essex. Both cvs. osmotically adjusted in response to salt stress; the maximal osmotic adjustment was 0.80 and 0.18 MPa in cv. Forrest and cv. Essex, respectively.

Carbon translocation was affected by shade in different tropical tree species differing in successional status and degree of shade tolerance. Plants of the early-successional shade-intolerant species Cecropia pachystachya and Schizolobium parahyba and of the late-successional shade-tolerant species Myroxylon peruiferum and Hymenaea courbaril were grown under full sun (FS) and natural shade treatments (NS) and assessed for [¹⁴C]-sucrose translocation. Most of the ¹⁴C was retained in the fed leaf after a 24 h translocation period. Under FS, the growing apical part of the plant was the most intense sink for most species. Shade affected growth and sink intensity differently in early and late successional species. Growth was more markedly affected in the early species. Whereas these continued to invest carbon into the growing apical part of the plant under shade conditions, the late successional species invested relatively more into other sinks.

In four genotypes of chickpea (Cicer arietinum L.) BG 362, BG 372, BG 329 and C235 the relationship between somatic embryogenesis of leaf explants and ethylene and methane evolution was studied. In BG 362, which was more embryogenic than other genotypes, a higher ethylene:methane ratio of 5.8:1 at day one after inoculation in the induction medium and a lower ethylene:methane ratio of 2.89:1 in the maturation medium was found. On the contrary, in BG 372 with the least embryogenic potential, a lower ethylene:methane ratio of 1.7:1 in the induction medium and a higher ethylene:methane ratio of 4:1 in the maturation medium was found. Thus, these ratios in induction and maturation stages seems to be markers for embryogenesis in leaf explants of chickpea.

Plants of bean (Vicia faba L. cv. Calvor 103) were salt-stressed with NaCl and CaCl₂ in concentrations inducing soil osmotic potentials (ψ_soil) from 0 to -1.2 MPa and were sprayed with proline (8.7 µM) and glycinebetaine (8.5 µM) solutions. Bean plants respond to increasing soil salinity by decreased leaf relative water content and osmotic potential. Salinity decreased the contents of dry mass, chlorophyll, soluble and hydrolysable sugars, soluble proteins and enhanced content of total free amino acids, Na⁺, Ca²⁺ and Cl^-. The ratio of K⁺/Na⁺ was decreased on salinization. The membranes of leaf discs from salt-stressed plants appeared to be less stable under heat stress (51 °C) than that of unstressed plants. The reverse was true for discs placed under dehydration stress (40 % polyethylene glycol 6000). Proline and glycinebetaine application reduced membrane injury, improved K⁺ uptake and growth. Also both solutes increased chlorophyll contents.

An improved version of a simple histochemical test for the in situ assaying the production of hydrogen peroxide in living plant tissue was demonstrated. The test solution containing 50 mM KI in a 4 % potato starch solution was directly applied to the fresh cut surface of the tissue to be tested. Incorporation of an enhancer potassium permanganate (1 % final concentration) into the test reagent resulted in a ten times greater hydrogen peroxide mediated oxidation of iodide ions to iodine, especially in the case when, e.g. suboptimal concentration of H₂O₂ is present or endogenous catalase decomposes the H₂O₂ in tissue as quickly as it is evolved. Subsequently, iodine is complexed by the starch to form a coloured product. H₂O₂ production by wound-induced oxidative burst or lignification can be easily discriminated due to the dual colour response.

Phosphorylase from tubers ofDioscorea dumentorum was fractionated by (NH₄)₂SO₄ precipitation and further purified to apparent homogeneity. Two forms of phosphorylase, Dd₁ and Dd₂ were obtained with different K_m, optimum temperatures and pH values. They obeyed Michaelis-Menten kinetics, they were activated by AMP, Mg²⁺ and Ca²⁺ but inhibited by ATP, ADP, ADP-glucose and Na₂SO₄. The molecular masses found for Dd₁ and Dd₂ were 102 000 and 195 000 respectively. SDS gel electrophoresis indicated that Dd₂ is a dimer and Dd₁ is a monomer. Both phosphorylase forms contained pyridoxal-5'-phosphate as their prosthetic group, which was essential for their activity. Dd₂ is more active in starch synthesis while Dd₁ is more active in starch degradation. The affinity of the enzyme forms for P_i is not a measure of the rate of glucan degradation. The decline in activities of the enzymes between pH 5.5 and 7.5 was due to unsuitable ionic forms of the enzymes and not due to irreversible denaturation. Both forms of phosphorylase showed double displacement reaction mechanism in direction of phosphorolysis, while in direction of synthesis, sequential mechanism was indicated. Indoleacetic acid activated the enzyme forms in direction of starch degradation but acted as an inhibitor in direction of glucan synthesis. The results of this study suggest that Dd₂ may have some synthetic function while Dd₁ has a degradative role. The effect of pH on the phosphorylase activity suggests histidine as the amino acid residue around the active site that might be involved in enzyme catalysis.

Chloris barbata Sw. andCyperus rotundus L. from a mercury contaminated site near a chloralkali plant were tested for relative tolerance to Hg by root-elongation method. The above two species from the Hg-contaminated site exhibited high tolerance to Hg compared to the same species from a non-contaminated site. Tolerance to Hg was higher inChloris barbata than inCyperus rotundus.

Fourteen days-old bean plants, grown on sand with Knop's nutrient solution were subjected to water stress (three days without irrigation). The stress led to a decrease in almost all lipid classes except phospholipids in the primary leaves. The content of palmitic acid increased, and that of the linolenic acid decreased. An increase of hexadecenoic acid in phospholipids was also observed. Rewatering for 24 h led to the recovery of the stressed plants including that of the photosynthetic apparatus, but the changes in the lipid composition were insignificant. The spraying of the plants before and after the water stress with 5 × 10-6 M solution of the phenylurea cytokinin 4-PU-30 alleviated negative effect of water stress on the lipid membrane composition permitting the plants to resist the harmful environment.

Aim of this work was to find the best source for obtaining high amount of copper amine oxidase (EC 1.4.3.6) that can be further used for analytical or industrial applications. The study focused on plant enzymes, because they occur in much higher content in the starting material than the enzymes from other sources, have higher specific activity and are also more thermostable. Presence of the amine oxidase was tested in extracts from 4 to 7-d-old seedlings of thirty-four various Fabaceae plants. Amine oxidases from nine selected plants were purified by general method involving ammonium sulfate fractionation, controlled heat denaturation, and three chromatographic steps. Kinetic properties of the amine oxidases purified were tested with a wide range of substrates and inhibitors and were found to be very similar. Best purification yield, and total and specific activities were obtained for the enzyme from grass pea (Lathyrus sativus) throughout all purification steps. Hence, the grass pea extract was chosen as a suitable candidate for massive production of the amine oxidase.