The dependence of luciferase activity in the homogenate of leaves of transgenic tobacco plants with chimeric firefly luciferase gene on ATP concentration and temperature was studied. The optimum ATP concentration was between 0.625 mM and 2.5 mM. The activity rapidly decreased if the homogenate was kept in 25°C and is completely lost during 30 min.

The rates of uptake of exogenous L[U-¹⁴C] aspartate and glutamate into tissues of vegetative growing tips ofFucus serratus and their metabolism were studied in the dark. In these non-photosynthetic conditions, aspartate was fixed and metabolically converted more rapidly than glutamate. Radioactivity from¹⁴C-aspartate was principally transferred into glutamate. On the other hand, metabolism of absorbed¹⁴C-glutamate was very slow and its rate did not increase during incubation time, but produced more diversified soluble radioactive compounds. Thus inF. serratus, glutamate principally seems to be in the dark more a temporary¹⁴CO₂ storage product coming from β-carboxylation than a rapidly turned over intermediate.

Water and K⁺ fluxes were examined in detopped plants ofZea mays L. (cv. White Horse Tooth), which were grown and exuded on half-strength Long Ashton nutrient solution containing the appropriate concentration of Zn²⁺ at 20 °C. In light-grown plants, 100 and 500 μM Zn²⁺ increased both water and K⁺ fluxes in detopped maize plants whereas 1 000 μM Zn²⁺ inhibited both fluxes. In the dark-pretreated plants, 1 000 μM Zn²⁺ in the medium stimulated K⁺ flux. The fluxes of K⁺, Zn²⁺, Ca²⁺ and Mg²⁺ were usually higher in detopped plants than in intact ones. At 1 000 μM Zn²⁺ in the exudation medium, Zn²⁺ concentration was higher in the xylem exudate of dark-pretreated plants than in roots of plants maintained in light. The results are discussed in relation to the influence of Zn²⁺ on the membrane permeability and transport in plants.

Following harvest, Kinnow mandarin (Citrus nobilis × Citrus deliciosa) fruits were variously treated with gibberellins (GA) and cytokinins. Ethylene caused marked chlorophyll (Chl) degradation and its effect was partially reversed by kinetin and benzylaminopurine (BAP) and to a lesser extent by GA₃ and GA₄₊₇. No appreciable accumulation of carotenoids (Car) occurred in these fruits irrespective of treatment. The loss of Chl during natural maturation was significantly reduced by cytokinins. Treatments with gibberellins alone or in combination with cytokinins were much less effective. The reduced loss of Chl in response to exogenously applied cytokinins may be probably related to a decline in its endogenous levels.

Chlorophyll (Chl) synthesis in isolated Scots pine embryos depended on exogenous application of cytokinin (CK) and Ca²⁺. At a constant benzyladenine (BA) level (4.4×10^-5 M) 10^-4 to 10^-2 M Ca²⁺ concentrations in mineral medium were optimum for Chl biosynthesis under both light and dark. At a zero or very low (10^-6 M) concentration of external Ca²⁺, Chl synthesis was relatively more Ca²⁺-dependent in embryos cultured in darkness than in the light, which suggested that the light: (a) stimulated the transport of Ca²⁺ from external sources to cytosol, and/or (b) interacted with Ca²⁺ directly in the pathway of Chl biosynthesis. The need of external Ca²⁺ was evidenced in experiments with modulators of Ca²⁺-transport systems. The reduction of the inward current of Ca²⁺ from readily accessible external sites by chelating agent (ethylene glycol-bis (beta-aminoethyl ether-N,N,N'N'-tetraacetic acid, EGTA) and Ca²⁺-channel blockers canceled the formation of Chl. The effect of EGTA depended on the level of external Ca²⁺. Inhibitory action of Ca²⁺-channel blockers depended on their kind and concentration: at the 10^-5 M concentration La³⁺>verapamil>nifedipine inhibited Chl formation. In the presence of Ca²⁺, the Ca²⁺-agonist A 23187 mimicked the BA effect and about 92% of Chl was synthesized as compared with the BA variant. Low concentrations of calmodulin antagonists reduced the amounts of Chl. Calmodulin was included in a second messenger system for BA action in promoting Chl biosynthesis in isolated Scots pine embryos.

The protein coding region of theE. coli DNA repair geneada combined with the CaMV 35S promoter has been transferred to tobacco by means ofAgrobacterium tumefaciens Ti plasmid. In transgenic plants having theada gene in a sense orientation, detectable amounts of O⁶-alkylguanine-DNA-alkyltransferase has been found whereas in non-transformed plants this activity is absent. Cell suspension cultures derived from the former plants showed lower sensitivity to the toxic (growth inhibiting) effects of the bifunctional alkylating agent 1-(2-chloroethyl)-1-nitroso-3-(aminomethyl-1,3-diazinylo)-methylurea compared with cell cultures derived from a control non-transformed plant or from transgenic plants harbouring theada gene in an opposite, non-sense orientation.

Increasing salinity induced a marked reduction in the plant growth, thoughPhaseolus seedlings tolerated salinity up to 120 mM NaCI. A great reduction in sugar and protein contents occurred with increasing salinity, whereas soluble nitrogen compounds and the relative contents of the photosynthetic pigments were increased in the treated plants. Increasing Ca concentration in the salinized medium appeared to improve the plant growth and to increase the contents of saccharides and proteins in the NaCl-treated plants. This suggests that Ca could be added to salinized media to overcome the deleterious effects of salinity on the growth and productivity of leguminous crop plants.

The antagonistic effects of some growth regulators [i.e. indol-3-yl-acetic acid (IAA), gibberellic acid (GA₃) or kinetin] on stress imposed by sea water on leaf area, pigment and photosynthetic activity in leaves of broad bean plants at different stages of development were investigated. Seed priming with GA₃ alleviated either partially or completely the effects induced by the two levels of sea water (10 and 25 %) used on leaf area at all experimental stages. However, IAA, GA₃ and kinetin inhibited leaf growth by themselves in almost all measurements. Seed pretreatment with kinetin alleviated the inhibition of pigment production in sea water-irrigated plants. Furthermore, GA₃ or kinetin nullified the deleterious effects imposed by irrigation with sea water particularly the high level (25 %) on photosynthetic¹⁴CO₂ fixation.

The fungal colonization of the angiosperm root parasiteCynomorium coccineum and the halophytic hostsLimonium delicatulum andArthrocnemum glaucum were investigated in a Mediterranean salt marsh in March 1992. The main fungal inhabitants on the leaves or shoot surface of the test plants wereAspergillus niger, Penicillium chrysogenum andCladosporium herbarum. The qualitative analysis of the fungal species associating the parasite, the hosts and the non-infected plants showed similar pattern. However, the total population exhibited quantitative differences coupled with the amount and the chemical composition of the exudates on plant surface and the quantity of transpired water. The fungal catch from the aerial shoot (inflorescence) of the parasite was higher than that collected from either the leaves or aerial shoots of non-infected or host plants. The fungal density on the leaves ofL. delicatulum was higher than those isolated from the aerial shoots ofA. glaucum. Infection byC. coccineum caused a marked drop in the total fungal population on leaves or shoot surfaces of the hosts as compared to the corresponding non-infected individuals. The stimulative effect of washings on spore germination of some isolated fungal species was matched with the density of fungi on the target plants.

Selected factors affecting somatic embryogenesis efficiency have been studied, namely genotype, explant type and its orientation in the medium, different basal media, different auxins for somatic embryo induction, and two ways of donor plant cultivation. The key role is played by genotype and auxin used, the minimum effect was observed due to basal media. In the series of subsequent experiments we have found the best combination of individual factors as follows: cv. Altona, 10 uM 2,4-D, L2 basal medium, central part of immature cotyledon as initial expiant oriented by adaxial side down on the agar medium, and field grown donor plants. This combination exhibited 100 % embryogenic explants with 5.43 ± 0.65 somatic embryos per expiant,i.e. somatic embryogenesis efficiency 5.43.

Multiple shoots (16-20 shoots per expiant) were induced from cotyledonary node region ofAnogeissus acuminata (Roxb. ex DC.) Guill. & Perr. on Murashige and Skoog's (MS) medium containing IAA 0.1 mg 1^-1 + BAP 1.5 mg 1^-1 and ascorbic acid 50 mg 1^-1, citric acid 25.0 mg 1^-1, arginine 25 mg 1^-1 and adenine sulphate 25 mg 1^-1. From the first node of seedling only 4-6 shoots per expiant were proliferated. Segments ofin vitro produced shoots were used as expiants for further multiplication of shoots upto 16 successive cultures at an interval of 4 week on MS medium with IAA 0.1 mg 1^-1 + BAP 1.0 mg 1^-1 and additives. The original cotyledonary expiant was repeatedly subcultured upto 4 times after harvesting crop of shoots, each time.In vitro produced shoots were rooted on half strength MS medium containing 0.5 mg 1^-1 IBA. Plantlets were transferred to pots. Other expiants (cotyledons, hypocotyl, and leaf) produced callus on medium containing auxins and cytokinins. The calluses differentiated into embryo like structures or roots on MS medium.

Floral induction in the long day plant spinach (Spinacia oleracea) has been shown to be accompanied by a thickening of plasmalemma. This change was observed at early evocation, in both shoot apices and leaves, as well as after inducing GA₃ treatment. To get further information on this thickening, plasma membranes from spinach leaves were isolated, in the present study, using aqueous two phase partitioning and the effect of variousin vitro treatments on their thickness was investigated. The average plasmalemma thickness was unaffected by Na⁺ and K⁺ ions. It was increased upon the effect of either Ca²⁺ or gibberellic acid. A thickening of plasmalemma was also observed when plasma membranes from vegetative plants were incubated with a cytosolic preparation from photoinduced plants. The results were discussed in relation with the plasmalemma modifications previously reported in spinach.

The effect of salinity on vacuolar pH was studied in carrot (Daucus carota L.) cells grown in liquid suspension culture either in the absence or presence of 150 mM NaCl. Both vacuolar and cytoplasmic pH were determined by several independent techniques. These techniques were NMR spectrometry, distribution of radioactive probes and spectrophotometric measurement of the absorbance changes of a naturally occurring vacuolar pH indicator. There was no difference in the cytoplasmic pH between cells grown in the presence or the absence of NaCl, but the vacuolar pH of cells grown in the presence of NaCl was higher by 0.38 to 1.05 pH units (depending on the technique that was used) than the vacuolar pH of cells grown in the absence of NaCl.

Ten major host-encoded pathogenesis-associated proteins have been found in extracts of intercellular sap of cucumber cotyledons infected with tobacco necrosis virus. By native and two-dimensional SDS-polyacrylamide gel electrophoresis, five major acidic virus-elicited proteins have been identified and classified on the basis of their molecular mass in three groups: group 1 included one protein of 14 kD; group 2, one protein of 28 kD and group 3, three proteins of 37-40 kD. Another five viruselicited proteins were basis, and were again classified in three groups: group 1, included two proteins of 15-16 kD; group 2, two proteins of 22-23 kD; group 3, one protein of 40 kD.

Several factors influencing reliability of the quantitative fluorimetric β-glucuronidase (GUS) assay in transgenic plant tissue have been investigated. We obtained linear dependence of fluorescence on both the duration of hydrolysis and the extract concentration. The stability of the enzyme in the homogenate was fairly high, the same as the stability of the substrate solution and of the final reaction product. The modification of the extraction/incubation buffer was proposed, resulting in several times higher activity in comparison with original procedure.

Transport of ions in young maize plants was affected by the temperature gradient (gradT). This influence was dependent on the age of plants and the solute concentrations in compartments at the two sides of plants. When gradT was increased, current, volume and heat flows rose during 20 min and then declined. On the other hand, the diffusive flow only declined. The character of the flow changes was given by the changes in permeability and reflection coefficients. In young plants under low solute concentration on both sides of plants, the permeability of the plant tissues shifted from positive to negative values. Under higher solute concentrations on both sides of the plants such phenomenon was not observed. At high gradT the reflection coefficients of the plant tissues were low, but they rose during levelling the temperature in the two compartments.

Vanadium bioaccumulation calculated as the ratio of its content in plant biomass to that in the substrate (Vbi-index) was studied in pea. Vbi was on average 11.275, 11.770 and 13.153 in the roots, and 0.809, 0.467 and 0.749 in the shoots of the cultivars Opal, Laser and Ramir, respectively. This indicates cultivar differences in vanadium uptake, and low translocation rates from roots to shoots. Vanadium (3 to 30 mg 1^-1) decreased shoot and root fresh and dry masses of the three cultivars. Seedlings of the cv. Opal were the most susceptible to higher concentrations of vanadium (20 to 30 mg 1^-1), whereas seedlings of cv. Laser were the most resistant.

NaCl salinity affected growth, the contents of chlorophyll, carotenoids, saccharides and some minerals (Na, K, Ca, Mg, P) in wheat (Triticum vulgaris L.) plants. Irrigation of soil with CaCl₂ or KCl greatly ameliorated the adverse effects of NaCl salinity. This counteraction was associated with an increase in contents of saccharides, proteins and Ca, Mg and P which might explain their role in osmotic adjustment.

Effects of benzothiazolium salts on chlorophyll (Chl) production inChlorella vulgaris (inhibition or stimulation) are closely connected with the applied effector concentration and with the character of the substituents.

Vitrification of stem explants of carnation was brought about by culturing in liquid medium. Cellulose and lignin levels were decreased in vitrified stems and leaves. Isolated cell walls of vitrified tissues were also characterized by low calcium content, low Ca²⁺/uronic acids ratio, low ratio of uronic acids to neutral sugars due to higher amounts of the latters. All these characteristics may account for the high wall plastic potential previously measured in vitrifying internodes.

Possibilities of adventitious buds induction on the cotyledons obtained from sterile seedlings ofAbies concolor xAbies grandis hybrid were investigated. The following variables influencing bud induction and their further development were studied: the effect of expiant age, the effect of different growth regulators and their concentrations and duration of their application. The most suitable expiants proved to be the cotyledons of 7 d old seedlings. The most efficient cytokinin was benzylaminopurine (S mg l^-1) in combination with napthaleneacetic acid (0.01 mg l^-1). The most optimal duration of treatment was 17 to 21 d culture of explants on induction medium. Shoot growth was achieved on basal medium to which 14 mg 1^-1 spermidine was added.