The suitability of the fluorescent dye rhodamine 123 for qualitative and quantitative determinations of the electrical potential difference (ΔΨ) in isolated pea (Pisum sativum L.) stem mitochondria was evaluated. A fluorescence quenching of rhodamine 123, as a consequence of dye uptake, occurred following mitochondria energization by both external and internal substrates. This quenching was associated to the generation of ΔΨ, because it was completely released by uncouplers and respiratory inhibitors. The conversion of the proton gradient (ΔpH) into ΔΨ, induced by nigericin or a permeant weak acid (phosphate), increased the quenching. The uptake of the probe was accompanied by 40 % of unspecific binding in coupled, but not in uncoupled, mitochondria. Rhodamine 123 quenching varied linearly with a K+-diffusion potential. ADP induced a transient and cyclic change of fluorescence which was associated to ATP synthesis. Consequently, rhodamine 123 did not influence oxygen consumption by mitochondria in both state 4 and 3, thus indicating that, at the concentrations assayed, the probe was not toxic. It is concluded that rhodamine 123, followed by fluorescence quenching, is a suitable probe to study the energetics of isolated plant mitochondria.

Light regulation of diurnal variation of sucrose-phosphate synthase (SPS) activity was studied in potato leaves during pre-tuber initiation stage. SPS activity was greatest at 14:00 and coincident with highest irradiance [2 000 µmol(PAR) m-2 s-1], and lowest at 18:00 during lowest irradiance (200 µmol m-2 s-1). In contrast, fructose-1,6-bisphosphatase (FBPase) activity was greatest at 12:00 (irradiance 1 700 µmol m-2 s-1). Inhibition of SPS activity by phosphorylation was least at 14:00 and highest at 18:00. Considerable inhibition was also observed at 10:00. SPS showed hyperbolic saturation kinetics in response to varying UDP-glucose concentrations. Diurnal variation of SPS activity was due to a change of affinity for its substrate (Km), but not due to change of Vmax. Phosphate (Pi) decreased the activation state of SPS. Inhibition by Pi was most severe under limiting UDP-glucose concentration. Sensitivity towards Pi inhibition at limiting substrate concentration was highest at 18:00 and lowest at 14:00. Change of tissue phosphate concentration followed the opposite pattern to that of diurnal variation of SPS activity and irradiance, being highest at 18:00 and lowest at 14:00. A dual mode of regulation is considered to exist for diurnal variation of potato leaf SPS: covalent modification and effector (Pi) regulation. Thus the change of tissue phosphate concentration may play a pivotal role in regulating potato leaf SPS.

Germination of two rice (Oryza sativa L.) cultivars (Ratna and IR 36) in the presence of 10 µM PbCl2 and 10 µM HgCl2 decreased germination percentage, germination index, shoot/root length, tolerance index and dry mass of shoots and roots. Mercury was more toxic than lead. Reduced glutathione (GSH), cysteine (Cys), ascorbic acid, and α-tocopherol alleviated the adverse effects of these metals on plants in the order GSH > Cys > ascorbic acid > α-tocopherol. The effects were more pronounced in tolerant cultivar IR 36 than in the relatively susceptible cultivar Ratna.

In vitro response of six tomato genotypes to different copper concentrations was studied. Cu was toxic to tomato explants at a relatively high concentration (100 µM), which reduced callus growth and shoot regeneration. Peto-86 followed by UC-97-3 were more tolerant to copper than the other genotypes. Cu (100 µM) induced the synthesis of eight new proteins (70.86 - 14.78 kD) in Peto-86 and six in Western Improve (46.43 - 14.78 kD) and UC-97-3 (77.69 - 14.78 kD). Cu-stress reduced the expression of some enzymatic bands of alcohol dehydrogenase and esterase, meanwhile, one peroxidase band at the locus Prx-1 was newly expressed under Cu-treatment.

The effects of cadmium and copper on in vitro growth of Bacopa monniera (L.) Wettst. was monitored. Cadmium (25 and 50 µM CdCl₂) inhibited plantlet growth and addition of 50 or 100 µm CuSO₄ partially alleviated this negative effect. Cadmium increased both protein and proline contents, but to a lesser extent with the additional supply of CuSO₄.

The activities of nitrate reductase and glutamine synthetase in leaves of greenhouse grown rose plants (Rosa hybrida cvs. Ilseta and Mercedes) grafted on various rootstocks were compared with those in leaves of non-grafted, own-root plants of these cultivars. The results obtained showed that the enzymatic activities as well as nitrate content in the leaves were altered by the grafting and by type of the rootstock used. These rootstock-imposed alterations differed between the two cultivars used in the study.

Direct differentiation of shoot buds from hypocotyl segments of 12-d-old seedlings of Tamarindus indica was obtained on Murashige and Skoog (MS) medium with or without growth regulators. The highest regeneration (66 %) and the maximum number of shoots (3 - 4) per explant were obtained from the explants on MS medium containing 6-benzylaminopurine (5 × 10-6 M). A maximum roots per shoot were produced on medium containing 3-indole butyric acid (5 × 10-6 M). The resulting plantlets were hardened and transferred to soil in pots where 75 % of them survived and resumed growth. Histological examination of explants suggests that the shoots were of de novo origin which would make this system suitable for transformation experiments.

It is a matter of controversy whether secondary wall deposition is dependent on lignification during the development of tracheary elements. To understand this, tracheary element differentiation was studied in the homogeneous calli obtained from the cotyledonary explants of Cucumis sativus subsequent to treatment with plant growth regulators, such as naphthalene acetic acid (NAA) and benzylamino purine (BAP), which are necessary for the induction of tracheary elements, along with metabolic blockers such as 2-aminoindan-2-phosphonic acid (AIP), 2,3,5-triiodobenzoic acid (TIBA) and nifedipine. Calli treated with AIP, a potential inhibitor of L-phenylalanine ammonia-lyase (PAL), have no PAL activity at any time during the culture period. There was a complete inhibition of lignification although secondary wall deposition was unaltered. Similar results were obtained using TIBA, an inhibitor of auxin transport, and nifedipine, a known calcium channel blocker. Thus the present study suggests that secondary wall deposition in the course of tracheary element differentiation need not to be dependent on lignification.

The effects of gibberellic acid (GA3) and N1-(2-chloro-4-pyridyl)-N2 phenylurea (4-PU-30) on maize seedling growth, photosynthetic parameters, and leaf protein composition were investigated. The agents used alone or in combination increased leaf growth and photosynthetic rate of the seedlings. Chlorophyll and total nitrogen contents in leaves as well as the quantity of individual protein fractions increased simultaneously. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of soluble proteins (albumins and globulins) revealed quantitative differences between 4-PU-30-treated plants and the other experimental variants. They differed in polypeptide composition associated with changes in soluble proteins and amino acids. However, GA3 did not induce similar changes in polypeptide composition of soluble proteins.

Seedlings of two common Mediterranean trees, Pinus halepensis L. and Quercus ilex L., were grown alone and together with seedlings of the same or of the other species in the same pot during one year to test the effects of intra- and inter-specific interference on terpene emission. Light, nutrients and water were amply supplied. There were higher emission rates in P. halepensis than in Q. ilex. The emission increased when the neighbour was a pine and decreased when the neighbour was a holm oak. Volatile organic compound and terpene emission rates followed inverse trends to foliar biomass or growth, which decreased when the neighbour was a pine.

The decrease in inorganic phosphate concentration in bean (Phaseolus vulgaris L. cv. Złota Saxa) roots induced decrease in respiration rate. The decrease observed in ATP pool in phosphate deficient (-P) roots was greater than it would result from the decline in respiration and possible involvement of alternative pathway, suggesting an increased energy utilization for growth and ion uptake. Indeed, relative growth rate was higher in -P plants until 12 d of culture and later dropped to the rate similar to the control. Net nitrate uptake rate was higher in -P plants than in +P plants at the beginning of phosphate starvation, then during the prolonged culture it decreased rapidly in -P plants and after 19 d it was 8 times lower than that in the control. The decline in ATP production during prolonged phosphate starvation influenced NO₃ ^- uptake more than root growth.

A simple and reliable method for extracting DNA has been developed for orchid species and hybrids. The high quality of DNA obtained is suitable for amplification via the polymerase chain reaction (PCR) for producing random amplified polymorphic DNA (RAPD) markers.

Solar UV-B radiation increased to 20 % over ambient level at Madurai was given to cowpea (Vigna unguiculata L. cv. Pusa-152) seedlings sufficiently supplied by potassium (0.88 mM K₂SO₄) and K-deficient (0.05 mM K₂SO₄). Leaf mineral composition was significantly changed due to both increased UV-B radiation and K-deficiency imposed independently or jointly for 12 d. A severe reduction in 23 kDa chloroplast protein was seen only in seedlings encountered combined stress.

Uptake mechanisms for neutral amino acids were investigated by expression of mRNA isolated from seedlings of Ricinus communis L. in Xenopus laevis oocytes. After injection of mRNA from root, hypocotyl and cotyledon currents elicited by saccharose and neutral amino acids ranged from 0.3 nA up to 2 nA depending on the respective substrate and the source of mRNA. These currents were due to expression of low affinity uptake mechanisms and the KM values found for amino acid induced charge flow range from 1 to 2 mM. The abundance and/or the specificity of the expressed mechanisms differ in the various tissues. Currents of similar magnitude were recorded for alanine and glutamine with mRNA isolated from root, hypocotyl and cotyledons. Serine and proline induced currents after injection of mRNA from hypocotyl and roots, in case of α-aminoisobutyric acid (AIB) induced currents were generally small with mRNA from all tissues tested. In addition, differential sensitivity of glutamine and AIB uptake in the high affinity range was evident towards the amino acid analogue 2-chloro-aminophenoxybutyric acid which indicated an additional set of carriers operating in the micromolar concentration range. The results suggest that multiple transporters for neutral amino acids exist in various tissues of the plant differing in specificity of charge flow and in sensitivity towards the inhibitor 2-chloro-aminophenoxybutyric acid.

Plants of Calamagrostis villosa were cultivated in nutrient solution alone or in association with a vesicular-arbuscular mycorrhizal (VAM) fungus Glomus etunicatum. They were supplied with two levels of inorganic phosphate (Pi; 0.1 or 1 mM) and with or without organic phosphate (dinatriumphenylphosphate, Po; 1 mM). Depression of growth and enhancement of root respiration of mycorrhizal plants in comparison with non-mycorrhizal plants were observed after 12 weeks of cultivation in a growth cabinet. Root colonization was not influenced by the higher phosphorus availability in contrast to the extraradical mycelium (ERM). The lengths of ERM hyphae both attached to the root surface and in the substrate were decreased substantially by higher phosphorus supply, irrespectively of its form.

Alfalfa (Medicago sativa L.) cell suspension cultures of Verticillium albo-atrum resistant and susceptible genotypes were established from leaf callus tissues. Treatment of cultures with conidia and heat-released elicitors of V. albo-atrum induced a large increase in the activity of phenylalanine ammonia-lyase, only in the cells of the resistant genotypes with a maximum after 12 h. In co-cultivation with the fungal conidia and resistant cell lines, the production of spores were inhibited.

The immature zygotic embryos of reciprocal maize hybrids (CHI-31 x GF1 and CHI-31 × GE2) were used as the initial material for induction of somatic embryogenesis in vitro. Histological analysis of somatic embryogenesis revealed high developmental variability. The arising formations were classified into 5 groups: A) somatic embryos phenotypically similar to zygotic embryos, B) polyembryos, C) formations with radicle but without meristematic plumule, D) formations with radicle without differentiated plumule, and E) formations with plumule without radicle. The formatioms A and B regenerated directly into plants. Plant regeneration from formations E required preculture on the rooting medium. Formations C and D failed to develope into plants possibly because of early loss of meristematic cell character during the embryo axis differentiation. The reverse sequence of radicle and plumule differentiation in somatic embryos in comparison with zygotic ones was noted. The epigenetic character of the scutellum, coleoptile, coleorhiza and leaves primordia development was discussed.

Sugar beet was grown in pots and the transport of 6-deoxy-d-glucose (6-dg) and of 2-aminoisobutyric acid (AIB) into leaf and root tissue segments was examined. The uptake of 6-dg into both tissues displayed two kinetic components, one apparently of diffusional nature, the other saturable (half-saturation constant 0.22 mM in leaves, 0.10 mM in roots; limiting rate 0.37 and 0.10 nmol mg-1 (fresh mass) h-1 for leaves and roots, respectively). It was suppressed by d-glucose and inhibited by diethyl-stilbestrol (DES), suloctidil (SUL) and 2,4-dinitrophenol (DNP), but little by D2O and not at all by sodium vanadate. It showed a temperature optimum at 30 °C and a pH optimum at 6.0 - 6.5 in both tissues. The uptake of AIB was also two-componental but even the specific component had an immeasurably high half-saturation constant although it was inhibited by high concentrations of AIB as well as by DES, SUL, DNP and D2O. However, vanadate stimulated the uptake by as much as 40% in both leaves and roots. The temperature optimum in leaves lay at 30 °C while in the roots no maximum was present; its pH optimum was at 5.5 in leaves and again no maximum was observed in roots. Both transport systems were unaffected by 0.1M NaCl or 0.1 M KCl and by the presence of cytokinin in the incubation vessel. However, spraying with the cytokinin N6-(m-hydroxybenzyl) adenosine decreased both the half-saturation constant and the limiting velocity of 6-dg uptake slightly in the leaves and substantially in the roots. Spraying had an insignificant effect on the uptake of AIB.

Age-induced changes in 1) nocturnal and diurnal acidity fluctuations that coincide with the ongoing environmental conditions, 2) the build up of abscisic acid (ABA) in plant roots and leaves during sunrise, midday, and sunset in all growing stages, 3) the changes in phosphoenolpyruvate carboxylase (PEPC) and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) activities as key enzymes of the photosynthetic pathways of C3 and CAM, 4) leaf water potential (ψ1), and 5) Km and Vmax for PEPC to express its activity and affinity, were studied in Mesembryanthemum nodiflorum during transition from C3 to CAM mode of CO2 fixation. The acidity during sunset in mature stage was higher than in earlier stages and reflected the impact of environmental conditions on physiological and metabolic changes. Moreover, the higher acidity during sunrise and sunset was observed during the senescence than the mature stage; this might be due to CO2 release and oxygen intake during senescence induced ethylene formation that lead to increased malic acid formation. The ABA concentration was high in M. nodiflorum leaves, but stomatal closure was insensitive to elevated ABA concentrations recorded. Vmax of PEPC, Km, and the affinity of PEPC during later stages indicated the ability of PEPC to fix CO2 taking up at night in CAM cycle of M. nodiflorum. Less affinity during sunrise indicated inhibitory effect of malate on PEPC during the release of CO2. The second peak of PEPC activity before sunset caused CO2 fixation. The RuBPCO was inactive at night. Slight increase in ABA during sunset, and night drop in air temperature and increase in relative humidity reduced markedly transpiration rate without decreasing ψ1.