Hypocotyls, cotyledons and true leaves of in vitro seedlings of 10 cucumber and melon genotypes resistant to downy and powdery mildew were cultured on several combinations of initiation and multiplication media to produce callus and subsequently cell suspensions as suitable sources for isolation and culture of protoplasts. Cotyledons of both species were shown to be the most responsive to variation in culture media. However, calli and cell suspensions derived from hypocotyls generally provided higher number of protoplasts by treatment with several enzymatic solutions. The protoplasts formed new cell walls after 12 h of culture in liquid culture medium and first cell division was observed 2 d later with more frequent divisions after one week of culture.

Plants of maize (Zea mays L.) were waterlogged for 7 d and irrigated (root application) or sprayed (foliar application) with 0, 1, 3 and 6 mg dm-3 of boron. The stability of leaf membranes was assessed by determining leakage of electrolytes from leaf discs exposed to heat (51 °C) and dehydration (40 % PEG). Leaf membranes were more stable to heat than to dehydration. The membranes of waterlogged plants were more injured than those of control plants. Waterlogging reduced contents of dry mass, chlorophyll, soluble proteins, total free amino acids and soluble sugars and leaf relative water content (RWC). Application of boron increased the stability of leaf membranes, chlorophyll, soluble sugars, soluble proteins, amino acids contents, leaf RWC and dry mass accumulation. Foliar boron application was more effective. Application of boron alleviated the deleterious effect of waterlogging.

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.

The expression of genes for synthesis of auxin (iaaM and iaaH) and cytokinins (ipt) was studied in tobacco plants transformed by two Agrobacterium tumefaciens strains C 58 and LBA 4404. The strain LBA 4404 carried binary vector plasmid pCB 1334 (ipt gene) and plasmid pCB 1349 (iaaM, iaaH and ila genes). Both plasmids carried reportered gene for npt II. Obtained plants expressed incorporated genes. New proteins with molecular masses of about 74, 40, 26, 25, 21 and 17 kDa for wild plasmid pTi C58; 60, 36, 31.5, 27, 26 and 17 kDa for binary vector plasmid pCB 1334 and 74, 49, 36, 31.5, 26 and 25 kDa for binary vector plasmid pCB 1349 were found in the patterns of soluble proteins. Significant changes in the content of chlorophylls, especially chlorophyll a, were detected in the plants carrying ipt gene and in plants transformed by the wild strain C58 of A. tumefaciens. Tobacco plants expressing ipt gene and genes from T-DNA of pTi C58 plasmid were dwarf, and in comparison to the controls, they had thicker stems, and the surface of the leaf blades was reduced to 20 - 50 %. Adventitious roots, growing from the stem, were typical for transformants overproducing auxins. Regenerants and transformants expressing genes from T-DNA of plasmid pTi C58 differed in the shape of the flowers and their fertility.

Effect of fluoride (10 and 50 mM) on the activities of sucrose metabolizing enzymes, alkaline inorganic pyrophosphatase, and transaminases in relation to the accumulation of free sugars, starch, and soluble protein was studied in detached ears of wheat cultured in a liquid medium. Culturing for 5 d in the presence of fluoride reduced the amount of grain starch whereas contents of total free sugars, particularly sucrose, and soluble protein increased. Fluoride inhibited the activities of soluble acid and neutral invertases, as well as sucrose synthase acting in the cleavage direction. Uptake of uniformly labelled ¹⁴C-sucrose or fructose was also drastically reduced by fluoride. Glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) activities also increased with fluoride addition in correspondence with an increase in soluble protein. Apparently, the wheat grain responds to fluoride-mediated disruption of carbon metabolism by a compensatory effect on nitrogen metabolism.

The influence of the applied photoperiod during floral stem development has been studied. A clear relationship between the length of the photoperiod, the frequency of light/dark switches, and the dynamic of growth and the subsequent morphology and architecture has been found. An increase in the day length (L/D = 16/8) or an increase in frequency of light/dark transitions (L/D = 7/7) stimulated the inflorescence growth. The two treatments induced the same enlargement in the early inflorescence, but the L/D = 7/7 condition causes a greater increase in the late one. The treatments induced also a diminution in the number of lateral inflorescences, principally with the high frequency light-on/light-off condition. Moreover than differences in the growth along the early inflorescence, at the level of the internode, a gradual growth was observed from the basal part to the upper part of the internode. Finally, the floral stem growth dynamic showed that it could be adapted to the environmental light/dark signalling by a good extension rate synchronisation.

Changes in ribonucleases (RNases) and glucose-6-phosphate dehydrogenase (G6P DH) activities, their content and subcellular localisation were studied in relation to virus multiplication in susceptible (cv. Samsun) or resistant (transgenic breeding line NCTG 83) tobacco plants infected with the potato virus Y^N (necrotic strain of PVY). Activities of RNases and G6P DH from diseased susceptible tobacco plants were markedly increased during the experimental period and significantly correlated with the multiplication curve of the PVY^N. In contrast, the activities of RNases and G6P DH were not changed after PVY inoculation of resistant breeding line NCTG 83 producing the CP mRNA of PVY. Changes in the content and in the subcellular localisation of RNases and G6P DH isozymes were also determined in mesophyll protoplasts isolated from healthy as well as PVY^N infected plants of both cultivars by differential centrifugation of broken protoplasts on day eight post inoculation (the culmination of multiplication curve of PVY and enhanced activity of both enzymes). The chloroplasts fraction from infected protoplasts showed an enhanced content of RNases (192.4% when compared with that from healthy control ones), and of G6P DH (174.4 %). The cytosol fraction from infected protoplasts contained slightly enhanced levels of G6P DH (117.4 %) and considerably enhanced levels of RNases (141.7 %). No significant differences in the activities, contents and subcellular localisation of RNases and/or G6P DH isozymes were observed in the resistant line NCTG 83. This is in accordance with no detectable contents of PVY.