We used chlorotetracycline fluorescence, alizarin staining and potassium pyroantimonate methods, as well as X-ray microanalysis, to demonstrate the differential localization of Ca2+ during pollen maturation. Level of loosely-bound Ca2+ ions was higher in the generative cell than in the vegetative cell of the mature pollen grain which is one of the symptoms of metabolic differentiation of the two sister cells.

Culture conditions for a fine dispersion of plated cells of Oryza sativa L. cv. IR 20, have been worked out. These plated cells developed microcalli containing large number of somatic embryos and subsequently plantlets. By using single cells and clusters of 2 - 4 cells, an efficient DNA-delivery by microprojectile bombardment into cells and its transient expression were assessed by employing a plasmid construct containing β-glucuronidase gene.

A concentration dependent decrease in root nitrate reductase (E.C.1.6.6.1) activity (NRA) by 0.1 to 2.0 mM lead acetate was noticed in three cultivars of Vigna radiata (L.) Wilczek (K851, MH8320 and ML337). Leaf NRA, on the other hand, increased significantly with increasing lead concentration which was more pronounced in cvs. MH8320 and ML337 than in cv. K851. Total nitrogen content of root and shoot was generally increased due to supply of the lead acetate, whereas the total nitrogen content of the colyledons was hardly affected in cvs. MH8320 and ML337 and decreased in cv. K851 during the early growth phase. The inhibition of root NRA could be alleviated by addition of inorganic salts (K2HPO4 and KNO3, but not CaCl2) or 6-benzylaminopurine (BAP) in the incubation medium. Lead mediated inhibition of root NRA was similar in light and dark grown seedlings, but lead induced increase in leaf NRA was more pronounced in the light than in the dark.

Media from embryogenic and non-embryogenic cell suspension cultures were analysed for protein content, electrophoretic protein patterns, glycoproteins and activity of peroxidases and β-glucosidases in order to characterize the physiological status of the cultures. On a dry mass basis the amount of extracellular proteins per cell was greater in embryogenic suspensions than in non-embryogenic suspensions. Non-embryogenic suspensions contained unidentified slimy compounds which were not present inembryogenic cultures. The extracellular Concanavalin A-specific glycoproteins gave different isoelectric focussing patterns and thus enabled embryogenic and non-embryogenic cultures to be differentiated. The extracellular peroxidase activity per cell dry mass was far greater in embryogenic than in non-embryogenic cultures. The isoenzymes differed in number and composition of the anionic bands. β-glucosidases were found in the same range of activity in both culture types, but the time course of enzyme activity during cultivation was significantly different. In the embryogenic culture the activity was correlated with dry mass increase, whereas in the non-embryogenic suspension the activity reached maximum during the linear growth phase. Polyphenoloxidase which was recently recognized as an intracellular marker for embryogenic stages was not released into culture media.

The ontogenetic changes in source-sink relation associated with the manipulation of reproductive sink at different positions on the plant in two okra cultivars (Arka Anamika and Pusa Makhamali) with distinct branching habit (cv. Arka Anamika tends to branch at the middle nodes unlike cv. Pusa Makhamali) were analysed. The cultivar differences for extension growth were nonsignificant except at treatment where all flowers and flower buds upto 8th mode were removed. The reproductive sink reductions resulted in a decrease of total dry matter accumulation per plant to an extent of 13 to 46% in Arka Anamika and 18 to 34 % in Pusa Makhamali. Fruits on the middle nodes appear to be prominent sinks for photo assimilates in cv. Arka Anamika. The reproductive sink manipulation did not result in any particular change in the trend of photosynthesis, but brought about a significant change in the partitioning of dry matter.

Shoot regenerations from hypocotyls and cotyledons of tomato(Lycopersicon esculentum Mill.) was inhibited by NaCl (100 and 150 mM).Shoot fresh and dry masses were also reduced. Addition of proline (100 mgdm-3) counteracted the inhibitory effect of NaCl. SDS-PAGE analyses of extracted proteins, revealed that in cultures grown in medium with 25 mM NaCl plus proline, extra polypeptides of Mr 190, 58, 45 and 26 kDa accumulated. As NaCl was increased in the medium a new protein of Mr 67 kDa also accumulated.

Source-sink relationship, which was influenced by both genotype and environmental factors, contributed to the variation in photosynthesis and photosynthate partitioning of wheat. Source reduction by partial defoliation increased leaf net photosynthetic rate (PN), and sink reduction decreased PN of irrigated wheat. However, the change in PN varied among genotypes. Source reduction enhanced photosynthate translocation into grain in irrigated wheat. However, the enhancement was more evident in cv. Lumai 215953 than incv. Lumai 15. Sink reduction had little effect on the translocation of photosynthate into grain in cv. Lumai 15, but decreased the translocation of photosynthate into grain and increased it into stem in cv. Lumai 215953. In rainfed, non-irrigated wheat, the source or sink manipulation influenced PN only slightly. The source reduction decreased the partitioning of photosynthates into the upper parts (including grains) of plant. However, very little effects of sink reduction on the production of photosynthates occurred in rainfed wheat. This showed that grain sink size was not a factor limiting the production of photosynthates, but controlled the partitioning of photosynthates. Sink reduction decreased photosynthate translocation into grains, and increased it into upper parts of rainfed wheat plant.

Friable calli were induced from mature excised shoots of Bambusa vulgaris on Murashige and Skoog's (MS) medium supplemented with 2.2 µM6-benzylamino-purine (BAP), 9.04 µM 2,4-dichlorophenoxyacetic acid and 14.76 µM indole-3-butyric acid (IBA) with 3 % (m/v) saccharose. Adventitious shoots with root hairs were achieved from calli on MS medium supplemented with 13.33 µM BAP and 1.23 - 2.46 µM IBA within 4 weeks of subculture. The frequency of shoot bud regeneration was better in the light incubated cultures than in the dark incubated cultures. Isolated shoots were rooted on liquid half-strength MS basal medium supplemented with 0.49 µM IBA and 2 % (m/v) saccharose. Histological observations confirmed the regeneration of shoot buds from calli. The rooted plantlets were successfully transferred to greenhouse.

A method for DNA extraction from Pyrrosia piloselloides (L.) Price and Pyrrosia longifolia (Burm.) Morton utilising hexadecyltrimethylammonium bromide is described. The genomic DNA isolated was found to be relatively pure and shown to be suitable for RAPD analysis.

The effect of five Azotobacter chroococcum strains and nitrogen content in nutrient media on callus growth of two Beta vulgaris L. cultivars were investigated, as well as the activity of nitrate reductase (NR), glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in inoculated callus tissue. On medium with full nitrogen content (1 N) the inoculation with A. chroococcum strain A2 resulted in the highest calli mass, while strains A8 and A14 maximally increased NR activity. On media with 1/8 N the highest effect on calli growth, GS and GDH activity had the strain A8. The strain A2/1 significantly increased callus proliferation on medium without N. Asymbiotic association between sugar beet calli and Azotobacter depended on genotype/strain interaction and was realised in presence of different nitrogen levels.

A floating sterile mutant of Ulva pertusa, which grows faster than the wild type and also during summer periods, was grown in the laboratory under "white light" (WR as reference), broad band isoquantic red (600 - 700 nm, RR) and blue (400 - 500 nm, BR) radiation. The observed specific growth rates at WR, BR, and RR were 8.6, 2.15, and 1.2 %, respectively. A stimulatory effect of BR on the growth at low irradiance (60 μmol m-2 s-1) was observed during the 15 d of culture. Around 42.1 % of total nitrogen was used by Ulva mutant in WR, but under BR and RR it was around 27.0 and 16.7 %, respectively. However, the utilization of nitrogen under BR was significantly higher than under RR. Considerable CO2 fluctuation in the light and dark periods was observed in all the cultures, and it was higher under WR, followed by BR and RR. The possible growth promoting effect of BR includes nitrogen uptake and the consumption of HCO3- which in turn leads to reduced air CO2 concentration in the medium.

The role of proline in imparting tolerance to salinity was investigated in Hydrilla verticillata, Najas indica and Najas gramenia. The plants were exposed to different concentrations of NaCl and artificial sea water (SWS) separately. The chlorophyll (Chl) a/b ratio decreased significantly in all the three plant species in both NaCl and SWS treatments, comparatively more in former than the latter. NaCl resulted in drastic decrease in this ratio in salt sensitive H. verticillata and N. indica, but in somewhat lesser decrease in salt resistant N. gramenia. Proline content increased at both NaCl and SWS treatments, especially at the latter. However, in H. verticillata proline content at 1.5 and 2.5 % NaCl decreased. It was concluded that proline cannot be used as a biochemical marker of salt tolerance in aquatic plants, however, the decrease in Chl a/b ratio in response to NaCl may be used as an index of salt sensitivity in this ecological group of plants.

Expression of the β-glucuronidase (GUS) reporter gene driven by the CaMV 35S, rolC, nos and mas promoters was assessed in the tips of 12 independent clones of transgenic sugar beet (Beta vulgaris) roots. Three questions were addressed: 1) expression pattern specific for a given promoter, 2) expression pattern variability, and 3) relationship between gene expression and cell differentiation. Characteristic patterns of tissue-specific expression were distinguished for each promoter. Striking differences, however, were found between some clones, bearing the same construct. Statistical analysis of the pattern variability proved that the variability is significantly lower within the construct than between constructs. rolC-GUS clones exhibited the lowest and CaMV 35S clones the highest pattern variability. Comparisons between the four promoters showed consistent GUS activity in areas playing a key role in tissue determination (the elongation zone) where cells switch from frequent mitosis and mostly isodiametrical growth, typical for the promeristem, to rapid elongation and differentiation. All of the clones were highly GUS-positive in the elongation zone of stele. Activity was commonly localised in the stele of the maturation zone for CaMV 35S, rolC and mas-GUS clones. CaMV 35S-GUS clones were highly active in the promeristem.

Transfer of chloroplasts from Solanum tuberosum into S. nigrum cell resulted in an atrazine sensitive cybrid plant. The shoots of this cybrid were bleached under atrazine stress. The cybrid displayed identical isoenzyme patterns that have been found in S. nigrum, and thus nuclei of the cybrid plant cells did not integrate any chromosomes or chromosome fragments from S. tuberosum nuclei. Under atrazine stress, differences in the isoenzyme expression were found in both the cybrid and original plants. In the peroxidase patterns, POX-4 was detected while POX-1 disappeared. Esterase patterns were less influenced, EST-4 was expressed in both plants under the influence of atrazine. Atrazine treatment of both cybrid and original plant shoots had specific effects on carbonic anhydrase, malate dehydrogenase, and glutamate oxaloacetate transaminase. The number and/or the staining intensity of bands corresponding to these isoenzymes decreased under atrazine stress more in cybrid plants (atrazine sensitive) than in S. nigrum (atrazine resistant).

The enzyme activities of the proline metabolic pathways were determined in control and satt-treated (140 M NaCl) calli derived from cotyledons of the domestic saltsensitive tomatoLycopersicon esculentum and the wild salt-tolerantL. pennellii. Glutamate, glutamine, asparagine, and aspartate levels increased in both genotypes under salt stress, while proline accumulation increased markedly only in the salt sensitive tomato. Activity of glutamine synthetase (GS) decreased in the salt-treated calli of the domestic species, whereas both NADH- and NADPH-glutamate synthase (GOGAT) activities increased; GS and NADPH-GOGAT decreased together in the salinized calli of the wild species. Decreasing ornithine levels were found due to NaCl in both tomato populations, while ornithine transaminase (OT) decreased in the wild type only. Increasing NADPH-Δ-pyrroline-5-carboxylate reductase (P5CR) and decreasing proline oxidase (Pro oxi) occurred in the salinized calli of the wild type. Conversely, Pro oxi and proline dehydrogenase (Pro dH) decreased highly in the salinized calli of the domestic population, while no significant changes in P5CR were found.

Gas exchange characteristics of a hemiparasiteMelampyrum arvense L. before and after attachment to the hostCapsella bursa pastoris (L.) Med. were compared. The net photosynthetic rates (P_N) on a leaf area basis were extremely low and in comparison to the value obtained for the host were about 15 % and 23 % for the unattached and attached hemiparasite, respectively. Also the concentration of photosynthetic pigments was low (as compared with the host the content of chlorophylls was about 33 % and 49 % and of carotenoids about 38 % and 36 % in the unattached and attached hemiparasite, respectively). On the other hand the rates of respiration were high (about 1.8 and 2.6 times higher in the unattached and attached hemiparasite, respectively, than in the host). In darkness stomatal conductance (g_S) of the host and the unattached hemiparasite was rapidly reduced to 10 % of the value obtained in light, g_S of the attached hemiparasite was decreased only by about 30%. A total reduction of g_S occurred at relative water content (RWC) of 85 %, 75% and 45 % for the unattached hemiparasite, the host, and the attached hemiparasite, respectively. The transpiration (E) rate in the preparasitic stage was very low, being 2.6 and 4.5 times smaller than in the host and the attached hemiparasite, respectively. In the attached hemiparasite WUE was 7.5 and 3 times poorer than in the host and in the preparasitic stage, respectively.

Wounding both cotyledons ofBidens pilosa (var.radiatus) induces the inhibition of hypocotyl growth. The wound signal is transmitted very rapidly from cotyledon to hypocotyl and can be visualized by the change in nucleotide pools. First we have shown that the irradiance of the plant can change the ATP level without plant wounding. Therefore, plants were harvested at the start of the light period. Under these conditions, we have determined in hypocotyl the levels of adenosine triphosphate (ATP), guanosine triphosphate (GTP) and non adenylic triphosphates (NTP), and adenylate energy charge (AEC) after wounding. We have observed a transient (2 min) increase in the ATP level followed by a decrease 5 to 30 min later. A similar result was obtained for the GTP level but with some delay. The GTP level increased in 5 min and then decreased after 60 min. For the NTP level the decrease is effective from 5 to 60 min after wounding. The calculation of AEC has shown that a very tight control in the level of ATP may be involved in response to wounding.

Changes in soluble protein profile and chlorophyll (Ch1) content in detached soybean leaves incubated in darkness or light were delayed by application of benzyladenine or indole-3-acetic acid and enhanced by abscisic acid. The degradation in light differed significantly from the degradation in darkness. Chl and proteins were lost at a higher rate in darkness than in light.

Suspension cultures of tomato cells were used to study the membrane injury by the toxic concentration of mercuric chloride. Assessment of electrolyte leakage, UV-absorbance of the tissue leachates, relative leakage ratio, injury index, membrane lipid peroxidation, lipoxygenase activity, α-amino nitrogen and total soluble carbohydrate contents showed the extent of membrane damage as a function of the increasing concentration of mercuric chloride. It is suggested that the selected parameters can be used as qualitative tests for determination of stress-induced membrane damage

Changes in the number of protoplasts, viability, protein and chlorophyll content were studied in tobacco mesophyll protoplasts cultivated either autotrophically in CPW medium with mannitol (MCPW) in the light or heterotrophically in CPW medium with glucose (GCPW) in the dark. The number and viability of protoplasts in the both cultivation media were unchanged. In MCPW in the light, the protein and chlorophyll content strongly decreased already after 12 h of cultivation, at 72 h of cultivation, values dropped to 23.6 % (proteins) and to 3.5 % (chlorophyll) in comparison with the initial content. In GCPW in the dark, the protein and chlorophyll contents decreased only slightly to 75 % (proteins) and to 57.7 % (chlorophyll).

In seedlings of the tropical tree speciesErythrina variegata Lam. andHardwickia binata Roxb. exposed to different acidic mist (H₂SO₄, pH 5, 3 and 2) for 5 d significant reduction in seedling growth, biomass accumulation and¹⁴CO₂ fixation were determined. In isolated chloroplasts a decrease in the activities of photosystem 2 and whole electron transport chain was observed only at pH 3 and 2, but no significant change in photosystem 1 activity was observed. SDS-PAGE analysis of crude leaf extracts of ribulose 1,5-bisphosphate carboxylase (RuBPC) indicated a significant loss of 55 and 15 kDa polypeptides at pH 2 inErythrina. The reduction in the RuBPC activity in seedlings grown under acidic mists correlated well with CO₂ fixation.