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).

Adventitious root formation in vitro in 1-mm stem slices cut from microshoots of apple cv. Jork 9 was studied using light and electron microscopy. When indole-3-butyric acid (IBA) had been added to the medium, starch grains accumulated during the first 24 h of culture in cells of the cambial region and in cells in the vicinity of vascular tissue and in the primary rays. This accumulation occurred only in the basal part of explants. After that, the nuclei in these cells were activated, and the density of the cytoplasm and the number of cell organelles increased, whereas starch was broken down. Cambium cells started to divide transversely and at 96 h, after several divisions, a continuous ring of isodiametric cytoplasmic cells had appeared around the xylem near the basal cutting surface. The cells in this ring were rich in cell structures, and did not contain large starch grains and a central vacuole. Root meristemoids regenerated from the portions of the ring that were localized in the primary rays. From the other cells in the ring, callus developed. The meristemoids did not grow into the direction of the epidermis as in shoots, but along the vascular bundles. After emergence from the cutting surface, the meristemoids were transformed into small, dome-like primordia. They developed a typical root apex with root cap, root ground meristem and tracheid connection with shoot vascular tissue.

We studied the influence of ethephon (2-chloroethylphosphonic acid) on the first stages of development of the main apex of Liatris spicata cv. Callilepis and its relation to the mobilization of soluble saccharides in the corm. Ethephon enhanced cell expansion during the first weeks of cultivation but did not affect the timing of flower initiation. Soluble saccharide concentration in the corm reached a maximum after sprouting, probably due to rapid growth of the shoot. There was no change in the concentrations of soluble saccharides immediately prior to flower initiation, which suggested that quantitative changes in these saccharides were not responsible for activating flower initiation. When corms were treated with ethephon there was a greater degree of soluble saccharide mobilization during the first weeks of cultivation than in untreated corms. However, this did not seem to be the direct result of the addition of this phytohormone and was more probably the consequence of cell growth being encouraged.

The content of cytokinins especially zeatin nucleotide decreased in shoots as a result of temperature increase. Simultaneously the cytokinins accumulated in roots. The changes in cytokinins distribution were followed by a decline of evapotranspiration after its initial temperature-induced uprising.

Developing fruits on lower nodes were major reproductive sink for photosynthates in bell pepper (Capsicum annuum L.). The removal of these fruits resulted in faster growth of other fruits on upper nodes. Sink manipulated plants have the higher rate of net photosynthetic rate at the later stages of plant development.

Four clones of Malus domestica Borkh. cv. Golden Delicious - namely Smoothee, Crielaard, Reinders and Golden B - were cultured in vitro from single-node microcuttings placed on solid medium under irradiance (PPFD) of 50 µmol m-2 s-1. After 9 months an average shoot proliferation of 5.3 was achieved; Crielaard showed the highest rate (7.1), followed by Golden B(5.4), Smoothee and Reinders (4.4). Proliferating shoots were then exposed to higher PPFD (80 µmol m-2 s-1) and different spectral composition of radiation using PMMA-B and PMMA-R/FR filters. High PPFD decreased the average proliferation rate to 4.5, in particular in Crielaard and Golden B, while it increased proliferation in Reinders. When a PMMA-R/FR filter was interposed, the mean proliferation rate slightly increased. PMMA-B filters decreased the overall proliferation rate to 3.0; only in Crielaard it was increased, but shoots were very small. Thus PPFD and spectral composition influenced in vitro shoot proliferation and growth and the responses were different among the clones.

The uptake of K⁺ by the leafy shoots of four submersed higher aquatic plants (Elodea canadensis, Ranunculus aquatilis, R. trichophyllus, and Callitriche hamulata) with different HCO₃ ^- affinity was measured in successive 2-h periods under the conditions of high or low photosynthetic rates (i.e. at pH 7.5 or 10). At pH 7.5 the uptake of K⁺ by species with the higher HCO₃ ^- affinity (E. canadensis, R. trichophyllus) was significantly faster than that by species with a lower HCO₃ ^- affinity (R. aquatilis, C. hamulata). In the former group of species, the K⁺ uptake rate at pH 7.5 was 1.7 - 3.5 times higher than at pH 10. At pH 10, the soft-water species, R. aquatilis, had the lowest net photosynthetic rate (PN) of the three HCO₃ ^- users but, in contrast to the relative hard-water species, R. trichophyllus, showed a small K⁺ efflux (47 nmol kg^-1 s^-1). Thus, K⁺ uptake by shoots was not strictly correlated with PN. A significant K⁺ efflux (73 - 86 nmol kg^-1 s^-1) occurred from all HCO₃ ^- users in darkness. The relatively low K⁺ uptake by the strict CO₂ user, C. hamulata, was quite independent of PN and light or darkness. It may be suggested that uptake of K⁺ by shoots of submersed plants depends on their HCO₃ ^- affinity.

Forty-one accessions belonging to ten genera of the tribe Triticeae representing both wild and cultivated species were analyzed by polymerase chain reaction (PCR). Of two consensus tRNA primers tested, one primer revealed characteristic amplification products for all of the species. A total of 35 tRNA markers were scored across all accessions. Five genus-specific and three species-specific markers were obtained. Genomic fingerprints were largely conserved within a genus. The phylogram obtained using parsimony has separated most of the accessions into their prevailing taxonomic species and genus groups. The phylogram showed close association among the three genera Secale, Triticum and Hordeum as expected. The Triticum-Secale relationship was closer than the Triticum-Hordeum or the Secale-Hordeum relationships. The tree also reflected the close associations among the forage grass species belonging to Leymus and Elymus. Thus tDNA-PCR helped to identify species and genera.

Plant regeneration was achieved from coleoptile tissue of wheat (Triticum aestivum L. cv. Kharachia-65). Coleoptiles (1.0 - 3.5 cm long) were excised from 2- to 5-d-old seedlings and cultured on Murashige and Skoog's (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D - 0.5, 2.5, and 5.0 mg dm^-3). Cream, friable callus was obtained after 6 weeks of inoculation. This callus was sub-cultured on MS medium supplemented with 2,4-D (2.5 mg dm^-3) and 5 % coconut water. After 6 weeks of sub-culturing white, cream or pale, friable, nodular callus was obtained. Plant regeneration occurred when this callus was sub-cultured on MS medium supplemented with 0.2 mg dm^-3 1-naphthalene acetic acid + 1.0 mg dm^-3 6-benzylaminopurine. For rooting, regenerated shoots or plantlets were transferred on MS medium supplemented with 0.5 mg dm^-3 indole-3-acetic acid. Rooted plantlets were directly transferred into pots and grown under field conditions. Seed setting invariably occurred in all plants.

Plants of teak (Tectona grandis L.f.) growing after transplantation to fertile soil for 5 - 6 months were subjected to water stress by withholding watering continuously for 3 weeks. The growth rates of both plants in height and developing leaves in length were unaffected during the first week after withholding watering, but they were decreased by about 50 % during the second week and became negligible during the third week of water stress treatment. The rate of leaf production and internodal elongation were also decreased in plants experienced 2 weeks of water stress. However, after rewatering, these plants regained growth potential and exhibited high rates of leaf expansion and plant growth comparable to those of well-watered plants. Diurnal course of net photosynthetic rate (PN) of plants subjected to water stress for 2 weeks was similar as that of well-watered plants. However, PN of plants subjected to water stress for 3 weeks was reduced in the afternoon. Similarly, stomatal conductance (gs) and transpiration rate (E) of plants experiencing 3-week water stress were decreased in the afternoon. Soon after rewatering, PN, gs and E reached similar values to those of well-watered plants.