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.

A reliable protocol for the regeneration of onion through repetitive somatic embryogenesis was established. Embryogenic callus was derived from mature seeds on Murashige and Skoog (MS) medium supplemented with 2 mg dm-3 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryos aroused on the surface of calli cultures and formed plantlets after the removal of 2,4-D or its substitution with 1 mg dm-3 kinetin (Kin). Reculturing the somatic embryos on 2,4-D containing medium led to secondary embryos formation. The embryogenic cultures which were preserved for five months on maintenance medium containing 2 mg dm-3 2,4-D + 0.5 mg dm-3 Kin have retained their ability for regeneration, while those kept on 2,4-D only, failed to form plantlets. Electrophoretic analysis of total soluble proteins revealed that the competence for successful conversion of somatic embryos into plantlets is associated with the expression of new set of proteins (112, 58 and 30 kD). The regenerated plants were successfully transferred to the soil.

During the germination, wheat and sorghum plants tolerated salinity up to 10 mM NaCl. Seedling growth and dry matter production remained more or less unchanged up to 10 mM NaCl in case of wheat and up to 5 mM NaCl in case of sorghum. The water content of test plant species exhibited nearly constant values irrespective of the salinity level applied. The proline and carbohydrate content increased with elevating NaCl, while free amino acids content decreased. Irrigating of seedlings with tryptophan had non significant effect at all salinity levels used.

Differences in anatomy and morphology of the kiwifruit leaves and leaf petioles might play a considerable role in the sex-determination. Three months after bud break (June), the kiwifruit leaves of both male and female plants, grown on the vegetative and generative shoots showed different leaf area (128.6 ± 13.45 cm2 in male and 104.5 ± 4.02 cm2 in female plants) and shape. The most frequently leaf shape was determined as "folium cordatum" and "folium rotundato-cordatum". Higher values of total leaf thickness of the female leaves (190 ± 3.84 µm) in comparison to male leaves (174 ± 3.52 µm) were estimated, resulting in the thicker adaxial leaf epidermis and especially in thicker palisade parenchyma in female leaves (136 ± 2.76 µm in comparison to 104 ± 1.61 µm in male leaves). Typically bifacial leaves were observed in both male and female leaves. Anomocytic stomata in hypostomatic leaves were found. The reticulate venation appears to be the main type of leaf venation. Stalked stellate multicellular trichomes on the abaxial leaf side were frequently observed in the leaves of both sexes. No important differences between male and female plants were found in the structures of vascular system in leaves and leaf petioles. Thus leaf thickness and surface morphology of adaxial leaf epidermis can be considered as important structural parameters in the sex determination.

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.

Thirty-day-old seedlings of two jute species (Corchorus capsularis L. cv. JRC 212 and C. olitorius L. cv. JRO 632) were subjected to short-term salinity stress (160 and 200 mM NaCl for 1 and 2 d). Relative water content, leaf water potential, water uptake, transpiration rate, water retention, stomatal conductance, net photosynthetic rate and water use efficiency of both jute species decreased due to salinity stress. The decrease was greater in C. olitorius than in C. capsularis and with higher magnitude of stress. Greater accumulation of Na+ and Cl- and a lower ratio of K+/Na+ in the root and shoot of C. olitorius compared with C. capsularis were also recorded. Pretreatment of seedlings with kinetin (0.09 mM), glutamic acid (4 mM) and calcium nitrate (5 mM) for 24 h significantly improved net photosynthesis, transpiration and water use efficiency of salinity stressed plants, the effect being more marked in C. olitorius. Among the pre-treatment chemicals, calcium nitrate was most effective.

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.

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.

The effect of concentration of zinc ions on chlorophyll (Chl) and dry matter contents in Senna occidentalis (L.) Link plants was studied under various availability of soil water. Diurnally, the internal Zn2+ content was predominantly affected by supplied Zn2+ and by daytime (Tm). Chl content, Chl a/b ratio, and Chl stability to heat were mainly affected by soil water content, and the dry matter content depended mostly on Tm.

The excised shoot tips of Beta vulgaris L. cvs. Ras poly, Gala and Lola were inoculated aseptically on Murashige and Skoog's (MS) medium with 0.25 mg dm-3 α-naphthaleneacetic acid (NAA) and 1.00 mg dm-3 6-benzylaminopurine (BAP). They produced multiple shoots at relatively high frequency. For shoot multiplication, a concentration of 1.00 mg dm-3 NAA and 0.25 mg dm-3 BA in the culture medium was found to be optimum for all the cultivars. MS medium supplemented with 3.0 mg dm-3 3-indolebutyric acid (IBA) and 0.02 mg dm-3 N-isopentenylaminopurine (2ip) was found most effective for rooting of micropropagated shoots. In vitro produced plantlets were successfully transferred to the soil. The percentage of survival was more than 95 % and micropropagated plants showed no morphological differences from those grown naturally.

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.

A non-destructive method of total vertical projections for estimating the length of rigid root systems, not introduced yet in plant sciences, is described. It is demonstrated on measuring less and more dense root systems of seedlings of Zea mays grown at hypoxic or control conditions. Photographs of six vertical projections (30° apart) of each root system were taken and evaluated. The method being based on proved mathematical formula offers unbiased estimation of the length of a rigid root system, curved in three-dimensional space, by non-destructive means. Thus, it is applicable during ongoing experimentation on plants grown in a solution culture. It was shown that less than 120 intersections between the root projection and test lines in one photograph ensured sufficient precision of the method and that the observer subjectivity could be overcome by presented instructions.

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.

Molecular markers were used to detect the influence of high concentrations of 2,4 dichlorophenoxyacetic acid (2,4-D) in the callusing media on DNA variations in regenerated rice plants. Restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD) and polymerase chain reaction (PCR) based RFLP analysis were carried out on 12 somaclones of Oryza sativa L. cv. B-370. In vitro culture induced DNA variations were detected in the regenerated plants but the effect of high auxin concentration in the medium could not be revealed. In a second study, fingerprinting of 15 semi-dwarf, high yielding somaclones of B-370 was carried out using RAPD technique. Amplification using 20 random primers produced a total of 167 DNA bands out of which 97 bands were polymorphic. A total of 32 unique DNA bands were detected across all the somaclones and they could be grouped based on their similarity to B-370. RAPD analysis helped to reveal similarity or differences among the somaclones while fingerprinting using additional RAPD markers was not successful.

Somatic hybridization of Brassica oleracea (cauliflower, cabbage, kohlrabi) and Armoracia rusticana (horseradish) protoplasts was performed to test the possibility of the formation of intergeneric hybrids. Out of three B. oleracea crops tested only cauliflower (B. oleracea var. botrytis) yielded viable hybrids when fused with horseradish protoplasts. Regenerated plants were asymmetric hybrids as revealed by morphological, karyological, and DNA analysis. In these plants, extensive elimination of A. rusticana genome occurred during subsequent cultivation. Using a chloroplast DNA probe no A. rusticana-specific hybridization signals were detected thus indicating a rapid elimination of horseradish cytoplasmic (chloroplast) components.

Mutagenesis provoked by exposure to increased concentration of antibiotics of five indigenous Rhizobium galegae strains resulted in the generation of several antibiotic-resistant mutants. The mutants differed from the wild type and one from another in respect to the nodulation capacity, the nitrogenase activity, the nodule ultrastructure, and the plant growth response. Galega plants inoculated with mutants resistant to streptomycin and rifampicin formed nodules with higher nitrogenase activity and accumulated more shoot dry biomass than plants inoculated with the parent strains. Resistance to kanamycin and nalidixic acid was associated with significant decrease of nitrogenase activity. A correlation between nitrogen-fixing efficiency and nodule infected cell ultrastructure was found. When the bacteroids occupied about 10 times higher area in infected cells of nodule than peribacteroid spaces and host cytosol had electron dense and homogenous structure, the nitrogenase activity was the highest.

In Kalanchoë blossfeldiana cv. Tom Thumb the initial rate of ATP-dependent H+-transport into tonoplast vesicles was stimulated up to three times if the H+-ATPase (EC 3.6.1.3) was energized a few minutes after pre-energization of the H+-PPase (EC 3.6.1.1). H+-PPase-activated ATP-dependent H+-transport was observed in plants of K. blossfeldiana cultivated in short day (SD) or long day (LD) conditions expressing different degrees of crassulacean acid metabolism (CAM). However, based on the higher activity and protein amount of H+-PPase and H+-ATPase present in the vacuolar membrane of SD plants the maximum H+-transport activity in the stimulated mode of the H+-ATPase was significantly higher in tonoplast vesicles of SD plants than of LD plants. Hence, a co-ordinated action of the H+-PPase and H+-ATPase at the tonoplast of Kalanchoë could allow a higher transport capacity at the vacuolar membrane when plants perform high CAM. Immunoprecipitation experiments with an antiserum raised against the A-subunit of the vacuolar H+-ATPase of Mesembryanthemum crystallinum L. showed that in SD and LD plants of K. blossfeldiana the H+-PPase was co-precipitated with the vacuolar H+-ATPase holoenzyme. The co-percipitation of the two transport proteins indicates a close structural localization of the H+-PPase and the A-subunit of the vacuolar H+-ATPase.

Total genomic DNA isolated from maize hygromycin B resistant cell line(hygr-G204) was used to transform the maize hygromycin B sensitive cell line(hygs-G204) to the hygr-phenotype using polyethyleneglycol treatment and the transformed calli were selected using hygromycin B. The primary transformant maize plants were regenerated and analysed at the molecular level using DNA hybridization, transgenome rescue and histochemical β-glucuronidase assay. The results indicated that genomic DNA-mediated transformation can lead to transfer, expression and stable integration of a DNA fragment into the host genome.

A screening of leaves of 25 taxa of angiosperms was made for their volatile toxicity against damping-off fungi. The volatile substances fromHyptis suaveolens andOcimum canum were toxic againstPythium aphanidermatum, P. debaryanum andRhizoctonia solani. The fungitoxicity of the leaves persisted for 15 d of storage. The volatile substances from the leaves ofO. canum were thermostable, while those fromH. suaveolens were thermolabile. The essential oils exhibited strong potency against the pathogens tested, non-phytotoxic nature to the host plants and superiority over commonly used synthetic fungicidesAgrosan G.N. andCaptan. The findings indicate the possibility to use these essential oils as potential natural fungicides in management of damping-off pathogens.

Accumulation of the extracellular proteins localized in intercellular spaces of barley primary leaves was examined after inoculation with powdery mildew (Erysiphe graminis f. sp. hordei) as biotic stress factor and after abiotic stresses such as heat shock, low temperature and heavy metal (Mg,Zn, Cu, Al, Cd and Co) treatment using native polyacrylamide gel electrophoresis. Six to eight major pathogen-induced proteins (bands on native gel) have been identified. Their accumulation at host-parasite incompatibility was more expressive than at compatibility interaction. Elevated temperature did not induce pathogenesis-related (PR) proteins while low temperature induced three of them. Cu, Al, Cd and Co induced accumulation pattern of extracellular proteins was very similar to that in powdery mildew inoculated leaves. Mg and Zn had no effect on the induction of protein accumulation in the intercellular spaces of leaves. Induction of PR proteins by different stresses indicated their general function in the resistance of plants to changing environment.

Ten-days-old bean plants (Phaseolus vulgaris L., cv. Cheren Starozagorski) were treated with simulated acid rain (pH 2.4, 2.2, 2.0 and 1.8). Anatomical changes in the primary leaves were studied 3, 48 and 168 h after a single treatment. This treatment induced: 1) change in the shape of palisade cells, contraction of their contact surfaces and expansion of spongy cells (pH 1.8, 3 h after treatment); 2) reduction of symplast connections among palisade cells and of apoplast in the spongy mesophyll (pH 1.8, 48 h after treatment); 3) destruction of adaxial epidermis and portions of palisade mesophyll, plasmolysis of spongy cells (pH 1.8, 168 h after treatment); 4) full destruction of mesophyll (pH 2.4, 2.2, 2.0 and 1.8, 168 h after treatment). The structure of abaxial epidermis was more stable than that of the adaxial one. With respect to anatomical parameters the studied species could be considered as comparatively resistant to acid rain.

An efficient protocol was developed for in vitro plant regeneration via somatic embryogenesis from cell suspension cultures of metal tolerant grass Echinochloa colona (L.) Link. Callus was obtained by culturing leaf base on MS medium supplemented with 0.5 mg dm^-3 of 6-benzylaminopurine (BAP) and 2.0 mg dm^-3 of 1-naphthaleneacetic acid (NAA). Cell suspensions were initiated and established in MS liquid medium containing 0.5 mg dm^-3 BAP, 1.0 mg dm^-3 NAA and 2.0 mg dm^-3 2,4-dichlorophenoxyacetic acid (2,4-D). A reduction in the concentration of 2,4-D to 0.5 mg dm^-3 induced formation of somatic embryos. The embryos developed and grew into normal plants in the presence of half strength MS medium without growth regulators. The regenerated plants were hardened in the greenhouse and subsequently grown in the open. This system may be also used for isolation and culture of protoplasts as a first step in somatic hybridization.

Changes in hexokinase particulate and soluble isozyme composition and activities in leaves of 65- and 115-d-old tobacco plants were determined by ion exchange chromatography on DEAE cellulose. During plant ageing, the activities of glucose and of fructose phosphorylating isozymes of particulate hexokinase decreased to 9.9 and 9.2 % of initial value, respectively. The activity of soluble hexokinase decreased to a lesser extent: that of glucose phosphorylating isozyme to 49.8 % and of fructose phosphorylating isozyme to 37.8 %. The activity of soluble fructokinase isozyme dropped to 34.8 %. Thus also the ratio of particulate and soluble isozymes was dependent on the age of leaf tissue.

The growth retardant paclobutrazol (PBZ) inhibited stem internode growth of in vitro cloned potato plants. The extent of growth inhibition caused by10-9 M PBZ in Murashige and Skoog medium was genotype-specific, varying between 10 - 60% of the stem growth of untreated controls in ten cultivars examined. An increase in percentage of de novo bud regenerating stem internode segments (SIS) as well as in the total number of buds per explant was observed in SIS taken from PBZ pretreated plants. PBZ applied directly into the regenerative media had no stimulatory effect on there generation process. We assume that the enhancing effect of PBZ on regeneration may be attributed to its interaction with cytokinin metabolism.

The physiological responses of alfalfa (Medicago sativa L. cv. Gilboa) to salinity (100 mM NaCl) and some inorganic nutrients (K+, Ca2+ and N as NO3-) were investigated. Salinity caused a substantial reduction in biomass, carbon assimilation rate, stomatal conductance, water use efficiency, leaf area, relative growth rate, NO3- content and nitrate reductase activity, whereas, transpiration rate was slightly affected. Inclusion of K+, Ca2+ and N as NO3- in plant nutrient medium in combination or alone brought about a marked stimulation in control plants and moderated the salinity caused reductions in NaCl treated plants. In addition, plants also exhibited differences in these parameters at two growth stages.

The ultrastructure of chloroplasts in the primary leaf of 10-d-old bean plants (Phaseolus vulgaris L., cv. Cheren Starozagorski) was studied 3, 5, 24, 48, 72 and 168 h after a single treatment with simulated acid rain (pH 2.4, 2.2, 2.0 and 1.8). Different changes in chloroplast structure till full destruction of organelles were traced. A determining factor for these changes was the histological localization of chloroplasts. In the chloroplasts of palisade parenchyma different degrees of expansion of thylakoids (3, 5, and 24 h after the single treatment), and conformational changes of the inner membrane system (48, 72 and 168 h) were observed. The chloroplasts of spongy parenchyma showed a significantly higher degree of structure resistance. The expansion of thylakoids was weak and did not depend on the duration of treatment. The ultrastructural changes of chloroplasts confirmed relative resistance of this species till pH 2.0.

Dry matter production of shoots and roots and the diurnal fluctuation of titratable acidity of single leaves were investigated in the CAM plant Agave attenuata during the first 70 d after germination. The plants were grown either in vermiculite sub-irrigated with a nutrient solution or in in vitro cultures on an inorganic nutrient agar. Two types of culture tube covers were used: either airtight closures or polypropylene caps with membranes permeable to air.
In the earliest ontogenetic phases of development (cotyledon and primary leaf stage), the plants were already able to carry out considerable nocturnal organic acid accumulation. In vitro cultivated plants, from the beginning of their development, were also capable of diurnal acid fluctuation, though of distinctly weaker activity than the pot plants. The mean relative growth rates (RGR) of pot culture plantlets approached a third of perennial herbaceous plants. Plantlets grown in in vitro culture reached only half to the one quarter of the RGR of pot plants. The reduced yield could be attributed to the low CO2 supply in the culture tubes and the less than optimal water and nutrient supply in the agar medium.

Changes in growth and anatomical structure of vascular tissues in stem, root and leaf of safflower plants grown in NaCl and CaCl2 solutions having different osmotic potentials (ΨS from 0 to -0.9 MPa) with addition of 0, 10 and 20 mg dm-3 zinc were studied. Shoot and root lengths, fresh and dry masses and fresh/dry mass ratio were lower in salt-stressed plants compared to unstressed plants. Salinity induced structural changes in stem, root and leaf tissues; few xylem vessels with smaller size were noticed in stressed plants. The higher concentration of Zn improved growth especially in roots and enhanced xylem formation in comparison to stressed plants grown at the same osmotic potential without Zn. Zn also protected xylem distructure by salinity in leaves.

Transient expression of the β-glucuronidase (GUS) gene introduced into Arabidopsis thaliana intact plants by T-DNA after vacuum infiltration of Agrobacterium tumefaciens was followed. The first incidence of GUS activity was found 2 - 3 d after treatment and a peak of activity one week after treatment in both A. thaliana races, Columbia and C24. GUS activity was sharply increased by cultivation of Arabidopsis plants at elevated temperature (29 °C) compared to cultivation at 25 °C. The density of inocula also influenced the GUS activity.

Eichhornia crassipes plants brought from the River Nile were cultured in jars containing river water supplemented with various concentrations of Cd, Pb, and Sr (0 to 100 μg cm-3), added simultaneously. Treatment continued for 20 d during which each cultivation solution was being replaced with fresh one every 3 d. The growth of Eichhornia was drastically retarded at heavy metal concentrations higher than 15 μg cm-3. At concentrations 15 or 25 μg cm-3, the accumulation of Cd and Pb to levels several times higher than those in control plants was found. More than 50 % of the uptaken metals were retained by roots alone. Leaves and leaf petiols received around 30 and 20 % of the accumulated metals, respectively. X-ray microanalysis indicated the presence of the three heavy metals in Ca oxalate crystals. Content of metals in the crystals increased progressively over time of exposure in a way similar to those in whole plant tissues. These results suggest a possible role for Ca oxalate crystalization in toxic heavy metal deposition and thus tolerance by Eichhornia.