Transformation of tomato (Lycopersicon esculentum Mill.) was carried out using disarmed Agrobacterium tumefaciens strain EHA 105 harboring a binary vector pBIG-HYG-bspA. The plasmid contains the bspA (boiling stable protein of aspen) gene under the control of a CaMV35S promoter and nopaline synthase (NOS) terminator, hygromycin phosphotransferase gene (hpt) driven by nopaline synthase promoter and polyadenylation signal of Agrobacterium gene7 as terminator and a promoterless gus gene. Very strong β-glucuronidase (GUS) expression was observed in transformed tomato plants but never in non-transformed (control). Since GUS expression was observed only in transformed plants, the possibility of the presence of endogenous GUS enzymes was ruled out. Possibility of false GUS positives was also ruled out because the GUS positive explants reacted positively to polymerase chain reaction (PCR) and PCR-Southern tests carried out for the presence of bspA gene, which indicated the integration of T-DNA in tomato genome. The promoterless GUS expression was hypothesized either due to leaky NOS termination signal of bspA gene or due to different cryptic promoters of plant origin. It was concluded that GUS expression was observed in the putative transgenics either due to the read through mechanism by the strong CaMV35S promoter or due to several cryptic promoters driving the gus gene in different transgenic lines.

The aim of this study was to investigate the protein alterations in the transgenic kiwifruit (Actinidia deliciosa) overexpressing a transcription factor gene, OSH1. Although transgenic plant with introduced OSH1 indicates suppression of gibberellin (GA)-20 oxidase activity, application of GA to transgenic kiwifruit could not completely recover plant morphology and protein profiles. Eleven proteins decreased in the transgenic kiwifruit detected by two-dimensional polyacrylamide gel electrophoresis showed homologies to kiwifruit hypothetical protein, osmotin I and photosynthesis related protein. These results suggest that introduction of an OSH1 into kiwifruit caused wide-range alterations at protein level and alterations of protein accumulation should be considered to evaluate the substantial equivalence of plants transformed by a transcription factor.

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.

Partial cDNAs sequences for arginine decarboxylase (Pvadc), S-adenosylmethionine decarboxylase (Pvsamdc) and spermidine synthase (Pvspds) were isolated from the bean Phaseolus vulgaris using primers designed from conserved regions of enzymes belonging to plant species. Sequence analysis showed that the Pvadc, Pvsamdc and Pvspds genes were most closely related to the orthologous genes from Glycine max, Phaseolus lunatus and Pisum sativum, respectively. The expression patterns of the genes, together with that of ornithine decarboxylase (Pvodc), were analysed in young and mature leaves, stems, roots, root tips, petals, stigma, ovaries, filaments and anthers of bean plants. Pvsamdc was found to be expressed at similar levels in all tissues. The other transcripts showed tissue specific expression. Pvadc was barely expressed in petals and not at all in roots tips, Pvspds was mainly expressed in roots, stigma and filaments, and Pvodc was detected only in roots.

Salix paraplesia was used as an experimental model to investigate the effect of short day photoperiod (SD) and low temperature (LT) on development of freezing tolerance and on endogenous abscisic acid (ABA) contents. We characterized differences in SD and LT-induced cold acclimation in three ecotypes from different altitudes. The results demonstrated that cold acclimation could be triggered by exposing the plants to SD or LT alone, and that a combination of the different treatments had an additive effect on freezing tolerance in all ecotypes studied. However, the high altitudinal ecotype was more responsive to SD and LT than the low altitudinal ecotype. Development of freezing tolerance induced by SD and LT was accompanied by changes in ABA contents which were ecotype-dependent. Although the stem had higher initial freezing tolerance, the leaves developed freezing tolerance more quickly than the stem and thus leaves may provide an interesting experimental system for physiological and molecular studies of cold acclimation in woody plants.

A protocol for in vitro propagation of Balkan endemic plant Salvia brachyodon Vandas from nodal segments was developed. 6-benzylaminopurine was more effective in axillary buds promotion when compared to thidiazuron. The rooting of regenerated shoots was induced by transferring them to the media supplemented with auxins. All tested auxins (indole-3-acetic acid, indole-3-butyric acid, and α-naphthaleneacetic acid) stimulated the rooting of S. brachyodon shoots. The acclimatization of in vitro rooted shoots was successful.

Explants obtained from in vitro-propagated plantlets of two potato cultivars, Shepody and Atlantic, were treated with five doses of γ-radiation (0, 2, 4, 6 and 8 Gy) to investigate the stimulating effects of low irradiation on the production and quality of microtubers in vitro. Microtubers of both cultivars treated with γ-radiation initiated 5 d earlier than in the non-irradiated control. The whole period of microtuberization was prolonged by 10 - 15 d with 4, 6 and 8 Gy irradiation treatment for cv. Atlantic. Irradiation of the plantlets (4 Gy) led to a significant increase not only in the microtuber number (116.7 and 34.5 % over the control) but also in the fresh mass (77.6 and 23.2 % in Shepody and Atlantic, respectively). Low dose irradiation (2 - 4 Gy) increased the starch content of microtubers. High doses (6 - 8 Gy) enhanced ascorbic acid and reducing sugar contents. 4 - 6 Gy doses also effectively increased the protein contents of microtubers.

The aim of the present work was to evaluate oxidative stress and plant antioxidant system of three contrasting bean (Phaseolus vulgaris L.) genotypes in the response to drought. Drought was imposed 14 d after emergence, by withholding water, until leaf relative water content reached 65 %. Water stress increased lipid peroxidation (LPO), membrane injury index, H₂O₂ and OH^⋅ production in leaves of stressed plants. Activities of the antioxidative enzymes superoxide dismutase (SOD) and ascorbate peroxidase (APOX) increased significantly under water stress in all the studied cultivars, while catalase (CAT) increased in cvs. Plovdiv 10 and Prelom, but decreased in cv. Dobrudjanski ran. Furthermore cv. Plovdiv 10 which had the highest APOX and CAT activities also showed the lowest increase in H₂O₂ and OH^⋅ production and LPO while cv. Dobrudjanski ran showed the lowest increases (and often the lowest values) in the antioxidant enzyme activities and the highest increases of H₂O₂ and OH^⋅ production, and LPO. On the basis of the data obtained we could specify cv. Plovdiv 10 and cv. Prelom as drought tolerant and cv. Dobrudjanski ran as a drought sensitive.

Proline accumulation in two different bean (Phaseolus vulgaris L.) cultivars, one drought-sensitive (Canario 60) and one drought-resistant (Pinto Villa) was investigated. Both tolerated salt concentrations up to 150 mM NaCl, but the sensitive Canario 60 did not survive at 400 mM NaCl. In response to salt stress, both cvs. accumulated proline in all the analyzed tissues, the lowest contents were detected in roots. Pinto Villa accumulated higher proline concentrations than Canario 60 only at 400 mM NaCl. The addition of polyamines or ornithine increased proline content in plant tissues without stress, while they decreased it under salt stress.

Explants of Chenopodium rubrum, a short-day plant, were decapitated and exposed to floral inductive treatment, and the extent of flowering of axillary buds was afterwards assessed. Isolated buds never responded to induction, whereas the presence of the petiole of the subtending leaf already assured a high degree of flowering. We may assume either that the petiole is the receptor organ of the photoperiodic signal or that its transporting role is indispensable.

There is increasing evidence that the sucrose normally added to the culture medium affects negatively the photosynthetic capacity of plantlets. At the same time, however, sucrose cannot be eliminated from the medium, as it is required for normal in vitro growth. We argue that this is true only under the conventional light conditions of growth-rooms. In the present paper irradiance of growth-rooms was increased 10 times and although the sucrose-inhibitory effect was found at high sucrose concentrations, it was possible to grow coconut (Cocos nucifera L.) plantlets without sucrose. Those plantlets showed both high photosynthetic capacity and comparable in vitro growth to those grown with sucrose in the medium under conventional growth-room irradiance. Nevertheless, the best growth was achieved under mixotrophic conditions where at high irradiance and moderate sucrose concentrations plantlets accumulated 27 % more biomass than plantlets grown without sucrose under high irradiance and 43 and 73 % more biomass than their counterparts at low irradiance with or without sucrose, respectively.

The effect of arsenic (32 - 96 µM) on the phosphorus content and Chl fluorescence was studied in soybean (Glycine max Merril) grown in the nutrient solution with and without phosphorus. The increased concentration of As led to the decrease in P content in plant organs. Parameters of Chl fluorescence of soybean leaves in the presence of these As concentrations did not show significant changes.

Two rice (Oryza sativa L.) cultivars, Koshihikari and Pokkali, were grown in solution culture at three concentrations of NaCl or Na₂SO₄ [0 (S0), 50 (S1), and 100 (S2) mmol dm^-3] and three N contents [0.7 (N1), 7 (N2) and 14 (N3) mmol dm^-3]. Salinity significantly decreased dry matter of both cultivars. Pokkali had better growth than Koshihikari under both saline and non-saline conditions. Applications of N enhanced development of shoot dry mass under S0 and S1 treatments up to N2. Under S2, N application had no effect on shoot dry mass of both cultivars. Root dry mass of both cultivars decreased with increasing N application at S1 and S2. Shoot and root NO₃-N content in both rice cultivars increased with increasing N concentration in the nutrient solutions. The absorption of NO₃-N was less in Koshihikari than Pokkali plants, and also was much less in Cl^- than SO₄ ^2- salinity suggesting the antagonism between Cl^- and NO₃ ^-. In addition a significant negative correlation between concentrations of NO₃-N and Cl^- in the shoots or roots was observed in both cultivars

Cytokinin-like activity of orotic acid and its 1-cyclohexyl and 1-phenyl derivatives was tested estimating the anthocyanin accumulation and inhibition of root formation in isolated buckwheat cotyledons (Fagopyrum esculentum Moench.). The anthocyanin content was stimulated most by 1-phenylorotic acid. Strong synergetic effect of the phenylurea cytokinin 4PU-30 was found.

The effect of elevated CO₂ concentration (CE) on leaf chlorophyll (Chl) and nitrogen (N) contents and photosynthetic rate (P_N) was evaluated during the post-flowering stages of rice grown at CE (570 ± 50 µmol mol^-1) in open top chamber (OTC), at ambient CO₂ concentration (∼ 365 µmol mol^-1) in OTC and at open field. Thirty-five day old seedlings were transplanted in OTCs or in field and allowed to grow till maturity. Chl and N contents were highest at the time of flowering and thereafter it started to decline. The rate of decline in Chl and N contents was faster in plants grown under CE mostly in later part of growth. Irrespective of treatment difference, flag leaf contained the highest amount of Chl and N than penultimate and third leaf. The higher P_N was observed in leaves under CE than in the leaves in other two growing conditions. Considering growth stage, P_N was the highest at flowering which reduced at the later part of growth due to degradation of Chl and N content of the leaf. Under CE it was 40.02 µmol m^-2 s^-1 at flowering and it reduced to only 14.77 µmol m^-2 s^-1 at maturity stage. The beneficial effect of CE in increasing leaf P_N may be maintained by applying extra dose of nitrogen at the later stages of plant growth.

Waterlogging of soybean plants (Glycine max L.) led to impaired symbiotic N₂ fixation and a marked decline in glutamine (Gln) concentration in xylem bleeding sap. Xylem Gln concentration increased during the growth cycle of the plant and was correlated with nodule formation. Treatments known to impair N₂ fixation, such as exposing the root system to pure N₂ gas or a mixture of Ar and O₂ (80:20; v/v), led to specific declines in xylem sap Gln. The decrease in Gln observed during waterlogging was also seen on transfer of nodulated plants to aerated hydroponics, where the decline was highly correlated with ureide content in the xylem sap. Upon flooding the nodulated root system, the specific decline in xylem sap Gln could be detected within 10 min and reached a minimum within 60 min, indicating that waterlogging has an immediate effect on N₂ fixation. It is concluded that xylem Gln arises directly from N₂-fixation and is a useful indicator of N₂ fixation activity of symbiotic soybean plants.

Thirty-six F₂ hybrid poplar (Populus trichocarpa × P. deltoides) clones were fumigated with ozone to record its effects on growth, correlate them with stomatal response and screen for ozone sensitivity. Fumigation was applied for 6 to 9 h each day for approximately 3 months at ozone concentrations of 85 to 128 μg g^-1 using open-top chambers. Height, diameter, number of leaves, stomatal conductance, transpiration rate, total biomass, biomass components and root/shoot ratios were reduced by ozone stress. Percent of leaf fall in ozone-treated plants was nearly three times higher than in control plants exposed to charcoal-filtered air. Leaf senescence, because of ozone exposure, did not appear to be associated with reduced biomass production. Some clones had a high percentage of leaf-fall with ozone exposure, but were able to maintain total biomass production near that of the control. Their response may be an example of an ability to adjust or compensate for ozone damage. There was no significant or consistent relationship between stomatal conductance and total biomass or the change in stomatal conductance as a result of ozone exposure and the change in total biomass. Taken together, these results suggest that effects of ozone on poplar growth cannot be solely correlated to changes in stomatal conductance, more physiological and biochemical parameters should be examined.

Morphological and anatomical features of Ebenus cretica leaflet, such as lanceolate shape, reduced size, dense cover with non-glandular hairs, epidermis of small cells, compact mesophyll, amphipleurous presence of palisade parenchyma, thick cuticle, development of numerous mesophyll phenol-storing cells and the amphistomatic type, disclose the xeromorphic character of the plant. In the island of Crete two ecotypes of E. cretica, ecotype A and ecotype C, are greatly extended. In ecotype A leaflets, the above features are more prominent than in ecotype C. This fact accomplished by physiological data favours the suggestion that plants of ecotype A are better adapted to the xerothermic environment of the island of Crete. This may be the reason that ecotype A occupies the major portion of the island and is predominant in the western and central regions. The distinction of ecotypes A and C, by evaluating the strategies these plants used in order to better adapt and the characteristics of their inflorescences may be used as a criterion for the selection of the most appropriate ecotype for application in floriculture and ornamental horticulture.

A regulatory hrpL non-virulent mutant of Erwinia amylovora is effective in controlling fire blight disease when inoculated on apple seedlings simultaneously with the pathogenic parental strain. Mechanisms involved in this protective effect were investigated. The use of two marker genes, uidA and lacZ, expressed in the hrpL mutant and the pathogenic strain, respectively, allowed to localize simultaneously the two inoculated strains in plant tissue. An anti-β-glucuronidase antibody was also used to detect the hrpL mutant. Both techniques indicated that the two strains localized mainly in separate areas of the leaf tissue. In addition, leaves infiltrated with the hrpL mutant exhibited a significant increase in peroxidase activity in contrast to a hrp secretion mutant known to be less effective in the protection. It is suggested that protection obtained with the hrpL mutant relies on the physical separation between the mutant and the parental strain after co-inoculation and the rapid and sustained activation of plant defense mechanisms in reactive tissue, i.e. not invaded by the virulent strain.

The use of two Agrobacterium tumefaciens strains for transformation of Triticum aestivum L. cv. Vesna was studied. Immature embryos, isolated 15 d after pollination, were co-cultivated with the super-binary LBA4404/pTOK233 and the binary AGL1/pDM805 vectors. While the transient GUS-intron expression was high (69.9 and 80.0 %), the number of plants regenerated on selective media containing hygromycin or phosphinotricin did not exceed 0.4 and 0.13 %, respectively. Nevertheless, the regenerated plants were fertile and produced seeds. The T₀ plants, as well as the T₁ seedlings, displayed the activity in the β-glucuronidase histochemical assay and a positive signal in PCR analysis for the presence of uidA gene sequences in their genomes. The data suggest that the transformation of wheat cv. Vesna with both Agrobacterium strains is feasible.

Seven plant species including three chenopods:Suaeda fruticosa, Kochia indica, Atriplex crassifolia and four grasses:Sporobolus arabicus, Cynodon dactylon, Polypogon monspeliensis, Desmostachya bipinnata, varied greatly in their seed germination and growth responses to soil moisture or salinity. The germination percentage of each species was significantly lower at soil moisture level of 25 % of water holding capacity than at the levels ranging from 50 to 125 %. Increase in salinity resulted in gradual decrease in seed germination of each species. Growth responses of species to salinity varied widely from significant decrease with slight salinity to stimulation up to salinity levels of 20 dS m^-2. Higher K⁺Na⁺ratios in plant shoots of all species compared to that in the root medium indicated selective K⁺uptake. Higher tolerance in chenopod species seems to be attendant on their ability for internal ion regulation.

In vitro protocols for plantlet regeneration and medium-term genotype conservation of eight wild species of Curcuma have been optimized. Both the phenomena were genotype-dependent and influenced significantly by type and concentration of cytokinins used. In general, benzyladenine (BA) was found superior to other cytokinins tested for plantlet regeneration and γ,γ-dimethylallylaminopurine (2iP) for conservation. Number of shoots per culture ranged from 1.3 to 7.2 and conservation period from 264 to 379 d. In 30-d-old cultures, highest frequency of shoot regeneration could be obtained in C. malabarica (7.2 shoots per culture) on MS + 11.4 μM zeatin. Curcuma sp. (unidentified wild species) could be conserved for maximum period (379 d) on MS + 24.6 μM 2iP followed by C. aromatica (363 d) on MS + 22.8 μM zeatin. The tissue culture-raised plantlets were morphologically similar to their parents. The in vitro-conserved plants multiplied rapidly in tissue cultures and produced normal rhizomes upon transfer to soil in net house.

Trichloroacetic acid (TCA) belongs to secondary atmospheric pollutants affecting the forest health. Distribution of [1,2-¹⁴C]TCA-residues and TCA biodegradation were investigated in 4-year-old nursery-grown trees of Norway spruce [Picea abies (L.) Karst.] in the whole plant/soil system. Radioactivity was monitored in needles, wood, roots and soil as well as in the air. During two weeks of exposure TCA was continuously degraded, especially in the soil. Estimates of radioactivity balance showed loss of radioactivity into the atmosphere in the form of ¹⁴CO₂; unincorporated [1,2-¹⁴C]TCA, chloroform, carbon monoxide and methane were not detected at all. TCA degradation to CO₂ was indicated also in the spruce needles. Moreover, it was found that soil litter contained [1,2-¹⁴C]TCA unavailable to microorganisms.

A full length cDNA (designated CrETR1) was isolated by polymerase chain reaction amplification of a cDNA library from periwinkle (Catharanthus roseus) cell cultures. CrETR1 cDNA encodes a polypeptide of 740 amino acids with a predicted molecular mass of 82 kDa. The deduced protein contains a hydrophobic ethylene-binding transmembrane region, a GAF domain, a third domain homologous to the histidine protein kinase domain of the prokaryotic two-component systems, and a fourth carboxyl-terminal domain homologous to the receiver domain of the response regulators, as found in the A. thaliana ethylene receptor ETR1. CrETR1 transcripts are strongly accumulated in petals and ovaries of C. roseus young plants whereas no significant changes are detected in cell cultures submitted to various stress or hormonal (including ethylene) treatments. The amount of the monoterpene indole alkaloid ajmalicine in the cells treated by ethylene is reduced after addition of inhibitors of histidine kinases showing a possible involvement of CrETR1 protein in the ethylene-related signalling pathway leading to alkaloid biosynthesis enhancement in C. roseus cell cultures.

An increase in exocarp peroxidase activity was observed in fruit at 5 to 30 days post pollination (DPP), and decreased at 40 and 50 DPP. Total peroxidase activity of the mesocarp was significantly lower than the exocarp in all developmental stages. Mesocarp peroxidase activity decreased consecutively from outer, to middle and, to inner tissue at every developmental stage. Total activity in the mesocarp peaked at 20 DPP. Native-PAGE of exocarp tissue showed at least two cathodic (basic) peroxidases and two anionic (acidic) peroxidases. The number of isozymes was greatest and bands most intense at 30 DPP. IEF-PAGE of the 5 to 50 DPP fruit exocarp showed at least 8 peroxidase isozymes (pI 4.6 to 9.6). Anion exchange chromatography showed only one peak of anionic peroxidase activity that was not evident until 15 DPP. This peak was greatest at 30 DPP and declined at 40 and 50 DPP. Cationic peroxidase isozymes appeared to be the predominant and most intense isoforms throughout fruit development. The changes in peroxidase activity corresponded to fruit formation and may be associated with susceptibility to fruit rot.

The lignin contents and anatomical structure of roots of wild cherry (Prunus avium L.) and pedunculate oak (Quercus robur L.) plantlets were compared to explain differences in response during transfer from in vitro to ex vitro conditions. Lignification of cell walls increased significantly in both oak and cherry roots during the period of acclimation and finally lignin content of root tissues of in vitro propagated plantlets reached the levels not significantly different from seedlings grown in soil. Later on when secondary tissues appeared, lignified secondary xylem constituted most of the tissues of both species. The most conspicuous interspecific difference in root structure was the presence of phi-thickenings in cortical layers just outer to endodermis in cherry roots cultivated ex vitro. Formation of phi-thickenings was avoided in vitro and their presence thus seems to be under environmental control. Suberised well established exodermis was present in roots of oak but not detected in those of cherry. Very early development of exodermis in oak roots, preceding suberisation of endodermis, was recorded in vitro but not in well aerated soil. While multilayered and well-developed cork occurred in oak, only thin walled and less suberised secondary dermal tissues were found in cherry.

A modified plant 7SL RNA gene from Arabidopsis thaliana designated AHIIA63M was introduced into potato plants via Agrobacterium-mediated transformation. No transgenic plants could be obtained using pGPTV-based binary vectors where AHIIA63M gene driven by polIII promoter was located close to the polII promoter of the selection gene. Special binary vectors with matrix attachment region (MAR) elements had to be used for transformation to insulate polII and polIII promoters within T-DNA. The level of AHIIA63M RNA in transgenic plants was lower than the levels of transcripts of transgenes driven by RNA polymerase II. The level of AHIIA63M transcript in transgenic potato plants was tissue specific. The highest expression was detected in roots and gynoecium and the lowest in tubers. Moreover, non-specific promoter activity within the MAR element was revealed. This activity contributed to AHIIA63M transcription. This is the first report of expression of a modified 7SL RNA gene in transgenic plants and promoter activity within the MAR element.

Corchorus olitorius plants treated by 5 μg cm^-3 of Cd, Pb, Al or Cu in hydroponic culture accumulated in leaves 190, 150, 350 and 325 μg g^-1(d.m.) of these metals, respectively, after 6 d of exposure. Exposure of Corchorus plants to tested metals resulted in a sharp rise in content of amino acids in leaf tissues, however the magnitude of accumulation was different from one metal to another. Presence of sulphur in the growth medium significantly increased uptake of Cd and Pb and cysteine (cyst) was more effective than K₂SO₄. Similarly, addition of salicylic acid (SA) in the growth medium significantly enhanced the ability of Corchorus plants to accumulate all these metals. Growth of Corchorus plants was significantly reduced by treatment with any of the four metals except Cu and added cyst, K₂SO₄ or SA alleviated the growth retarding effect of metals.