Effects of long-term sodium chloride salinity (100 and 200 mM NaCl; ECe = 6.85 and 12.3 dS m^-1) were studied in tolerant (Kharchia 65, KRL 19) and susceptible (HD 2009, HD 2687) wheat genotypes. NaCl decreased relative water content (RWC), chlorophyll content (Chl), membrane stability index (MSI) and ascorbic acid (AA) content, and increased the contents of hydrogen peroxide, thiobarbituric acid reactive substances (TBARS), and activities of superoxide dismutase (SOD), ascorbate peroxidase (APOX) and glutathione reductase (GR). Kharchia 65 showed lowest decline in RWC, Chl, MSI and AA content, lowest increase in H₂O₂ and TBARS contents and higher increase in SOD and its isozymes, APOX and GR, while HD2687 showed the highest decrease in AA content, highest increase in H₂O₂ and TBARS contents and smallest increase in activities of antioxidant enzymes. KRL 19 and HD 2009 showed intermediate response both in terms of oxidative stress and antioxidant activity.

This study reports survival and physiological responses of micropropagated Ceratonia siliqua L. cvs. Galhosa and Mulata plants during ex vitro acclimatization under ambient (AC; 330 μmol mol^-1) or elevated (EC; 810 μmol mol^-1) CO₂ concentration and a photosynthetic photon flux density of 125 μmol m^-2 s^-1. CO₂ enrichment during acclimatization did not improve survival rate that was around 80 % for both treatments. Eight weeks after ex vitro transplantation, photosynthetic capacity and apparent quantum yield in acclimatized leaves were higher in comparison with those in in vitro-grown leaves, without any significant difference between CO₂ treatments. Chlorophyll content increased after acclimatization. However, EC led to a decrease in the total amount of chlorophyll in new leaves of both cultivars, compared to those grown at AC. Soluble sugars and starch contents were not markedly affected by growth EC, although starch had significantly increased after transfer to ex vitro conditions. EC induced an increase in the stem elongation and in the effective life of leaves, and a decrease in the number of new leaves.

Various ecophysiological investigations on carnivorous plants in wet soils are presented. Radial oxygen loss from roots of Droseraceae to an anoxic medium was relatively low 0.02 - 0.07 μmol(O₂) m^{- 2} s^-1 in the apical zone, while values of about one order of magnitude greater were found in both Sarracenia rubra roots and Genlisea violacea traps. Aerobic respiration rates were in the range of 1.6 - 5.6 μmol kg^-1 (f.m.) s^-1 for apical root segments of seven carnivorous plant species and 0.4 - 1.1 μmol kg^-1 (f.m.) s^-1 for Genlisea traps. The rate of anaerobic fermentation in roots of two Drosera species was only 5 - 14 % of the aerobic respiration. Neither 0.2 mM NaN₃ nor 0.5 mM KCN influenced respiration rate of roots and traps. In all species, the proportion of cyanide-resistant respiration was high and amounted to 65 - 89 % of the total value. Mean rates of water exudation from excised roots of 12 species ranged between 0.4 - 336 mm 3 kg^-1 (f.m.) s^-1 with the highest values being found in the Droseraceae. Exudation from roots was insensitive to respiration inhibitors. No significant difference was found between exudation rates from roots growing in situ in anoxic soil and those kept in an aerated aquatic medium. Carnivorous plant roots appear to be physiologically very active and well adapted to endure permanent soil anoxia.

A high frequency adventitious shoot regeneration protocol was developed for henbane (Hyoscyamus niger L.) using thidiazuron (TDZ). Hypocotyl, cotyledon and stem explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of N⁶-benzylaminopurine and TDZ. MS medium supplemented with 16 μM TDZ was the most effective for providing 100 % regeneration frequency associated with a 19.53 shoots per hypocotyl explant. Plantlets were rooted on MS medium supplemented with different concentrations of indole-3-butyric acid (IBA) and α-naphthaleneacetic acid. High rooting and survival was achieved using half strength MS medium supplemented with 8 μM IBA.

Tobacco leaf disc explants were inoculated with Agrobacterum tumefaciens strain GV2260 carrying p35S GUS-INT to determine the influence of different co-cultivation temperatures (18 - 26 °C), periods (24 - 96 h) and media (solid and liquid) on transformation efficiency. Kanamycin-resistant shoots developed on leaf discs inoculated with Agrobacterium after 4 weeks of culture initiation. Regenerated shoots were excised and rooted in the basal medium supplemented with 100 mg dm ^-3 kanamycin and 250 mg dm ^-3 augmentin. The rooted plantlets were finally transferred to compost and confirmed by GUS assay and PCR analysis. The highest transformation frequency was achieved from the explants co-cultivated with A. tumefaciens in liquid medium for 48 h at 22 or 24 °C.

Floral phenology, pollen quality and seed set of Plantago crassifolia plants, grown in the presence of increasing NaCl concentrations, were studied to test how this Mediterranean halophyte responded to salt stress during the reproductive phase of its life cycle. "Reproductive success" was maximal in plants grown in non-saline conditions, or in the presence of 100 mM NaCl, but it was negatively affected by higher salinities, due to a progressive reduction of pollen fertility, seed set, and seed viability.

The effects of NaCl (0, 50, 100, 150 and 200 mM) on growth, water relations, glycinebetaine, free proline, ion contents, stomata number and size of Kochia prostrata (L.) Schard were determined. Shoot and root fresh and dry matter, root and shoot length, relative growth rate, net assimilation rate, relative water content, water use efficiency, soluble sugars and glycinebetaine contents were not changed at low NaCl concentrations, but they were significantly decreased at 200 mM NaCl. The K⁺, Mg²⁺ and Ca²⁺ contents, water potential, chlorophyll a+b and carotenoides contents, and stomata number and size were reduced already at low concentrations of NaCl. In contrast, the Na⁺, Cl^- and proline contents increased several times with increasing NaCl concentration. Kochia prostrata is a salt tolerant species, the optimal growth of this plant occurred up to 150 mM NaCl. The mechanisms of salt tolerance in the plant may be balance among ion accumulation and production of glycinebetaine, proline, soluble sugars for maintenance of pressure potential.

The activities of sucrose phosphate synthase (SPS), sucrose synthase (SUSY), neutral invertase (NI) and soluble acid invertase (SAI) were measured in callus cultures of four Mexican sugarcane cultivars (Saccharum spp.) with a different capacity to accumulate sucrose in stem parenchyma cells. The results indicated that sucrose accumulation in callus was positively correlated to the activity of SPS and SUSY and negatively to the activity of SAI and NI while SPS explained most of the variation found for sucrose accumulation and NI least.

In order to establish an efficient system for in vitro plant regeneration of a short day plant Chenopodium rubrum L. and a long day plant Chenopodium murale L., optimum culture conditions for somatic embryogenesis were investigated. The effects of different growth regulators, their combination and their concentrations on somatic embryos induction in different explant types (root, hypocotyl, cotyledon and leaf) were tested. Somatic embryogenesis was induced in both plants on Murashige and Skoog (MS) medium supplemented with sucrose (3 %), agar (0.7 %) and 1 - 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D) as the sole growth regulator. The largest embryogenic capacity was found in root explants of Chenopodium rubrum on 1 μM 2,4-D and in basal parts of cotyledons in C. murale plants on 10 μM 2,4-D.

Shoot cultures of Alyssum markgrafii O.E. Shulz, endemic nickel hyperaccumulating species of central Balkan, were established and maintained on Murashige and Skoog medium supplemented with 0.2 mg dm^-3 benzyladenine (BA). Nickel in form of NiCl₂ . 6 H₂O was supplemented at 22 different concentrations ranging from 0.0001 to 15 mM but none of them was lethal to cultures. High Ni²⁺ concentrations (10 mM or more) arrested shoot growth which, upon transfer to Ni-free medium, commenced via axillary bud proliferation. Shoots that developed from axillary buds through the subculture manifested increased tolerance to Ni²⁺ expressed as shoot elongation. Shoot multiplication and dry biomass production decreased with increase of Ni²⁺ in medium. Only the accumulation of Ni²⁺ in tissues increased with Ni²⁺ content of the medium. Apart from shoot cultures, high Ni²⁺ accumulation was registered in undifferentiated callus cultured on medium with 0.5 mg dm^-3 BA and 0.5 mg dm^-3 naphthylacetic acid. Highest content of accumulated Ni was 2.37 μg g^-1 (d.m.) in shoots and 2.65 μg g^-1 (d.m.) in callus, both measured on medium with 15 mM Ni²⁺.

Based on the NH₂-terminal sequence of three PR-10 isoforms previously identified in Lupinus albus leaves and a conserved amino-acid region in the PR-10 proteins from leguminosae, a pair of oligonucleotides was designed and used to amplify the corresponding cDNA fragment from a L. albus leaves cDNA library. A fragment of DNA of 200 bp was isolated from the polymerase chain reaction (PCR) mixture and subsequently used to screen the cDNA library. A cDNA coding for a PR-10 protein of 158 amino acid residues was cloned and sequenced. Subsequent studies involving Northern and Western blot analysis have shown that the PR-10 protein isoforms are differentially expressed during the development of the healthy lupin plant. High mRNA and protein contents were detected in roots and hypocotyls of both 7- and 20-d-old plants. In young leaves, the mRNA and protein contents were low and increasead in mature leaves. Tissue printing experiments with root sections suggest that the proteins are extracellular and are mainly associated with the vascular tissues in mature roots.

Experiments were performed to optimize the macronutrients concentrations for in vitro rooting of Ceratonia siliqua micropropagated shoots. Several dilutions of Murashige and Skoog (MS) medium were tested: full-strength MS, half-strength MS (1/2MS), and 1/2MS + full N. The frequency of in vitro rooting was enhanced when the 1/2MS was used (50 % rooted shoots). Mature leaves from 20 - 30 year-old carob trees and from 2 year-old micropropagated plants were collected and the concentrations of macronutrients (N, P, K, Ca, Mg) assessed. Based on the mineral composition of the leaves a new medium was formulated and compared with the previous ones showing an increment of the rooting frequency to 80 %. Moreover, shoots rooted in the new medium did not show symptoms of apical necrosis that occurred in the other tested media.

We describe the multi-step regeneration system of medicinal plant Schisandra chinensis (Turcz.) Baill. The seeds were pre-treated with 0.005 μM thidiazuron. Subsequently the zygotic embryos of the early heart stage were cultured on medium with 50 μM of 2,4-dichlorophenoxyacetic acid (2,4-D) and after three weeks the embryogenic calli were transferred to a medium with 10 μM of 2,4-D and 4 μM of 6-benzyladenine and were sub-cultured at the 4-week intervals. Abscisic acid (30 μM) and polyethyleneglycol (3 %) significantly influenced the synchronization of development of the somatic embryos (SEs) to the globular stage. The following culture on a medium without growth regulators resulted in full developed cotyledonary stage SEs. Indole-3-butyric acid (0.05 μ) contributed to their rapid conversion to plantlets.

Potted two-year-old lemon trees [Citrus limon (L.) Burm. f.], cv. Verna grafted on sour orange (C. aurantium L.) rootstock, growing in greenhouse, were subjected to drought for 33 d. Control plants were daily irrigated at field capacity. Values of sap flow (SF) were compared with transpiration (E) rates measured gravimetrically. The results underlined the robustness and high sensitivity of the compensation heat-pulse technique for estimating transpiration on a wide range of SF. Good direct correlations between E and SF rates on an instantaneous and daily basis were obtained in both treatments. On a daily basis, a common calibration curve can be used for both irrigation treatments. On an instantaneous basis, changes in SF were matches by similar changes in E in both treatments, although the relationships between these parameters presented different intercepts in each treatment. Sap flow rates were influenced by weather conditions in trees growing in non-limiting soil water conditions. This makes it possible to evaluate the significance of any sap flow measurement in relation to the reference value calculated for the vapour pressure deficit at the time the measurement was taken.

The effects of zirconium ascorbate (Zr-ASC), a water-soluble complex of Zr, were examined on wheat seedlings (Triticum aestivum L. cv. MV. 20). Hydroponically grown plants were exposed to 10, 33, 55, 100 and 550 µM Zr-ASC (Zr₁₀, Zr₃₃etc.). After 9 d of treatment inhibition of germination, retarded root and shoot growth, and increased activities of antioxidant enzymes (guaiacol peroxidase, ascorbate peroxidase, and glutathione reductase) showed that Zr-ASC was only harmful at and over a concentration limit of 100 µM. Chlorophyll (Chl) content of plants was only decreased by Zr₅₅₀. Zr-ASC at lower concentrations was beneficial for plant development: Zr₁₀ and Zr₃₃ enhanced root elongation, Zr₅₅ induced about 30 % increase in the total Chl content, while the activity of antioxidant enzymes was not elevated indicating that no oxidative stress was generated by the intracellularly accumulated Zr⁴⁺ ions.

The aim of this work was to assess the responses of seedlings of five species of Nicotiana genus to red and far red radiation. N. acuminata exhibits positive photoblastic behaviour and germination was completely inhibited under far red and darkness. In N. glauca germination was reduced under far red and darkness, but the other species showed neutral behaviour. The hypocotyl elongation was inhibited in N. glauca and N. tabacum under white and far red radiation. In N. langsdorffii and N. debneyi hypocotyl was elongated under far red radiation. Only in N. acuminata red radiation promote greater hypocotyl elongation than dark condition. On the phylogenetic tree obtained from restriction analysis N. glauca and N. acuminata are grouped in one branch, while the other species, N. langsdorffii, N. debneyi and N. tabacum, are grouped in the other branch cluster. Moreover, the N. debneyi behaviours under different radiation treatments were similar to those of N. tabacum. These two species are allopolyploid members of the genus Nicotiana, as also was confirmed by this study.

It is suggested that cathodal esterase isoenzyme (E₁) might be used as a marker for the VA 1 gene on 7 S maize chromosome and for the "B" linkage group of Tripsacum dactyloides in Zea mays L. ×T. dactyloides L. hybrids. The latter genic zones have a regulatory effect on fertility and on the apomictic mode of reproduction.

The effects of three cytokinins, kinetin 4.5 μM (Kin), 6-benzylaminopurine 4.5 μM (BA) and N-phenyl-N'1,2,3- thiadiazol-5-yl-urea 4.5 μM (TDZ), and the effects of different treatment duration on the regeneration of adventitious shoots from quince (Cydonia oblonga Mill.) leaves were studied. In a first experiment, leaves treated with Kin for 0, 8, 16 and 24 d were transferred to BA or TDZ-containing growth medium. In a second experiment TDZ applied for 0, 4, 8, 12, 16 and 24 d was followed by BA. All treatments included 0.5 μM α -naphthaleneacetic acid (NAA). In the sequence Kin-BA, the production of adventitious shoots decreased and reddish-coloured nodular structures (RNS) of meristematic appearance increased with increasing duration of Kin treatment, while somatic embryo formation was optimal at 8 d. In the Kin-TDZ sequence, shoot production was initially pronounced, but it declined with increasing duration of the Kin treatment, while the number of roots, somatic embryos and RNS increased. TDZ-BA treatments induced marked shoot production, which gradually increased with increasing duration of TDZ treatment. The presence of TDZ and a long treatment duration appeared to be very important factors in inducing caulogenesis. Kin appeared to be effective in shoot induction but not in shoot development; the results of this work demonstrate that RNS were adventitious shoots blocked at an early developmental stage on account of insufficient cytokinin activity. BA was less effective than TDZ in inducing shoot regeneration. Finally, both Kin and BA applied after 2,4-D treatment promoted somatic embryo induction.

The effect of NaCl on the growth and activity of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were investigated in the seedlings of four potato cultivars (Agria, Kennebec; relatively salt tolerant, Diamant and Ajax; relatively salt sensitive). The shoot fresh mass of Agria and Kennebec did not changed at 50 mM NaCl, whereas in Diamant and Ajax it decreased to 50 % of that in the controls. In Agria and Kennebec, SOD activity increased at 50 mM NaCl, but no significant changes observed in Diamant and Ajax. At higher NaCl concentration, SOD activity reduced in all cultivars. CAT and POD activities increased in all cultivars under salt stress. Unlike the other cultivars, in Ajax seedlings, APX activity increased in response to NaCl stress. We also observed new POD and SOD isoenzyme activities and changes in isoenzyme compositions under salt stress. These results suggest that salt-tolerant potato cultivars may have a better protection against reactive oxygen species (ROS) by increasing the activity of antioxidant enzymes (especially SOD) under salt stress.

A protocol of high frequency shoot organogenesis and plant establishment from stem derived callus has been developed for Tylophora indica (Burm. f.) Merrill. - an endangered medicinal plant. Callus was developed on Murashige and Skoog (MS) medium supplemented with 10 μM 2,4,5-trichlorophenoxy acetic acid (2,4,5-T). Multiple shoot induction was achieved from the surface of the callus after transferring onto shoot induction medium. The highest rate (80 %) of shoot multiplication was achieved on MS medium containing 5.0 μM kinetin. The developed shoots rooted best on half-strength MS medium supplemented with 0.5 μM indole-3-butyric acid (IBA). The in vitro raised plantlets with well developed shoot and roots were acclimatized successfully and grown in greenhouse.

Treating androgenic embryos of Aesculus hippocastanum L. with low temperatures (6 °C) improved their germination and regeneration into plantlets. The embryos derived from anther cultures showed better results than those derived from microspore cultures.

We have cloned and characterized the expression of Crocus sativus AGAMOUS1 (CsAG1), a putative C-type MADS-box gene homologous to AGAMOUS (AG) from a triploid monocot species crocus (Crocus sativus L.). The typical domain structure of MIKC-type plant MADS proteins was identified. Phylogenetic analysis of the deduced amino acid sequence indicated that the isolated gene forms a clade with the AGAMOUS homologs from the monocots Hyacinthus orientalis and Phalaenopsis equestris. A differential splicing event altering the amino acid sequence at the C terminus was identified, leading to the formation of two mRNAs differing ten nucleotides in size. The presence of both differentially spliced transcripts was restricted only to mature crocus flowers and particularly to stamens and carpels.

Lead nitrate at concentration of 150 mg dm^-3 inhibits root growth of Lupinus luteus seedlings by bout 20 %, which is accompanied by an increase of K⁺ leakage from the root cells. Non-denaturing isoelectric focusing in polyacrylamide slab gel has shown that lead stimulates the activity of most lipoxygenase isoenzymes and induces one additional isoenzyme with pI 6.9.

A Na⁺/H⁺ antiporter catalyzes the transport of Na⁺ and H⁺ across the tonoplast membrane. We isolated a vacuolar Na⁺/H⁺ antiporter cDNA (SsNHX1) clone from a euhalophyte, Suaeda salsa. The nuclear sequence contains 2262 bp with an open reading frame of 1665 bp. The deduced amino acid sequence is similar to that of AtNHX1 and OsNHX1 in rice, with the highest similarities within the predicted transmembrane segments and an amiloride-binding domain. Northern blot analysis shows that the expression of the S. salsa gene was increased by salt stress. The results suggest that the SsNHX1 product is likely a Na⁺/H⁺ antiporter and may play important roles in the salt tolerance of S. salsa.

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.