Transformation of fenugreek (Trigonella foenumgraecum) was carried out with A281 oncogenic strain of Agrobacterium tumefaciens using root, cotyledon and hypocotyl explants excised from 1-week-old seedlings, which showed that the plant was highly susceptible to transformation. Tumors (calli) were selected on 50 mg dm^-3 kanamycin. They were analyzed for β-glucuronidase (GUS) expression. Presence of uidA (gus) gene, was confirmed by polymerase chain reaction (PCR) amplification.

Tobacco leaf discs were transformed with a plasmid pBIBnNHX1, containing the selectable marker neomycin phosphotransferase gene (nptII) and Na⁺/H⁺ vacuolar antiporter gene from Brassica napus (BnNHX1), via Agrobacterium tumefaciens-mediated transformation. Thirty-two independent transgenic plants were regenerated. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that the BnNHX1 gene had integrated into plant genome and Northern blot analysis revealed the transgene expression at various levels in transgenic plants. Transgenic plants expressing BnNHX1 had enhanced salt tolerance and could grow and produce seeds normally in the presence of 200 mM NaCl. Analysis for the T₁ progenies derived from seven independent transgenic primary transformants expressing BnNHX1 showed that the transgenes in most tested independent T₁ lines were inherited at Mendelian 3:1 segregation ratios. Transgenic T₁ progenies could express _BnNHX1 and had salt tolerance at levels comparable to their T₀ parental lines. This study implicates that the BnNHX1 gene represents a promising candidate in the development of crops for enhanced salt tolerance by genetic engineering.

1-Methylcyclopropene (1-MCP) applied alone did not influence significantly the chlorophyll and carotenoid content of the older leaves of Arabidopsis thaliana (L.) Heynh., but retarded the senescence of the younger ones (6^th and 7^th leaf nodes). However, 1-MCP effectively blocks the ethylene induced senescence of excised rosette leaves. The preliminary application of 1-MCP (3 h in advance to the treatment by Ethrel) almost totally eliminated the ethylene action. Similar trend was also observed after simultaneous application of Ethrel and 1-MCP, and the effects of both treatments on the chlorophyll and carotenoid destruction are comparable.

Strawberry plants (Fragaria×ananassa Duch.) cvs. Nyoho and Toyonoka were exposed to temperatures of 20, 33, and 42 °C for 4 h, and protein patterns in leaves and flowers was analyzed by 2-dimensional polyacrylamide gel electrophoresis and immunoblotting. In leaves and flowers of both cultivars, the content of most proteins decreased, but a few new proteins appeared in response to heat stress. These heat shock proteins (Hsps) were detected in the range of 19 - 29 kDa in leaves, and 16 - 26 kDa in flowers. The intensity of a 43 kDa protein spot increased in response to heat stress in Nyoho flowers, but not in Toyonoka flowers. The peaHsp17.7 antibody recognized one band at approximately 26 kDa in leaves, and two bands at approximately 16 and 17 kDa in flowers of both cultivars. These results show that the effects of heat stress on Hsp synthesis in strawberry plants differ between plant organs and between cultivars.

In grapevine (Vitis vinifera L.) leaf chlorophyll (Chl) a and Chl b and carotenoid contents were higher in plants grown at low photon flux densities (PFD) than in those grown at medium and high PFD. The highest Chl a variable to maximum fluorescence ratio F_v/F_m was observed in plants grown at medium PFD while the minimum fluorescence F₀ was highest in those at high PFD. In isolated thylakoids, both high and low PFD caused marked inhibition of whole chain and photosystem 2 (PS2) activities. The artificial exogenous electron donor diphenyl carbazide significantly restored the loss of PS2 activity in low PFD leaves.

Diurnal changes of sucrose-phosphate synthase (SPS) activity in different seasons were measured in Prosopis juliflora (Swartz) DC. leaves. SPS activity showed large variations with two distinct peaks, one around 09:00 and another at 21:00. The diurnal pattern was apparently not due to circadian rhythm since the activities were directly related to the changes in environmental parameters (irradiance, temperature, and leaf to air vapour pressure deficit) in different seasons. During the day, the enzyme showed changes in kinetic properties, differential sensitivity to allosteric modulators, differential response to ATP concentration, to concentration of endogenous sucrose, and to protein phosphatase inhibitors. These results taken together indicate the modulation of SPS in synchrony with photosynthesis and suggest the existence of multiple levels of modulation, presumably as an adaptive response to changing environmental extremes.

Understanding the molecular basis of polarity induction in plant cells is a research aspect that extends from signal perception and transduction to morphogenesis. A gradient of cytoplasmic ion fluxes generated through ion channels plays a crucial role in subsequent events leading to polar growth. Convincing evidence is now available implicating temporal and spatial distribution of Ca²⁺ in cytoplasm, generated by localized activity of calcium channels, as the early biochemical events associated with polarity induction. Ion channel antagonists are common tools for studying ion channel structure and function. Coupled with a fluorescent dyes, calcium channel antagonists (phenylalkylamine and dihydropyridine), have been used to localize L-type calcium channels. Additionally, the advent of Confocal Laser Scanning Microscopy has made possible the visualization of Ca²⁺ channels in plant cells. Persisting problems of dye loading and their cellular compartmentation have been addressed by developing a variety of experimental protocols. Present article highlights the current state of our understanding of these concepts, methodologies and their applications in different aspects of plant development.

Oat (Avena sativa L.) plants cultured in soil and hydroponic culture were treated with cadmium [0.154 mg g^-1 (dry soil) and 100 μM CdSO₄, respectively] for 21 d and growth rate and various biochemical processes were studied. Applied cadmium reduced plant growth and chlorophyll content. Changes in activity of enzymes involved in C, N and S metabolism and in guaiacol peroxidase activity were observed. In particular, O-acetylserine sulphydrylase (OASS; EC 4.2.99.8) activity was increased by Cd exposure in both growth conditions, probably as a resistance mechanism to cadmium based on the production of phytochelatins. Results show that both field and hydroponic conditions represent suitable systems for investigating Cd effects on plant growth and metabolism.

Maize (Zea mays ssp. mays) and eastern gamagrass (Tripsacum dactyloides) are known for their susceptibility to chilling injuries. Their hybrid (Z. mays × T. dactyloides) showed higher tolerance to low temperatures (-2 °C) in the field than its parents. Exposure to 5 °C for 2 or 3 d reduced the variable to maximal chlorophyll fluorescence ratio (F_V/F_M), an indicator of the maximum photochemical efficiency of the photosystem 2, and the variable to minimal fluorescence ratio (F_V/F₀) more in maize and eastern gamagrass than in hybrid plants. Chlorophyll contents for rewarming plants (25 °C for 3 d) were lower than before chilling in both parents while values for hybrid plants were similar. Electrolyte leakage was higher in chilled than control plants but it did not show significant differences among genotypes. Our data suggest that hybrid plants have higher capacity to recover from chilling injury in controlled conditions than their parents.

The effect of applied arachidonic acid, prostaglandin (PGE₁) and various sterols and combinations of arachidonic acid + sterols, on flowering of Pharbitis nil were ascertained by using a tissue culture technique. It was found that arachidonic acid, PGE₁ stigmasterol, testosterone, cholesterol, stigmasterol + arachidonic acid, β-sitosterol + arachidonic acid and cholesterol + arachidonic acid all caused earlier flowering. Four inhibitors of prostaglandin biosynthesis (gentisic acid, acetylsalicylic acid, salicylic acid and oleic acid), inhibited flowering completely. The results confirm that the compounds tested could possibly play a role in the flowering of P. nil.

The effect of acute ozone (O₃) fumigation on isozyme patterns of superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) in mature (ML) and young leaves (YL) of two poplar clones, contrasting in O₃-sensitivity was analysed. Untreated leaves of both the O₃-sensitive (O₃-S) clone Eridano of Populus deltoides×P. maximowiczii and the O₃-resistant (O₃-R) clone I-214 of P.×euramericana showed four distinct SOD isoforms with a relative mobility (R_f) of 0.54 (MnSOD), 0.60 (Cu/ZnSOD), 0.65 (unidentified), and 0.71 (Cu/ZnSOD). After O₃-fumigation the activity of the SOD isoforms showed only quantitative variations with respect to control plants. In ML of untreated O₃-R plants seven POD isoforms (R_f= 0.13, 0.19, 0.34, 0.59, 0.64, 0.70 and 0.75) were found, while in YL one isoform (R_f= 0.34) was undetected. Only three POD isoforms in both ML and YL of untreated O₃-S plants were resolved. The electrophoretic pattern of POD in O₃-S leaves was greatly modified by acute O₃-fumigation with the appearance of new isoforms in both YL and ML and the disappearance of an isoform (R_f= 0.13) in YL. Additionally, O₃-exposure induced the appearance of two APX isoforms in YL (R_f= 0.66 and 0.70), and one isoform in ML (R_f= 0.70) of the O₃-S clone. By contrast, the activity of the three APX isoformes (R_f= 0.64, 0.70 and 0.76) detected in O₃-R leaves showed only quantitative variation with respect to untreated plants. From these data it is concluded that: 1) in these poplar hybrids antioxidant enzyme activity is developmentally regulated and greatly affected by acute O₃ stress treatments and 2) the different enzymes activity displayed by the two poplar clones, especially for POD and APX isoformes, could partly explain their distinct O₃-sensitivity.

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.

Thirteen flavonoid aglycons, contained in the strongly allelopathic epicuticular exudates of Dittrichia viscosa, were investigated for their effects on lettuce seedling radicle growth. Concerning radicle length and mass, variable results were obtained, with most of the substances having no effect, some being inhibitory and some even promotive. Shoot mass was slightly reduced in four cases. Seed germination rates, root hair and lateral root formation were not affected either. Three of the compounds (namely quercetin 3,3-dimethylether, naringenin and eriodictyol) induced a strong ageotropic response in radicle growth.

Net photosynthetic rate (P_N), transpiration rate (E), and stomatal conductance (g_s) during water stress and after rehydration were measured in Phaseolus vulgaris, Beta vulgaris, and Zea mays. Immediately before imposition of water stress by cessation of watering, plants were irrigated with water (control), 100 μM abscisic acid (ABA), and/or 10 μM N⁶-benzyladenine (BA). In all three species, application of ABA decreased g_s, E, and P_N already 1 h after application. However, during water stress g_s, E, and P_N in plants pre-treated with ABA remained higher than in plants pre-treated with water. Positive effects of ABA application were observed also after rehydration. In contrast, the effects of pre-treatment with BA were species-specific. While in bean plants BA application ameliorated negative effect of water stress, only very slight effects were observed in maize, and in sugar beet BA even aggravated the effects of water stress.

Carob tree (Cerafoma siliqua L.) micropropagated shoots were rooted on half-strength Murashige and Skoog medium, supplemented with different types and concentrations of sugars, in order to determine the effects of sugar composition and concentration on in vitro rooting and in vivo establishment of the plantlets. Among the various sugars tested, the best rooting response was obtained with 145 mM sucrose, both in terms of rooting frequency and index of rooting. The use of filter-sterilized rather that autoclaved fructose increased root number and root length. Sugar treatment during rooting slightly influenced plantlet survival and growth during acclimatization. A reduction in the glucose concentration during rooting was beneficial for plantlet acclimatization.

The in vitro cultures of apple rootstock MM 106 produced the highest fresh mass (FM) when 0.1 mM B was included in the culture medium. By increasing B concentration of the culture medium from 0.1 to 6.0 mM, FM and contents of B, P, Ca, and Mg in explants increased, whereas K, Fe, Mn, and Zn contents decreased. SPAD units of leaves characterizing chlorophyll contents declined as B concentration of the culture medium increased from 0.1 to 6.0 mM.

Nodal explant cultures from field-grown five jojoba genotypes (EC 99690, EC 99692, EC 99692, EC 267779 and EC 171284; male and female plants), could be established on Murashige and Skoog (MS) medium. The nodal explants of different genotypes as well as sex elicited differential requirements of N⁶-benzyladenine (BA) for optimum shoot regeneration and medium-term conservation. Female nodal explants of EC 99692 produced maximum shoots (10 shoots per explant) followed by male of EC 171284 (9.3 shoots per explant) on MS + 10 μM BA. The pulse treatment of 50 μM indole-3-butyric acid for 20 min caused in vitro rhizogenesis in 44 - 67 % cultures of various genotypes tested. A significant difference was observed for the conservation period of male and female cultures of all the genotypes. MS + 10 μM BA supported the shoot cultures of EC 99690, EC 99691 and EC 267779 for maximum conservation period, while MS + 5 μM BA proved to be optimum for conserving the shoots of EC 99692 and EC 171284. Generally, the female shoot cultures of genotypes survived for longer period than male ones.

Direct shoot bud differentiation was achieved in leaf segments of Achras sapota cv. Cricket Ball inoculated on Schenk and Hildebrandt's medium supplemented with 5.0 μM thidiazuron and 8.88 μM benzylaminopurine. Leaves from middle part of the shoots and segments obtained from middle portion of leaf showed highest potential to regenerate shoot buds. Histological examination of developing shoot buds showed their de novo regeneration with clear vascular connection with the mother tissues.

Plant regeneration through indirect somatic embryogenesis was attempted from leaf, internode, node and shoot-tip derived callus of Leptadenia reticulata. Somatic embryos at the highest frequency was induced on Murashige and Skoog (MS) medium supplemented with 8.87 μM benzyladenine (BA) and 2.46 μM indole-3-butyric acid (IBA). From different explants, only shoot-tip and node explant derived calli induced somatic embryos. Transfer of the embryogenic callus to suspension cultures of the same concentration of growth regulators facilitated the development of embryos. Suspension cultures with reduced concentration of BA (2.22 μM) either alone or in combination with 0.49 μM IBA fostered maturation of embryos. Half-strength MS solid medium with 1.44 μM GA₃ and BA (0.22 or 0.44 μM) facilitated conversion of embryos into plantlets at higher rate compared to that on with BA alone. About 77 plantlets were recovered from 10 mg callus. Plantlets transferred to small cups and subsequently to field survived in 80 %. All the plantlets established in the field exhibited morphological characters similar to that of the mother plant.

Fatty acid ω-3 desaturase (FAD) is the key enzyme catalyzing the formation of trienoic fatty acids. We utilized an Arabidopsis FAD7 gene and the seven independent transgenic rice plants harbouring 1 to 3 copies of this gene were generated. The expression of FAD7 mRNA was different among independent transgenic lines regardless of the copy number. The total linolenic acid (18:3) contents reduced by about 7 - 32 % in transgenic rice plants but the linoleic acid (18:2) content increased accordingly. With or without wounding treatments, the jasmonate content was higher in transgenic lines than in wild-type rice plant. The transgenic lines overproducing jasmonate also showed increased expression of PR1b mRNA and allene oxide synthase inresponse to wounding.

Hypocotyl, cotyledon and cotyledonary node explants of Calendula officinalis L were cultured on Murashige and Skoog (MS) media supplemented with various concentrations of thidiazuron (TDZ), kinetin (KIN), α-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA) to induce adventitious shoot regeneration and micropropagation. The highest frequency of adventitious shoot regeneration was achieved from hypocotyl and cotyledon explants on MS media supplemented with 0.75 mg dm^-3 TDZ and either 0.25 or 0.50 mg dm^-3 IBA. Efficient in vitro clonal propagation was also induced from cotyledonary nodes on a range of media supplemented with 0.75 mg dm^-3 TDZ and 0.05 mg dm^-3 NAA or 2 mg dm^-3 KIN and 1 mg dm^-3 NAA. Regenerated shoots were excised and rooted in MS medium supplemented with 1 mg dm^-3 NAA. The rooted plantlets were finally transferred to pots.

The effects of drought and rehydration on tea seedlings were significant. After five days of drought imposition the contents of chlorophylls, carotenoids, ascorbate and glutathione, and activities of guaiacol peroxidase and glutathione reductase decreased. Simultaneously, contents of proline, H₂O₂ and superoxide anion, lipid peroxidation and activities of catalase and superoxide dismutase increased. These parameters recovered to different degrees during subsequent rehydration.

Enzyme changes in non-rooted (treated with Fe-EDTA) and rooted (treated with Fe-EDDHA) stems of rootstock GF-677 (Prunus amygdalus×P. persica) during adventitious root formation in vitro have been recorded. The first roots appeared approximately after 12 d on the rooting medium. By contrast to non-rooted stems, rooted stems showed a maximum of soluble peroxidase activity on the 9^th day, of ionically bound peroxidase to cell wall on the 6^th and 12^th day and of catalase on the 6^th and the 15^th day. A time course study of changes of soluble peroxidases isoenzymes showed that there was a band visible only in the rooted stems and also a new band appeared three days before the emergence of roots.

Thermotolerance of pearl millet (Pennisetum glaucum cv. ICMV-94133) and maize (Zea mays cv. Golden) was assessed at germination and vegetative stage. Final percentage of germinated seeds and rate of germination (number of days to 50 % germination) decreased due to high temperature (45 °C) similarly in the both species. In contrast, at the vegetative stage, high temperature (38/27 °C) caused a significant reduction in shoot dry mass of maize, whereas this attribute remained almost unchanged in pearl millet. Relative growth rate and net assimilation rate (NAR) increased significantly in pearl millet due to high temperature, but in contrast, in maize NAR was slightly reduced. Concentrations of N, P, and K in the shoots of both species increased at high temperature, but N accumulation was more pronounced in pearl millet than in maize. High temperature caused a marked increase in both shoot and root Ca²⁺ concentration in maize, but it did not affect that of pearl millet. S concentration in the shoots of maize decreased significantly due to high temperature, whereas that in pearl millet remained unaffected. Shoot Na⁺ concentration of both species was not significantly affected by high temperature. High temperature caused a significant increase in uptake of N, P, and K⁺ in pearl millet, but the uptake of Ca²⁺, Mg²⁺, Na⁺ and S remained unaffected in this species. In contrast, in maize, a significant increase in uptake of K⁺ and Ca²⁺, and a decrease in uptake of N, S, Mg²⁺, and Na⁺ were found at high temperature. Overall, maize showed lower tolerance to high temperature compared with pearl millet.