Apical parts of stems of Brassica napus L. var. oleifera cv. Gorczanski (winter rape) and cv. Mlochowski (spring rape), grown in vitro, were subjected to direct electric current (DC) of different polarity, duration and voltage. The positive orientation of DC, i.e. anode attached to the apical part and cathode to the medium, markedly enhanced the differentiation of the apical meristem in winter rape. The reverse polarity was without effect. DC treatment of positive polarity resulted in spring rape in transition of all explants to generative state while 70 % of non-treated plants remained at vegetative stage. Even negative orientation of DC brought about a rise in percentage of flowering plants with regard to control. The developmental effects of DC were dependent only to a low degree or not at all on duration and voltage of the treatment.

The structure and development of Myrtus communis L. secretory cavities has been studied in young and expanded leaves, using light and scanning electron microscope. Secretory cavities are continuously formed during leaf development, but in mature leaves the rhythm of their appearance shows steep decrease. Each secretory cavity is developed from a single epidermal cell, which undergoes a periclinal division followed by anticlinal and several oblique cell divisions. The lumen of the secretory cavity is initiated by cell wall separation, i.e., schizogenously. The secretory cells line the cavity, where the secreted material is collected. Secretory cavities are covered by modified epidermal cells, which do not seem to form any special aperture. Essential oils seem to be discharged after mechanical treatment of the leaf.

Fe deficiency was imposed in Citrus sinensis L. cultivars Valencia and New Hall grafted on C. aurantium and Swingle citrumelo rootstocks by the absence of Fe (-Fe) or by the presence of bicarbonate in the Hoagland nutrient solution. In Fe-deprived leaves total and active Fe concentration, and peroxidase and catalase activities were decreased while the ratios carotenoids/chlorophylls, P/Fe, and K/Ca were increased. Fe(III) chelate reductase activity was induced in (-Fe)-treated roots whereas it was depressed in bicarbonate-treated roots.

The effect of five antibiotics: carbenicillin, chloramphenicol, cefotaxime, kanamycin and hygromycin on the organogenesis from callus cultures of Coryphantha elphantidens (Lem.) Lem. have been studied. Carbenicillin and cefotaxime stimulated shoot regeneration from callus. All antibiotics under study suppressed rooting of in vitro formed shoots. After five sequential subcultures on kanamycin supplemented medium, antibiotic resistant callus was obtained. To study the impact of kanamycin on resistant callus, total protein content was also studied. Selected callus showed a remarkable increase in callus mass. Antibiotic resistant plants have been selected by screening callus pieces on kanamycin supplemented media. Total protein content increased with subsequent subcultures in kanamycin resistant callus. The kanamycin selected shoots withstood the stability test after 2 months on antibiotic free medium. Plants were raised from the callus, which formed roots in 20 mg dm^-3 kanamycin, which was under study.

The effects of photoperiod on the development of in vitro grown plantlets of yam (Dioscorea alata L.), were investigated. Plantlets were transplanted into pots, acclimatizated until they reached vegetative stages V₁ (3 leaves) or V₂ (8 leaves), and then grown under 12-h or 16-h photoperiod. The formation and development of underground tubers was only induced under 12-h photoperiod. Tuber initiation was not related to the initial vegetative stage of plants, and the tubers were visible at about 18 - 24 d. On the contrary, a 16-h photoperiod inhibited tuber formation and stimulated vine and leaf growth. The total dry matter production and the number of leaves per plant of V₁ stage plants were 50 and 30 % lower respectively, after 44 d under 12-h compared to 16-h photoperiod. These parameters were not influenced by photoperiod in V₂ stage plants. Consequently, the effect of 12-h photoperiod on dry matter of V₁ plants was attributed to a source limitation related to the early initiation of tuberization. The transfer of plants grown under 12-h to 16-h photoperiod stopped tuber growth and starch accumulation. On the other hand, it stimulated the shoots and the roots to grow.

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.

The effect of two N-forms (NH₄⁺ and NO₃^-) and NaCl on pattern of accumulation of some essential inorganic nutrients was examined in sunflower (Helianthus annuus L.) cv. Hisun 33. Eight-day-old plants of were subjected for 21 d to Hoagland's nutrient solution containing 8 mM N as NH₄⁺ or NO₃^., and salinized with and addition of NaCl to the growth medium had no significant effect on total leaf N. However, root N of NH₄-supplied plants decreased significantly with increase in NaCl concentration, whereas that of NO₃-supplied plants remained unaffected. There was no significant effect of NaCl on leaf or root P, but the NO₃-supplied plants had significa concentration of leaf P than that of NH₄-supplied plants at varying salt treatments. Salinity of the rooting med did not show any significant effect on Na⁺ concentrations of leaves or roots of plants subjected to two differen N. NH₄-treated plants generally had greater concentrations of Cl^- in leaves and roots and lower K⁺ content in leaves than NO₃-supplied plants. Ca²⁺ concentrations of leaves and roots and Mg²⁺ concentrations of leaves decreased in NH₄-supplied plants due to NaCl, but they remained unaffected in NO₃-treated plants.

In vitro root culture of yellow wort (Blackstonia perfoliata (L.) Huds.) was initiated on Murashige and Skoog (MS) medium. In the presence of benzylaminopurine (BAP) numerous adventitious buds formed, which developed into shoots. Presence of indole-3-butyric acid (IBA) in media significantly decreased number of buds, but increased development of lateral roots. On hormone-free medium shoots successfully rooted and developed flowers and viable seeds that formed another generation. Shoot cultures of B. perfoliata inoculated with suspension of Agrobacterium rhizogenes strain A4M70GUS developed hairy roots at 3 weeks and they were cultured on hormone-free MS medium. Spontaneous shoot regeneration occurred in 3 clones.

Abscisic acid (ABA) and salicylic acid (SA) were sprayed on leaves of wheat genotypes C 306 and Hira at 25 and 40 d after sowing under moderate water stress (-0.8 MPa) imposed by adding PEG-6000 in nutrient solution. ABA and SA increased the activities of superoxide dismutase, ascorbate peroxidase, glutathione reductase, and catalase in comparison to unsprayed control plants. Both ABA and SA treatments decreased the contents of hydrogen peroxide and thiobarbituric acid reactive substances, a measure of lipid peroxidation, compared to unsprayed plants. The beneficial effect of increase in antioxidant enzymes activity and decrease in oxidative stress was reflected in increase in chlorophyll and carotenoid contents, relative water content, membrane stability index, leaf area and total biomass over control plants. The lower concentrations of ABA (0.5 mM) and SA (1.0 mM) were generally more effective than higher concentrations.

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.

The effects of nitrogen [75 and 150 kg (N) ha^-1] and elevated CO₂ on growth, photosynthetic rate, contents of soluble leaf proteins and activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and nitrate reductase (NR) were studied on wheat (Triticum aestivum L. cv. HD-2285) grown in open top chambers under either ambient (AC) or elevated (EC) CO₂ concentration (350 ± 50, 600 ± 50 μmol mol^-1) and analyzed at 40, 60 and 90 d after sowing. Plants grown under EC showed greater photosynthetic rate and were taller and attained greater leaf area along with higher total plant dry mass at all growth stages than those grown under AC. Total soluble and Rubisco protein contents decreased under EC but the activation of Rubisco was higher at EC with higher N supply. Nitrogen increased the NR activity whereas EC reduced it. Thus, EC causes increased growth and P_N ability per unit uptake of N in wheat plants, even if N is limiting.

Young plants of Lotus creticus creticus growing in a hydroponic culture were submitted to 0, 70 and 140 mM NaCl treatments for 28 d and the growth and ecophysiological characteristics of these plants have been studied. The growth of Lotus plants was not affected by salinity when applied for a short period (about 15 d); however, 140 mM NaCl induced a decrease in shoot RGR at the end of the treatment. The root growth was not decreased, even it was stimulated by 140 mM NaCl. The osmotic adjustment of Lotus plants at 70 and 140 mM NaCl maintained constant pressure potential, avoiding the visual wilting. For a similar leaf water potential, cuticular transpiration of salinized plants was lower than in control plants due to the salinity effect on the cuticle. Moreover, the presence of hairy leaves (60 and 160 trichomes per mm² in young and adult leaves, respectively) allows keeping almost 81 % of sprayed water and absorbing the 9 % of the water retained, decreased the epidermal conductance to water vapour diffusion.

Oxygen evolution and chlorophyll a fluorescence transients of two barley (Hordeum vulgare L) cultivars subjected to polyethylene glycol induced osmotic stress was examined. The relative water content of the plants was used as a measure of their water status. The results suggested that although dehydration was considerable, photosystem 2 was weakly affected by the osmotic treatment.

A rapid and efficient procedure is outlined for in vitro clonal propagation of an elite cultivar of jewel orchid (Anoectochilus formosanus). Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with 1 mg dm^-3 benzyladenine or 1 - 2 mg dm^-3 thidiazuron (TDZ). Addition of activated charcoal (1 g dm^-3) to the TDZ containing medium promoted multiple shoot formation (11.1 shoots per explant). However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 medium supplemented with 2 % sucrose and 0.5 g dm^-3 activated charcoal. Rooting was induced in 100 % of the regenerated shoots in the same media. The plantlets were acclimatized and established in greenhouse.

The effects of different cadmium concentrations [17 mg(Cd) kg^-1(soil) and 72 mg(Cd) kg^{- 1}(soil)] on Cannabis sativa L. growth and photosynthesis were examined. Hemp roots showed a high tolerance to Cd, i.e. more than 800 mg(Cd) kg^-1(d.m.) in roots had no major effect on hemp growth, whereas in leaves and stems concentrations of 50 - 100 mg(Cd) kg^-1(d.m.) had a strong effect on plant viability and vitality. For control of heavy metal uptake and xylem loading in hemp roots, the soil pH plays a central role. Photosynthetic performance and regulation of light energy consumption were analysed using chlorophyll fluorescence analysis. Seasonal changes in photosynthetic performance were visible in control plants and plants growing on soil with 17 mg(Cd) kg^-1(soil). Energy distribution in photosystem 2 is regulated in low and high energy phases that allow optimal use of light and protect photosystem 2 from overexcitation, respectively. Photosynthesis and energy dissipation were negatively influenced by 72 mg(Cd) kg^-1(soil). Cd had detrimental effects on chlorophyll synthesis, water splitting apparatus, reaction centre, antenna and energy distribution of PS 2. Under moderate cadmium concentrations, i.e. 17 mg(Cd) kg^-1(soil), hemp could preserve growth as well as the photosynthesis apparatus, and long-term acclimation to chronically Cd stress occurred.

Pollen embryogenesis was successfully induced in Solanum nigrum L. (2n=6×=72). Stimulation of androgenesis expressed as the frequency of androgenic responsive anthers was observed after 10 and 20 mM ethyl methanesulphonate (EMS), 10 and 20 mM sodium azide (NaN₃) and 0.2 mM N-nitroso-N-methylurea (MNU) treatment applied on seeds for 24 h. The frequency of androgenesis on the medium with sucrose was higher than on the medium with maltose. Androgenic regenerants originated also in the anthers collected from donor plants where survival after mutagenic treatment was lower than 50 %. Green haploid (3x), aneuploid (to 8x) and dihaploid (6x) plants were obtained. The high frequency of aneuploids among androgenic plants is explained by cell division irregularities in microsporial calli.

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.

Direct somatic embryogenesis from ray floret explants of five chrysanthemum cultivars has been obtained within 12 - 15 d on Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA). Scanning electron microscopic observation also confirmed the direct origin of somatic embryos from explants. Somatic embryos developed asynchronously on the adaxial surface of explants. Among the five cultivars tested, Birbal Sahani was best responding (40 % explants responded on 4 mg dm^-3 2,4-D and 2 mg dm^-3 BA supplemented medium). Precocious germination of somatic embryos was noticed on the same medium. The best sucrose concentration in the medium was found to be 60 g dm^-3 where 70 % explants responded while 55 % embryogenic response was obtained on medium supplemented with 400 mg dm^-3 inositol. Plants developed from somatic embryos were transferred to soil and produced true-to-type flowers.

Chrysanthemum morifolium cv. Regol Time plants were found to be infected with Chrysanthemum B carlavirus (CVB). They were made CVB-free by using meristem tip culture, chemotherapy and thermotherapy. The plants were indexed by biological assay, double antibody sandwitch enzyme linked immunosorbent assay (DAS-ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR). The maximum number of virus-free plants (26.7 %, as indexed by RT-PCR) was obtained with 2-thiouracil at 0.04 g dm^-3 concentration. Only 12 % plants were found to be virus-free, after being kept at 38 °C for 30 d. For indexing CVB in chrysanthemums, RT-PCR was found to be the most reliable method.

The special conditions during in vitro culture result in the formation of plantlets of abnormal morphology, anatomy and physiology. After ex vitro transfer, these plantlets might easily be impaired by sudden changes in environmental conditions, and so need a period of acclimatization to correct the abnormalities. This review is focused upon contemporary information on the changes in leaf structure, water relations and photosynthesis during acclimatization of plantlets to ex vitro conditions. It also describes some ways of improving plant survival and for the speeding up of acclimatization.

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.

The effects of treatment with NaCl (3, 100 and 300 mM) for 1, 2, 3 and 7 d on plant growth and ion accumulation were analyzed in 2-week and 8-week-old Annona muricata and A. squamosa plants. Fresh mass and root growth inhibition were directly related to the increase in salinity, particularly for A. squamosa. Two-weeks old seedlings were sensitive to 100 and 300 mM NaCl particularly after 7 d, whereas 8-week-old plants were shown to be more resistant to NaCl even at 300 mM NaCl. Na⁺ and Cl^- mostly accumulated in young leaves. Our results suggest that A. squamosa is more sensitive than A. muricata to salt stress and that older seedlings of both species are more tolerant than younger seedlings.

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.

A simple histo-cytochemical method, combining Evans blue staining to assess cell death and in vivo 3,3'-diaminobenzidine uptake for H₂O₂ localisation, has been used to evaluate O₃ damages in leaf tissues of three Phaseolus vulgaris L. cultivars (Cannellino, BLF, Saxa) with different sensitivity to the pollutant. Bean plants were exposed to a single pulse of O₃ (150 ± 10 mm³ m^-3 × 3 h) and leaves were examined at different time-span after fumigation. Cannellino proved to be the most sensitive, showing chlorotic spots 2 h after fumigation and chlorotic lesions 24 h later. In BLF, necrotic spots appeared 4 h after fumigation and reddish necrotic lesions (bronzing) developed in further 24 h. Saxa remained symptomless up to 10 d of observation, thus appearing tolerant. The early appearance of symptoms in Cannellino correlated with H₂O₂ accumulation in leaf tissues and consequent extensive cell death, involving both palisade and spongy mesophyll. H₂O₂ accumulation was observed also in BLF, though to a lesser extent and dead cells were rare at 2 h after fumigation. However, they increased in number 24 h later, forming small groups in the palisade mesophyll. These groups further enlarged in the next 24 h, again involving only palisade mesophyll. In Saxa leaves, H₂O₂ accumulation was found only in the epidermal cells, though the number of dead cells was very similar to BLF, at least up to 24 h after fumigation. However, in Saxa, dead cells have been always found singly scattered through the palisade mesophyll, or forming very small groups around substomatal cavity, thus remaining invisible at a macroscopic level.

The effect of a short heat treatment in combination with different culture medium composition on the efficiency of in vitro induced androgenesis in Silene latifolia ssp. alba was studied. The heat shocks (33 and 37°C) were applied for 1, 3, and 5 d. The best androgenic response was observed at 25°C and after a one-day treatment at 33°C. All other treatments reduced androgenic response. Among different media compositions tested, the most satisfactory results were obtained on BMS medium supplemented with 6-benzylaminopurine (0.5 mg dm^-3) and sucrose. The green, albino and chimeric, only female, plants were regenerated. Flow cytometry of 110 regenerants identified haploids, mixoploids (n+2n and 2n+4n) and dihaploids.

The purpose of this research is micropropagation of Ginkgo biloba L. Apical and nodal meristems removed from plantlets or apical buds from a tree were used as explants. Meristems produced an extensive callus and single or rare multiple shoots on Murashige and Skoog medium with different growth regulators and endosperm extract (En) obtained from mature seeds of the same species. For successful root production it was necessary to transfer the shoots to a rooting medium with En.

Stable transformation of Coffea canephora P. was obtained by particle bombardment of embryogenic tissue. Leaf explants were cultured on medium supplemented with 5 µM isopentenyl-adenosine to induce direct embryogenesis. Explants with somatic embryos were transferred to half strength MS medium with 9 µM 2,4 dichlorophenoxyacetic acid. After 2 weeks, the explants with somatic embryos and embryogenic tissue were bombarded with tungsten particles (M-25) carrying the plasmid pCambia3301 (containing the bar and uidA genes) using a high pressure helium microprojectile device. The bombarded explants were submitted to selection on medium containing 5 µM ammonium glufosinate herbicide as selective agent. After 6 months, putative transgenic embryos were transferred to a growth regulator-free medium for germination. The regenerated plantlets were β-glucuronidase (GUS) positive whereas no GUS activity was observed in non-transgenic controls. Incorporation of the bar gene into the genome was confirmed by PCR and Southern blot analysis of the regenerated transformed plants. Greenhouse grown transgenic coffee plants were found to withstand the recommended level of the herbicide Finale™ for weed control.

Tobacco plants infected with the potato virus Y (PVY) were studied during the acute-infection period. The control enzymes of metabolic pathway of host's RNA degradation tending to biosynthesis of PVY-RNA, its coarse/fine regulation and content of host's RNA were monitored. Activities of ribonucleases, phosphomonoesterases and phosphodiesterases in both the crude homogenates and the partially purified enzyme preparations from the diseased leaves were markedly increased when compared to the tissues from healthy plants. The curves of enzyme activities positively correlated with the multiplication curve of the PVY and negatively correlated with the decreased contents of host's RNA. The enzyme activity in homogenate samples did not significantly differ from the corresponding purified enzyme preparations.

Young maize plants, grown hydroponically, were supplied with 1/10 the optimal amount of iron (0.75 mg dm^-3). Foliar treatments with solutions, containing N⁶-benzyladenine (BA), indole-3-acetic acid (IAA) or (2-chloroethyl)-trimethylammoniumchloride (CCC) were conducted after chlorosis had been well manifested. Changes in growth, chlorophyll content, rate of photosynthesis, catalase and peroxidase activities in leaves, and the contents of Fe, Cu, Zn, Mn, and P in leaves were recorded. Growth regulators improved (CCC, IAA) or aggravated (BA) the physiological state of chlorotic plants. Their effect might be explained by changes in Fe transport towards the leaves, by increased efficiency of Fe utilization, and by effects on plant metabolism not involving Fe.