Plants were in vitro regenerated from leaf callus of Desmodium affine and D. uncinatum. Leaf explants were induced to form callus when aseptically cultured on Murashige and Skoog medium (MS) supplemented with 6 mg dm-3 6-benzylaminopurine (BAP) in combination with 1 mg dm-3 naphthaleneacetic acid (NAA). Regeneration of shoots was induced when callus was cultured on MS medium supplemented with 6 mg dm-3 BAP and 0.01 mg dm-3 NAA. Roots regenerated in high frequency when differentiated shoots were subcultured on MS medium supplemented only with 0.01 mg dm-3 NAA. The regenerated plantlets were successfully grown in pots. Calli from D. incanum failed to regenerate shoots.

The inoculation of mycorrhizal maize plants with three isolates of microaerophilic diazotrophic bacteria obtained from the mycelium of arbuscular mycorrhizal fungi associated with three grasses (Arrhenatherum elatius - bacterial isolate ARR, Agropyrum repens - isolate AGR and Poa annua - isolate POA) caused no increase in nitrogen content in plant biomass. The inoculation with bacterial isolate ARR resulted in the decreased plant growth. Bacterial isolate AGR decreased the percentage of the root length colonized by arbuscular mycorrhizal fungus Glomus fistulosum. The inoculation with both mycorrhizal fungus and isolate POA increased significantly the concentration of phosphorus in plant shoots compared to uninoculated control.

Seedlings of two common Mediterranean trees, Pinus halepensis L. and Quercus ilex L., were grown alone and together with seedlings of the same or of the other species in the same pot during one year to test the effects of intra- and inter-specific interference on terpene emission. Light, nutrients and water were amply supplied. There were higher emission rates in P. halepensis than in Q. ilex. The emission increased when the neighbour was a pine and decreased when the neighbour was a holm oak. Volatile organic compound and terpene emission rates followed inverse trends to foliar biomass or growth, which decreased when the neighbour was a pine.

A culture system was developed which allows continuous production of adventitious buds. Segments of seedlings germinated on media supplemented with different growth regulator combinations were used as explants. Cultivation was carried out in two phases, which alternate permanently: (I) without and (II) with growth regulators. Thus it was possible to establish meristematic tissue cultures from 5-10% of the seedlings tested. They have produced buds now for over 3 years with multiplication rates of about 1.5 within 5-6 weeks.

Direct differentiation of shoot buds from hypocotyl segments of 12-d-old seedlings of Tamarindus indica was obtained on Murashige and Skoog (MS) medium with or without growth regulators. The highest regeneration (66 %) and the maximum number of shoots (3 - 4) per explant were obtained from the explants on MS medium containing 6-benzylaminopurine (5 × 10-6 M). A maximum roots per shoot were produced on medium containing 3-indole butyric acid (5 × 10-6 M). The resulting plantlets were hardened and transferred to soil in pots where 75 % of them survived and resumed growth. Histological examination of explants suggests that the shoots were of de novo origin which would make this system suitable for transformation experiments.

Increasing salinity induced a marked reduction in the plant growth, thoughPhaseolus seedlings tolerated salinity up to 120 mM NaCI. A great reduction in sugar and protein contents occurred with increasing salinity, whereas soluble nitrogen compounds and the relative contents of the photosynthetic pigments were increased in the treated plants. Increasing Ca concentration in the salinized medium appeared to improve the plant growth and to increase the contents of saccharides and proteins in the NaCl-treated plants. This suggests that Ca could be added to salinized media to overcome the deleterious effects of salinity on the growth and productivity of leguminous crop plants.

Growth and Na⁺, K⁺, Cl^-, proteins, sugars and proline concentrations were measured in three triticale genotypes M2A, DF99 and Asseret grown on nutrient solution with or without 75 mM NaCl. In saline conditions, leaf area of the three triticales was reduced by 50 % and dry to fresh mass ratio increased. Total protein concentration was diminished by 10 %. K⁺ concentration decreased whereas Na⁺ and Cl^- accumulated in roots and shoots of salt-stressed plants. This ion accumulation was greater in roots of Asseret than in roots of the other triticales. Soluble sugar concentration increased in M2A and Asseret and decreased in DF99. Proline concentration increased in M2A and DF99 and decreased in Asseret. Osmotic adjustment was essentially realized by Na⁺ and Cl^- uptake. Non-reducing sugars and proline contributed too, but to a lesser extent.

Exposure of coffee to low temperatures caused growth inhibition, changes in metabolic rates, and membrane alterations. Root tissue exposed to 10 °C evolved significantly lower rates of metabolic heat compared with controls grown at 25 °C, and the values were closely associated with the observed root growth inhibition. Electron paramagnetic resonance spectra of intact tissues showed that the spin probe 5-doxylstearic acid was capable to intercalate within the cellular membrane lipids. Indeed, at the depth of the 5th carbon atoms of the alkyl chains, the nitroxide radical detected more rigid membranes in seedlings exposed to 10 °C compared with 25 °C treated samples. Ascorbate peroxidase and catalase activities did not show appreciable changes under chilling conditions, while guaiacol peroxidase activity increased 55 % compared to the control. On the other hand, glutathione reductase activity decreased, in parallel to a significant decline in the capacity to reduce triphenyl-tetrazolium. Our results showed a marked correlation between lipid peroxidation and root tissue damage, which seemed to be associated with increased membrane rigidity.

Total nonstructural saccharides (TNS) content in young plant ofAlopecurus pratensis was always above 4% of dry matter even at several types of stress treatment (nitrogen deficiency, low irradiance). TNS content was in negative correlation with concentration of total nitrogen in all cases. Positive correlation was found between the TNS content in plants and relative increase in their root growth rate.

Effects of N⁶-benzyladenine, kinetin, zeatin, N⁶-benzyladenosine, kinetin riboside, zeatin riboside, jasmonic acid, indole-3-acetic acid, indole-3-butyric acid, indole-3-propionic acid, abscisic acid and gibberellic acid on proliferation of hyphae of arbuscular mycorrhizal (AM) fungus Glomus fistulosum were studied under axenic conditions in vitro. The growth of intraradical hyphae of G. fistulosum was fully suppressed by 30 µM indole-3-acetic acid, but a perceptible decrease in the proliferation of the hyphae was observed already at 3 µM. Because such concentration is near the concentrations common in root tissues in vivo, the effect may be biologically significant. Similar effect was also observed for Glomus mosseae. Inhibitory effects of abscisic acid and cytokinins occurred only at very high, non-physiological concentrations. Ribosylated cytokinins showed stronger inhibition effects than their non-ribosylated counterparts. No stimulation of proliferation of hyphae by any plant hormone tested was observed.

Spring oilseed rapeBrassica napus L. ssp.oleifera cv. HM-81 was transformed with T_L-DNA of the Ri plasmid of the agropine strainAgrobacterium rhizogenes 15834. Selfed progenies (R₂ and R₃ generations) were studied for changes in values of growth characteristics and fatty acids contents. Transformants are 'homozygous' for T_L-DNA. Both generations of transformants differed significantly from the nontransformed control plants in reduced length, lower number of pods per plant, lower total mass of seeds and the higher number of branches. The contents of palmitic, linoleic and linolenic acids were significantly higher in transformants when compared with the control. On the contrary, the contents of both stearic and oleic acids were in most of transformants significantly lower. Only traces of erucic acid (less than 0.05 % ) were found, both in transformed and nontransformed plants.

The greatest growth of wheat and rape plants in vitro was reached on media with 5 or 9 % sucrose, respectively. The highest efficiency for transfer of these plants to ex vitro conditions was found at the same sucrose concentrations. The content of endogenous non-structural saccharides (glucose, fructose, sucrose, starch and fructans) increased with increasing sucrose concentration in the medium up to 10 %.

The growth and nodulation ofTrifolium alexandrinum were compared at six levels (0 - 1.2 % NaCl) of salinity. Dry mass of shoots and roots, 14 and 20 weeks after the commencement of salinity treatment, increased at low levels of salinity (0.1 - 0.2 % NaCl) but decreased with higher NaCl concentrations (0.4 - 1.2 %). Nodulation occurred at NaCl concentrations up to 0.8 %. Nodule mass decreased with increasing salinity levels. The nodule size remained unaffected at NaCl concentrations up to 0.4 % but was reduced at higher concentrations.

A precursor in tetrapyrrole biosynthesis, 5-aminolevulinic acid (ALA), was applied via presowing soaking in Vigna catjung, V. mungo, and V. radiata. ALA increased plant growth and influenced dry matter accumulation in leaves, stems, and pods through increased chlorophyll content and photosynthetic CO₂ absorption. At harvest, ALA treated plants had increased number of pods per plant, seeds per pod, 100 seed dry matter, biological yield, and the harvest index. Therefore, pretreatment of seeds with optimal concentration of ALA is recommended for improving the growth and productivity of tropical legumes.

It is a matter of controversy whether secondary wall deposition is dependent on lignification during the development of tracheary elements. To understand this, tracheary element differentiation was studied in the homogeneous calli obtained from the cotyledonary explants of Cucumis sativus subsequent to treatment with plant growth regulators, such as naphthalene acetic acid (NAA) and benzylamino purine (BAP), which are necessary for the induction of tracheary elements, along with metabolic blockers such as 2-aminoindan-2-phosphonic acid (AIP), 2,3,5-triiodobenzoic acid (TIBA) and nifedipine. Calli treated with AIP, a potential inhibitor of L-phenylalanine ammonia-lyase (PAL), have no PAL activity at any time during the culture period. There was a complete inhibition of lignification although secondary wall deposition was unaltered. Similar results were obtained using TIBA, an inhibitor of auxin transport, and nifedipine, a known calcium channel blocker. Thus the present study suggests that secondary wall deposition in the course of tracheary element differentiation need not to be dependent on lignification.

Direct somatic embryogenesis from shoot apical meristems of pea is described. Somatic embryos were induced directly (without callus intervention) from meristematic tissues grown on a medium supplemented with 2.5 µM picloram. Within 4 to 5 weeks, fully morphologically developed somatic embryos were obtained. Somatic embryos originated from apical as well as from basal parts of meristem explants. The initiation and development of somatic embryos was asynchronous, basal somatic embryos developed more quickly than apical ones. Abundant secondary embryogenesis was observed after isolation of primary somatic embryos and culturing them on media for germination. Morphologically normal somatic embryos germinated on medium without growth regulators; the conversion rate was increased by application of 10 µM thidiazuron (TDZ). TDZ was also able to induce shoot bud regeneration on embryos without differentiated a shoot apex, allowing to germinate up to 78 % of all harvested somatic embryos with various morphology. The protocol was successfully tested in 47 out of 48 P. sativum and P. arvense cultivars as well as in two wild peas (P. elatius, P. jomardi).

The antagonistic effects of some growth regulators [i.e. indol-3-yl-acetic acid (IAA), gibberellic acid (GA₃) or kinetin] on stress imposed by sea water on leaf area, pigment and photosynthetic activity in leaves of broad bean plants at different stages of development were investigated. Seed priming with GA₃ alleviated either partially or completely the effects induced by the two levels of sea water (10 and 25 %) used on leaf area at all experimental stages. However, IAA, GA₃ and kinetin inhibited leaf growth by themselves in almost all measurements. Seed pretreatment with kinetin alleviated the inhibition of pigment production in sea water-irrigated plants. Furthermore, GA₃ or kinetin nullified the deleterious effects imposed by irrigation with sea water particularly the high level (25 %) on photosynthetic¹⁴CO₂ fixation.

Mungbean (Vigna radiata L. Wilczek cv. Sujata and cv. K851) seedlings were grown in paper towelins in dark under 0, 0.5, 1, 2 and 3 % (m/v) NaCl salinity. Germination percentage, shoot and root length, fresh mass of both cultivars decreased with salinity. Total soluble saccharides and proline accumulated in the root and shoot of salt stressed seedlings. The proline accumulation in the root was four to five times higher than that of the shoot of NaCl treated etiolated mungbean seedlings.

Maize (Zea mays L. cv. Ganga-5) seedlings were grown in the presence of ferulic acid (0.5 - 3.0 mM) for 8 d. Treatment with ferulic acid considerably decreased shoot and root length, increased the activity of peroxidase, catalase and indole-3-acetic acid (IAA) oxidase and decreased the activity of polyphenol oxidase. The increased activity of peroxidase correlated with pronounced increase in content of lignin and phenolic compounds

Changes in soluble protein profile and chlorophyll (Ch1) content in detached soybean leaves incubated in darkness or light were delayed by application of benzyladenine or indole-3-acetic acid and enhanced by abscisic acid. The degradation in light differed significantly from the degradation in darkness. Chl and proteins were lost at a higher rate in darkness than in light.

Ears of uniculm wheat (Triticum aestivum L. cv. Gigas) grown in liquid medium for 11d absorbed more solution when the saccharose concentration was 2 % than when it was 10 %. When the ears were grown in 6 % saccharose solution, the rate of uptake from the solution was between that from the 2 and 10 % saccharose medium. Dry mass per grain increased with the saccharose concentration in the medium and the reduced uptake of solution did not decrease the moisture percentage of the grain. The culture of ears decreased pH of the solution with 2 % saccharose more than with 10 %. Addition of 0.5% chloramphenicol to the culture solution had no adverse effect on grain mass; it prevented contamination of the solution and maintained a higher pH

The plants of mustard (Brassica juncea L.) were treated with 0, 25 and 50 ΜM gibberellic acid (GA₃) at three fully developed leaf stage (30 d after sowing). Effect of GA₃ on carbonic anhydrase activity, photosynthetic rate, leaf area index and dry mass was studied at 50, 70 and 90 d after sowing. At harvest 1000 seed mass, pod number and seed yield were recorded. GA₃ treatment (50 ΜM) enhanced all the characteristics studied.

Induction and growth of soybean callus cultures were influenced by NaCl, especially at the highest concentration tested (150 mM). Protein content was raised as NaCl was increased in the Murashige and Skoog medium. Total sulfhydryl group (-SH) and glutathione (GSH) concentrations were also increased in NaCl treated cultures. The affinity (Km) of glutathione reductase (GR) for oxidized glutathione (GSSG) was gradually increased as NaCl level was raised in the medium. The GSH/GSSG ratio was raised significantly as the result of GR activity. The increase in GR activity may constitute an adaptive response of soybean callus to NaCl.

Secondary somatic embryogenesis and shoot organogenesis from primary somatic embryos of Papaver somniferum L. are described. The embryos became malformed, the root meristem expressed dividing activity without position-dependent cell differentiation, causing abnormal development or arrested growth of primary somatic embryos. The adventitious shoots regenerated from embryo hypocotyl, but secondary somatic embryos had an epidermal origin close to the root meristem. The regeneration occurred without hormonal treatment, indicating endogenous nature of triggering signals. These signals are probably related to the integrity loss of morphogenetic steps during development of primary somatic embryos, which appeared to induce an activation of cells competent to regeneration.

Zygotic embryos of Karwinskia parvifolia, isolated from seeds obtained from different regions of Mexico, were cultured on Woody Plant Medium (WPM) supplemented with 0.06 µM indole-3-acetic acid, 0.03 µM gibberellic acid, and 2 µM 6-benzylaminopurine. The growth of embryos and multiplication of shoots from stem segments were achieved. Rooting of excised shoots could be initiated on basal WPM medium with prolonged subculture period to 2 months, or on WPM medium supplemented with 10 µM 1-naphthaleneacetic acid. Multiplication capacity of shoots and rooting of K. parvifolia differed in dependence on the origin of explant material. The shoot multiplication was much lower than that of Karwinskia humboldtiana. The rooting depended on the origin of K. parvifolia seeds. The regenerated plants were successfully transferred to glasshouse.

Four clones of Malus domestica Borkh. cv. Golden Delicious - namely Smoothee, Crielaard, Reinders and Golden B - were cultured in vitro from single-node microcuttings placed on solid medium under irradiance (PPFD) of 50 µmol m-2 s-1. After 9 months an average shoot proliferation of 5.3 was achieved; Crielaard showed the highest rate (7.1), followed by Golden B(5.4), Smoothee and Reinders (4.4). Proliferating shoots were then exposed to higher PPFD (80 µmol m-2 s-1) and different spectral composition of radiation using PMMA-B and PMMA-R/FR filters. High PPFD decreased the average proliferation rate to 4.5, in particular in Crielaard and Golden B, while it increased proliferation in Reinders. When a PMMA-R/FR filter was interposed, the mean proliferation rate slightly increased. PMMA-B filters decreased the overall proliferation rate to 3.0; only in Crielaard it was increased, but shoots were very small. Thus PPFD and spectral composition influenced in vitro shoot proliferation and growth and the responses were different among the clones.

Media from embryogenic and non-embryogenic cell suspension cultures were analysed for protein content, electrophoretic protein patterns, glycoproteins and activity of peroxidases and β-glucosidases in order to characterize the physiological status of the cultures. On a dry mass basis the amount of extracellular proteins per cell was greater in embryogenic suspensions than in non-embryogenic suspensions. Non-embryogenic suspensions contained unidentified slimy compounds which were not present inembryogenic cultures. The extracellular Concanavalin A-specific glycoproteins gave different isoelectric focussing patterns and thus enabled embryogenic and non-embryogenic cultures to be differentiated. The extracellular peroxidase activity per cell dry mass was far greater in embryogenic than in non-embryogenic cultures. The isoenzymes differed in number and composition of the anionic bands. β-glucosidases were found in the same range of activity in both culture types, but the time course of enzyme activity during cultivation was significantly different. In the embryogenic culture the activity was correlated with dry mass increase, whereas in the non-embryogenic suspension the activity reached maximum during the linear growth phase. Polyphenoloxidase which was recently recognized as an intracellular marker for embryogenic stages was not released into culture media.

Effects of water stress at different stages of plant growth on leaf relative water content (RWC), osmotic potential (Ψos) and changes in contents of chlorophyll, abscisic acid (ABA), zeatin riboside (t-ZR), ethylene and proline in six cultivars of French bean (Phaseolus vulgaris L.) were studied. Under water stress, Ψos and RWC were highest in cv. Contender and lowest in cvs. IIHR-909 and Sel-2. The increase in contents of ABA and proline was marked in cv. Contender followed by cv. UPF-626. Decrease in t-ZR and chlorophyll contents was prominent in cv. IIHR-909. Ethylene production surged in all the cultivars under 4- and 8-d stress and declined under 12-d stress.

When the proper stimuli are given, somatic plant cells may form adventitious embryos, roots or shoots. The three pathways of regeneration show apparent similarities. They consist of three analogous phases: 1) dedifferentiation (during which the tissue becomes competent to respond to the organogenic/embryogenic stimulus), 2) induction (during which cells become determined to form either a root, a shoot or an embryo), and 3) realization (outgrowth to an organ or an embryo). The first phase may involve a period of callus growth (indirect regeneration), but often cells present in the explant become competent without cell division or without cell division at a large scale (direct regeneration). In an explant, only very few cells show the organogenic/embryogenic response. In direct regeneration, the three regenerative pathways start from cells in different tissues. This is most obvious when the different types of regeneration occur in the same explant. The hormonal trigger for the dedifferentiation phase is a general one, probably auxin. During the induction phase, each pathway requires specific hormonal triggers. During the realization phase, hormones should be absent or at low concentration. The successive steps in the regeneration process coincide with events on the molecular and biochemical levels, but so far no coherent picture has emerged. In particular during the early stages of regeneration, research on these levels is hampered by a technical problem, viz., the very low proportion of cells that participate in the process of regeneration. New methods may overcome this problem.

The involvement of ethylene in shoot formation in vitro was studied in one year old lavandin (Lavandula officinalis Chaix x Lavandula latifoliaVillars) callus. A peak in ethylene evolution characterized thenon-regenerating leaf callus line, as compared to the shoot-forming calyxcallus line, on the growth medium provided with 2,4-dichlorophenoxyacetic acid (1 mg dm-3) and kinetin (0.5 mg dm-3). After one year in culture, calyxcallus attained the capacity to grow on auxin-reduced media, showing decreased ethylene production and faster shoot bud emergence, when transferred onto the regeneration medium, supplemented with 10 mg dm^-3 benzyladenine. Shoot formation was also inhibited by addition of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid, indicating an involvement of ethylene in the failure of regeneration.