Somatic embryos were induced from immature cotyledons and immature embryonal axis ofArachis hypogaea L. on L-6 basal medium supplemented with NAA, picloram or 2,4-D at 5-50 mg 1^-1. Immature embryonal axis produced a higher number of somatic embryos in comparison with immature cotyledons. The highest number of responding cultures was produced on medium supplemented with NAA (50 mg 1^-1), while the highest average number of somatic embryos per culture was produced on medium with 2,4-D (10 or 20 mg 1^-1) and picloram (30 mg 1^-1) from cotyledons. The somatic embryos developed into plants on basal medium supplemented with activated charcoal and about 100 plants were successfully transferred to the field.

Increasing salinity of growth medium induced a reduction in growth and transpiration rate. The concentrations of chlorophylls and carotenoids were increased in most cases in broad bean leaves while in pea plants they remained more or less unchanged with the rise of salinization up to 80mM NaCl. Thereabove a significant decrease in these contents was observed. A stimulation of the net photosynthetic rate of pea was observed at the lowest levels of NaCl but at the highest levels inhibitory effect was recorded. In broad bean all salinization levels inhibited photosynthetic activity, but dark respiration of both plant species was stimulated. The content of Na⁺ in the roots and shoots of both species increased at increasing salinity. In broad bean, Ca²⁺ concentration in shoots and K⁺ and Ca²⁺ contents of roots increased at increasing salinization, while in pea plants, the content of K⁺ and Ca²⁺ was almost unaffected by salinity. Salinity induced an increase in the content of these ions in pea roots. Mg²⁺ content in shoots and roots of both broad bean and pea decreased at increasing salinity except in roots of pea, where it was generally increased.

Two staining methods for aspartate aminotransferase were compared after electrophoretic resolution of its isozymes in polyacrylamide gels. The first one uses L-aspartic acid and Fast Blue BB salt (classical method), the second uses L-cysteine sulfinic acid and a redox system with phenazine methosulfate and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide. The seeds of pea, horse bean and soybean were used as a model plant source of the enzyme. The staining method with L-cysteine sulfinic acid is very reliable and more sensitive than the Fast Blue BB method and allows detection at very low isozyme activities in the gel.

Stem internodes with axillary buds were excised from 5-year old trees ofFicus benjamina cv. Exotica. The effect of 6-benzylaminopurine (BAP), gibberellic acid (GA₃), indole-3-acetic acid (IAA), naphthaleneacetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D) on shoot growth and proliferationin vitro was investigated. Multiple shoots were developed after 3-4 weeks from stem internodes with axillary buds incubated in Murashige and Skoog (MS) medium supplemented with phloroglucinol (PG) and BAP. Optimum shoot proliferation took place in the presence of 1.0 mg l^-1 BAP. Shoots obtained could be elongated in a medium with 0.5 mg l^-1 GA₃ prior to their rooting. The root initiation was successfully induced on MS medium either with IAA at 0.5-0.1 mg l^-1 or in plant growth regulator-free medium. All rooted plantlets were subsequently transferred to a peat, humus and perlite mixture in a culture room with high humidity and covered with plastic bags. After one month the plantlets were established for growing in a greenhouse.

Addition of either abscisic acid (ABA) or kinetin at 10^-6 M to salinized media (20-120mM NaCl) induced remarkable effects on growth ofPhaseolus vulgaris plants. Whereas ABA inhibited the plant growth and the rate of transpiration, kinetin induced stimulation of both parameters. Moreover, ABA increased proline and phosphorus concentrations in the salinized plants whilst kinetin decreased them.
ABA induced stimulation of the transport of K, Ca and Cl from root to shoot, accumulation of K, Na and Cl in root cells and inhibits the transport of Na and accumulation of Ca. Kinetin appeared to inhibit the transport and accumulation of Na and Cl, transport of K, and stimulates the accumulation of K and Ca as well as the transport of Ca. The highest influence of both ABA and kinetin was mostly observed when these hormones were used in combination with the highest concentration of NaCl (120 mM) in the medium.

A method of cultivation and effectiveness of different light sources and light regimes in photoperiodic induction of flowering in non-rosette long-day plantChenopodium murale L. ecotype 197 are described. Under the described conditions of cultivation 5 days, of continuous light produced by incandescent bulbs (TESLA 74 3x40 W, red 4.9 μWcm-²nn-¹, far-red 7.4 μWcn-2nm-¹, blue 0.25 μW cm-²nn-¹) induced flowering in the majority of plants.

In comparison with the parental cv. Finale the 'RisfixC' supernodulator exhibited higher, continuously increasing nodule number and fresh mass accumulation, but substantially lower individual nodule fresh mass, leghemoglobin concentration, and specific acetylene reduction activity of nodule tissue. There were no substantial differences between Finale and 'RisfixC' in total acetylene reduction, nodule leghemoglobin accumulation per nodulated root, total and specific CO₂ evolution from nodulated roots and gross CO₂ respiratory costs of acetylene reduction. The 'RisfixC' also exhibited a substantially lower plant dry mass production (by 30%), but nitrogen concentration in shoots and carotenoid concentration in leaf tissue were significantly higher by 33 and 14%, and the chlorophylla+b content insignficantly higher than in the parental cultivar. In contrast, the nodulation mutant 'Risnod29', exhibited a somewhat higher nodule fresh mass accumulation (by 21%) and individual nodule fresh mass (by 23%), total and specific acetylene reduction (by 49 and 19%) and a somewhat more rapid plant dry mass accumulation compared with the cv. Finale.

Potato spindle tuber viroid (PSTVd, severe strain) was isolated from tobacco plants transformed with a dimeric infectious expression construct which was maintained for a long time inAgrobacterium tumefaciens. Afterin vitro hybridization of PSTVd to complete minus transcripts of PSTVd (lethal), the resulting heteroduplexes were analyzed by electrophoresis under native conditions. Electrophoretic analysis revealed an appearence of electrophoretically distinguishable heteroduplexes, suggesting an accumulation of mutated sequence variants of PSTVd had occurred in the plant transformants. TGGE analysis of PSTVd cDNA, re-cloned from the original expression vector pCB1413 to plasmid pUC18 confirmed the accumulation of mutations in the cDNA and the instability of this sequence inA. tumefaciens maintained at 4°C for 2.5 years. One of these point mutations was identified by sequencing the PSTVd cDNAs isolated from the individualE. coli colonies. This transition (GC→AT) was localized at the position of 81 in the PSTVd genome, causing the change of C to U in PSTVd plus RNA. Transformation of tobacco with the freshly prepared expression vector containing the dimeric sequence of PSTVd lethal KF440-2 lead to the propagation of PSTVd electrophoretically identical to that derived from the original sequence and maintained in the tomato by a conventional infection.

Different behaviour of small groups of stomata on a single leaf blade (stomatal patchiness) is reviewed. The occurrence of stomatal patchiness depends on plant species, age, leaf position, environmental conditions,etc. The possibility of errors in conventional evaluation of stomatal and non-stomatal (biochemical) limitations of photosynthesis resulting from patchy stomatal closure is analysed. The consequences of stomatal patchiness for leaf and plant photosynthesis and water economy are discussed. A brief survey of the techniques currently used for detection and quantification of stomatal patchiness is presented.

Phytotoxic micromycetes appear to be responsible for the apple replant disease (ARD). This was suppressed by the inoculation of apple-tree seedlings with some species of vesicular-arbuscular mycorrhizal (VAM) fungi-Glomus fasciculatum andG. macrocarpum. After the inoculation, growth of apple-tree seedlings improved in dependence on the type of soil, on VAM fungus species and on the ARD appearance. After 12-month cultivation, plant biomass (height, shoot and root dry masses) was markedly increased by inoculation withG. fasciculatum. Similarly, the numbers of colony forming units per unit soil (CFU) of phytotoxic micromycetes and of diazotroph bacteria (associative dinitrogen-fixing bacteria) in the rhizosphere was affected; CFU of phytotoxic micromycetes decreased, whereas CFU of the genusAzospirillum was higher. These bacteria could also serve as antagonists against phytotoxic micromycetes. It is also suggested that the ratio of CFU of diazotroph bacteria to CFU of phytotoxic micromycetes can be used as an indicator of the degree of ARD. It may be assumed that the use of some VAM fungi can replace the chemcial treatment of the soil with ARD.

Axillary meristems of short day plantChenopodium rubrum are localized as caulinar, foliar or axillar. The localization of axillary meristems and axillary buds of 14 day old plants varied in similar pattern as in other plant species so far investigated: after several nodes with foliar axillary meristems the caulinar ones were produced. However, unlike in other species, in C.rubrum a very high percentage of caulinar meristem is produced also on the first node. In this case, like in the case of its later differentiation at higher nodes, the formation of caulinar meristem is confined also to the vegetative state. It was found that the caulinary position coincides with higher responsiveness to photoperiodic induction. The developmental significance of such behaviour is discussed.

Photosynthesis inhibition in algae (Chlorella) and plant (spinach) chloroplasts by quaternary ammonium salts of heptacaine {N-[2-(2-heptyloxyphenylcarbamoyloxy)-ethyl]-N-alkylpiperidinium bromides} depended on the alkyl chain length of the alkyl substituent and showed good correlations with theoretical hydrophobic fragment constants as well as with experimentally determined physico-chemical parameters, namely extraction constants and surface activities.

Flow cytometry was used to analyse the DNA content of nuclei isolated from intact plant tissues and from callus and cell suspension cultures invitro. Cell nuclei were isolated either mechanically (chopping, syringing) or by a hypotonic lysis of isolated protoplasts. Although both methods gave similar results, a slight shift to lower ploidy levels was observed after protoplast isolation from intact tissues and calli. No differences were observed if the two methods were compared using cell suspension cultures. The results showed that flow cytometry is a rapid method of nuclear DNA content analysis in intact plant tissues and variousin vitro cultures.

Pistia stratiotes is used for the epuration of domestic sewage in the Biyem Assi phytopurification station. During the process, Fe²⁺, Mn²⁺, Zn²⁺ and Pb²⁺ are absorbed in substantial amounts by the plant. These metals modify the H⁺/K⁺ exchange system at the root level. H⁺ efflux is inhibited by Fe²⁺ and by Zn²⁺ and enhanced by Mn²⁺ and Pb²⁺. K⁺ influx is inhibited by Fe²⁺, by Zn²⁺ and by Pb²⁺ and enhanced by Mn²⁺. It is shown that the purification capacity ofPistia stratiotes can vary with the composition of the heavy metals in the surrounding medium.

The interactive effects of certain phytohormones (GA₃, IAA or kinetin) and drought on plant-water relations and mineral accumulation of the three crop plants; maize, cowpea and broad bean, were studied. Phytohormone applications were capable of counteracting to some extent, the adverse effects of drought on transpiration, stomatal frequency, and leaf area.

A series of binary vector plasmids derived from the T-DNA of theAgrobacterium tumefaciens strain C58, carrying the five plant morphoregulatory genes 1, 2, 4, 5 and 6b in different combinations, was used in the transformation ofNicotiana tabacum leaf discs. Protein patterns of the transgenic tobacco analysed through SDS-PAGE have shown changes in the polypeptides with M_r: ∼120, 60, 55, 43 and 27 kDa (for tobacco with transgene 4); ∼60, 55, 43, 26-25, 21, 18 kDa (for tobacco with transgenes 1, 2 and 5); ∼70, 60, 26, 25, 18 kDa (for tobacco with transgene 5); ∼60, 55, 48, 26, 18 kDa (for tobacco with transgenes 4, 5, 6b); ∼60, 55, 22 and 18 kDa (for tobacco with transgene 6b); ∼60, 55, 43, 26 and 18 kDa (for transgenes 5, 6b); ∼60, 55, 22, 18 and 16 kDa (for transgenes 4 and 6b). All types of transgenic plants showed quantitative changes in protein content. Mendelian segregation ratio to kanamycin resistance in the progeny of transgenic tobacco clones in the R1 generation was 3∶1 except in transgenic tobacco carrying transgenes 1, 2 and 5.

Plant regeneration through somatic embryogenesis was obtained in chickpea (Cicer arietinum L.) using immature cotyledons and immature embryonal axes as explants. 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3.5-6-trichloropicolinic acid (picloram) and 3.6-dichloro-0-anisc, acid (dicamba) in concentrations 1, 2, 5, 10 mg dm^-3 were found better than naphthaleneacetic acid (NAA) (10-20 mg dm^-3) for the induction of globular and heart-shaped somatic embryos. The embryos developed upto the dicotyledonary stage on medium supplemented with saccharose, mannitol and silver nitrate (AgNO₃) and developed further into plantlets on medium containing gibberellic acid (GA₃) and abscisic acid (ABA). The frequency of somatic embryogenesis was dependent on the genotype and auxins used.

Somatic embryogenesis was induced in immature zygotic embryos of pea (Pisum sativum L.), synthetic auxins α-naphthalene acetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-amino-3,5,6-trichloropicolinic acid (picloram, PIC) being used. Only one (line HM-6) of 46 genotypes tested exhibited good potential for somatic embryogenesis. 2,4-D was found as the best somatic embryo inductor. Three different ways of somatic embryo conversion have been described. Plantlets from individual somatic embryos were micropropagated as somaclones and subsequently rooted. A sterile morphological mutant has been found within a group of fertile plants of T₀-generation. Sufficient amount of T₁-seeds is available for somaclonal variation studies.

The efficiency ofAgrobacterium-based transformation technique in oilseed rape and cauliflower was influenced by cultivar specificity, donor plant age and explant type. Marked differences in demands for plant hormone contents in the regeneration medium were recorded already among different types of nontransformed explants. The highest regeneration capacity was recorded with stem and leaf segments isolated from one-month-old aseptically grown plants. The regeneration was markedly species-dependent. Regeneration of transformed plants from stem segments and thin layers isolated from field-grown oilseed rape plants (at the most 2% of regenerating explants) and from oilseed rape hypocotyls (0.8% of regenerating explants) and cauliflower (1.2% of explant regenerated transformed shoots) was achieved after disarmedAgrobacterium treatment. Hypersensitive reaction of explants could be prevented by using prolongedin vitro precultivation and delayed application of the selective agent.

Structural adaptations to increased transport activities were investigated in the cells of vascular parenchyma at the site of the lateral root junction, in non-stressed plant roots. Typical transfer cells were differentiated in dicotyledonousHelianthus tuberosus and in two different genotypes ofH. annuus, the cv. IBH166 and a decorative form. In the representatives of monocotyledonous, no structural adaptations occurred in the roots ofHordeum vulgare but small and rare cell wall protuberances were found in xylem and phloem ofZea mays inbred line VIR17. Some degree of cell wall labyrinth differentiation was seen in xylem and typical transfer cells were found in phloem of the roots of the maize hybrid CE380. The capability of vascular parenchyma to differentiate transfer cells did not depend on species, genotype, or on the growing conditions withHelianthus. On the other hand, the development of the structural adaptations in monocotyledonous representatives depended on both the species and the genotype. This capability may be linked with the taxonomic and evolutionary position of plant species.

Floral induction in the long day plant spinach (Spinacia oleracea) has been shown to be accompanied by a thickening of plasmalemma. This change was observed at early evocation, in both shoot apices and leaves, as well as after inducing GA₃ treatment. To get further information on this thickening, plasma membranes from spinach leaves were isolated, in the present study, using aqueous two phase partitioning and the effect of variousin vitro treatments on their thickness was investigated. The average plasmalemma thickness was unaffected by Na⁺ and K⁺ ions. It was increased upon the effect of either Ca²⁺ or gibberellic acid. A thickening of plasmalemma was also observed when plasma membranes from vegetative plants were incubated with a cytosolic preparation from photoinduced plants. The results were discussed in relation with the plasmalemma modifications previously reported in spinach.

Sugar uptake by potato tuber discs was studied. Discs were used "fresh" or after a 24-h ageing period. It was shown that ageing increases (by 3 to 4 times) the rate of glucose and sucrose uptake. Sucrose uptake by fresh tissues was insensitive to the presence of glucose or fructose while a competitive effect was observed after ageing. This indicates the development of an invertase activity, which was inhibited by tris-buffer. Sucrose and glucose uptake by aged discs was dependent on cellular metabolism as shown by the sensitivity to low temperature and metabolic inhibitors (NaCN, DNP, CCCP). Involvement of thiol groups was demonstrated by the inhibition with NEM and PCMBS. Orthovanadate, which decreases phosphate uptake by 85 % (Poderet al. 1986) did not produce any effect on glucose and sucrose uptake by aged tissues. Fusicoccin produced only a slight stimulation (15 %). These results argue in favour of the involvement of specific ATPases in ion and sugar uptake. No involvement of a redox system was observed. ABA and BAP inhibited the uptake induced by ageing but had no effect on the endogenous sugar content. BAP would act by its effect on the amount of ATP while ABA would act at the membrane level. The results are discussed in relation to the mechanisms of transfer of glucosyl groups and to the transport of sucrose by the symplasmic pathway. Reçule

The protein coding region of theE. coli DNA repair geneada combined with the CaMV 35S promoter has been transferred to tobacco by means ofAgrobacterium tumefaciens Ti plasmid. In transgenic plants having theada gene in a sense orientation, detectable amounts of O⁶-alkylguanine-DNA-alkyltransferase has been found whereas in non-transformed plants this activity is absent. Cell suspension cultures derived from the former plants showed lower sensitivity to the toxic (growth inhibiting) effects of the bifunctional alkylating agent 1-(2-chloroethyl)-1-nitroso-3-(aminomethyl-1,3-diazinylo)-methylurea compared with cell cultures derived from a control non-transformed plant or from transgenic plants harbouring theada gene in an opposite, non-sense orientation.

The rates of canopy apparent photosynthesis (P_C) and canopy respiration (R_c) were studied during vegetation season in two erectophile and two planophile hybrids of maize (Zea mays L.) grown at two canopy densities [7.5 plants m^-2 (HD) and 4.5 plants m^-2 (LD)]. Large differences in P_C, R_c, R_C/P_C, leaf area index (LAI), dry matter accumulation and grain yield were found among hybrids and plant densities. Variations in P_C and R_C were associated mainly with changes in LAI. There was also found change in P_C per unit LAI with time. The average R_C/P_C was 28.9 % for all treatments throughout the vegetation season. P_C and R_C per unit dry matter were higher in LD than in HD and decreased throughout the measurement period. The HD stand had higher P_C and yield in hybrids with erectophile foliage, whereas LD stand had higher P_C after male tetrad and got higher yield in hybrids with planophile foliage. Only R_C in hybrids of the two foliage types was higher under HD than under LD throughout the vegetation period.

Phenylalanine labelled by¹⁴C was administered to the cultured cells and the intact flowers ofCarthamus tinctorius, and the biosynthetic activity of carthamin in these two materials was compared. The cultured cells took up positively the fed substrate, but they could not incorporate the label into carthamin, while incorporation of the radioactivity from phenylalanine into the red pigment occurred in the intact flowers. The activities of polyphenol-oxidizing enzymes were screened in the cell cultures and the intact tissues from the herbal plant. Polyphenol-oxidizing enzymes were operative normally in the mother explant, whereas their activity patterns changed altogether in the cultured cells, where kurenamin, a new reddish pigment, is produced actively. The data are discussed in terms of the phenotypic changes in the polyphenol metabolism of the cultured cells propagated under restricted culture conditions.

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.

Several factors influencing reliability of the quantitative fluorimetric β-glucuronidase (GUS) assay in transgenic plant tissue have been investigated. We obtained linear dependence of fluorescence on both the duration of hydrolysis and the extract concentration. The stability of the enzyme in the homogenate was fairly high, the same as the stability of the substrate solution and of the final reaction product. The modification of the extraction/incubation buffer was proposed, resulting in several times higher activity in comparison with original procedure.

Relative competition among various plant parts for N during water stress,i.e. nitrogen distribution index (NDI) was determined in relation to specific nitrogenase activity (SNA) and nodule and soil nitrogen in both indeterminate (H-77-216) and determinate (ICPL-151) types of pigeonpea (Cajanus cajan L.) under greenhouse conditions. Two levels of water stress,i.e. moderate (soil Ψ_w) -0.77 MPa) and severe (soilΨ_w -1.34 MPa) were created by witholding the irrigation at vegetative (40 DAS) and flowering (70 DAS) stages. At vegetative stage under moderate stress the highest NDI was in nodules of cv. H-77-216 and in leaf of cv. ICPL-151. Under severe stress both the cultivars showed negative values of NDI, with maximum loss of N from root and nodules. Cultivar ICPL-151 behaved differently at flowering and vegetative stages. Very high loss of N from different plant parts was seen at flowering under severe stress. All the plant parts showed gain in N during rehydration. Loss and gain in N at both the stages under stress and rehydration respectively, correlated with available N in soil. Specific nitrogenase activity (SNA) and nodule N were maximum at moderate stress and related with NDI values of leaf and nodules.

The fungal colonization of the angiosperm root parasiteCynomorium coccineum and the halophytic hostsLimonium delicatulum andArthrocnemum glaucum were investigated in a Mediterranean salt marsh in March 1992. The main fungal inhabitants on the leaves or shoot surface of the test plants wereAspergillus niger, Penicillium chrysogenum andCladosporium herbarum. The qualitative analysis of the fungal species associating the parasite, the hosts and the non-infected plants showed similar pattern. However, the total population exhibited quantitative differences coupled with the amount and the chemical composition of the exudates on plant surface and the quantity of transpired water. The fungal catch from the aerial shoot (inflorescence) of the parasite was higher than that collected from either the leaves or aerial shoots of non-infected or host plants. The fungal density on the leaves ofL. delicatulum was higher than those isolated from the aerial shoots ofA. glaucum. Infection byC. coccineum caused a marked drop in the total fungal population on leaves or shoot surfaces of the hosts as compared to the corresponding non-infected individuals. The stimulative effect of washings on spore germination of some isolated fungal species was matched with the density of fungi on the target plants.