Responses of wheat (Triticum aestivum L.) to various concentrations of NaCl and levels of drought were followed. With the rise of NaCl or drought, or NaCl and drought together, growth was retarded. The water content of shoots and roots was mostly unchanged. The chlorophyll and carotenoid contents were increased in plants subjected to salinity or drought or both. Only high salinity level induced a considerable decrease in net photosynthetic rate (P_N) and dark respiration rate (R_D). P_N and R_D were decreased with the decrease of soil moisture content. The content of Na⁺ in the shoots and roots of wheat plants increased with increasing salinity or decreasing soil moisture content or both treatments. Considerable variations in the content of K⁺, Ca²⁺ or Mg²⁺ were induced by the NaCl, drought or both treatments.

Arabidopsis thaliana tumors were induced by octopine strain ofAgrobacterium tumefaciens B6S3 and its derivatives with modified T-DNA. Flowering shoots appeared sponta-neously onin vitro cultivated tumors and set seeds. R₁ and R₂ progeny of octopine synthesizing plants segregated in opine synthesis activities 3:1 and 15:1. Octopine synthase activity showed absolute linkage with agropine synthesis in most lines. In R₃ and R₄ progenies, the fraction of octopine synthase and agropine synthesis positive plants was lower than expected, but Mendelian segregation was restored if plants were cultivated on medium with 5-azacytidine. The most probable mechanism of disapearance of opine synthesis is cytosine methylation. The effect of 5-azacytidine lasted for at least next two generations.

Leaves of micropropagated highbush blueberry (Vaccinium corymbosum) cv. 'Bluetta' have been observed during the acclimatization phase. In vitro-developed leaf cells were circular and small, the spongy parenchyma was discontinuous and disorganized and formed by 1-2 layers of cells with large intercellular spaces and the palisade to spongy mesophyll thickness ratio was 1:1.5. After rooting ex vitro, the first leaves formed under natural conditions showed substantial changes in the anatomical characteristics. After 6 months, the plants produced leaves similar to those in field-grown plants. The palisade cells were rectangular, the spongy parenchyma was formed by 3-4 layers of cells and the intercellulars were around the stomata. Leaves from field-grown plants lost 24 % of water during 150 min after excision while leaves from in vitro shoots lost about 50 % of water in the same time. Leaves from in vitro shoots showed a higher number of smaller stomata (361 per mm²), with the guard cells forming a circular ring; the stomata frequency in field-grown leaves was 241 per mm² and the guard-cells were elliptical.

An improved method for the isolation of leaf mesophyll protoplasts ofSolanum tuberosum L. cvs. Xenia and Bintje, their culture and regeneration to plants is described. The improvements involve the culture of nodal stem explants on Schenk and Hildebrandt (SH) medium supplemented with AgNO₃ (7.5 mg 1¹) and Alar 85 (5 mg 1^-1), and the pre-treatment of donor plants for 2-3 weeks at a short photoperiod (6 h), a low irradiance (17 (μmol m^-2s^-1) and a low temperature (20 °C). Regeneration of cell wals was observed within 2-3 d and cell divisions within 7-12 d after resuspending freshly isolated protoplasts in Sidorov et al. (SW) medium. The resulting microcalli were cultured on solid media containing zeatin (1-2,5 mg 1^-1) and/or NAA (0.1 and/or 0.01 mg 1^-1) and subsequently on medium with BAR (0.25 mg1^-1) and GA^-3 (0.1 mg 1¹) for shoot regeneration.

A method for micropropagation ofDalbergia sissoo has been developed. Single node segments obtained from coppice shoots of a mature tree (20 - 25 year old) produced 3-4 shoots per explant on Murashige and Skoog (MS) medium containing 4.4 x 10^-6 M benzylaminopurine (BAP) and 4.4 × 10^-7 M of Β-naphthoxy acetic acid (NOA) (shoot multiplication medium) within 4 weeks. Thein vitro regenerated shoots were 3 - 4 cm in length and provided 2 to 3 culturable nodal segments which on shoot multiplication medium again produced 3-4 shoots. Following this procedure 18-24 shoots were produced from single nodal segment within 60 d. 80 % of the shoots directly produced five roots when they were firstly treated with MS medium supplemented with 10^-5 M indole-3-butyric acid (IBA) and subsequently transferred to half strength liquid MS medium containing 1 % activated charcoal followed by half strength liquid MS free hormones, vitamins and activated charcoal. Thein vitro raised plants were hardened for survival after transplantation to soil by exposing them to various humidity conditions, gradually from higher to low, with nearly 100 % transplant success.

In pea, maize and tomato plants a hitherto undescribed L-tryptophan dehydrogenase activity (TDH) has been detected. This enzyme catalyzes the reversible formation of indolepyruvic acid (IPyA) from L-tryptophan (L-trp). TDH and L-glutamate dehydrogenase (GDH), related enzymes in their mode of action, could be separated by gel chromatography. Enzymatic activity of TDH was sustained by both pyridine coenzymes NAD/NADP. With pea TDH the coenzyme NAD displays, at optimum pH 8.5 and at room temperature, only about 40-70 % of the activity of NADP. The amination of IPyA is catalysed more actively than the deamination of L-trp. L-trp/IPyA, L-glu/ketoglutarate, L-ala/pyruvate reacted as dehydrogenase substrates; L-phe/ phenylpyruvate, D-trp and D-phe did not react with pea enzyme extracts. A considerable similarity between the active centres of TDH and GDH has been found using inhibitors: absence of heavy metals, presence of a carbonyl group, indispensibility of bivalent ions for the enzyme activity. Pea TDH and GDH were distinctly inhibited by sodium azide. For the activity of TDH the presence of SH groups is less important than for GDH. The TDH activity in the investigated plants was lower than the GDH activity. The possible role of TDH in the regulation of the IPyA pool is discussed.Doc. RNDr. PhMr. M. Kutáček died on 28 November, 1989. The final form for print was prepared by dr. Ivana Machdckovd of the same Institute, who will also answer the reprint requests. Received June 6, 1990; accepted October 10, 1990

Drought created by withholding the irrigation at 30 and 45 d after sowing significantly decreased relative water content (RWC) and osmotic potential (ψ_s) ofVigna radiata (L.) Wilczek cv. MH-83-30 nodules. Potassium fed plants showed higher RWC, whereas ψ_s was further declined irrespective of soil moisture levels. The nitrogenase activity and leghemoglobin content of nodules markedly decreased under drought and nodules of potassium fed plants showed better recovery after rehydration. The proline content significantly increased under drought but declined upon reirrigation. Also, the C, N and K contents of nodules significantly declined under drought.

Sampling of natural cocksfoot (Dactylis glomerata L.) populations was carried out on the O Morrazo peninsula in NW Spain, characterized by a strong moisture gradient. The plants were kept in greenhouse under standard conditions. Nevertheless, they differ in height of plants, length and width of flag leaves, panicle size, stomatal density and size as well as in flowering period. The effects of two levels of soil water deficit and two levels of salinity on photosynthetic rate were tested. One population was exceptionally well adapted to its original environment with great tolerance to water deficit and salinity

The effect of crude volatile oils from the leaves ofEucalyptus globulus andE. citriodora and the pure terpenes - cineole and limonene from these oils, (in vapour form) was studied onPhaseolus aureus, Lens esculentum, Hordeum vulgare and Avena sativa. The parameters like germination of seeds, seedling growth, values of cell survival, and content of water and chlorophyll of the crops formed the system of bioefficacy study. The allelopathic impact of the oil vapours from the eucalypt tree becomes evident from the negative response of the parameters studied. The impact of the E. citriodora oil vapours compared to that ofE. globulus oil or the pure terpenes was seen to be relatively greater in almost all parameters under investigation. A strong reciprocal correlation that exist between the concentration and the seedling growth or the water content of the crops under study supports the dose linked allelopathic phenomenon. It is suggested that oil vapours ofEucalyptus exert their effect through impairing the respiratory as well as photosynthetic ability of the target plants.

The changes in ubiquitin mRNA level in intact plants of both susceptible (cv. Spunta) and partially resistant (cv. Pampeana-INTA) potato cultivars after inoculation with low doses (l0³ sporangia cm^-3) ofPhytophthora infestons were studied after 72 h of treatment. Inoculation leads to 5-fold accumulation of potato ubiquitin transcripts in both cultivars. This result supports the connection between ubiquitin expression and defense reaction in plants.

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.

Plants regenerated from callus cultures derived from leaf discs and mesophyll protoplasts ofPetunia hybrida cv. Rose of Heaven exhibit a high frequency of genetic and chromosomal variation. Of twelve leaf disc-derived plants examined, only three had the normal diploid chromosome number (2n=14) while seven were tetraploid and two were aneuploid (16 and 27 chromosomes). Of seventeen plants derived from two protoplasts, none had the diploid chromosome number. Most had 28 chromosomes, one 29, two 27, one 26 and one had variable numbers (14-28) in different root tip cells. In all cases aneuploidy was associated with developmental abnormality. In addition, heritable differences in growth, morphology and flower pigmentation were observed in callus-derived tetraploids and diploids, including one diploid which differed from parent plants in at least four characters. These results are discussed in terms of the importance ofPetunia in genetics research and for studies of somaclonal variation.

Decapitated pending vines are the most responsive parts of vanilla plants. Inflorescence localization along training vines showed a decreasing gradient from the decapitated end to the base in sun locality and was irregular and showed opposite gradient in shade locality. Soluble and ionic peroxidase activities, determined in the leaves and in the internodes, showed a marked peak during the end of June and a gradient along training vines which was inverse to the gradient of inflorescence localization and opposite between sun or shade conditions, supporting the utility of peroxidases as spatial and temporal biochemical markers of flowering processes.

Esterase isoenzyme spectra of different organs of seedlings and field-grown plants of fiber flax (Linum usitatissimum L., cv. Belinka) were studied by electrophoresis in polyacrylamide gel for estimating ontogenetic variability of gene expression. Formation of individual isoesterases depended on the type of tissue and the stage of its development. Isoesterases characteristic of exclusively one or some tissues of the same developmental stage were revealed simultaneously with basic esterase isoforms active in all analysed parts of seeds, seedlings and field-grown plants. The revealed changes of esterase isoenzyme spectrum during germination show tissue and time specificity of the endogenous regulation of genes controlling their formation.

Transpiration rate, stomatal frequency, growth, contents of pigments, saccharides, total nitrogen, proteins and some nutritive elements (K, Ca, Mg, P) of radish plants were significantly lowere dwith the rise in salinization levels using NaCl. Spraying radish shoots with proline solution (200 g m-³) counteracted the above adverse effects, especially at low and moderate salinity.

The interaction between native and introduced fungi and their effect on plant growth and mineral uptake were studied. The host plants wereLygeum spartum andAnthyllis cytisoides, the introduced fungus wasGlomus fasciculatum. The four soils used were selected from disturbed and contaminated by mining activities areas. Inoculated and uninoculated plants were grown in the unsterilized and sterilized soils (with and withouth native microflora, respectively). Plants inoculated withG. fasciculatum were higher and had higher tissue P concentration than uninoculated plants, especially inA. cytisoides. However, this inoculation was not effective in unsterilized substrates, suggesting a competition between introduced and native fungi. Concentration of mineral elements other than P varied depending on the host plant and soil. Decrease in Fe, Cu, Mn, Zn and Pb was observed in mycorrhizalA. cytiosides plants and a slight increase in Zn concentration was noted in mycorrhizalL. spartum plants. The study showed that the type of soil and their populations of native endophytes have a considerable effect on plant response to mycorrhizal symbiosis, especially in disturbed soils.

The C₄ speciesFlaveria trinervia is obviously better adapted to saline environments than the C₃ speciesF. pringlei. Treatment with 100 mM NaCl diminished crop growth rate inF. pringlei by 38% but not inF. trinervia. Under saline conditions, more assimilates were invested in leaf growth inF. trinervia but not inF. pringlei. Electrolyte concentration inF. trinervia in control and salt treated plants is lower than inF. pringlei. Fluorescence data do not indicate a damage of PS 2 charge separation in both species. Whether the C₄ photosynthetic pathway inF. trinervia is responsible for the improved salt tolerance compared toF. pringlei remains an open question.

In six rice cultivars the relative amounts of 55, 26-28 and 15 kDa polypeptides were mostly lower in plants grown for 15 d under low irradiance than in plants grown under saturating irradiance. This decline in the polypeptides, especially in those related to light-harvesting chlorophyll proteins, was accompanied with a decrease in the whole chain electron transport and the photosystem 2 activity.

The content of lead in greening etiolated barley leaves remained the same, regardless the time of incubation of excised leaves in the presence of lead ions (8-24 h). The lead deposits have not been detected within mesophyll cells, but were found in intercellular spaces of mesophyll, in guard cells and in cuticle covering stomata. This suggests that lead may be transported in the leavesvia transpiration stream. Lead reduced the content of chlorophyll, especially chlorophyllb content and the average number of grana, whereas in the presence of kinetin the content of chlorophyll increased. In the combined treatment (lead + kinetin) kinetin diminished the inhibitory effect of lead on the chlorophyll content. The number of chloroplasts in mesophyll cells remained unchanged after lead treatment, whereas kinetin alone or applied together with lead increased the average chloroplasts number. The thylakoids system in chloroplasts of kinetin and kinetin + lead treated plants was similar to that observed in control, although the grana number was smaller. Both lead and kinetin increased the content of condensed chromatin in nuclei.

The effects of salinity (0, 50, 100, 150, and 200 mM NaCl) and heat-shock (42°C) and their interactions on germination, seedling growth, and some relevant metabolic changes of two cultivars (cv. Giza 155 and cv. Stork) of wheat (Triticum vulgaris L.) were studied. Germination studies indicate that plants tolerated salinity up to 100 mM NaCl. The lengths of roots and shoots and their water content, as well as fresh and dry matter yield of cv. Giza 155 seedlings remained more or less unchanged up to 100 mM NaCl and of cv. Stork up to 50 mM NaCl. Salinity induced progressive increase in soluble carbohydrates, soluble proteins and proline in cv. Giza 155 and in soluble proteins, proline and other free amino acids in cv. Stork. However, under the higher salinity levels, in cv. Giza 155 increase in soluble carbohydrates was accompanied by lose in other free amino acids, whereas in cv. Stork an opposite effect was obtained. Heat-shock treatment (42°C for 24 h) induced a significant decrease in the final germination percentage, the shoot and root lengths, fresh matter yield and the water content. The dry matter yield of the two cultivars was considerably increased as compared with the corresponding treatments with NaCl only. Heat-shock treatment resulted in a significant increase, in the amount of soluble carbohydrates and proline in salt treated seedlings of both cultivars. The pattern of changes in amino acids was opposite to that of soluble proteins, indicating that the increase in soluble proteins was at the expense of other amino acids in cv. Giza 155 andvice versa in cv. Stork.

Patchouli (Pogostemon cablin Benth.) plants grown under shade (LI) showed an increased height due to internodal elongation, leaf area, leaf area index, and chlorophyll (Chl)b, Chl (a+b) and carotenoid (Car) contents compared to the plants grown in sunlight (HI). The number of branches and green leaves decreased under LI treatment, with a marginal variation in the patchouli oil yield. A comparison between the relative contents of photosynthetic pigments indicated that Chlb and Car accumuled preferentially over Chla in the LI grown plants.

The effects of Cu²⁺ on growth, chlorophyll and other ion contents ofKoeleria splendens originated from Cu-contaminated soil have been investigated in nutrient solution. The most evident Cu²⁺ effects concern the root growth, especially the root length. Since in plants grown under lower Cu²⁺ concentrations (4 and 8 μM) root elongation, biomass, chlorophyll, Mg²⁺, Fe²⁺, Ca²⁺ and K⁺ content were increased compared with the control, the development of an adaptive mechanism ofK. splendens to Cu²⁺ is suggested. High Cu²⁺ concentration (160 μM) caused a significant reduction in root length and biomass as well as a decreased rate of chlorophyll biosynthesis. The reduction of growth can be correlated with the toxic effect of Cu²⁺ on photosynthesis, root respiration and protein synthesis in roots. 160 μM Cu²⁺-treatment had a negative influence on the concentrations of Ca²⁺, Fe²⁺, Mg²⁺ and K⁺ and a positive influence on the Cu²⁺ concentration in the plant tissues. Loss of nutrients similar to the senescence response suggests that excess of Cu²⁺ leads to the progressive senescence of the plants. Our results demonstrate the existence of an adaptive mechanism ofK. splendens under low Cu²⁺ concentrations, while high Cu²⁺ quantities cause disturbances in plant function.

Molybdenum appliedin vivo to winter wheat (Triticum aestivum L., cv. Sadovo 1) grown on acid soil had cryoprotective effect. In Mo-treated plants the amount of digalactosyl diacylglycerol in the leaves increased during cold acclimation and that of monogalactosyl diacylglycerol remained relatively unchanged. An opposite tendency was observed in untreated plants. The most pronounced effect of freezing temperature on lipid changes was observed with respect to the different rate of phospholipid degradation in both kinds of plants. The amount of phospholipids at -5°C in Mo-treated plants increased while in untreated ones it significantly decreased. In general, the changes observed in Mo-treated plants corresponded to those shown for frost resistant plants.

Influence of inoculation with efficient rhizobia or nitrate fertilization in alleviating salinity (NaCl, CaCl₂ and Na₂SO₄) stress was investigated in sand culture experiments. Shoot dry mass declined beyond salinity level corresponding to electrical conductivity (EC) 5.6 dS m^-1 in control or in inoculated plants and after EC 7.4 dS m^-1 in nitrate fed ones. Root growth was more sensitive and decreased at EC 3.3 dS m^-1. Nitrate reductase activity in leaves reduced at EC 3.3 dS m^-1 but in inoculated and nitrate fed plants it reduced at EC 5.6 dS m^-1. Na⁺ accumulation increased at EC 5.6 and 7.4 dS m^-1 in roots and, shoots, respectively. In inoculated and nitrate fed plants Na⁺ content in roots increased at EC 7.4 dS m^-1. Content of Ca²⁺ increased slightly only in shoots and content of K⁺ was unaffected. Besides inoculation, application of small doses of nitrogen should prove beneficial for legume cultivation in saline soils.

The effect of 80 mM NaCl on the structure and ultrastructure of root cells ofPhaseolus vulgaris plants has been investigated. Roots of plants treated with NaCl were shorter and had less secondary roots than control plants. In control plants, epidermal cells were isodiametric and uniformly placed forming a thin layer, whereas in stressed plants, the shape and disposition of epidermal cells was less regular. The cortical cells of control plant were round-shaped and distributed allowing large and well defined intercellular spaces, whereas stressed plants presented irregular cells, which were interdigitated, thus decreasing the volume of the intercellular spaces. Presence of 80 mM NaCl did not result in significant differences in the number, shape or distribution of the cell organelles. Membrane vesiculation was often observed in cells of NaCl treated plants. Addition of 80 mM NaCl to the growth medium considerably increased the leakage of solutes from intact plant roots back to the solution especially K⁺ and Ca²⁺.

Nodules of faba bean (Vicia faba L. cv. Giza 3) plants grown in pots containing clay-loam soil for 90 d have an active nitrate reductase (NR), while the leaves did not show detectable activity. Spraying the plant with increasing concentrations of Al³⁺ or Cd²⁺ (0-1000 μM) significantly inhibited the nodules NR activity, the decline being more pronounced in Cd²⁺ treatment. The specific activity of glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) were more prominent in the 60- than in 90-d-old plants; GOT was always higher than GPT. Furthermore, GOT was more sensitive to Al³⁺ and Cd²⁺ treatments and its activity was significantly decreased when the metal concentration increased. Also, Cd²⁺ proved to be more effective than Al³⁺ in suppressing the GOT activity in the nodules, with less significant effect observed in the leaves. In contrast, GPT was hardly affected by the various metal treatments, particulary in the leaves.

The effects of 4 or 8 drought cycles on four grass species,Cenchrus pennisetiformis, Leptochloa fusca, Panicum turgidum, andPennisetum divisum were assessed in a pot experiment. There were significant differences between the species in biomass production under water stress.C. pennisetiformis andP. turgidum produced significantly greater fresh and dry matter thanP. divisum and especially thanL. fusca. L. fusca had the lowest andP. divisum highest osmotic potentials compared with the other species after the completion of 4 or 8 drought cycles. Osmotic adjustment (difference between osmotic potential of droughted/rehydrated plants and control plants) was highest inL. fusca. The stomatal conductance was significantly decreased with increased drought stress inC. pennisetiformis. The elasticity ofC. pennisetiformis, P. turgidum andP. divisum increased with increase in number of drought cycles, whereas that ofL. fusca remained unchanged.L. fusca andP. turgidum had the lowest leaf hydration of all species after 8 drought cycles. The chlorophyllsa andb in all species remained unaffected by drought treatments. The proline content ofC. pennisetiformis andL. fusca increased significantly with increased drought stress, whereas that ofP. turgidum remained unaffected after 4 or 8 drought cycles.L. fusca synthesized great amount of leaf soluble proteins during 8 drought cycles, whereasP. divisum had low protein content after 4 drought cycles. The protein contents ofC. pennisetiformis andP. turgidum remained unaffected after 8 drought cycles. The leaf epicuticular wax ofL. fusca increased consistently with increased drought stress, but leaf wax ofP. divisum increased only at the highest drought stress and that ofC. pennisetiformis andP. turgidum increased after 4 drought cycles. On the basis of these results it was established thatC. pennisetiformis andP. turgidum were the most tolerant,P. divisum intermediate, andL. fusca the most sensitive to drought stress. The osmotic adjustment did not positively correlate with the degree of drought resistance.