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

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

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.

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.

Wounding both cotyledons ofBidens pilosa (var.radiatus) induces the inhibition of hypocotyl growth. The wound signal is transmitted very rapidly from cotyledon to hypocotyl and can be visualized by the change in nucleotide pools. First we have shown that the irradiance of the plant can change the ATP level without plant wounding. Therefore, plants were harvested at the start of the light period. Under these conditions, we have determined in hypocotyl the levels of adenosine triphosphate (ATP), guanosine triphosphate (GTP) and non adenylic triphosphates (NTP), and adenylate energy charge (AEC) after wounding. We have observed a transient (2 min) increase in the ATP level followed by a decrease 5 to 30 min later. A similar result was obtained for the GTP level but with some delay. The GTP level increased in 5 min and then decreased after 60 min. For the NTP level the decrease is effective from 5 to 60 min after wounding. The calculation of AEC has shown that a very tight control in the level of ATP may be involved in response to wounding.

The effects of fusaric acid, a phytotoxin produced byFusarium pathogens, on the metabolism of isolated maize root mitochondria and on maize seed germination and seedling growth were investigated. The phytotoxin inhibited basal and coupled respiration when succinate and α-ketoglutarate were the substrates. Coupled respiration dependent on NADH was inhibited, but basal respiration was not. Consistently, succinate cytochromec oxidoreductase activity was decreased whereas NADH cytochromec oxidoreductase was not affected. The ATPase activities of carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone stimulated mitochondria and of freeze-thawing disrupted mitochondria were inhibited. These results indicate that the phytotoxin impairs the respiratory activity of maize mitochondria by at least three mechanisms: (1) it inhibits the flow of electrons between succinate dehydrogenase and coenzyme Q, (2) it inhibits ATPase/ATP-synthase activity and (3) it possibly inhibits α-ketoglutarate dehydrogenase. Seed germination and seedling growth were also affected by fusaric acid with the most pronounced effect on root development. These effects can possibly contribute to the diseases ofFusarium- infected plants

Agrobacterium tumefaciens binary vectors carrying kanamycin resistance gene and either C58 T-DNA gene 4 for cytokinin synthesis or genes 1 and 2 for auxin synthesis were constructed and used for transformation of a short-day tobacco Maryland Mammoth. Kanamycin resistant plants were regenerated from a small fraction of transformed tissue and the presence of T-DNA in their genome was verified by Southern blotting. The level of endogenous cytokinin in plants transgenic for gene 4 and the level of endogenous IAA in those transgenic for genes 1 and 2 increased by more than 100 %. A number of morphological characteristics distinguish them from untransformed controls.

Buthionine sulfoximine (BSO) specifically inhibits γ-glutamylcysteine synthetase and decreases a cellular level of glutathione (GSH) in maize seedling roots. Exogenous GSH restores Zn-phytochelatins synthesis in BSO-treated maize plants.

Phaseolus vulgaris L. seedlings were grown under different air humidities simulating conditions during micropropagation (very high humidity duringin vitro cultivation and low air humidity after transferex vitro). The functional stomatal apparatus developed after a short period of growth at low air humidity at the beginning of plant ontogeny or after transfer from high to low air humidity, but not in plants grown steadily under high air humidity. The ability of stomata to regulate gas exchange was not persistent and disappeared after transfer of plants from low to high humidity.

Amphistomatous C₃ (Nicotiana tabacum L., Datura stramonium L.) and C₄ (Sorghum saccharatum Pers. and Zea mays L.) species were examined to find how (if at all) their inherent differences in water-use economy are reflected in apparent cuticular transpiration or vice versa. Transpiration efficiency (TE) was calculated from steady state photosynthesis (A) and transpiration (E) rates estimated for the upper side of the leaf after light induction of stomata opening. Apparent cuticular transpiration ('E_c) was measured as the part of transpiration which was not eliminated by convective counteraction of the air stream passing across the amphistomatous leaf: total pressure difference (AP) across the leaf was increased and the minimal value of E_ΔPτ0 was taken as the apparent cuticular transpiration rate ('E_c). 'E_c was treated relative to E at AP equal to zero (E_GDP=0), E'_cr. Measurements were carried out under two leaf-air vapour pressure differences (VPD).
E_r (i.e. E_GDPτ0/E_GDP=0) versus GDP patterns differed qualitatively between the investigated C₃ and C₄ plants. TE increased and 'E_cr decreased from tobacco, stramony, maize to sorghum for both VPD of air. 'E_cr and TE were approximately linearly related, the slope being dependent on VPD. The increase in VPD resulted in larger E and slightly smaller epidermal conductance (g) at GDP equal to zero. Both E'_cr and E'_cr decreased markedly at the same time especially, for species with high TE. The results were considered as an indirect confirmation that E'_c values estimated by the technique used reflect species-specific differences in external peristomatal and cuticular vapour loss, at least in a relative sense.

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.

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.

Limitations on photosynthesis, characterized by leaf CO₂ exchange, chlorophyll fluorescence, and thylakoid structure, were studied under environmental conditions simulating culturein vitro. These were simulated by growingPhaseolus vulgaris plants in nutrient solution under high relative humidity of air (>90%), and CO₂ concentrations (c_a) that decreased with the development of photosynthetic activities during plant ontogeny (1200 to 300 mg m^-3). The ontogeny of such model plants was more rapid, primary leaves reached photosynthetic maturity 2 to 3 d earlier and their life span was 7 to 14 d shorter than in control plants. Their photosynthetic activityin situ was limited, after reaching "photosynthetic maturity", similarly to plants grownin vitro. When measured under optimal conditions, however, 50 to 70% higher net photosynthetic rates (P_N) were found in leaves of different ages as compared with plants grown under c_a of 700 mg m^-3 and a lower air humidity (30-35%). This increase in P_N was associated with a high conductance for CO₂ transfer by adaxial and abaxial epidermes. In model plants, the dark respiration rate (R_D) was almost twice that in the control, while the photorespiration rates were similar to controls; CO₂ compensation concentration was about 50% of that in controls. The ratios P_N/R_D were similar in control and in model plants. Chlorophylla+b content in leaves of the model plants was lower than that in the control plants. Grana extent increased with plant age in the model plants while it decreased in the control ones. In both the stromal and granal membranes of the chloroplasts in model plants, a marked accumulation of carotenoids occurred independent of age. The ratio of variable to maximal fluorescence, F_v/F_m, did not differ in the model and the control plants. In the control plants, photochemical quenching (q_P) slightly increased with plant age and was not affected by CO₂ concentration present during measurement. In the model plants, q_P increased with elevated CO₂ concentration in young plants and decreased in saturating CO₂ concentrations in older plants. Nonphotochemical quenching (q_NP) was lower in the model plants and increased under CO₂ saturating conditions. Vitality index, Rfd, was markedly lower in the model plants than in the control ones and a decline was found in saturating CO₂ concentration.

The influence of increasing copper concentrations on seed germination, seedling survival and radicle length ofMinuartia hirsuta, Silene compacta, Alyssum montanum andThlaspi ochroleucum was studied. Seed germination was highly affected by the higher Cu²⁺ concentrations (80 and 160 μM), while lower Cu²⁺ concentrations seemed to be necessary for seed germination, even for the plants originated from non-Cu²⁺-rich soils (i.e. A. montanum). Nevertheless, plants originated from Cu²⁺-rich soils (M. hirsuta, S. compacta) showed a higher demand of Cu²⁺ for rapid seed germination. Cu²⁺ at higher concentrations severely reduced growth rate of radicle, especially inA. montanum andT. ochroleucum. These data clearly indicate the reduced suitability of the above mentioned plant species for reclamation on Cu²⁺ soils. Lower Cu²⁺-concentrations had no influence on seedling survival inM. hirsuta andS. compacta, but a progressive reduction of a number of survived seedlings with increasing Cu²⁺ concentration was found, that was more pronounced inA. montanum andT. ochroleucum.

Changes in growth and phosphorus content in plants and seeds of fenugreek with increasing cadmium concentration was evaluated. Root length and shoot length ranged from 11.63 to 27.72 and from 9.70 to 54.78 cm, respectively. With the increasing Cd²⁺ concentration there was a significant decrease in root and shoot length, and fresh mass. Various phosphorus fractions of shoot decreased with increasing Cd²⁺ concentration except lipid P and nucleic acid P which increased at 65 and 95 d after sowing and protein P only increased at vegetative stage. In seeds (60 d after flowering) lipid P increased except at 2.5 μg(Cd²⁺) g^-1 (soil) while protein P decreased.

The effects of NaCl and polyethylene glycol (PEG) on superoxide dismutase (SOD) and peroxidase (P) activities, lipid peroxidation (LP) and proline content in seeds and leaves of drought tolerant (FC-506 and MS-100) and drought sensitive (MS-612 and MS-13) sugar beet cultivars were examined. After PEG and NaCl treatment in tolerant cultivars both in seeds and leaves SOD activity mainly increased, though P activity increased only in leaves of tolerant cultivars. In drought sensitive cultivars the decrease of SOD and P activity was mostly observed. LP increased in seeds and leaves of all examined cultivars. The proline content increased in the leaves of examined cultivars and was significantly higher in drought tolerant plants. On the other hand, in the seeds only slight increase in proline content was found. The results obtained indicated that drought tolerance could be correlated with high proline content and enzymatic defense against lipid peroxidation.

Proline content, ion accumulation, cell wall and soluble peroxidase activities were determined in control and salt-treated calli (150 nM NaCl) and whole plants (30 mM NaCl) of two rice cultivars (salt sensitive cv. IKP and salt tolerant cv. Aiwu). Under salinity, the highest accumulation of Na⁺, Cl^- and proline occurred in calli, roots and younger leaves of cv. IKP, coupled with the highest decrease in K⁺ content; accumulations of Na⁺ and Cl^- were restricted to older leaves in cv. Aiwu. Relative growth rates of calli and roots or shoots from both cultivars were not linked to peroxidase activities. High concentrations (1 M) of exogenously applied glycerol did not inhibitin vitro activities of soluble peroxidase extracted from control and salt-treated calli or plants. Conversely, 35-55% (in cv. IKP) or 60-80% (in cv. Aiwu) of soluble peroxidase activities were found in presence of isosmotic proline concentration. There were no differences between proline and glycerol effects onin vitro cell wall peroxidase activities.

Several morphological characteristics differed when wheat (Triticum aestivum L. cv. Doha 88) was grown under a cool (10 °C), warm (20 °C), and hot (30 °C) regime. Development of leaves was linearly related to shoot meristem temperature, and the time between appearance of successive leaves on the main culm was independent of growth temperature. Area and dry mass of leaves and roots increased exponentially with time, and variations between growth temperature regimes were reduced when plants were compared at a similar developmental age. In isolated thylakoids thermal stability of photosystem 2 and of whole electron transport chain was enhanced with the increase in growth temperature. Therefore this cultivar is able to acclimate to contrasting temperature regimes.

Rates of whole chain and photosystem 2 activities in chloroplasts isolated fromVigna unguiculata L. seedlings grown under ultraviolet-B (UV-B) enhanced radiation were less affected by 3, 6 and 9 mM CdCl₂ for 60 min at 0 °C in the dark than the rates in chloroplasts from control plants grown under normal irradiation. The results are in agreement with changes in contents of chloroplast 55, 47, 43, 33, 29, 27-25, 23 and 17 kDa polypeptides that were significantly lowered at 3, 6 and 9 mM CdCl₂ only in chloroplasts from control plants. On the other hand, in the simultaneous treatment of chloroplast isolated from control plants the UV-B supported the inhibitory effect of all applied concentrations of CdCl₂. The photosystem 1 activity was only marginally affected in the all experimental variants.

Lotus corniculatus has been transformed byAgrobacterium tumefaciens harbouring therolABC, rolAB, and 35S-rolC constructs. Growth, shoot and root morphology, cytokinin and auxin concentrations in the shoots, chlorophyll content in the leaves, nodulation and nodule nitrogenase activity of clonal progenies of selected transformants, in which the transferredrol sequences were proved, were compared with control plants.
The most striking alterations were observed in the 35S-rolC transformants (dwarfism, shoot and root branching, shorter internodes, delayed nodule formation, decreased nitrogenase activity, reduced content of leaf pigments) in comparison with either therolAB, rolABC, or controls. TherolAB androlABC transformants were very similar to the control plants in their morphology as well as in most other traits measured.

The barley (Hordeum vulgare L.) seeds were germinated in the non-saline conditions after 12 h imbibition in 2 % of NaCl solution. The results of treatment were: (l) the membrane system in meristematic cells of root tips developed well; (2) many profiles of endoplasmic reticulum and Golgi bodies appeared; and (3) the quantities of amoeboid plastids and amoeboid mitochondria increased. Thus the inhibitory effects of short-term NaCl stress on plants were reversible, and simultaneously NaCl treatment enhanced the metabolic activities in cells. The amoeboid form may be an adaptive form of plastids or mitochondria to an enhanced metabolic activity.