The interactive effects of salinity stress (40, 80, 120 and 160 mM NaCl) and ascorbic acid (0.6 mM), thiamin (0.3 mM) or sodium salicylate (0.6 mM) were studied in wheat (Triticum aestivum L.). The contents of cellulose, lignin of either shoots or roots, pectin of root and soluble sugars of shoots were lowered with the rise of NaCl concentration. On the other hand, the contents of hemicellulose and soluble sugars of roots, starch and soluble proteins of shoots, proline of either shoots or roots, and amino acids of roots were raised. Also, increasing NaCl concentration in the culture media increased Na⁺ and Ca²⁺ accumulation and gradually lowered K⁺ and Mg²⁺ concentration in different organs of wheat plant. Grain soaking in ascorbic acid, thiamin or sodium salicylate could counteract the adverse effects of NaCl salinity on the seedlings of wheat plant by suppression of salt stress induced accumulation of proline.

The effects of 2-aminopurine, olomoucine, 6-dimethylaminopurine (inhibitors of cyclin-dependent kinases; CDK), and sodium vanadate (a potent inhibitor of protein phosphatases) on DNA endoreduplication were investigated during elongation and differentiation of the primary roots in Pisum sativum. When compared with the untreated control plants, at least one additional round of DNA replication was evidenced to occur within most cells, the majority of which have attained 4C DNA level, and a considerably greater portion of cells represented the endopolyploid state with nuclear DNA content approximating the 8C value. It is concluded that cellular commitment to DNA endoreduplication may appear not only as a consequence of suppression imposed directly upon CDK activity, but also as an indirect output connected with the decreased activity of cdc25 protein phosphatase, an enzyme necessary to turn the switch on for appropriate conformation of the CDK/cyclin B complex. By calculating the absorption profiles of Feulgen-stained nuclei, specific phosphorylation-dependent changes in chromatin condensation of endopolyploid cells have been revealed. It is proposed that acquisition of a certain critical level of chromatin condensation constitutes a prerequisite for additional rounds of DNA synthesis in plants.

Spirodela punctata was cultivated on phosphate-deficient medium (-P_i) with racemic 1-amino-2-phenylethylphosphonic acid (PheP) as a source of P_i. The growth of duckweed was inversely correlated with PheP concentration. The growth of plants on medium -P_i with 0.1 M PheP was accelerated whereas with 0.001 mM PheP was slower than in -P_i control. PheP at low concentrations decreased loss of chlorophyll in comparison with -P_i plants. Content of anthocyanins decreased but activity of the extractable constitutive phosphatases of pH 6.0 and pH 7.5 increased along with increasing concentration of PheP in the medium. We suggest that S. punctata does not break P-C bonds but probably PheP interrupts processes involved in the regulation of P_i-starvation response.

Loss of chlorophyll (Chl) and carotenoids (Car) of leaves and changes in Chl fluorescence emission and polarisation, malondialdehyde (MDA) accumulation, and 2,6-dichlorophenol indophenol (DCPIP) photoreduction in chloroplasts of wheat seedlings grown under different irradiance and subsequently exposed to high irradiance stress (HIS; 250 W m^-2) were studied in mature and senescent primary wheat leaves. Faster rate of pigment loss was observed in leaves of moderate irradiance (MI; 15 W m^-2) grown plants, compared to high irradiance (HI-1 and HI-2; 30 and 45 W m^-2) ones when exposed to HIS. A relatively lower loss of Car in the plants grown in HI-1 and HI-2 exposed to HIS suggests HI adaptation of these seedlings. The slower rate of increase in the ratio of Chl fluorescence emission (F₆₈₅/F₇₃₅) also may suggest photoprotective strategy of HI grown seedlings. There was a positive correlation between MDA accumulation and Chl fluorescence polarisation. The DCPIP photoreduction activity in chloroplasts isolated from HI-1 and HI-2 grown plants exposed to HIS showed slower loss of electron transport activity compared to MI grown plants. These observations suggest that plants grown under higher irradiance have capacity to manage the excess quanta better than those grown under lower irradiance.

The effects of increasing relative humidity on the growth and salt tolerance of two melon (Cucumis melo L.) cultivars, Revigal C-8 (salt sensitive) and Galia (salt tolerant) was investigated. One month after germination, the plants were exposed for 15 d to 0 (control) and 80 mM NaCl, under relative humidity (RH), 30 and 70 %. The growth of the whole plant, leaf, stem and root of cv. Revigal C-8 was increased with increasing RH. On the other hand, cv. Galia showed an increase in root growth with increasing RH only under the NaCl treatment. Under salinity, most of the Na⁺ was withheld in the stems. An increase in RH in the NaCl treatment significantly decreased Na⁺ and Cl^- concentrations in leaves of cv. Revigal C-8, while it had no effect on their concentrations in cv. Galia. In both cultivars, increasing RH under NaCl condition significantly decreased water contents in leaves and stems, and increased osmotic potential in roots. The amount of the root exudate of cv. Galia was significantly decreased with increasing RH, while it was not affected in cv. Revigal C-8. Under the NaCl treatment, cv. Galia had significantly higher leaf osmotic potential than cv. Revigal C-8 at both relative humidities and higher amount of root exudate at 30 % RH.

To test whether different nitrogen form (nitrate or ammonium) in substrate can alter the response to elevated partial pressure of CO₂ (pCO₂) plants of perennial ryegrass (Lolium perenne cv. Bastion) were grown from seeds in growth chambers under pCO₂ of either 35 Pa (ambient, CA) or 70 Pa (elevated, CE) in a hydroponic system (with nutrient and pH control) for 24 d. Nitrogen was supplied as ammonium, nitrate or an equimolar mixture of both N forms. Under CE plants grew faster than their counterparts under CA during the first 14 d but after 23 d of cultivation stimulation disappeared. Despite the strong positive effect of mixed forms of N on plant growth, the beneficial effect of CE was similar to that in the other two N treatments. However, the almost alike final growth response to CE had different underlying mechanisms in different N treatments. Plants supplied with nitrate as a sole source of nitrogen had lower leaf mass ratio but much higher specific leaf area compared to plants supplied with ammonium. The decrease in the content of leaf organic N (per unit of structural dry mass) under CE was found only in leaves of plants supplied with ammonium on day 14. Nevertheless, the available form of N evidently contributes to changes of leaf N content under CE. The high levels of N and non-structural saccharides in plants supplied with ammonium at CE suggest that the CO₂ response of these plants was controlled by factors other than amount of available carbon and nitrogen.

Nodal explants excised from 18 to 20-year-old female plants of Simmondsia chinensis if cultured on Murashige and Skoog's medium supplemented with 20 μM N⁶-benzyladenine (BA) differentiated an average of 2.7 ± 0.4 shoots in 11.5% explants. The percentage of nodal explants inducing multiple shoots enhanced significantly if in vitro raised shoots were used as source of explants. Nearly 100% cultures differentiated an average of 4.7 ± 2.0 shoots per explant on the same medium. Nearly 85% of the shoots induced roots when a pulse treatment of 50 μM indole-3-butyric acid (IBA) was given prior to their transfer to semi-solid MS medium containing 10 μM IBA + 0.5% activated charcoal + 1 μM BA. Plantlets were gradually hardened in Soilrite and acclimatized to soil.

Photosynthetic parameters were compared in three types of transgenic tobacco plants: ipt-transgenic plants with slightly elevated endogenous cytokinin (CK) content, Pssu-ipt-transgenic plants with markedly increased CK content, and zmp-transgenic plants with slightly elevated CK content accompanied by elevated auxin content. Slightly increased CK content promoted net photosynthetic rate (P_N) in both ipt- and zmp-transgenic plants, and chlorophyll (Chl) and carotenoid contents in zmp-transgenic plants. Morphology, growth characteristics, stomatal conductance, and parameters of Chl a fluorescence kinetics were similar in control and transgenic plants with slightly higher CK content. No significant effect of increased level of endogenous auxin (indole-3-acetic acid) on development of zmp-transgenic plants and measured parameters was found. Pssu-ipt-transgenic plants with highly increased CK content revealed suppressed root development, wilting of plants, and depression of P_N and stomatal conductance; however, Chl content was slightly increased and parameters of Chl a fluorescence kinetics did not indicate damage to photosynthetic apparatus.

Influence of increased K supply on growth and nutrient content in pearl millet (Pennisetum glaucum) under severe water stress was assessed in a pot experiment under glasshouse conditions. Nineteen-day-old plants of two lines, ICMV94133 and WCA-78 were subjected for 30 d to 235, 352.5, and 470 mg(K) kg^-1(soil) and two water regimes (100 and 30% field capacity). Increasing soil K supply did not alleviate the adverse effect of water deficit on the growth of two lines of pearl millet. Accumulation of N and K in the shoots of both lines was higher under water deficit than that under well-watered conditions, but such effect was not observed for P or Ca.

Abscisic acid (ABA) content and relative water content (RWC) in second fully expanded leaves of cold hardened plants and in dehydrated leaves of freezing tolerant barley (Hordeum vulgare L. cv. Lunet) were compared. ABA content and RWC in leaves did not change during the first day of cold hardening. On the contrary, dehydration of leaves led to a decrease of RWC and to an increase of ABA content.

Fully expanded leaves of tomato (Lycopersicon esculentum) growing with either complete or nitrogen-deficient nutrient solution were analysed for leaf water status, gas exchange and chlorophyll fluorescence during the vegetative and reproductive phases. N-deficiency did not affect leaf water relations but did decrease light saturated photosynthetic rate as well as stomatal conductance in the vegetative stage. A lower variable to maximum fluorescence ratio (Fv/Fm) was found in N-limited plants which also showed an increase in leaf starch content and in starch to sucrose ratio. The inhibition of photosynthesis and the alteration of photosynthates partitioning were responsible for the growth reduction in N-stressed plants. During the reproductive phase the limitation of photosynthesis may be due to a large accumulation of starch which determines both a decrease in the carbon demand from the sinks and a decrease in CO2 conductance in the mesophyll.

Pyruvate kinase is an important enzyme of glycolytic pathway that also functions in providing carbon skeleton for fatty acid biosynthesis. It has been purified to near homogeneity from Ricinus communis, Selenastrum minutum, Cynodon dactylon, Brassica campestris and B. napus, and characterised. Partially purified preparations are reported from several other sources. A phosphoenolpyruvate (PEP) phosphatase accompanies pyruvate kinase. In plants, two isozymes of pyruvate kinase are reported, namely cytosolic and plastidic. Isoforms of cytosolic pyruvate kinase have also been reported from spinach. In most cases pyruvate kinase is a tetrameric protein and the molecular mass lies between 200 to 250 kDa. The pH optimum is in the range of 6.2 to 7.5. It requires both Mg²⁺ and K⁺ for maximum activity. ATP, citrate, and oxalate inhibit pyruvate kinase in most cases. A sequential compulsory ordered mechanism of binding of substrates to the enzyme has been proposed.

Effect of the benzothiodiazole (BTH) pre-treatment was monitored during the acute infection stage in the susceptible and the hypersensitive tobacco plants infected with the tobacco mosaic virus (TMV). Dynamic changes in the contents of chlorophyll, the total proteins, and the pathogenesis related proteins (PR-proteins), and activities of ribonucleases (RNase), phosphomonoesterase (PME), phosphodiesterase (PDE), and glucose-6-phosphate dehydrogenase (G6P DH) were studied. Neither the protein nor the chlorophyll contents were significantly changed by the TMV infection and/or the BTH treatment. The BTH pre-treatment caused a substantial reduction in the multiplication of TMV in the locally-infected leaves of the hypersensitive cultivar Xanthi-nc (to 15.1%). A lesser decrease (to 50.3%) was observed in the locally-infected leaves of susceptible cultivar Samsun. But in the systemically-infected leaves of this cultivar, only a 4-d delay in the multiplication of TMV was found. In the locally-infected leaves of both cultivars, the activities of the RNase, PME, PDE and G6P DH were sharply increased during the acute phase of TMV multiplication (when compared with the healthy plants) and the curves of these activities correlated with the multiplication curves of TMV. The BTH alone also strongly enhanced the activities of these enzymes early after application. Only low additional increases in some enzymes and even slight declines in the others were observed when the inoculation of leaves of cultivar Xanthi-nc followed the pre-treatment with the BTH. No inhibition of the enzymes was observed when the direct effect of different concentration of the BTH (1 - 1000 μM) was examined in vitro during a measurement of the activity. The analysis of intercellular proteins by PAGE under native conditions shows the similar spectrum of the proteins extracted from either the BTH-treated or the TMV-infected tobacco cv. Xanthi-nc.

Tomato (Lycopersicon esculentum Mill. cv. PKM 1) plants growing under field conditions were exposed for 15 d to solar radiation with UV-B component (280 - 320 nm) enhanced to 6.3 kJ m-2 d-1. This simulated a 15% stratospheric ozone depletion over Madurai (9° 50' N latitude). Lipid peroxidation in the leaves of UV-B treated plants was 32% higher compared to the control. Superoxide dismutase (SOD) and catalase activities registered parallel promotion by 126 and 50 %, respectively, in the UV-B treated plants. Further, the contents of total phenols and anthocyanins in the leaves have also been enhanced by 40 and 156%, respectively. On the contrary, polyphenol oxidase activity demonstrated a 58 % inhibition in the leaves of UV-B treated plants. While anthocyanins and phenols are proposed to act as antioxidants, the reduction in polyphenol oxidase activity may maintain the turnover of phenols in the UV-B treated plants.

Artichoke (Cynara scolymus L.) leaf size and shape, glandular and covering trichomes, stomatal density, stomata shape, pore area and epicuticular waxes during micropropagation stages were studied by scanning electron microscopy (SEM) and morphometric analysis with the aim to improve the survival rate after transfer to greenhouse conditions. Leaves from in vitro shoots at the proliferation stage showed a spatular shape, ring-shaped stomata, a large number of glandular trichomes and juvenile covering hairs, but failed to show any epicuticular waxes. Leaves from in vitro plants at the root elongation stage showed a lanceolated elliptic shape with a serrated border, elliptical stomata, decreased pore area percentage, stomatal density, and mature covering trichomes. One week after transfer to ex vitro conditions, epicuticular waxes appeared on the leaf surface and stomata and pore area were smaller as compared to in vitro plants. Artichoke acclimatization may be improved by hormonal stimulation of root development, since useful morphological changes on leaves occurred during root elongation.

Callus induction was obtained on Murashige and Skogg agar medium with 45 μM 2,4-dichlorophenoxyacetic acid under dark at 25°C. Among the four explant types investigated, the best callus induction was obtained from two-week old fronds to which a surgical incision was applied in the basal (meristematic) region. This treatment resulted in 89.11% of fronds producing callus which continued to proliferate for another 24 months. To obtain plant regeneration pieces of calluses were transferred onto Murashige and Skoog agar medium containing 22 μM indole-3-acetic acid and 4.6 μM kinetin and maintained under 16-h photoperiod (irradiance of 30 μmol m^-2 s^-1) at 23°C. Green fronds formed on all callus pieces. The regenerated fronds were later transferred onto Wang medium where they formed roots. The regenerated Lemna minor L. plants obtained through indirect organogenesis did not differ morphologically from individuals forming the stock collection.

Plants subjected to stress show accumulation of proline and other amino acids. The role played by accumulated amino acids in plants varies from acting as osmolyte, regulation of ion transport, modulating stomatal opening, and detoxification of heavy metals. Amino acids also affect synthesis and activity of some enzymes, gene expression, and redox-homeostasis. These roles played by amino acids have been critically examined and reviewed.

Potted two-year-old lemon plants (Citrus limon (L.) Burm. fil.) cv. Fino, growing under field conditions were subjected to drought by withholding irrigation for 13 d. After that, plants were re-irrigated and the recovery was studied for 5 d. Control plants were daily irrigated maintaining the soil matric potential at about -30 kPa. Young leaves of control plants presented higher leaf conductance (g1) and lower midday leaf water potential (Ψmd) than mature ones. Young leaves also showed higher leaf water potential at the turgor loss point (Ψtlp) than mature leaves. In both leaf types g1 decreased with increased vapour pressure deficit of the atmosphere. From day 1 of the withholding water, predawn and midday leaf water potentials (Ψpd and Ψmd) decreased, reaching in both cases minimum values of -5.5 MPa, with no significant differences between mature and young leaves. Water stress induced stomatal closure, leaf rolling and partial defoliation. No osmotic adjustment was found in response to water stress in either leaf type, but both were able to enhance the cell wall elasticity (elastic adjustment). After rewatering, leaf water potential recovered quickly (within 2 d) but g1 did not.

Sugar cane (Saccharum officinarum L. cv. Copersucar SP80-3280) seedlings were grown in nutrient solution with varying concentrations (0, 2 and 5 mM) of cadmium chloride for 96 h. Leaves were analysed for catalase (CAT), glutathione reductase (GR) and superoxide dismutase (SOD) activities. Although a clear effect of CdCl₂ on plant growth was observed, the activity of SOD was not altered significantly. However, the CAT activity decreased as the concentration of CdCl₂ increased. GR exhibits a significant increase in activity at 2 and 5 mM CdCl₂. CAT and SOD isoenzymes were further characterised by analysis in non-denaturing PAGE. Activity staining for SOD revealed up to seven isoenzymes in untreated control and 2 mM CdCl₂ treated plants, corresponding to Cu/Zn-SOD isoenzymes. At 5 mM CdCl₂, only six Cu/Zn-SOD isoenzymes were observed. No Fe-SOD and Mn-SOD isoenzymes were detected. For CAT, one band of activity was observed.

Treatment of spruce, fir and oak trees with herbicides, which may be one of the forest damage inducing agents, caused pronounced changes in the contents and distribution of indole-3-acetic acid (IAA) and cytokinins (CKs) one year after treatment, i.e. at the time of the first microscopically visible damage in treated trees. In Picea pungens IAA content increased in the terminal buds by about 105 % and in the apical buds of the first order branches by 220 %. The same was true for young sprouts of Abies nordmanniana, while in leaves of oak trees IAA content was decreased by 15 % after glyphosate treatment and by 30 % after 2,4-dichlorophenoxyacetic acid (2,4-D) treatment. Another striking feature was a significantly decreased content of IAA in the lower parts of roots in Picea pungens (50 % of the control), which is accompanied by an increase in IAA content in the middle part of the roots (130 %). On the other hand, the IAA content of both sprouts and roots of A. nordmanniana was significantly increased after herbicide treatment.
In P. pungens, the content of free cytokinins (sum of zeatin, zeatin riboside, isopentenyladenine and isopentenyladenosine) decreased due to herbicide treatment. The strongest decrease was seen in roots, especially in their upper and middle parts (the average reduction of cytokinin content in roots was 63 %). In the above-ground organs the reduction was seen namely for isopentenyladenine and isopentenyladenosine, while the abundance of zeatin riboside was, on the other hand, higher in treated plants. In Quercus robur leaves, the total content of cytokinins also decreased, namely after glyphosate treatment. In consequence of these changes, CK/IAA ratio decreased pronouncedly in all organs of herbicide-treated trees, with the exception of oak leaves treated by 2,4-D.

Putrescine effect on nitrate reductase activity, organic nitrogen and protein contents, and plant growth under Cd or Pb (0.1 - 2 mM) and salinity (5 and 100 mM NaCl) stresses was examined in Indian mustard (Brassica juncea L. cv. RH-30) seedlings. Cd or Pb and salinity inhibited nitrate reductase activity and decreased organic nitrogen and protein contents in leaf tissue. The increased nitrate reductase activity induced by putrescine was correlated with increased organic nitrogen and protein contents and growth of plants.

To determine the allelopathic potential of root exudate from early developmental stage of rice (Oryza sativa L), 6-d-old seedlings of eight cultivars were grown with 3-d-old alfalfa (Medicago sativa L.), cress (Lepidium sativum L.) or lettuce (Lactuca sativa L.) seedlings in Petri dishes under controlled condition. All rice cultivars (cv. Norin 8, Kamenoo, Nipponbare, Kinuhikari, Koshihikari, Sasanishiki, Yukihikari and Hinohikari) inhibited growth of roots, shoots and fresh mass of alfalfa, cress and lettuce seedlings. Effectiveness of cv. Koshihikari was the greatest and more than 60% inhibition was recorded in all bioassays, followed by that of cv. Norin 8 of which effectiveness was more than 40%.

Proline accumulation in leaves of barley (Hordeum vulgare L. cv. Alfa) seedlings treated with 150 mM NaCl was promoted in the light and suppressed in the dark. The light/dark changes of proline content was enhanced with each 12 h light/12 h dark cycle and the proline content increased steadily. Root and shoot concentrations of Na⁺ and Cl^- in salt treated plants increased about 10 to 25 times as compared to the control. The content of these ions and the content of malondialdehyde were higher in the shoot of seedlings exposed to salt stress for 4 d in the light in comparison with the seedlings exposed to NaCl for 4 d in darkness. Light stimulated both ions and proline accumulation in the leaves and has no effect in the roots. Oxygen uptake was higher in the seedlings kept 4 d in the light which have higher endogenous free proline content. Chlorophyll fluorescence measurements showed that the photochemical activity of PS 2 slightly decreased as a result of salt stress and was not influenced by light regimes during plant growth.

Dog rose (Rosa canina L.) plants in the bloom stages of flowering were sprayed by indole-3-acetic acid (IAA) in concentrations of 0.06 and 0.60 mM and gibberellic acid (GA₃) in concentrations of 0.60 and 1.50 mM. Ascorbic acid, total sugar, reducing sugar and carotenoid contents gradually increased, while the protein content remained unchanged and the content of phenolic substances decreased during hypanthium development. Ascorbic acid, total sugar, reducing sugar and carotenoid contents increased in hypanthium sprayed by GA₃ and IAA. However, IAA and GA₃ applications (except low concentrations) decreased contents of phenolic substances. IAA and GA applications might be a good way to produce the high quality hypanthium in R. canina.

Effects of four culture media on callus induction, regeneration and number of plants per unit culture were studied with mature seeds from five indica rice genotypes as explants. Based on the morphology, the calli were classified into four types as I to IV. Type I and type II are most suited to initiate suspension cultures or as target material for transformation. Number of plants regenerated per unit culture, formation of easily dissociating cell clusters and frequency of type I and type II calli were highest on NBKNB medium. Thus NBKNB medium is suitable for in vitro culture of even the hitherto recalcitrant indica genotypes.

Diurnal and seasonal changes of net photosynthetic rate (P_n) and the efficiency of photosystem 2 (F_v/F_m) were measured on two perennial species growing on a soil catena in semi-arid south-east Spain. Stipa tenacissima, a tussock grass, grows on shallow soil at the top of the catena and Retama sphaerocarpa, a leguminous shrub, grows in the valley bottom. A linear relationship was found between light saturated photosynthetic rates (P_max) and diffusive leaf conductance (g_l) in both Retama and Stipa indicating that the intercellular CO₂ concentration (c_i) was maintained constant in both species diurnally. Relatively high values of calculated c_i in Retama cladodes suggested that was not the primary limitation to carbon assimilation. F_v/F_m for the two species when well watered was around 0.8. Although Retama cladodes maintained this value throughout the year, F_v/F_m decreased to a minimum of 0.43 in Stipa leaves, at the end of the dry season. Our data suggest that plants in the Rambla Honda can substantially reduce transpiration without reducing photosynthetic rates to the same extent by closing their stomata, because P_n is reduced primarily by high respiration, decreased mesophyll conductance and by photoinhibition or permanent damage of photosystem 2.