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.

Multiple shoot regeneration from the cut plumular ends of embryo axes of chickpea (Cicer arietinum L.) was evaluated on Murashige and Skoog medium having different concentrations of thidiazuron (TDZ) (0.1 to 10.0 mg dm^-3) 6-benzylaminopurine (BAP) (0.5 and 1.0 mg dm^-3), kinetin (0.5 and 1.0 mg dm^-3) or zeatin (2.0 and 4.0 mg dm^-3). TDZ (0.2 mg dm^-3) was found to be the most effective cytokinin as it produced multiple shoots in 100 % of the explants from genotypes C235, ICC5166, ICC12269, ICC4951, ICC11531, BG256 and a local cultivar. Shoots were elongated on growth regulator-free medium, and rooted on growth regulator-free medium containing 1/4 MS salts + full vitamins + 3 % sucrose. Plantlets formed were acclimatized for 12 - 15 d in MS medium with a gradual reduction in sucrose concentration and transferred into pots filled with soil and kept in the field; this resulted in more than 70 % survival. The plants developed normally and produced fertile flowers and set seeds. Low temperatures, maximum 19.0 °C, and minimum 8.2 °C, during the first 15 d of transfer favoured survival on transfer to pots.

Salinity induced changes in α-amylase activity in three cotton cultivars (NIAB-Karishma, NIAB-86 and K-115) was studied during germination and early seedling growth under controlled conditions. The increase in NaCl concentration resulted in the decrease in α-amylase activity and break down of starch into reducing and non-reducing sugars in all cultivars, however, it was more pronounced in NIAB-86. K-115 showed highest germination followed by NIAB-Karishma and NIAB-86.

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.

Vegetative fragments of the subtidal macroalga Gracilaria edulis (Gmel.) Silva cultured under field conditions at Thonithurai were subjected in the laboratory to UV-B radiation (280 - 320 nm). UV-B inhibited the accumulation of chlorophyll and phycobiliproteins, and lowered agar yield (23 to 43%) and its gel strength (22 to 36%) under 12 to 72 h exposure. The longer the exposure to UV-B radiation the more significant impact on pigments and phycocolloid properties of Gracilaria edulis was observed.

Using the whole plant and model systems, we demonstrate that the aluminum ions (Al³⁺) stimulate phenolic-dependent lipid peroxidation. Lipid peroxidation in barley (Hordeum vulgare L. cv. Donor) roots was 30 % higher under AlCl₃ treatment than without Al. Major decomposition product of lipid peroxidation was 4-hydroxynonenal (4-HNE) but not thiobarbituric acid reactive substances (TBARS), a widely used markers for lipid peroxidation. Similarly, AlCl₃ stimulated lipid peroxidation of soybean liposomes in the presence of chlorogenic acid (CGA) and H₂O₂/horseradish peroxidase system which can oxidize phenolics. Al³⁺ was found to enhance lipid peroxidation induced by oxidized CGA. Intermediates of lignin biosynthesis in plants, including p-coumaric acid, ferulic acid, sinapic acid and coniferyl alcohol, also showed similar effects. These results suggest that Al³⁺ has a potential to induce oxidative stress in plants by stimulating the prooxidant nature of endogenous phenolic compounds.

An integrated approach has been developed for targeted retrieval of microsatellite markers from selected regions of the field bean (Vicia faba L.) genome. The procedure is based on a combination of advanced physical and genetic mapping techniques and includes the following steps: 1) flow-sorting of metaphase chromosomes, 2) construction of microsatellite-enriched chromosome-specific DNA libraries, 3) isolation of polymorphic microsatellite sequences from the libraries, 4) testing chromosome specificity of the microsatellites using polymerase chain reaction with purified fractions of individual chromosome types, and 5) integration of chromosome-specific markers into a genetic map. Several strategies for isolation of microsatellite clones were tested, including direct screening of non-enriched libraries with single or mixed probes and screening of the libraries after one or two rounds of enrichment. Finally, the usefulness of this approach was demonstrated by the retrieval of novel markers from a selected portion of the largest field been chromosome (No. 1).

Response of two spring wheat (Triticum aestivum L.) cultivars, salt tolerant SARC-I and salt sensitive Potohar, to different concentrations of NaCl was examined under glasshouse conditions. Eighteen-day-old plants of both the lines grown in sand culture were irrigated with 0 (control), 80, 160 or 240 mM NaCl in full strength Hoagland's nutrient solution. Shoot fresh and dry masses, and leaf area per plant of SARC-I at the vegetative stage, were significantly greater than those of cv. Potohar at higher salt concentrations, however, relative growth rate (RGR) of cv. Potohar was significantly higher than that of SARC-I. SARC-I had higher net photosynthetic rate (P_N), stomatal conductance (g_s) and transpiration rate (E) than cv. Potohar at the vegetative stage, but the cultivars did not differ significantly in water-use efficiency (P_N/E), intrinsic water use efficiency (P_N/g_s), and intercellular/ambient CO₂ concentration ratio. At the grain development stage, SARC-I had significantly higher P_N and g_s in the flag leaf than cv. Potohar under salinity. SARC-I was superior to cv. Potohar with respect to number of grains per spike, number of grains per spikelet, mean grain mass, and grain yield per plant at all NaCl concentrations.

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.

Ammonium ion accumulation in detached rice leaves treated with phosphinothricin (PPT), an inhibitior of glutamine synthetase (GS), was investigated in the light and darkness. PPT treatment increased NH₄⁺ content and induced toxicity in rice leaves in the light but not in darkness, suggesting the importance of light in PPT-induced NH₄⁺ toxicity in detached rice leaves. PPT treatment in the light resulted in a decrease of activities of the cytosolic form of GS and the chloroplastic form of GS. The photosynthetic electron transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea reduced NH₄⁺ accumulation induced by PPT in the light. In darkness, PPT-induced NH₄⁺ accumulation and toxicity were observed in the presence of glucose or sucrose.

The effects of NaCl on putrescine (Put) content and diamine oxidase (DAO) activity in roots of rice seedlings were examined. NaCl treatment lowered the content of Put and increased the activity of DAO in roots. Our current results indicate that Cl^- is not required for NaCl-induced decline in Put content and increase in DAO activity in roots. Put content in roots of rice seedlings exposed to NaCl is possibly regulated by DAO activity.

The individual and interactive role of calcium and abscisic acid (ABA) in amelioration of water stress simulated by polyethylene glycol (PEG) 6000 was investigated in two contrasting wheat genotypes. PEG solution (osmotic potential -1.5 MPa) was applied to 10-d-old seedlings growing under controlled conditions and changes in photosynthetic rate, activities of ribulose-1,5-bisphosphate carboxylase and phosphoenolpyruvate carboxylase, water potential and stomatal conductance were observed in the presence of 0.1 mM ABA, 5 mM calcium chloride, 1 mM verapamil (Ca²⁺ channel blocker), and 1 mM fluridone (inhibitor of ABA biosynthesis). ABA and calcium chloride ameliorated the effects of water stress and the combination of the two was more effective. The two genotypes varied for their sensitivity to ABA and Ca²⁺ under stress. As was evident from application of their inhibitors, ABA caused more alleviation in C 306 (drought tolerant) while HD 2380 (drought susceptible) was more sensitive to Ca²⁺.

Kiwifruit plants (Actinidia deliciosa cv. Hayward) were grown in Hoagland nutrient solution with calcium nitrate, potassium nitrate, ammonium nitrate or ammonium chloride as the nitrogen source. Plants grown in the solution with nitrate nitrogen displayed a higher oxalate content, greater shoot length and leaf area, and higher content of ascorbic acid and NO₃^- ions in the leaves. Plants grown in the solution with ammonium nitrate, and particularly with ammonium chloride, showed low oxalate content, low content of ascorbic acid and NO₃^-, high content of Cl^- and Na⁺, low shoot length and leaf area. Oxalate formation appeared to be connected with the assimulation of nitrate, more precisely with nitrate reduction, while ammonium nitrogen assimilation did not induce the synthesis of oxalic acid.

Nostoc rivulare was grown in batch cultures under controlled CO₂ and NO₃^- concentrations to modulate the photosynthetic source:sink relationship. Increasing CO₂ supply accelerated the accumulation of chlorophyll (Chl) a, i.e., supplemental CO₂ combined with double concentrations of NO₃^- more than doubled the amounts of Chl a relative to those of the original medium. Photosynthetic oxygen evolution and respiratory oxygen uptake were both enhanced by elevated CO₂ and NO₃^-. Contents of soluble sugars and starch (total non-structural saccharides) as well as insoluble saccharides (structural fraction) were affected by altering CO₂-NO₃^- combinations. Uptake as well as reduction of either NO₃^- or NO₂^- was inhibited by CO₂ deprivation. Expanding the sink size via increasing NO₃^- supply enhanced photosynthesis and thus the sink (NO₃^-) acted as a feed forward stimulator of the source (photosynthesis). The regulatory role of nitrate on photosynthesis was most influential in CO₂-deprived cultures since it could enhance photosynthesis to higher levels than CO₂-supplemented, nitrate-free cultures.

We determined the characteristics of phytochelatin synthase from tobacco (Nicotiana tabacum cv. Bright Yellow-2) cells, especially the conditions for the enzyme stability. From the results, we proposed the optimum assay conditions of the enzyme activity.

Seasonal dynamic of total nonstructural saccharides (TNS) and individual saccharides (starch, sucrose, glucose, fructose, fructans) was followed in rhizomes and stem bases of Calamagrostis epigeios (L.) Roth at two types of meadows communities in the South Moravia (Czech Republic): cnidion and molinion alliances, which differ in their water regime. The TNS were formed mainly by fructans and starch, while glucose, sucrose and fructose were low. The amount of TNS in rhizomes and stem bases of plants from wet cnidion site was higher than in plants from drier molinion site. The seasonal trends of all saccharides were similar in the both sites. During growing season (June to October) the main storage sugar was fructan (18 - 21 % of dry biomass). At the beginning of September the content of fructan decreased to 10 - 12 % and simultaneously the content of sucrose increased from 1 to 3 %. This may increase frost resistance. The content of TNS in the stem bases was lower than in the rhizomes. During winter time the stem bases contained 2 to 2.5 % of sucrose. Plant height and aboveground biomass were also higher in molinion site.

Using preparative isoelectric focusing, fast performance liquid chromatography, and polyacrylamide gel electrophoresis accumulation of several Al-induced cytoplasmic proteins was described in barley roots. Two of them, 27 and 28 kDa polypeptides were isolated by continuous-elution electrophoresis system in sufficient quantities for their further characterisation.

Araucaria bidwillii Hook. and Araucaria cunninghamii Don D. are two species of conifers whose seeds belong to different physiological categories: A. bidwillii seeds are recalcitrant, while A. cunninghamii seeds are orthodox. The extraction of enzymes and storage proteins was carried out from A. bidwillii and A. cunninghamii megagametophytes. The endopeptidase activities of both species were assayed with azocasein and with haemoglobin; the exopeptidase activities were detected by various N-carbobenzyloxy-dipeptides and L-leucine p-nitroanilide. The use of appropriate proteinase inhibitors, i.e. pepstatin A, ethylenediaminetetraacetic acid and trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, showed the presence of aspartic and metallo proteinases and the absence of the cysteine ones both in A. bidwillii and in A. cunninghamii ungerminated seeds. Since the results do not show differences between the types of enzymes in the ungerminated araucarian seeds and those present in some ungerminated angiosperm seeds (barley, wheat, maize, rice, buckwheat), we conclude that their physiological role is similar. The electrophoretical analyses of soluble and insoluble storage proteins of A. cunninghamii showed patterns similar to those found in other gymnosperms, while the storage protein patterns of A. bidwillii seeds were rather atypical.

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 present investigations were conducted to ascertain the influence of Pb₂₊ and Cd₂₊ both individually and in combination on selected enzymes of tricarboxylic acid (TCA) cycle. All the enzymes of TCA cycle examined (α-ketoglutarate dehydrogenase, isocitrate dehydrogenase, succinate dehydrogenase, malate dehydrogenase) were affected deleteriously by Pb₂₊ as well as Cd₂₊ and these metals in combination gave more or less an additive effect.

The present study was carried out to investigate the degree of leaf wetness and its capacity to retain water droplets in relation to leaf morphological characteristics of Valeriana jatamansi J. grown under open and shade habitats. Leaves developed in open habitats had less wettability but higher capacity to retain water droplets and more number of stomata than shade leaves. A significant positive correlation of contact angle (ϑ) were noticed with trichome length, droplet retention and wax content.

Transient expression of β-glucuronidase (GUS) in zygotic embryo axes of two cotton (Gossypium hirsutum L.) cultivars NHH-44 and DCH-32 was induced by Agrobacterium mediated transformation or by particle bombardment. For Agrobacterium transformation, disarmed A. tumefaciens strain GV 2260/p35SGUSINT was used. In cv. NHH-44, the maximum frequency of transient expression (14.28 %) was achieved on spotting Agrobacterium paste on the apical regions of the split embryo axes. The method resulted in a transformed callus line, which showed strong GUS activity. Integration of NPTII gene was confirmed by Southern analysis. Transgene expression by particle bombardment was achieved with p35SGUSINT and pIBGUS plasmids independently. The maximum frequency of GUS expression in 29.16 % explants was observed in cultivar NHH-44 with gold microcarriers (1.1 µm) when bombarded once with rupture disc of 7586 kPa at target cell distance of 6 cm. A transformed callus line was obtained when explants were bombarded with p35SGUSINT and cultured on Murashige and Skoog's medium supplemented with B₅ vitamins, 0.1 mg dm^-3 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea, 0.01 mg dm^-3 α-naphthaleneacetic acid, 3 % glucose + 50 mg dm^-3 kanamycin. High GUS activity was observed in callus tissue as well as in somatic embryo like structures achieved in liquid shake cultures.

Micropropagated shoots of Stackhousia tryonii were exposed (individually or in combination) to indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and 1-naphthalene acetic acid (NAA) at concentrations 1, 2 or 4 g dm^-3 with the view to induce rooting under ex vitro conditions. The treated microshoots were grown in a mist room for four weeks and assessed for survival, rooting percentage, number of roots and root length. The results showed that IBA at 2 g dm^-3 was most effective in inducing roots. Mixing of two or more auxins markedly reduced rooting percentage indicating antagonistic effects. The results demonstrated the potential of combining ex vitro rooting and hardening in one step, with view to reducing costs of multiplying plants via micropropagation.

For most plant species growing in polluted areas no mutagenicity assays are available. We have studied the possibility of using the alkaline protocol of the Comet assay as a method for detecting induced DNA damage in wildly growing weeds. The monofuctional alkylating agent ethyl methanesulphonate (EMS) was applied on leaves of 10 weed species (ordered according to the diameter of the nuclei): Arabidopsis thaliana, Convolvulus arvensis, Bellis perennis, Urtica dioica, Lamium album, Chenopodium rubrum, Plantago media, Poa annua, Taraxacum officinale, and Agropyron repens. With increasing concentrations of EMS (2 to 10 mM) the DNA damage, expressed by the averaged median tail moment values, significantly increased in nuclei of all weeds studied. Using the Head Extent parameter of the Komet version 3.1, we have measured the diameter size of the nuclei of the 10 weed species either immediately after the isolation of the nuclei or after 20 or 45 min of treatment with alkaline buffer (pH > 13). According to the increase of the diameter of the nuclei (including the formed "halo") resulting from the to alkaline buffer treatment, electrophoretic conditions (unwinding and electrophoresis time) for the Comet assay can be selected for the individual weed species.

The effect of brassinosteroids (BRs) on catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7) and superoxide dismutase (SOD, EC 1.15.1.1) activity in tomato leaf discs was analyzed at 25 and 40 °C. Tomato leaf discs were preincubated for 24 h in Petri dishes with 24-epibrassinolide (EBR) or a polyhydroxylated spirostanic analogue of brassinosteroids (MH5). Both concentrations (10.60 and 2.12 nM) of EBR and MH5 stimulated the activity of SOD at 25 and 40 °C, the MH5-stimulated increase of this enzyme activity was greater. Peroxidase activity was unaffected at 25 °C, while at 40 °C this activity was enhanced by both compounds. The changes in catalase activity markedly depended on the structure BRs, doses and temperature. The results suggest a possible role of EBR and MH5 in the reduction of cell damage produced by heat stress due to induction of enzymatic antioxidants.