Habituated sugar beet calli have been characterized as having a deficiency in some tetrapyrrole containing compounds. However, peroxidases might be dissociated from the other tetrapyrrole containing compounds. When light-cultured normal and habituated calli were transferred to darkness their peroxidase activity reduced and increased, respectively, indicating that habituation could not strictly be characterized by a deficiency in peroxidase content but rather by a different regulation of its activity. This regulation could be mediated through soluble effectors which act as potential peroxidase inhibitors and/or by a differential expression of the peroxidase isoenzyme patterns which were present in these tissues in both light and darkness. The different peroxidase activity and the nature of acidic and basic peroxidase isoenzymes in normal and habituated tissues could explain the different features of both types of cultures.

Rice (Oryza sativa L.) seedlings were grown under NaCl stress. The leaf growth of resistant cv. Damodar was less affected than that of the susceptible cv. Jaya. The leaf protein content showed no distinct cultivar or age dependent differences under NaCl salinity. There was a significant increase in chlorophyll (Chl) and carotenoid (Car) contents of 25-d-oldseedlings of both cv. Jaya and cv. Damodar. However, Chl and Car content of 15-d-old seedlings of cv. Jaya decreased and that of cv. Damodar increased, under NaCl stress.

Excised embrya and subsequently divided embrya of Podophyllum peltatum were cultured on the Murashige and Skoog medium supplemented with different growth regulators, because traditional methods of breaking seed dormancy failed. The growth of excised embrya was stimulated by 1 or 0.1 mg dm-3gibberellic acid (GA3), 0.1 mg dm-3 GA3 + 0.2 mg dm-3 kinetin (kin), or 0.2mg dm-3 kin. GA3 (1 mg dm-3) showed the best effect; after 5 weeks the plantlets had 1.5 - 2 cm long cotyledons, 5 - 6 cm long roots, 88 % of embrya germinated and developed further. The addition of 0.5 mg dm-3 zeatin + 0.2 mg dm-3 naphtaleneacetic acid (NAA), 0.2 mg dm-3 NAA, and 1 mg dm-3 kinetin inhibited the growth of embrya. 1 mg dm-3 kinetin + 0.1 mg dm-3 NAA, 0.1 mg dm-3 zeatin and 0.2 mg dm-3 6-benzylaminopurine resulted in a compact appearance of plantlets and a lower germination rate. Divided embryo cultures produced plantlets via somatic embryogenesis which occurred only on the 2,4-dichlorophenoxyacetic acid containing media. The maturation of somatic embrya was observed on media without any auxin.

Changes in growth and anatomical structure of vascular tissues in stem, root and leaf of safflower plants grown in NaCl and CaCl2 solutions having different osmotic potentials (ΨS from 0 to -0.9 MPa) with addition of 0, 10 and 20 mg dm-3 zinc were studied. Shoot and root lengths, fresh and dry masses and fresh/dry mass ratio were lower in salt-stressed plants compared to unstressed plants. Salinity induced structural changes in stem, root and leaf tissues; few xylem vessels with smaller size were noticed in stressed plants. The higher concentration of Zn improved growth especially in roots and enhanced xylem formation in comparison to stressed plants grown at the same osmotic potential without Zn. Zn also protected xylem distructure by salinity in leaves.

We present a simple and rapid technique for the double staining of plant peroxidase and other proteins in the same polyacrylamide gel using the principle of iodide oxidation followed by Coomassie Blue counterstain. The colored bands of peroxidase isozymes and proteins are easily distinguishable. An additional benefit of the method is the use of the low cost chemicals, as well as it eliminates the need for a potentially hazardous reagents frequently used in the detection of peroxidase isozymes.

Electrophoretic seed protein patterns of 18 samples belonging to 14 species and 4 sections of Lathyrus are treated by principle component analysis (PCA). The morphological ground and karyological structure data of these samples are also discussed in the light of sectional and groups delimitation. The species under study of section Cicercula are separated into 3 groups and L. hirsutus referred to the most primitive species within Lathyrus species. This agrees with their previous grouping delimination based on morphological characters and with chromosomal features such as karyotype structure. L. aphaca referred to the most advanced species in this genus, which agrees with the modification of this morphological characteristics and reduction in chromosome criteria. Section Nissolia hasan intermediate position between section Cicercula and section Aphaca.

Protocols were developed for plant regeneration from callus induced in mature embryos of rice. Somaclonal variation was scored by genome mutation, chromosome mutation and plasmon mutation in R₀, R₁ and R₂ plant progenies. The frequency of haploids and diploids appeared in the ratio of 20:33. Variation in the chromosome number in callus cells was found to be high and age dependent. Different types of chlorophyll deficient mutants including albinos appeared in R₂ plant progeny where gene mutation frequency was the highest (52.4 %). The results revealed that a high frequency of somaclonal variation is possible to generate by tissue culture techniques.

Wounding-induced extracellular pH shifts were characterized previously in excised segments of maize (Zea mays L.) coleoptiles. In the present study it is demonstrated that similar pH shifts also occur in Triticum aestivum L., Secale cereale L., Hordeum vulgare L., Avena sativa L., Sorghum durra (Forsk.) Stapf, and Setaria italica (L.) Beauv., with characteristic quantitative differences between the species. Indole-acetic acid induces pronounced drops of the medium pH in all species except Setaria italica.

A non-destructive method of total vertical projections for estimating the length of rigid root systems, not introduced yet in plant sciences, is described. It is demonstrated on measuring less and more dense root systems of seedlings of Zea mays grown at hypoxic or control conditions. Photographs of six vertical projections (30° apart) of each root system were taken and evaluated. The method being based on proved mathematical formula offers unbiased estimation of the length of a rigid root system, curved in three-dimensional space, by non-destructive means. Thus, it is applicable during ongoing experimentation on plants grown in a solution culture. It was shown that less than 120 intersections between the root projection and test lines in one photograph ensured sufficient precision of the method and that the observer subjectivity could be overcome by presented instructions.

Cadmium-treated pea plants exhibited PEPC and NADP-ME activities, titratable acidity, and malate concentration in the leaves similar to controls. The PEPC activity in the roots of Cd-treated plants decreased by about 40 %, and NADP-ME increased more than twice. The titratable acidity remained similar as in the leaves, but the malate content diminished by about 30 %. The application of 500 g(d.m.) m-3 biomin and a combination of biomin and algae suspensions to Cd-treated plants brought about an increase in the titratable acidity and in the malate concentrations in the leaves and the roots.

After an electrophoretic separation of proteins from Euglena gracilis and dry seeds of Phaseolus vulgaris in native conditions in polyacrylamide gels, gels were incubated in mixtures containing NAD+, Mg-ATP2-, glucose 6-phosphate, G6P dehydrogenase, and either phenazine ethosulfate and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (PES/MTT) or phenazine methosulfate and nitro blue tetrazolium (PMS/NBT) as coupled redox system for NAD+ kinase activity detection. In the presence of PES/MTT, 4 bands were revealed for E. gracilis, among which two corresponded to NAD+ kinase activity, the other corresponding to a NAD+ reductase activity due to alcohol dehydrogenase (ADH). In the presence of PMS/NBT, only the bands of NAD+ kinase activity were revealed. With Phaseolus vulgaris, 3 bands of ADH were always revealed in both mixtures, and only the use of PMS/NBT allowed the detection of NAD+ kinase as a fourth band. With both materials, NAD+ reductase staining in gels was intensifed in the presence of GTP or ATP and even further with ADP or GDP. The results demonstrate that: 1) the NAD+ kinase and NAD+ reductase are two distinct enzymes; 2) the NAD+ reductase corresponds to ADH.

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.

A modification of the procedure of extraction of cell wall solution for enzymatic determination of ascorbic acid and its reduction level in the apoplast of leaf cells is proposed. The modification consists in infiltration of leaves with citric acid/sodium phosphate buffer, pH 3, instead of customarily used neutral solutions. In acidic media autooxidation of ascorbic acid is effectively suppressed, so that infiltration could be performed at laboratory temperatures. Using polyacrylamide gel electrophoresis and infiltration solutions of pH down to 1.5 it is shown, that at pH 3 the extracted fluid is not contaminated with intracellular substances if appropriate vacuum and centrifugation forces are used. The modification is shown to be more effective for leaves ofPhaseolus than for those ofSpinacia. In cell walls of mature leaves of these species the concentration of ascorbic acid was found to be around 1 mM, with reduction level up to 0.90. The role of ascorbic acid in cell walls as ozone scavenger is discussed

Wheat leaves(Triticum aestivum L. cv. San Agustin INTA) were detached at the moment they had reached maximum expansion, put in tubes containing water and left in darkness. Under these conditions, leaf protein content decreased mainly as consequence of an increased rate of breakdown. In the range of 0 to 72 h after detachment, western blot analysis of leaf protein extracts displayed both similar proportions of total protein and quality of ubiquitin conjugates. Northern blot analysis of leaf RNA extracts revealed a 1.6 kb ubiquitin mRNA transcript which increased 3.5-fold after 48 h of treatment. Thus wheat leaves maintain both their ability for the ubiquitination of proteins and the transcription of ubiquitin mRNA at stages of senescence in which rates of protein breakdown are increased. The results suggest that the ubiquitin-dependent proteolytic pathway contributes to leaf protein breakdown during senescence

A hypothesis of existence of a general adaptation syndrome (GAS), in which different types of stress evoke similar coping mechanisms, resulting in adaptations, is tested for plants. As stress coping mechanisms, oxy-free radical scavengers and antioxidants, osmoregulation, the role of abscisic acid, jasmonates, nitric oxide, synthesis of heat shock proteins and phytochelatins as heavy metal detoxifiers are discussed.

Standardized laboratory techniques for the vegetative growth of the duckweedSpirodela polyrhiza (Lemnaceaé), and for formation as well as germination of their turions were described. Increasing photon fluence rates of blue or red light increased the yield of turions. A specific stimulating effect of blue light was demonstrated under autotrophic but not under mixotrophic conditions. Therefore the spectral composition of light is not important in mixotrophic formation of turions whereas in autotrophic formation light sources with a higher portion of blue light are recommended. Dark-grown (etiolated) turions showed accelerated germination and higher germination percentage in comparison with light-grown turions after induction by a single red light pulse. This difference was overcome in continuous red light by speeding up the germination response of light-grown turions. Use of Petri dishes (8 cm³ nutrient solution) instead of Erlenmeyer flasks (50 cm³ nutrient solution) retarded germination response. Especially for long term experiments the use of Erlenmeyer flasks is recommended. Storage of turions for 72 h at 25 ‡C following at 5 ‡C in darkness after-ripening resulted in a decreased lag phase of the light-induced germination both after induction by a single light pulse and in continuous light.

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.

Qualitative and quantitative differences in cytokinin levels of gram seeds were observed during germination of control and 6-benzylaminopurine (BAP; 4.44 x 10^t-5 M) treated seeds. Zeatin riboside (ZR), zeatin (Z), dihydrozeatin (DHZ), isopentenyladenosine (iPA) and isopentenyladenine (iP) were determined using high pressure liquid chromatography. In the control seeds the ZR content increased markedly only after 24 h of germination. In BAP treated seeds, the increase in ZR content was stronger than in control seeds and it persisted during further germination. Z content increased during germination in both control and treated seeds. After 48 h of germination iP appeared in control and especially in treated seeds.

Maize (Zea mays L. cv. Ganga-5) seedlings were grown in the presence of ferulic acid (0.5 - 3.0 mM) for 8 d. Treatment with ferulic acid considerably decreased shoot and root length, increased the activity of peroxidase, catalase and indole-3-acetic acid (IAA) oxidase and decreased the activity of polyphenol oxidase. The increased activity of peroxidase correlated with pronounced increase in content of lignin and phenolic compounds

Concentration of cytokinins (CKs) of zeatin-type (Z) and isopentyladenine-type (iP) were determined using an enzyme-linked immunosorbent assay (ELISA) technique in xylem sap and bulk leaf extract of the hemiparasiteMelampyrum arvense before and after attachment to the host (Capsella bursa pastoris). In all the samples Z-type were dominant, though the iP-type was also frequent. The results also demonstrate that in comparison with the unattached hemiparasite after attachment to the host the concentration of Z-type in xylem sap increased about 92 and 182 times in the light and dark phase, respectively, and that of the iP-type about 34 times in the two phases. The concentration of Z-type and iP-type in leaves expressed per dm³ of cell water was at the level of 10^-7 M and 10^-8 M, respectively in the unattached hemiparasite. After attachment, the concentration of CKs increased to 10^-5 M and 10^-6 M for Z-type and iP-type, respectively. In xylem sap the concentration of Z-type was at the level of 10^-9 M and 10^-7 M and in the unattached and attached hemiparasite, respectively. The concentrations of iP-type were 10^-10 M and 10^-8 M for the unattached and attached hemiparasite, respectively. The diurnal oscillation of CK concentration was evident in the unattached hemiparasite, but after attachment their level was nearly constant and independent of the diurnal cycle.

In two selected nitrogen-fixing cyanobacterial speciesAnabaena fertilissima andAnabaena variabilis pH, irradiance and different inocula sizes considerably modified the toxic effect of 50 % effective concentration (EC 50) dose of the pesticideFuradan (carbofuran 75 DB). Maximum growth and chlorophyll (Chl)a content ofA. fertilissima was observed in the pH range of 8 - 9 and that ofA. variabilis at pH 7-8, while at acidic pH (5 - 6) and at pH above 9 these parameters were considerably retarded. Toxicity of the EC 50 dose ofFuradan was increased further at pH 5-6, whereas reduction in the toxicity to the test cyanobacteria was observed at pH 7.8 - 9.0. The experimental organisms grew comparatively better and synthesized higher amount of Chl a at an irradiance of 12.5 than at 7.5 or 2.5 W m^-2. The toxicity of EC 50 dose of the pesticide gradually decreased with the increasing irradiance. The toxic effect ofFuradan was larger when the initial cyanobacterial population concentration was low andvice versa.