Suspension cultures of tomato cells were used to study the membrane injury by the toxic concentration of mercuric chloride. Assessment of electrolyte leakage, UV-absorbance of the tissue leachates, relative leakage ratio, injury index, membrane lipid peroxidation, lipoxygenase activity, α-amino nitrogen and total soluble carbohydrate contents showed the extent of membrane damage as a function of the increasing concentration of mercuric chloride. It is suggested that the selected parameters can be used as qualitative tests for determination of stress-induced membrane damage

Changes in the number of protoplasts, viability, protein and chlorophyll content were studied in tobacco mesophyll protoplasts cultivated either autotrophically in CPW medium with mannitol (MCPW) in the light or heterotrophically in CPW medium with glucose (GCPW) in the dark. The number and viability of protoplasts in the both cultivation media were unchanged. In MCPW in the light, the protein and chlorophyll content strongly decreased already after 12 h of cultivation, at 72 h of cultivation, values dropped to 23.6 % (proteins) and to 3.5 % (chlorophyll) in comparison with the initial content. In GCPW in the dark, the protein and chlorophyll contents decreased only slightly to 75 % (proteins) and to 57.7 % (chlorophyll).

Tephrosia purpurea Pers. was found to accumulate high proline content in dry habitat. The proline content was higher in shoots, especially in leaves, than in roots. Pod walls and young seeds showed the highest proline content. The proline content of young leaves was higher than that of mature and old leaves. During leaf senescencein vitro proline content increased rapidly upto 6 h and further decreased in leaves as well as in leachate. High proline content seems to be positively related with 'survival capability' of this plant.

In seedlings of the tropical tree speciesErythrina variegata Lam. andHardwickia binata Roxb. exposed to different acidic mist (H₂SO₄, pH 5, 3 and 2) for 5 d significant reduction in seedling growth, biomass accumulation and¹⁴CO₂ fixation were determined. In isolated chloroplasts a decrease in the activities of photosystem 2 and whole electron transport chain was observed only at pH 3 and 2, but no significant change in photosystem 1 activity was observed. SDS-PAGE analysis of crude leaf extracts of ribulose 1,5-bisphosphate carboxylase (RuBPC) indicated a significant loss of 55 and 15 kDa polypeptides at pH 2 inErythrina. The reduction in the RuBPC activity in seedlings grown under acidic mists correlated well with CO₂ fixation.

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.

The influence of macronutrients and growth regulators on in vitro shoot proliferation and rooting of an East Spanish population ofMyrtus communis L. were studied. Preincubation of field material on a medium without mineral salts prevented the browning from phenolic exudates. For multiplication, nodal segments of 5 mm fromin vitro produced shoots were cultured on Murashige and Skoog (MS), Schenk and Hildebrandt (SH) and Heller (H) media (full strength or diluted to 1/2 or 1/4), with 6-benzylaminopurine (BAP) at concentrations 4.4, 13.3 and 22.2 ΜM or kinetin (K) at concentrations 4.7, 14.0 and 23.2 ΜM. The optimum shoot proliferation was on quarter-strength MS medium with 4.4 ΜM BAP, whereas the maximum number of nodal segments was produced on half-strength MS medium with 4.4 ΜM BAP. Rooting of shoots was obtained by adding 2.5 - 24.6 ΜM indole-3-butyric acid (IBA) and broad range of macronutrients; Lloyd and McCown (WPM) and Gresshoff and Doy (GD) media both full strength or diluted to 1/2 were optimum. No rooting was obtained in the presence of α-naphthaleneacetic acid (NAA).

Plants of two wheat (Triticum aestivum L.) cultivars differing in salt tolerance were grown in sand with nutrient solutions. 35-d-old plants were subjected to 5 levels of salinity created by adding NaCl, CaCl₂ and Na₂SO₄. Growth reduction caused by salinity was accompanied by increased Na⁺ and Cl^- concentrations, Na⁺/K⁺ ratio, and decreased concentration of K⁺. The salt tolerant cv. Kharchia 65 showed better ionic regulation. Salinity up to 15.7 dS m^-1 induced increased uptake of Na⁺ and Cl^- but higher levels of salinity were not accompanied by further increase in uptake of these ions. Observed increases in Na⁺ and Cl^- concentrations at higher salinities seemed to be the consequence of reduction in growth. Uptake of K⁺ was decreased; more in salt sensitive cultivar. This was also accompanied by differences in its distribution.

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

Potato spindle tuber viroid (PSTVd, severe strain) was isolated from tobacco plants transformed with a dimeric infectious expression construct which was maintained for a long time inAgrobacterium tumefaciens. Afterin vitro hybridization of PSTVd to complete minus transcripts of PSTVd (lethal), the resulting heteroduplexes were analyzed by electrophoresis under native conditions. Electrophoretic analysis revealed an appearence of electrophoretically distinguishable heteroduplexes, suggesting an accumulation of mutated sequence variants of PSTVd had occurred in the plant transformants. TGGE analysis of PSTVd cDNA, re-cloned from the original expression vector pCB1413 to plasmid pUC18 confirmed the accumulation of mutations in the cDNA and the instability of this sequence inA. tumefaciens maintained at 4°C for 2.5 years. One of these point mutations was identified by sequencing the PSTVd cDNAs isolated from the individualE. coli colonies. This transition (GC→AT) was localized at the position of 81 in the PSTVd genome, causing the change of C to U in PSTVd plus RNA. Transformation of tobacco with the freshly prepared expression vector containing the dimeric sequence of PSTVd lethal KF440-2 lead to the propagation of PSTVd electrophoretically identical to that derived from the original sequence and maintained in the tomato by a conventional infection.

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.

Supply of 100 μM spermidine (Spd) in the nutrient solution containing 10 mM nitrate as the sole nitrogen source, increased growth of roots and shoots, total nitrogen content andin vivo orin vitro nitrate reductase (NR) activity of leaves of 10-d oldLeucaena leucocephala seedlings. Spd and the cytokinins benzyladenine or kinetin also increased growth, total nitrogen andin vivo NR activity of isolated cotyledons. The synergistic effects of nitrate, kinetin and Spd in increasing NR activity, indicate that the Spd acted at different level than the nitrate or cytokinin.

The addition of NaCl to cadmium had significant synergistic effect on the wheat root and shoot fresh mass, relative growth rate and net assimilation rate, while showed no significant effects on the dry mass production, leaf area, leaf area ratio, leaf mass ratio and specific leaf area. Additive depression of the rate of photosynthesis and the stomatal conductance was recorded, while no significant effect on the transpiration rate was observed. The Cd stress disturbed the mineral nutrition of the wheat plants either directly or indirectly, NaCl markedly reduce the uptake and internal concentration of K and Ca in the shoot. The combination of cadmium and NaCl showed no additive effects on the content of ions in the root as well as in the shoot of wheat plants.

Hybridization between the nearly extinct speciesDiplotaxis siettiana andBrassica juncea is prevented because of strong prefertilization barriers. Use of mentor pollen ofD. siettiana irradiated with 1000 Gy gamma radiation before the incompatible pollination led to fertilization. 5 d after pollination 17% ovules showed entry of pollen tubes, 10 d after pollination 27% ovules showed small globular embryos which grew no further. No embryos were found in control pollinations. Thus , use of irradiated mentor pollen brings about fertilization in this difficult cross and hybrids can be obtained if embryos are rescued.

The short term uptake of phosphate involving 10 min absorption followed by 5 min desorption, both at 30 °C, in the concentration range 1.0×10^-9 to 7.5×10^-2 M KH₂PO₄ by fresh and washed maize (Zea mays L. cv. Ganga Safed-2) roots can be described by a single isotherm having five phases (0 and I-IV) with regularly spaced kinetic constants. Almost identical kinetics were observed in both fresh and washed maize roots. The kinetics of phase 0 in the concentration range 1.0×10^-9-3.0×10^-5 M. was sigmoidal in fresh maize roots, however, in washed tissue exhibited 2 phases termed here as 0a and 0b. 0a covered the concentration range 1.0×10^-9-5.0×10^-6 M and 0b 6.0×10^-6-3.0×10^-5 M. In the concentration range 1.0×10^-4-7.5×10^-2 M four distinct phases, termed as I, II, III and IV were evident in both fresh and washed maize roots. Each phase obeyed Michaelis-Menten kinetics. The values of K_m and V_max have been estimated for each phase. The uptake isotherm was accompanied by discontinuous transitions.

The induction of sister chromatid exchanges (SCEs) inVicia faba root-tip cells after short-term (2 h) and long-term (24 h) treatments with alkylating agents (N-methyl-N-nitrosourea, ethyl methanesulphonate) and maleic hydrazide was studied. The primary roots were treated with mutagens before or after 5-bromodeoxyuridine (BrdU) incorporation into DNA and the influence of mutagen application on SCE induction in the cells with non- and BrdU-substituted chromosomal DNA. On the contrary, application of maleic hydrazide after the incorporation of BrdU into DNA strongly increased the rate of SCEs. The lowest limit concentrations of mutagens capable of significantly increasing SCE frequency in the cells with non-substituted DNA after the long-term treatment were estimated.

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 effect of L-proline, glycinebetaine, L-and D-asparagine on rice seedling growth under NaCl stress was investigated. Glycinebetaine and L-asparagine were not effective in reducing NaCl inhibition in shoot growth of rice seedlings. L-Proline and D-asparagine were found to be able to reduce shoot growth inhibition under NaCl stress. However, L-proline, glycinebetaine, D- and L-asparagine further enhanced NaCl inhibition of root growth.