Modifications of water status, growth rate and antioxidant system in two wheat cultivars as affected by salinity stress and salicylic acid.

Salicylic acid (SA) has an important role in drought-tolerance in wheat (Triticum aestivum L.) but its relevance to the salinity-tolerance is not well understood. In the present study, possible roles of SA and salinity responses were examined using two wheat cultivars i.e., drought-tolerant Sakha-69 and drought-sensitive Gemaza-1, exposed to 150 mM NaCl. Parameters were determined for growth i.e. fresh or dry mass (FM, DM), osmotic concentration (OC) of organic/inorganic solute, leaf relative water content (LRWC), photosynthesis pigment content (PPC), and selective antioxidant system (AOS) enzyme/molecule that might be involved in the stress remediation. Sakha-69 exhibited salinity tolerance greater than Gemaza-1 and SA ameliorated their salinity stresses like drought stress, suggesting that a common tolerant mechanism might be involved in the stresses. Salinity decreased root growth by 44-52% more strongly than shoot (36-41%) in FM or those in DM (32-35%). SA ameliorated root growth (40-60%) more efficiently than shoot (6-24%) for DM/FM. These results suggested that salinity and SA might target sensitive roots and hence influencing shoot functions. In fact, salinity reduced PPC by 10-18%, LRWC by 16-28%, and more sensitively, OC of inorganic solutes (K+, Ca2+, Mg2+) in shoot (19-36%) and root (25-59%), except a conspicuous increase in Na+, and SA recovered all the reductions near to control levels. SA and salinity increased additively most parameters for OC of organic solutes (sugars and organic acids) and AOS (glutathione and related enzyme activities), like drought responses. However, SA decreased the Na+ and proline contents and catalase activity in a counteracting manner to salinity. It is concluded from this experiment that SA-mediated tolerance might involve two mechanisms, one specific for minerals in root and the other related to drought/dehydration tolerance governed in the whole module systems.

Uranium accumulation and its phytotoxicity symptoms in Pisum sativum L.

Environmental contamination by uranium (U) and other radionuclides is a serious problem worldwide, especially due to, e.g. mining activities. Ultimate accumulation of released U in aquatic systems and soils represent an escalating problem for all living organisms. In order to investigate U uptake and its toxic effects on Pisum sativum L., pea plantlets were hydroponically grown and treated with different concentrations of U. Five days after exposure to 25 and 50 µM U, P. sativum roots accumulated 2327.5 and 5559.16 mg kg-1 of U, respectively, while in shoots concentrations were 11.16 and 12.16 mg kg-1, respectively. Plants exposed to both U concentrations showed reduced biomass of shoots and reduced content of photosynthetic pigments (total chlorophyll and carotenoids) relative to control. As a biomarker of oxidative stress, lipid peroxidation (LPO) levels were determined, while antioxidative response was determined by catalase (CAT) and glutathione reductase (GR) activities as well as cysteine (Cys) and non-protein thiol (NP-SH) concentrations, both in roots and shoots. Both U treatments significantly increased LPO levels in roots and shoots, with the highest level recorded at 50 µM U, 50.38% in shoots and 59.9% in roots relative to control. U treatment reduced GR activity in shoots, while CAT activity was increased only in roots upon treatment with 25 µM U. In pea roots, cysteine content was significantly increased upon treatment with both U concentrations, for 19.8 and 25.5%, respectively, compared to control plants, while NP-SH content was not affected by the applied U. This study showed significant impact of U on biomass production and biochemical markers of phytotoxicity in P. sativum, indicating presence of oxidative stress and cellular redox imbalance in roots and shoots. Obtained tissue-specific response to U treatment showed higher sensitivity of shoots compared to roots. Much higher accumulation of U in pea roots compared to shoots implies potential role of this species in phytoremediation process.

Physiological changes of a green alga (Micractinium sp.) involved in an early-stage of association with Tetrahymena thermophila during 5-year microcosm culture.

Endosymbioses between phototrophic algae and heterotrophic organisms are an important symbiotic association in that this association connects photo- and heterotrophic metabolism, and therefore, affects energy/matter pathways and cycling in the ecosystem. However, little is known about the early processes of evolution of an endosymbiotic association between previously non-associated organisms. In previous studies, we analyzed an early process of the evolution of an endosymbiotic association between an alga and a ciliate by using a long-term culture of an experimental model ecosystem (CET microcosm) composed of a green alga (Micractinium sp.), a bacterium (Escherichia coli), and a ciliate (Tetrahymena thermophila). The results revealed that an algal type, isolated from 5-year cultures of the microcosm, prolonged the longevity of the ancestral and derived clones of T. thermophila in the absence of bacteria, suggesting that a cooperative algal phenotype that benefited the ciliate had evolved in the microcosm. Here, we investigated the physiological changes of the derived Micractinium clones that benefited Tetrahymena, focusing on the release of carbohydrates by and abundance of photopigments in the ancestral and 2 derived algal clones (SC10-2 and SC9-1) isolated from inside Tetrahymena cells. Analyses using HPLC revealed that the algal isolates released glycerol and sucrose at higher concentrations per cell and also contained higher levels of photopigments per cell at pH 7.2, in comparison with the ancestral strain. These phenotypic characters were considered responsible for the increased longevity of Tetrahymena cells, and thus supported the cooperator alga hypothesis.

Changes in the water status and osmotic solute contents in response to drought and salicylic acid treatments in four different cultivars of wheat (Triticum aestivum).

Salicylic acid (SA) controls growth and stress responses in plants. It also induces drought tolerance in plants. In this paper, four wheat (Triticum aestivum L.) cultivars with different drought responses were treated with SA in three levels of drain (90, 60, 30% of maximum field capacity) to examine its interactive effects on drought responses and contents of osmotic solutes that may be involved in growth and osmotic adjustment. Under drought condition, the cultivars Geza 164 and Sakha 69 had the plant biomass and leaf relative water content (LRWC) greater than the cultivars Gemaza 1 and Gemaza 3. In all cultivars, drought stress decreased the biomass, LRWC, and the contents of inorganic solutes (Ca, K, Mg) and largely increased the contents of organic solutes (soluble sugars and proline). By contrast, SA increased the biomass, LRWC and the inorganic and organic solute contents, except proline. Correlation analysis revealed that the LRWC correlated positively with the inorganic solute contents but negatively with proline in all cultivars. SA caused maximum accumulations of soluble sugars in roots under drought. These results indicated that SA-enhanced tolerance might involve solute accumulations but independently of proline biosynthesis. Drought-sensitive cultivars had a trait lowering Ca and K levels especially in shoots. Possible functions of the ions and different traits of cultivars were discussed.

Mannose accommodation of Vigna angularis cells on solid agar medium involves its possible conversion to sucrose mediated by enhanced phosphomannose isomerase activity.

Mannose is an unusable carbon source for many plants. In our study we compared the effects of mannose and sucrose on growth and sucrose levels in azuki bean (Vigna angularis) cells grown in liquid media and in solid media. The suspension cells grew actively in a liquid medium containing 90 mM sucrose but not in that containing 90 mM mannose, where the intracellular sucrose levels were reduced to 20% or less of those in sucrose-grown cells. These results suggested that the limited conversion of mannose to sucrose resulted in cell growth inhibition. When sucrose-grown suspension cells (1 x 10(5)) were transferred onto agar medium containing mannose, they grew little initially, but, after a month lag period, they started to form many callus colonies at a high apparent variation rate (1.3 x 10(-3)). Time-course studies for sugar and enzyme analysis revealed that the mannose-accommodated cells were capable of converting mannose to sucrose, with enhanced phosphomannose isomerase activity. The mannose-accommodated cells actively grew in liquid medium with sucrose but lost their ability to grow with mannose again, suggesting a specific trait of callus culture for mannose utilization. The possible differences in the metabolic activities and other physiological characteristics are discussed between callus and suspension cells.

Different effects of galactose and mannose on cell proliferation and intracellular soluble sugar levels in Vigna angularis suspension cultures.

Plant cells utilize various sugars as carbon sources for growth, respiration and biosynthesis of cellular components. Suspension-cultured cells of azuki bean (Vigna angularis) proliferated actively in liquid growth medium containing 1% (w/v) sucrose, glucose, fructose, arabinose or xylose, but did not proliferate in medium containing galactose or mannose. These two latter sugars thus appeared distinct from other sugars used as growth substrates. Galactose strongly inhibited cell growth even in the presence of sucrose but mannose did not, suggesting a substantial difference in their effects on cell metabolism. Analysis of intracellular soluble-sugar fractions revealed that galactose, but not mannose, caused a conspicuous decrease in the cellular level of sucrose with no apparent effects on the levels of glucose or fructose. Such a galactose-specific decrease in sucrose levels also occurred in cells that had been cultured together with glucose in place of sucrose, suggesting that galactose inhibits the biosynthesis, rather than uptake, of sucrose in the cells. By contrast, mannose seemed to be metabolically inert in the presence of sucrose. From these results, we conclude that sucrose metabolism is important for the heterotrophic growth of cells in plant suspension-cultures.

Effect of copper on acid phosphatase activity in yeast Yarrowia lipolytica.

Acid phosphatase (APase) activity of the yeast Yarrowia lipolytica increased with increasing Cu2+ concentrations in the medium. Furthermore, the enzyme in soluble form was stimulated in vitro by Cu2+, Co2+, Ni2+, Mn2+ and Mg2+ and inhibited by Ag+ and Cd2+. The most effective ion was Cu2+, especially for the enzyme from cultures in medium containing Cu2+, whereas APase activity in wall-bound fragments was only slightly activated by Cu2+. The content of cellular phosphate involving polyphosphate was decreased by adding Cu2+, regardless of whether or not the medium was rich in inorganic phosphate. Overproduction of the enzyme stimulated by Cu2+ might depend on derepression of the gene encoding the APase isozyme.

Copper extrusion after accumulation during growth of copper-tolerant yeast Yarrowia lipolytica.

The Cu2+-tolerant yeast Yarrowia lipolytica accumulated Cu2+ until the late logarithmic phase. Thereafter, Cu2+ was temperature-dependently extruded into phosphate-limited culture medium containing high concentrations of heavy metal ions but not into 10 mM 2-(N-morpholino)ethane sulfonic acid (MES) buffer (pH 6.0). Peptone in the culture medium played an important role in the extrusion, which proceeded even when peptone was substituted with cysteine or histidine, but not with any other amino acid tested.

Characteristics of copper tolerance in Yarrowia lipolytica.

We discovered that a mutant strain of the dimorphic yeast Yarrowia lipolytica could grow in the yeast form in high concentrations of copper sulfate. The amount of metal accumulated by Y. lipolytica increased with increasing copper concentrations in the medium. Washing with 100 mM EDTA released at least 60% of the total metal from the cells, but about 20-25 micromol/g DW persisted, which represented about 30% of the soluble fraction of cultured cells. The soluble fraction (mainly cytosol) contained only about 10% of the total metal content within cells cultured in medium supplemented with 6 mM copper. We suggest that although a high copper concentration induces an efflux mechanism, the released copper becomes entrapped in the periplasm and in other parts of the cell wall. Washing with EDTA liberated not only copper ions, but also melanin, a brown pigment that can bind metal and which located at the cell wall. These findings indicated that melanin participates in the mechanism of metal accumulation. Culture in medium supplemented with copper obviously enhanced the activities of Cu, Zn-SOD, but not of Mn-SOD.

Relief of arsenate toxicity by Cd-stimulated phytochelatin synthesis in the green alga Chlamydomonas reinhardtii.

In most photosynthetic organisms, inorganic arsenic taken up into the cells inhibits photosynthesis and cellular growth. In a green alga, Chlamydomonas reinhardtii, 0.5 mM arsenate inhibited photosynthesis almost completely within 30 min. However, in cells acclimated with a sublethal concentration (0.05 to 0.1 mM) of Cd, the inhibition of photosynthesis at 30 min after the addition of arsenate was relieved by more than 50%. The concentrations of arsenic incorporated into the cells were not significantly different between the Cd-acclimated and the non-acclimated cells. The Cd-acclimated cells accumulated Cd and synthesized phytochelatin (PC) peptides, which are known to play an important role in detoxification of heavy metals in plants. By the addition of an inhibitor of glutathione (an intermediate in the PC biosynthetic pathway) biosynthesis, buthionine sulfoximine, cells lost not only Cd tolerance but also arsenate tolerance. These results suggest that glutathione and/or PCs synthesized in Cd-acclimated cells are involved in mechanisms of arsenate tolerance.

Changes in the levels of phytochelatins and related metal-binding peptides in chickpea seedlings exposed to arsenic and different heavy metal ions.

Phytochelatin-related peptides were analyzed in chickpea plants exposed to six different heavy-metal ions. Cadmium and arsenic stimulated phytochelatin and homophytochelatin synthesis in roots but other metals did not. These metals, however, caused an overall increase in the precursors, glutathione, homoglutathione and cysteine. These changes may be different biochemical indexes for heavy-metal contamination.

Impacts of fly-ash on soil and plant responses.

Coal combustion produces carbon dioxides, SO x, NO x and a variety of byproducts, including fly-ash, flue gas and scrubber sludge. Fly-ash consists of minute glass-like particles and its deposition on leaves inhibits the normal transpiration and photosynthesis of plants. Fly-ash also affects the physicochemical characteristics of soil because it is generally very basic, rich in various essential and non-essential elements, but poor in both nitrogen and available phosphorus. The massive fly-ash materials have been a potential resource for the agricultural activities as well as the other industrial purposes. Practical value of fly-ash in agriculture as an 'effective and safe' fertiliser or soil amendment can be established after repeated field experiments. Here remains to be disclosed the biological processes and interactions due to 'lack and excess' of the fly-ash exposures along with abiotic and biotic factors. These may involve the symbiotic fixation of nitrogen and the biological extraction of metals following immobilisation of toxic heavy metal ions, as well as other neutralisation and equilibration processes during weathering. Nitrogen-fixing plants with an apparent heavy metal-tolerance can be helpful as the early colonisers of fly-ash dumps and nearby areas.

Possible roles of phytochelatins and glutathione metabolism in cadmium tolerance in chickpea roots.

The possible roles of phytochelatin (PC) and glutathione (GSH) in the heavy metal detoxification in plants were examined using two varieties (CSG-8962 and C-235) of chickpea ( Cicer arietinumL.). The seedlings were grown for 5 days and the roots were treated with 0-20 micro M CdSO(4) for 3 days. The CSG-8962 seedlings exhibited more Cd-tolerant characteristics than did the C-235, where the roots, rather than shoots, suffered from more toxic effects by Cd. Both the seedlings synthesized the large amounts of PCs and homo-phytochelatins (hPCs) in roots, but only a little in shoots in response to Cd. The Cd treatments also caused a marked increase in the levels of GSH and cysteine in both the root and shoot tissues, suggesting that Cd may activate the GSH biosynthesis and, hence, enhance PC synthesis in the plants. Such a Cd-sensitive PC synthesis in chickpea plants does not explain the difference in Cd sensitivity in the varieties, but can be used as a biochemical indicator for Cd contamination in various environments. In the chickpea plants, possible PC-dependent and independent mechanisms for Cd tolerance are discussed.