Accumulation of inorganic and organic osmolytes and their role in osmotic adjustment in NaCl-stressed vetiver grass seedlings

The accumulation of inorganic and organic osmolytes and their role in osmotic adjustment were investigated in roots and leaves of vetiver grass (Vetiveria zizanioides) seedlings stressed with 100, 200, and 300 mM NaCl for 9 days. The results showed that, although the contents of inorganic (K⁺, Na⁺, Ca²⁺, Mg²⁺, Cl⁻, NO₃⁻, SO₄²⁻ and H₂PO₃⁻)) and organic (soluble sugar, organic acids, and free amino acids) osmolytes all increased with NaCl concentration, the contribution of inorganic ions (mainly Na⁺, K⁺, and Cl⁻) to osmotic adjustment was higher (71.50–80.56% of total) than that of organic solutes (19.43–28.50%). The contribution of inorganic ions increased and that of organic solutes decreased in roots with the enhanced NaCl concentration, whereas the case in leaves was opposite. On the other hand, the osmotic adjustment was only effective for vetiver grass seedlings under moderate saline stress (less than 200 mM NaCl).     

Effects of water deficit on photosynthetic rate and osmotic adjustment in tetraploid wheats

Osmotic adjustment, accumulation of soluble saccharides, and photosynthetic gas exchange were studied in five durum wheat (Triticum turgidum L. var. durum) and one wild emmer wheat (Triticum turgidum L. var. dicoccoïdes) cultivars of contrasting drought tolerance and yield stability. Soil water contents (SWC) were 100, 31, 20, and 12 % of maximum capillary capacity. Under mild water stress (SWC 31 to 20 %), osmotic adjustment capacity and high accumulation of saccharides were found in cv. Cham1, a high yielding and drought tolerant cultivar, and in var. dicoccoïdes, while lowest values were noted in the durum wheat landraces Oued-Zenati and Jennah-Khotifa. Under more severe water stress (SWC 12 %), the cv. Cham1 maintained higher net photosynthetic rate (PN) than other genotypes. The observed changes in the ratio intercellular/ambient CO2 concentration (ci/ca) indicated that under mild and severe water stress, the decrease in PN was mainly due to stomatal and non-stomatal factors, respectively.     

Changes in free amino acids and osmotic adjustment in leaves of water-stressed bean

Bean ( Phaseolus vulgaris L. cv. Topcrop) plants were raised in a growth chamber in small pots or flower boxes and kept at full water regime until the full development of primary leaves (14–16 days). Both potted and flower box-grown plants were subjected to a gradually increased water stress of about 60–70 kPa day⁻¹ leaf water potential (stressed plants) or full water regime (control). The water potential, osmotic potential and turgor pressure in freshly detached primary leaves and osmotic potential at zero turgor were calculated using pressure/volume curves. Changes in free amino acids and amides were also measured in parallel trials. Water relation parameters documented that in the stressed leaves there was moderate osmotic adjustment, which was more evident in the potted plants. If considered 0% ionised, the accumulation of free amino acids and amides (μmol g⁻¹ H₂O) accounts for a van't Hoff's value of about 10.2 kPa in the small pot-grown and 5.5 kPa in the box-grown plants. The values are twice as high if considered 100% ionised. Proline accumulation accounted for about 6.4% of the pool enlargement in the potted plants and 22.3% in the flower box plants.     

Relationship between osmotic stress and polyamines conjugated to the deoxyribonucleic acid-protein in wheat seedling roots

The contents of covalently conjugated polyamines (CC-PAs) and noncovalently conjugated polyamines (NCC-PAs) to deoxyribonucleic acid-protein (DNP) isolated from wheat (Triticum aestivum L.) seedling roots under osmotic stress were detected. Results showed that after osmotic stress treatment for 7 d, the levels in NCC-spermine (NCC-Spm) and NCC-spermidine (NCC-Spd) of drought-tolerant Yumai No. 18 cv. increased more markedly than that of drought-sensitive Yangmai No. 9 cv., while the NCC-putrescine (NCC-Put) could not be statistically detected in two cultivars. Exogenous Spm treatment alleviated osmotic stress injury to Yangmai No. 9 cv. seedlings, coupled with marked increases of NCC-Spm and NCC-Spd levels of this cultivar. Under PEG osmotic stress, the concomitant treatment of drought-tolerant Yumai No. 18 cv.seedlings with methylglyoxyl-bis (guanylhydrazone) (MGBG), an inhibitor of S-adenosylmethionine decarboxylase (SAMDC), aggravated osmotic stress injury to this cultivar, coupled with market decreases of the NCC-Spm and NCC-Spd levels. The levels in CC-Put and CC-Spd of drought-tolerant Yumai No. 18 cv. increased more markedly than that of drought-sensitive Yangmai No. 9 cv. Under osmotic stress. The treatment of drought-tolerant Yumai No. 18 cv. seedlings with phenanthrolin (o-Phen), an inhibitor of transglutaminase (TGase), aggravated osmotic stress injury to this cultivar, coupled with a reduction of sum contents of CC-Put+CC-Spd. These results suggested that NCC-Spm and NCC-Spd, together with CC-Put and CC-Spd, in DNP of roots could enhance tolerance of the wheat seedlings to osmotic stress.     

Relationship between osmotic stress and polyamines conjugated to the deoxyribonucleic acid-protein in wheat seedling roots

Polyamines (PAs) are biologically ubiquitous aliphatic amines that are implicated in many aspects of growth, development, sex differentiation, ripeness and senescence of plants[1―6]. It has been well documented that PAs are closely associated with plant …     

Ratio of free to bound polyamines during maturation in mung-bean hypocotyl cells

Free- and bound-polyamine levels were estimated in successive segments of the mung-bean hypocotyl. Three aliphatic polyamines (putrescine, spermidine and spermine) were found in proportions which depended on the state of maturation. In young cells, most of the polyamines were located in the protoplasm whereas in older cells they were mostly bound to the cell walls. Spermidine was always the main bound polyamine, and putrescine, the main free polyamine.Abbreviation EDTA ethylenediaminetetraacetic acid     

Changes in osmotic water permeability of the eel gills during seawater and freshwater adaptation

Summary Changes in osmotic water permeability of the isolated gills of the Japanese eel,Anguilla japonica were studied during transfer to seawater or to fresh water. The water permeability increased gradually during the course of seawater transfer and attained a maximal level after 2 weeks. The water permeability of the freshwater eel gills was reduced when calcium ions were added to the incubation medium at a concentration of 1 mM, where-as no effect of the ion was observed on the gills of the seawater-adapted eel even at a higher concentration (10 mM). In contrast to seawater transfer, the water permeability decreased to a low level almost immediately (3 h) after transfer from seawater to fresh water. The acute reduction of the water permeability was also seen in the gills of the hypophysectomized eel after transfer to fresh water.The gradual increase in the gill water permeability during seawater transfer is correlated with an increase in the number of chloride cells. In scanning electron microscopy, chloride cells of seawater-adapted eel gills exhibit a pit-like structure, which was larger than in the freshwater eel. On transfer from seawater to fresh water, the pit diameter became smaller within 6 h. Hypophysectomy did not affect the change in gill surface structures during transfer to fresh water. The junctions between the chloride cells of seawater eel gills are reported to be of the leaky type. The parallel change in osmotic water permeability and in pit size of the chloride cells during seawater or freshwater transfer or after hypophysectomy suggests that these cells could provide a major route of water as well as ion movement.This paper is a portion of a thesis presented to Hokkaido University by t. Ogasawara in partial fulfilment of the requirements for Doctor of Fisheries     

Changes of free, soluble conjugated and bound polyamine titers of jojoba explants under sodium chloride salinity in vitro

Jojoba (Simmondsia chinensis L.) single node explants were cultured in a basal medium supplemented with 17.8 microM 6-benzyladenine and four levels of sodium chloride concentration (0, 56.41, 112.82 and 169.23 mM). The free, the soluble conjugated and the insoluble bound forms of polyamines (PAs) (putrescine (Put), spermidine (Spd) and spermine (Spm)) were determined monthly during a 3-month proliferation stage. Free Put and Spd were found in higher levels in the control treatment, while Spm content was higher in the salt treatments. All soluble conjugated PAs were found to be in lower concentrations in explants growing on medium supplemented with salt, while the opposite was true for the insoluble bound PAs. It appeared that certain PAs and PAs forms could play a significant role in the adaptation mechanism of jojoba under saline conditions.     

Ectomycorrhizal fungal species and strains differ in their ability to produce free and conjugated polyamines

Production of free and conjugated polyamines by one strain of Laccaria proxima (Boud.) Maire, three strains (H, O, K) of Paxillus involutus (Batsch) Fr., and one strain of Pisolithus tinctorius was studied in vitro. Spermidine (Spd) was the main polyamine in the 4-week-old mycelium of all the fungi. It was mainly present in the free form, but it also occurred in conjugated forms. Paxillus involutus strain H released large amounts of free putrescine (Put), and the Pisolithus tinctorius released a compound probably related to cadaverine (Cad). On the other hand, these two fungi contained less conjugated polyamines than the other fungi. In addition to the amounts, the forms (perchloric acid soluble and insoluble) of conjugated polyamines in the mycelium varied between species and strains. L. proxima contained nearly as much insoluble conjugated Spd as free Spd, whereas Paxillus involutus strains O and K contained relatively large amounts of soluble conjugated Spd. The results suggest that ectomycorrhizal fungal species and strains differ in their ability and need to produce conjugated polyamines. The small amounts of soluble conjugated polyamines found in the culture filtrates indicate that some specific conjugated polyamines may be involved in polyamine translocation across the plasma membrane.     

Salt tolerance of Beta macrocarpa is associated with efficient osmotic adjustment and increased apoplastic water content

The chenopod Beta macrocarpa Guss (wild Swiss chard) is known for its salt tolerance, but the mechanisms involved are still debated. In order to elucidate the processes involved, we grew wild Swiss chard exposed to three salinity levels (0, 100 and 200 mm NaCl) for 45 days, and determined several physiological parameters at the end of this time. All plants survived despite reductions in growth, photosynthesis and stomatal conductance in plants exposed to salinity (100 and 200 mm NaCl). As expected, the negative effects of salinity were more pronounced at 200 mm than at 100 mm NaCl: (i) leaf apoplastic water content was maintained or increased despite a significant reduction in leaf water potential, revealing the halophytic character of B. macrocarpa; (ii) osmotic adjustment occurred, which presumably enhanced the driving force for water extraction from soil, and avoided toxic build up of Na⁺ and Cl^– in the mesophyll apoplast of leaves. Osmotic adjustment mainly occurred through accumulation of inorganic ions and to a lesser extent soluble sugars; proline was not implicated in osmotic adjustment. Overall, two important mechanisms of salt tolerance in B. macrocarpa were identified: osmotic and apoplastic water adjustment.     

Inheritance of osmotic adjustment to water stress in three grain sorghum crosses

Water stress is one of the major constraints to the grain yield of sorghum in tropical and sub-tropical areas of the world. Osmotic adjustment has been widely proposed as a plant attribute that confers adaptation to water stress. The inheritance of osmotic adjustment to water stress was investigated in a series of generations derived from the three possible bi-parental crosses between two inbred sorghum lines with a high capacity for osmotic adjustment (Tx2813 and TAM422; high-OA lines) and one with a low capacity (QL27; low-OA line). Broad-sense heritability on a single-plant basis was generally found to be high. Analysis of segregation ratios by the mixture method of clustering identified two independent major genes for high osmotic adjustment. The line Tx2813 possessed a recessive gene which is given the symbol oa1; the line TAM422 possessed an additive gene which is given the symbol OA2. There was some evidence that there may be other minor genes which influence the expression of osmotic adjustment in these crosses as two putative transgressive segregants, with higher osmotic adjustment than the parents, were identified from the cross between Tx2813 and TAM422. Populations of recombinant inbred lines were developed and characterised for osmotic adjustment for two of the crosses (QL27 x TAM422, low-OA x high-OA; Tx2813 x TAM422, high-oal x high-OA2). These will be used to conduct experiments which test hypotheses about the contribution of the high-osmotic-adjustment genes to the grain yield of sorghum under a range of water-stress conditions.     

Modeling of plant adaptation to climatic drought induced water deficit

Soil moisture flux to root surface is considered the main determining factor of the transpiration intensity of plants. This assumption is valid not only in optimal plant physiological conditions without any physical barrier for the evaporation from the leaves, but in climatic drought as well, when high usable soil water amount cannot supply the evapo-transpiration intensity of plant. A new algorithm we built up describing the plant adaptation in climatic drought when stoma’s closure and reduction of plant’s potential evapo-transpiration (PET) starts. The adaptation algorithm of Doorenbos et al. (1978) is developed further defining that soil moisture content initiating the stomata’s closure. The critical soil moisture content is varying according to the PET, and drought tolerance of plant. If soil moisture content is less than the critical one, the plant evapo-transpiration (ET) can be highly different in the drought tolerance plant groups. The new drought tolerance algorithm is applied to maize field plots on chernozem soil of the experimental station of the Debrecen University, in East Hungary. Simulated soil water storages are compared to measured ones of a field plot treatment in five consecutive years. The soil moisture content profiles are measured with a BR-150 capacitance probe (Andrén et al. 1991). Differences between measured and simulated soil water storages are not significant in 2003. Simulations indicate low soil water storages in autumn of 2006, and in the first half of 2007 predicting the low maize production realized in 2007. The new plant adaptation algorithm can be used for a climate and soil moisture content sensitive irrigation control as well. The maize production is an illustrative biohydrological example of water flow through the soil-plant-atmosphere continuum.     

Solutes contributing to osmotic adjustment in cultured plant cells adapted to water stress

Osmotic adjustment was studied in cultured cells of tomato (Lycopersicon esculentum Mill cv VFNT-Cherry) adapted to different levels of external water potential ranging from −4 bar to −28 bar. The intracellular concentrations of reducing sugars, total free amino acids, proline, malate, citrate, quaternary ammonium compounds, K⁺, NO₃⁻, Na⁺, and Cl⁻ increased with decreasing external water potential. At any given level of adaptation, the maximum contribution to osmotic potential was from reducing sugars followed by potassium ions. The sucrose levels in the cells were 3- to 8-fold lower than reducing sugar levels and did not increase beyond those observed in cells adapted to −16 bar water potential. Concentrations of total free amino acids were 4- to 5-fold higher in adapted cells. Soluble protein levels declined in the adapted cell lines, but the total reduced nitrogen was not significantly different after adaptation. Uptake of nitrogen (as NH₄⁺ or NO₃⁻) from the media was similar for adapted and unadapted cells. Although the level of quaternary ammonium compounds was higher in the nonadapted cells than that of free proline, free proline increased as much as 500-fold compared to only a 2- to 3-fold increase observed for quaternary ammonium compounds. Although osmotic adjustment after adaptation was substantial (up to −36 bar), fresh weight (volume increase) was restricted by as much as 50% in the adapted cells. Altered metabolite partitioning was evidenced by an increase in the soluble sugars and soluble nitrogen in adapted cells which occurred at the expense of incorporation of sugar into cell walls and nitrogen into protein. Data indicate that the relative importance of a given solute to osmotic adjustment may change depending on the level of adaptation.     

Response of photosynthetic performance,water relations and osmotic adjustment to salinity acclimation in two wheat cultivars

Experiment was conducted to identify the impacts of the salinity acclimation process on the photosynthetic efficiency, osmotic adjustment, membrane integrity, and yield components in two wheat cultivars differing in their salinity tolerance. The design of the experiment was factorial randomized block, where genotype is factor 1 and acclimation treatments represent factor 2. Genotypes were grown from emergence to 30 days after sowing (DAS) by irrigating with tap water [electrical conductivity (EC) of 0.776 dS m^?1]. Thereafter, both the genotypes were divided into two groups and exposed to either irrigation with sublethal level of salinity EC of 2.09 or 3.76 dS m^?1 for 21 days. At booting stage (65 DAS), both groups were subjected to lethal level of salinity stress EC of 12 dS m^?1 for 21 days, followed by irrigation with tap water till maturity. Non-acclimated plants were irrigated with tap water from emergence to 65 days, then directly irrigated with lethal level of salinity for 21 days, followed by irrigation with tap water till maturity. The control plants were continuously irrigated with tap water from emergence until maturity. The non-acclimated plants had decreased electron transport rates at the donor and acceptor side of PSII and PSI in Giza 168, and decreased electron transport rates at PSII acceptor side in Sakha 8 compared to control plants. In both genotypes, the non-acclimated plants had decreased chlorophyll a, b, carotenoid, proline and total soluble sugar concentration, relative water content, membrane stability index, yield and yield components compared with acclimated plants. While, osmotic potential and lipid peroxidation showed an opposite trend. Overall, acclimation treatment (EC of 2.09 dS m^?1) during vegetative stage alleviated the inhibitory effects of lethal level of salinity stress at booting stage through enhanced photosynthetic efficiency and osmotic adjustment, resulting in increased membrane integrity, biomass production and grain yield than in non-acclimated plants.     

Changes in free polyamines and gene expression during peach flower development

Free polyamine contents and expressions of five genes encoding for polyamine biosynthetic enzymes were investigated at four different stages during peach (Prunus persica L. Batsch cv. Akatsuki) flower development. Fresh mass of peach flowers increased, accompanied by reduction in contents of total polyamines and putrescine/spermidine ratio due to decrease in putrescine content. Spermidine, the largest fraction, and spermine, the least part, underwent minor change. Expressions of the five key genes involved in polyamine biosynthesis during flower development did not parallel the changes in free polyamines.     

Contributions of soluble carbohydrates to the osmotic adjustment in the C4 grass Setaria sphacelata: a comparison between rapidly and slowly imposed water stress

Photosynthetic carbohydrate content in Setaria sphacelata var. splendida under rapidly and slowly induced water deficit and its contribution to osmotic adjustment were studied. In short-term stress experiments, a decrease in the total content of sucrose (Su) and starch (St) was observed in leaf discs submitted to stress. An increase in the ratio between free hexoses and sucrose was found in stressed leaves, but no significant differences were found in the amount of free hexoses nor in the ratio between soluble and insoluble sugars. In long-term stress experiments, a higher amount of soluble sugars and a lower amount of starch were found in stressed leaves, when compared to the control. The ratios of free hexoses to sucrose and of soluble to insoluble sugars were also higher in stressed leaves. The contribution of the accumulation of soluble sugars to osmotic adjustment was absent in rapidly stressed leaves and was of minor importance in slowly stressed leaves.     

Fluctuation in free and conjugated polyamines in Scots pine seedlings after changes in temperature and daylength

Free, soluble and insoluble conjugated polyamines from the needles, roots and stem of five month old Scots pine (Pinus sylvestris L.) seedlings inoculated with Suillus variegatus (Fr.) O. Kuntze and seedlings without inoculation were analysed during decrease in daylength and temperature. Temporary changes in free, soluble and insoluble conjugated polyamine pools caused by a decrease in daylength or temperature were observed. Inoculation of pine seedlings affected significantly the polyamine levels of five month old pine seedlings. The roots of inoculated seedlings contained significantly higher levels of free and soluble conjugated purtrescine and free, soluble conjugated and insoluble conjugated spermidine than the roots of noninoculated seedlings. The needles of inoculated seedlings contained significatly higher concentrations of free putrescine and soluble conjugated spermidine but lower amount of free spermine than the needles of noninoculated seedlings. The stems of inoculated seedlings contained higher concentrations of free putrescine but lower amounts of insoluble conjugated spermine. Changes in polyamine levels in noninoculated seedlings were observed after shortening of the daylength, whereas in inoculated ones changes were induced mainly by the decrease in temperature. The possible role of polyamines in the initial stage of cold hardening process is discussed.     

Economic incentive for applying vetiver grass to remediate lead, copper and zinc contaminated soils

The application of vetiver grass (Chrysopogon zizaniodes) for phytoremediation of heavy metal contaminated soils can be promoted by economic return through essential oil production. Four levels of lead (0, 500, 2000, and 8000 mg kg(-1) dry soil), copper (0, 100, 400, and 1600 mg kg(-1) dry soil) and zinc (0, 400, 1600, and 6400 mg kg(-1) dry soil) were used to study their effects on vetiver growth, essential oil composition and yield. This study also investigated the effect of nitrogen concentrations on vetiver oil yield. Vetiver accumulated high concentrations of Pb, Cu and Zn in roots (3246, 754 and 2666 mg kg(-1), respectively) and small amounts of contaminants in shoots (327, 55, and 642 mg kg(-1), respectively). Oil content and yield were not affected at low and moderate concentrations of Cu and Zn. Only the application of Pb had a significant detrimental effect on oil composition. Extraction of vetiver essential oils by hydrodistillation produced heavy metal free products. High level of nitrogen reduced oil yields. Results show that phytoremediation of Cu and Zn contaminated soils by vetiver can generate revenue from the commercialization of oil extracts.     

Effect of common sage plant treatment with polyamines on the contents of proline and free and conjugated polyamines under oxidative stress

Common sage (Salvia officinalis L.) plants grown in water culture to the stage of 4–5 true leaves were treated with paraquat (PQ) ( 1 ml of the solution containing 0.1 μM PQ in 0.05% Tween 80), putrescine (Put), spermidine (Spd), or Spermine (Spm) (all polyamines (PA) at the concentration of 0.5 mM in 0.05% Tween 80) or PA plus PQ for 6, 12, 24, 36, or 48 h. Under normal conditions, treatment with individual PA did not induce any changes in the content of free proline. Under oxidative stress induced by PQ, oxidation of proline and free PA by ROS resulted in their spending and demanded restoration of free PA due to hydrolysis of their insoluble conjugates. As distinct from treatment with PQ only, its combination with PA in the light was accompanied by proline accumulation. Treatment with Put plus PQ induced accumulation of intracellular Put and its soluble and insoluble conjugates. Treatments with Spd and Spm in combination with PQ resulted similarly in the increase in the levels of their intracellular soluble and insoluble conjugates. In treatments with these high-molecular PA, polyamine oxidase was activated and diaminopropan formation was observed. It seems likely that, for restoration of high-molecular PA homeostasis, their degradation by polyamine oxidase or production of insoluble conjugates is induced. Thus, the amount of free and conjugated particular PA is under strict control and is maintained at a definite level. A decrease in the content of a particular free PA induces primarily a decrease in the content of its soluble and then insoluble conjugates. The data obtained demonstrate the effects of exogenous PA on the content of intracellular proline under oxidative stress but do not allow a conclusion about direct regulation of its content by PA.     

水分胁迫及复水过程中小麦抗氧化酶的变化

对两个抗旱性不同的小麦品种进行水分胁迫和复水处理,研究其抗氧化酶活性的响应。在水分胁迫下,陇春-20的相对含水量高于优鉴-24,复水24h后,优鉴-24的相对含水量恢复较快且高于陇春-20。水分胁迫下,优鉴-24中H2O2含量增加迅速,而且各阶段含量均高于陇春-20,复水后两个品种的H2O2含量都下降,这表明优鉴-24在水分胁迫时受到更严重的氧化胁迫。采用温和胶电泳结合抑制剂实验发现小麦有3条Mn—SOD,一条Fe—SOD和Cu／Zn-SOD同工酶带,CAT同工酶有3条谱带。在水分胁迫和复水期间,优鉴-24的SOD和CAT活性高于陇春-20,随着水分胁迫程度的增加,两个品种的SOD和CAT活性都增强,复水后,优鉴-24的SOD活性继续增强,而陇春-20的Mn—SOD—3活性略微降低,Fe—SOD和Cu／Zn—SOD活性略微升高,陇春-20的CAT活性降低。水分胁迫诱导了Mn—SOD—1在优鉴-24及Mn—SOD-2和Fe—SOD在陇春-20中的表达。