Growth metabolism of wheat under drought stress at the jointing-booting stage

AimsThe aim of this study was to investigate the effects of drought stress on primary, secondary metabolites and metabolic pathways in the leaves of wheat, these parameters were evaluated to determine the physiological adaptive mechanisms by which wheat tolerates drought stress at the jointing-booting stage.MethodsA pot experiment was carried out in rain-proof shelter. The relative growth rate, photosynthetic characteristics and metabolism seedlings exposed to stresses lasting 12 days at jointing-booting stage were measured.Important findings The results displayed that the photosynthesis decreased under drought stress, causing the decreases of relative growth rate and dry matter mass. Profiles of 64 key metabolites produced by wheat including organic acids, amino acids, carbohydrates, purine, etc. were examined, 29 of them were changed significantly under drought stress. Principal component analysis (PCA) showed that 64% variations can be explained by the two principal components. One-way ANOVA analysis results revealed that long term drought stress decreased malic acid, citric acid and aconitic acid significantly, indicating inhibited tricarboxylic acid cycle. We further found that prolonged drought stress led to accumulation of progressive amino acids (proline, serine, valine) and carbohydrates (myo-inositol, fructose, clucose) in wheat leaves and depletion of transamination products (asparagine, glutamine, γ-aminobutyric acid). These results imply wheat may enhance its drought tolerance mainly by increasing amino acid biosynthesis and glycolysis under water-deficit conditions. Our findings suggest that drought condition altered metabolic networks including transamination, the tricarboxylic cycle, gluconeogenesis/glycolysis, glutamate-mediated proline biosynthesis, and the metabolisms of choline, pyrimidine and purine. This study provides new insights into the metabolic adaptation of wheat to drought stress and important information for developing drought-tolerant wheat cultivars.     

拔节孕穗期小麦干旱胁迫下生长代谢变化规律

采用盆栽试验模拟干旱胁迫(土壤相对含水量40%-45%)在小麦(Triticum aestivum)拔节孕穗期胁迫12天, 测定其生长速率、光合特征及关键代谢产物含量, 以探讨干旱胁迫对拔节孕穗期小麦叶片初生及次生代谢产物的影响及其涉及的代谢途径, 讨论小麦生长代谢变化规律及应答机制。研究表明: 干旱胁迫使小麦叶片气孔受限制导致光合速率下降; 使叶绿素含量下降直接影响光系统II活性, 最终导致生长率降低。检测出的初级代谢产物组包括有机酸、氨基酸、碳水化合物、嘧啶和嘌呤等64个代谢产物, 其中29个代谢产物在干旱胁迫下发生明显的变化。主成分分析(PCA)结果显示全部样本均分布在95%的置信区间内, 两个主成分得分为64%。单因素方差分析结果表明, 干旱胁迫导致苹果酸、柠檬酸、乌头酸等参与三羧酸(TCA)循环的代谢产物消耗明显, 且引起大部分氨基酸(如脯氨酸、丝氨酸、缬氨酸)和碳水化合物(肌醇、果糖、葡萄糖)大量积累的同时转氨基代谢(天冬酰胺、谷氨酰胺和γ氨基丁酸)产物消耗, 研究证明干旱胁迫明显地促进小麦叶片的糖酵解和氨基酸合成途径, 但抑制了TCA循环和转氨基反应, 加速氨基酸代谢网络向脯氨酸合成转变过程。这些结果表明干旱胁迫引起了转氨基反应、TCA循环、糖酵解/糖异生、谷氨酸介导的脯氨酸合成, 以及嘧啶和嘌呤等代谢网络系统广泛的变化, 说明小麦在合成大量的氨基酸和碳水化合物类物质的同时也消耗了大量的能量, 暗示了糖异生到脯氨酸合成的转变。     

盐胁迫环境下不同抗盐性小麦品种幼苗长势和内源激素的变化

采用两种浓度NaCl溶液,对不同抗盐性小麦品种德抗961(抗盐性强)和泰山9818(抗盐性弱)萌发期幼苗进行胁迫处理,观察其幼苗长势和内源激素含量变化.结果表明,盐胁迫抑制小麦幼苗生长,抗盐性弱的泰山9818受抑制较重.苗、根ABA含量随盐胁迫浓度增加而提高,泰山9818的增幅高于德抗961.苗、根IAA含量随盐胁迫浓度增加而降低,但德抗961的IAA含量高于泰山9818,说明盐胁迫下抗盐性强的品种具有较高IAA合成量.2品种GA3含量变化因盐胁迫浓度而异.在低盐胁迫下抗盐性强的品种苗中GA3含量提高以适应盐胁迫利于苗的生长,在高盐胁迫下2品种GA3含量降低.盐胁迫使苗中ZR含量增加,且德抗961的苗中ZR含量高于泰山9818,而根中ZR含量则前者低,说明盐胁迫下抗盐性强的品种可迅速将根部合成的ZR向苗中转移,促进苗的生长.2品种IAA/ABA、GA3/ABA比值随盐胁迫浓度增加和时间延长而下降,德抗961 IAA/ABA比值大于泰山9818.在盐胁迫下,抗盐性强的品种协调自身激素平衡的能力较强可能是其生长受抑制较小的重要原因.     

Membrane permeability and micro- and macroelement accumulation in spring wheat cultivars during the short-term effect of salinity- and PEG-induced water stress

The comparative responses of ten spring wheat cultivars to water stress were investigated. Wheat plants were cultured under hydroponics conditions (Hoagland nutrient) to the stage of three-leaf seedlings. Then, the water medium was supplemented with PEG (drought) or NaCl (salinity) to obtain a water status equal to −1.5 MPa. After a 2-day treatment, the changes in the following parameters were determined: fresh and dry weight, macro- and microelement accumulation, membrane injury (electrolyte leakage, lipid peroxidation) and fatty acid content of the phospholipid fraction of plasmalemma (in comparison to plants not stressed, taken as a control). Generally, the plants were more significantly influenced by water stress stimulated by PEG than by NaCl treatment, as compared to the plants cultivated in the control media. The results of the decrease in water content in leaves and electrolyte leakage from cells corresponded well with the intensity of lipid peroxidation (determined by malondialdehyde—MDA-content) and were chosen for the selection of investigated genotypes for tolerance to both stresses. The more tolerant genotypes exhibited the opposite changes in phospholipid fatty acid unsaturation for two applied stresses i.e. NaCl treatment caused a decrease in unsaturation whereas in PEG-treated plants an increase in unsaturation was observed. These changes were reversed for less tolerant plants, i.e. NaCl treatment influenced an increase in fatty acid unsaturation whereas in PEG-treated plants a decrease in unsaturation was measured. The ratio of U/S (unsaturated to saturated fatty acids) correlated with the total amount of accumulated macroelements. The content of Mg, Ca and S in leaves of plants undergoing both stress factors (NaCl and PEG) dropped whereas the K and P content increased in leaves of wheat seedlings cultured on media containing NaCl only. For microelements, a decrease in the accumulation of these nutrients was detected in all investigated seedlings. However, a greater reduction in the level of these elements occurred in seedlings grown on media with PEG in comparison to those grown on NaCl containing media.     

抗旱性不同的冬小麦幼苗对渗透胁迫的生理效应

抗旱性不同的冬小麦幼苗在渗进胁迫下叶片相对透性、Pro含量和在总游离氨基酸中的Pro比例均增加,但品种间变化幅度有明显差别。在严重胁迫下,抗旱性弱的品种济南13,烟农15和鲁麦5号的叶片相对透性增加大于抗旱性强的品种昌乐5号,秦麦3号和山农587;而Pro含量和在总游离氨基酸中Pro比例的增加小于抗旱性强的品种、叶片Pro累积与RWC呈显著负相关。     

抗旱性不同的冬小麦幼苗对渗透胁迫的生理反应

抗旱性不同的冬小麦幼苗在渗透胁迫下叶片相对透性、Pro 含量和在总游离氨基酸中的Pro 比例均增加,但品种间变化幅度有明显差别.在严重胁迫下,抗旱性弱的品种济南13.烟农15和鲁麦5号的叶片相对透性增加大于抗旱性强的品种昌乐5号、秦麦3号和山农587;而Pro 含量和在总游离氨基酸中Pro 比例的增加小于抗旱性强的品种.叶片Pro 累积与RWC 呈显著负相关.     

接种两种固氮菌增强小麦幼苗抗渗透胁迫及生长能力

苗期是小麦(Triticum aestivum)物质和能量积累的关键时期, 苗期干旱影响小麦的后期群体建成。利用田菁茎瘤固氮根瘤菌(Azorhizobium caulinodans) ‘ORS571’与巴西固氮螺菌(Azospirillum brasilense) ‘Yu62’浸种侵染小麦和聚乙二醇(PEG)模拟渗透胁迫, 研究渗透胁迫下接菌小麦种子的发芽状况; 利用固氮菌涂抹小麦幼苗叶部, 测定PEG模拟渗透胁迫下小麦幼苗根体积、叶片相对含水量、脯氨酸含量及可溶性蛋白含量, 探究固氮菌增强小麦幼苗抗渗透胁迫的能力。结果表明, 接种混合固氮菌后在渗透胁迫下小麦种子的发芽率明显提高; 在渗透胁迫下叶部涂抹固氮菌小麦的根体积、叶片相对含水量、脯氨酸含量及可溶性蛋白含量明显升高, 表明接种固氮菌可提高小麦幼苗的抗渗透胁迫能力, 且混合固氮菌对小麦幼苗叶片的增强效果优于单一固氮菌。     

An integrative proteome analysis of different seedling organs in tolerant and sensitive wheat cultivars under drought stress and recovery

Roots, leaves, and intermediate sections between roots and leaves (ISRL) of wheat seedlings show different physiological functions at the protein level. We performed the first integrative proteomic analysis of different tissues of the drought‐tolerant wheat cultivar Hanxuan 10 (HX‐10) and drought‐sensitive cultivar Chinese Spring (CS) during a simulated drought and recovery. Differentially expressed proteins (DEPs) in the roots (122), ISRLs (146), and leaves (163) showed significant changes in expression in response to drought stress and recovery. Numerous DEPs associated with cell defense and detoxifications were significantly regulated in roots and ISRLs, while in leaves, DEPs related to photosynthesis showed significant changes in expression. A significantly larger number of DEPs related to stress defense were upregulated in HX‐10 than in CS. Expression of six HSPs potentially related to drought tolerance was significantly upregulated under drought conditions, and these proteins were involved in a complex protein–protein interaction network. Further phosphorylation analysis showed that the phosphorylation levels of HSP60, HSP90, and HOP were upregulated in HX‐10 under drought stress. We present an overview of metabolic pathways in wheat seedlings based on abscisic acid signaling and important protein expression patterns.     

Metabolic responses of wheat roots to alkaline stress

Aims The aim of this study was to investigate the effects of alkaline stress on primary, secondary metabolites and metabolic pathways in the roots of wheat (Triticum aestivum). The results were used to evaluate the physiological adaptive mechanisms by which wheat tolerated alkali stress.Methods A pot experiment was carried out in the greenhouse. For each plastic pot, five wheat seeds were planted. After germination, seedlings were allowed to grow under controlled water and nutrient conditions for two months, then seedlings were exposed to alkaline stress (NaHCO₃-Na₂CO₃) for 12 days. The relative growth rate (RGR), absolute water content (AWC), metal elements, free cations and metabolites were measured.Important findings The alkaline stress caused the reduction of RGR and AWC. Alkaline stress caused a rapid increase of Na content with the concurrent decrease in K and Cl content, resulting in inhibited metal element accumulation and an ionic imbalance. In the present study, alkaline stress strongly enhanced Ca accumulation in wheat roots, suggesting that an increased Ca concentration can immediately trigger the salt overly sensitive (SOS)-Na exclusion system and reduce Na-associated injuries. Also, 70 metabolites, including organic acids, amino acids, sugars/polyols and others, behaved differently in the alkaline stress treatments according to a GC-MS analysis. The metabolic profiles of wheat were closely associated with alkaline-stress conditions. Alkaline stress caused the accumulation of organic acids, accompanied by the depletion of sugars/polyols and amino acids. Organic acids could play a central role in the regulation of intracellular pH by accumulating vacuoles to neutralize excess cations. Glycolysis and amino acid synthesis in roots were inhibited under salt stress while prolonged alkaline stress led to a progressive tricarboxylic acid (TCA) cycle. The severe negative effects of alkaline stress on sugar synthesis and storage may reflect the toxic levels of Na⁺ accumulating in plant cells in a high-pH environment, implying that the reactive oxygen species detoxification capacity was diminished by the high pH. A lack of NO₃^- in wheat roots can decrease synthase enzyme activities, limiting the synthesis of amino acids. Under salt stress, the TCA cycle and organic acid accumulation increased, but glycolysis and amino acid synthesis were inhibited in roots. Thus, energy levels and high concentrations of organic acids may be the key adaptive mechanisms by which wheat seedlings maintain their intracellular ion balance under alkaline stress.     

Impact of osmotic stress on physiological and biochemical characteristics in drought-susceptible and drought-resistant wheat genotypes

In this study, the seedlings of two wheat cultivars were used: drought-resistant Chinese Spring (CS) and drought-susceptible (SQ1). Seedlings were subjected to osmotic stress in order to assess the differences in response to drought stress between resistant and susceptible genotype. The aim of the experiment was to evaluate the changes in physiological and biochemical characteristics and to establish the optimum osmotic stress level in which differences in drought resistance between the genotypes could be revealed. Plants were subjected to osmotic stress by supplementing the root medium with three concentrations of PEG 6000. Seedlings were grown for 21 days in control conditions and then the plants were subjected to osmotic stress for 7 days by supplementing the root medium with three concentrations of PEG 6000 (D1, D2, D3) applied in two steps: during the first 3 days of treatment ?0.50, ?0.75 and ?1.00 and next ?0.75, ?1.25 and ?1.5 MPa, respectively. Measurements of gas exchange parameters, chlorophyll content, height of seedlings, length of root, leaf and root water content, leaf osmotic potential, lipid peroxidation, and contents of soluble carbohydrates and proline were taken. The results highlighted statistically significant differences in most traits for treatment D2 and emphasized that these conditions were optimum for expressing differences in the responses to osmotic stress between SQ1 and CS wheat genotypes. The level of osmotic stress defined in this study as most suitable for differentiating drought resistance of wheat genotypes will be used in further research for genetic characterization of this trait in wheat through QTL analysis of mapping population of doubled haploid lines derived from CS and SQ1.     

Plant growth, metabolism and adaptation in relation to stress conditions. XXVII. Can ascorbic acid modify the adverse effects of NaCl and mannitol on amino acids, nucleic acids and protein patterns in Vicia faba seedlings?

The adverse effects of either NaCl or mannitol on amino acids, protein patterns and nucleic acids in Vicia faba seeds were investigated. The exogenous addition of 4 mM ascorbic acid to the stressing media in which the broad bean seeds were germinated in combination with either the ionic (NaCl) or osmotic (mannitol) stressor induced significant protective changes in the total amount and in the relative composition of amino acids in general and in proline, glycine, glutamic, aspartic, alanine and serine in particular. It also induced changes in nucleic acids (RNA and DNA) content. These changes occurred throughout the entire period of the experiments (12 days). Separate administration of NaCl or mannitol enhanced the occurrence of particular novel proteins that were not detected in control bean seeds (water medium). Protein banding patterns of broad bean seedlings treated with NaCl or mannitol in combination with 4 mM ascorbic acid showed different de novo protein bands, with different molecular weights, at different stages of seedlings growth, with lower levels or a nearly complete absence of the major stress proteins. The pattern of changes for amino acids and nucleic acids and the range of protein bands extracted from the variously treated broad bean seedlings indicate a positive role of ascorbic acid in the alleviation of the damage effects induced by NaCl and mannitol. The importance of this role in the stress tolerance of broad beans is discussed.     

The Positive Correlations Between the Activity of Superoxide Dismutase and Dehydration Tolerance in Wheat Seedlings

Damage to crops by drought is still a serious problem in large areas of the world. Considerable research has been undertaken to discover the mechanisms of drought injury and drought resistance of plants. However, the critical features of drought injury have not yet been identified. In the past ten years a free radical hypothesis has been suggested to account for subcellular damage caused by severe environments. Superoxide (oxygen radical) is normally produced in hydrated tissues. It is controlled by free radical scavenging reactions. One such scavenger is the enzyme superoxide dismutase (SOD). Under water stress, production of excess free radicals may occur in dehydrated plant tissues and this probably damages the membranes by causing peroxidation of the lipid components. So far few studies have been done to determine if drought injury is correlated with the free radical mechanism. In the present study, the SOD activities in wheat seedlings under water stress have been investigated by measuring the photoreduction of nitro blue tetrazolium using a spectrometric method. Meanwhile, the viabilities of wheat seedlings during drying were followed by tetrazolium test. These results provided information on the relationship between SOD activity and the dehydration tolerance of the plant. Results indicated that SOD activity changed with the time after germination. The activity of SOD of 24 h seedlings was 1.9 times higher than those of 72 h seedlings based on fresh weight. SOD activity in shoot was also higher than in root. These results were consistent with the results obtained from rating of the viabilities of seedlings during drying. The 24h seedlings were more tolerant of dehydration than 72 h seedlings and root were more sensitive of drought than shoot. In addition, shoot and root tips showed the higher SOD activities than non-tip region and they also showed a higher survival ability upon dehydration. In dehydration and subsequent rehydration, SOD activity, different from many other enzymes in plants, increased rather than declined during drying. After rehydration SOD activity returned to nearly the original level. Therefore, the positive correlations were found to exist between SOD activity and dehydration tolerance. It is reasonable to suggest that SOD enzyme may play a protective role against damage caused by free radicals which may be produced excessively during dehydration in wheat seedling.     

Effects of drought stress and selenium supply on growth and physiological characteristics of wheat seedlings

The paper studied the effects of drought stress, selenium (Se) supply and their combination on growth and physiological characteristics of wheat (Triticum aestivum L. cv Shijiazhuang NO. 8) seedlings. The experimental design included two water treatments (well-watered, 75% of maximum field capacity; drought stress, 30% of maximum field capacity) and two Se levels (0; 0.5 mg/kg) to determine whether Se can modify the negative impacts of drought stress on seedling growth and physiological traits. Drought stress caused a marked decline in growth parameters and soluble protein content, whereas it induced an increase in root activity, proline content and the activities of peroxidase (POD) and catalase (CAT) of leaf tissue. On the other hand, Se supply induced an increase in biomass accumulation only under well-watered condition. Under drought stress, Se supply increased free proline content, root activity and the activities of POD and CAT in leaf tissue, but did not significantly affect on growth parameters. These results implied that drought stress brought harmful effects on wheat seedlings, and that Se supply was favorable for biomass accumulation of wheat seedlings under well-watered condition. However, it did not significantly affect on biomass accumulation under drought stress, although it increased root activity and activities of some antioxidant index in experimental periods.     

The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat

It emerged recently that there is an inter‐relationship between drought and ultraviolet‐B (UV‐B) radiation in plant responses, in that both stresses provoke an oxidative burst. The purpose of this investigation was to compare the effects and interaction of drought and UV‐B in wheat and pea. The absence of changes in relative leaf water content (RWC) after UV‐B treatments indicate that changes in water content were not involved. RWC was the main factor resulting in reduced growth in response to drought. Increases in anthocyanin and phenols were detected after exposure to UV‐B. The increases do not appear to be of sufficient magnitude to act as a UV‐B screen. UV‐B application caused greater membrane damage than drought stress, as assessed by lipid peroxidation as well as osmolyte leakage. An increase in the specific activities of antioxidant enzymes was measured after UV‐B alone as well as after application to droughted plants. Proline increased primarily in drought‐stressed pea or wheat. Proline may be the drought‐induced factor which has a protective role in response to UV‐B. The physiological and biochemical parameters measured indicate the UV‐B light has stronger stress effectors than drought on the growth of seedlings of both species. The two environmental stresses acted synergistically to induce protective mechanisms in that pre‐application of either stress reduced the damage caused by subsequent application of the other stress.     

Wheat (Triticum aestivum L.) growth enhancement by Azospirillum sp. under drought stress

Plant growth promoting rhizobacteria (PGPR) can enhance plant growth by alleviating soil stresses. Although previously investigated, some new interesting details are presented regarding the alleviating affects of Azospirillum sp. on wheat growth under drought stress in this research work. We hypothesized that the isolated strains of Azospirillum sp. may alleviate the adverse effects of drought stress on wheat (Triticum aestivum L.) growth. Three different strains of Azospirillum lipoferum (B1, B2 and B3) were used to inoculate wheat seedlings under drought. During the flowering stage the seedlings were subjected to three drought levels with five different time longevity, including control. Pots were water stressed at 80% (S0), 50% (S1) and 25% (S2) of field capacity moisture in a 25 day-period. Soil and plant water properties including water potential and water content, along with their effects on bacterial inoculum and wheat growth, were completely monitored during the experiment. While stress intensity significantly affected bacterial population and wheat growth, stress longevity only affected wheat water potential and water content. Compared to uninoculated treatments strain B3 (fixing and producing the highest amounts of N and auxin, respectively, with P solubilizing and ACC-deaminase activities) increased wheat yield at S1 and S2 by 43 and 109%, respectively. However, strain B2 (producing siderophore) was the most resistant strain under drought stress. The results of this experiment may elucidate the more efficient strains of Azospirillum sp. for wheat inoculation under drought stress and the mechanisms by which they alleviate the stress.     

INFLUENCE OF He-Ne LASER IRRADIATION ON PROTECTIVE ENZYME ACTIVITIES AND LIPID PEROXIDATION IN WHEAT SEEDLINGS BY DROUGHT STRESS DAMAGE

 用He-Ne激光(5.23 mW·mm^–2)处理经5%、10%、15% PEG6000胁迫的小麦幼苗, 分析了干旱胁迫条件下激光处理对小麦幼苗保护酶活性及脂质过氧化作用的影响。适度干旱胁迫的小麦幼苗经He-Ne激光辐照后, 丙二醛(MDA)含量和超氧自由基(O₂^–.)产生速率显著降低(p<0.05), 而过氧化物酶(POD)活性和抗坏血酸(AsA)、谷胱甘肽(GSH)含量却显著增加(p<0.05)。总体上看, 5%和10% PEG6000胁迫的小麦幼苗经激光辐照3 min后抗旱性增强。     

Influence of water stress on endogenous hormone contents and cell damage of maize seedlings

Phytohormones play critical roles In regulating plant responses to stress. We Investigated the effects of water stress Induced by adding 12% (w/v) polyethylene glycol to the root medium on the levels of abscisic acid (ABA), indole-3-acid (IAA), zeatin (ZT), and gibberellin3 (GA3) in maize leaves. The results suggested that water stress had significant effects on the four hormone levels. There was a transient increase in the IAA content during the initial stage of adaptation to water stress in maize leaves, but it dropped sharply thereafter in response to water stress. ABA content increased dramatically in maize leaves after 24 h of exposure to water stress, and then the high levels of ABA were maintained to the end, The contents Of ZT and GA3 rapidly declined in maize leaves subjected to water stress. The effects of water stress on chlorophyll content, electrolyte leakage and malondialdehyde levels in maize leaves were also studied. The variation of cell damage was negatively correlated with ZT and GA3 levels in maize leaves under water stress. Thus, we explored the roles of ZT and GA3 on the growth of maize seedlings under water stress by exogenous application. It is possible that both ZT and GA3 were effective in protecting maize seedlings from water stress, which would be of great importance for the improvement of drought tolerance in maize by genetic manipulation.