Differential metabolic responses of perennial grass Cynodon transvaalensis×Cynodon dactylon (C₄) and Poa Pratensis (C₃) to heat stress

Differential metabolic responses to heat stress may be associated with variations in heat tolerance between cool‐season (C₃) and warm‐season (C₄) perennial grass species. The main objective of this study was to identify metabolites associated with differential heat tolerance between C₄ bermudagrass and C₃ Kentucky bluegrass by performing metabolite profile analysis using gas chromatography‐mass spectrometry. Plants of Kentucky bluegrass (Poa Pratensis‘Midnight’) and hybrid bermudagrass (Cynodon transvaalensis×Cynodon dactylon‘Tifdwarf’) were grown under optimum temperature conditions (20/15°C for Kentucky bluegrass and 30/25°C for bermudagrass) or heat stress (35/30°C for Kentucky bluegrass and 45/40°C for bermudagrass). Physiological responses to heat stress were evaluated by visual rating of grass quality, measuring photochemical efficiency (variable fluorescence to maximal fluorescence) and electrolyte leakage. All of these parameters indicated that bermudagrass exhibited better heat tolerance than Kentucky bluegrass. The metabolite analysis of leaf polar extracts revealed 36 heat‐responsive metabolites identified in both grass species, mainly consisting of organic acids, amino acids, sugars and sugar alcohols. Most metabolites showed higher accumulation in bermudagrass compared with Kentucky bluegrass, especially following long‐term (18 days) heat stress. The differentially accumulated metabolites included seven sugars (sucrose, fructose, galactose, floridoside, melibiose, maltose and xylose), a sugar alcohol (inositol), six organic acids (malic acid, citric acid, threonic acid, galacturonic acid, isocitric acid and methyl malonic acid) and nine amino acids (Asn, Ala, Val, Thr, γ‐Aminobutyric acid, IIe, Gly, Lys and Met). The differential accumulation of those metabolites could be associated with the differential heat tolerance between C₃ Kentucky bluegrass and C₄ bermudagrass.     

Comparative proteomic and metabolomic analyses reveal mechanisms of improved cold stress tolerance in bermudagrass (Cynodon dactylon (L.) Pers.) by exogenous calcium

As an important second messenger, calcium is involved in plant cold stress response, including chilling (<20 °C) and freezing (<0 °C). In this study, exogenous application of calcium chloride (CaCl₂) improved both chilling and freezing stress tolerances, while ethylene glycol‐bis‐(β‐aminoethyl) ether‐N,N,N,N‐tetraacetic acid (EGTA) reversed CaCl₂ effects in bermudagrass (Cynodon dactylon (L.) Pers.). Physiological analyses showed that CaCl₂ treatment alleviated the reactive oxygen species (ROS) burst and cell damage triggered by chilling stress, via activating antioxidant enzymes, non‐enzymatic glutathione antioxidant pool, while EGTA treatment had the opposite effects. Additionally, comparative proteomic analysis identified 51 differentially expressed proteins that were enriched in redox, tricarboxylicacid cycle, glycolysis, photosynthesis, oxidative pentose phosphate pathway, and amino acid metabolisms. Consistently, 42 metabolites including amino acids, organic acids, sugars, and sugar alcohols were regulated by CaCl₂ treatment under control and cold stress conditions, further confirming the common modulation of CaCl₂ treatment in carbon metabolites and amino acid metabolism. Taken together, this study reported first evidence of the essential and protective roles of endogenous and exogenous calcium in bermudagrass response to cold stress, partially via activation of the antioxidants and modulation of several differentially expressed proteins and metabolic homeostasis in the process of cold acclimation.     

Effects of Cadmium Exposure on Growth and Metabolic Profile of Bermudagrass [Cynodon dactylon (L.) Pers.]

Metabolic responses to cadmium (Cd) may be associated with variations in Cd tolerance in plants. The objectives of this study were to examine changes in metabolic profiles in bermudagrass in response to Cd stress and to identify predominant metabolites associated with differential Cd tolerance using gas chromatography-mass spectrometry. Two genotypes of bermudagrass with contrasting Cd tolerance were exposed to 0 and 1.5 mM CdSO₄ for 14 days in hydroponics. Physiological responses to Cd were evaluated by determining turf quality, growth rate, chlorophyll content and normalized relative transpiration. All these parameters exhibited higher tolerance in WB242 than in WB144. Cd treated WB144 transported more Cd to the shoot than in WB242. The metabolite analysis of leaf polar extracts revealed 39 Cd responsive metabolites in both genotypes, mainly consisting of amino acids, organic acids, sugars, fatty acids and others. A difference in the metabolic profiles was observed between the two bermudagrass genotypes exposed to Cd stress. Seven amino acids (norvaline, glycine, proline, serine, threonine, glutamic acid and gulonic acid), four organic acids (glyceric acid, oxoglutaric acid, citric acid and malic acid,) and three sugars (xylulose, galactose and talose) accumulated more in WB242 than WB144. However, compared to the control, WB144 accumulated higher quantities of sugars than WB242 in the Cd regime. The differential accumulation of these metabolites could be associated with the differential Cd tolerance in bermudagrass.     

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.     

Comparative effects of salt and alkali stresses on growth, osmotic adjustment and ionic balance of an alkali-resistant halophyte Suaeda glauca (Bge.)

Effects of salt and alkali stresses on growth, osmotic adjustment and ionic balance of Suaeda glauca (Bge.), an alkali-resistant succulent halophyte, were compared. The results showed that alkali stress clearly inhibited the growth of S. glauca. Moreover, the concentrations of Na⁺ and K⁺ both increased with increasing salinity under both stresses, suggesting no competitive inhibition between absorptions of Na⁺ and K⁺. The mechanism underlying osmotic adjustment during salt stress was similar to alkali stress in shoots. The shared essential features were that organic acids, betaine and inorganic ions (dominated by Na⁺) mostly accumulated. On the other hand, the mechanisms governing ionic balance under both stresses were different. Under salt stress, S. glauca accumulated organic acids and inorganic anions to maintain the intracellular ionic equilibrium, but the anion contribution of inorganic ions was greater than that of organic acids. However, the concentrations of inorganic anions under alkali stress were significantly lower than those under salt stress of the same intensity, suggesting that alkali stress might inhibit uptake of anions, such as NO₃ ^– and H₂PO₄ ^–. Under alkali stress, organic acids were the dominant factor in maintaining ionic equilibrium. The contribution of organic acids to anions was 74.1%, while that of inorganic anions was only 25.9%. S. glauca enhanced the synthesis of organic acids, dominated by oxalic acid, to compensate for the shortage of inorganic anions.     

Warming and drought differentially influence the production and resorption of elemental and metabolic nitrogen pools in Quercus rubra

The process of nutrient retranslocation from plant leaves during senescence subsequently affects both plant growth and soil nutrient cycling; changes in either of these could potentially feed back to climate change. Although elemental nutrient resorption has been shown to respond modestly to temperature and precipitation, we know remarkably little about the influence of increasing intensities of drought and warming on the resorption of different classes of plant metabolites. We studied the effect of warming and altered precipitation on the production and resorption of metabolites in Quercus rubra. The combination of warming and drought produced a higher abundance of compounds that can help to mitigate climatic stress by functioning as osmoregulators and antioxidants, including important intermediaries of the tricarboxylic acid (TCA) cycle, amino acids including proline and citrulline, and polyamines such as putrescine. Resorption efficiencies (REs) of extractable metabolites surprisingly had opposite responses to drought and warming; drought treatments generally increased RE of metabolites compared to ambient and wet treatments, while warming decreased RE. However, RE of total N differed markedly from that of extractable metabolites such as amino acids; for instance, droughted plants resorbed a smaller fraction of elemental N from their leaves than plants exposed to the ambient control. In contrast, plants in drought treatment resorbed amino acids more efficiently (>90%) than those in ambient (65–77%) or wet (42–58%) treatments. Across the climate treatments, the RE of elemental N correlated negatively with tissue tannin concentration, indicating that polyphenols produced in leaves under climatic stress could interfere with N resorption. Thus, senesced leaves from drier conditions might have a lower nutritive value to soil heterotrophs during the initial stages of litter decomposition despite a higher elemental N content of these tissues. Our results suggest that N resorption may be controlled not only by plant demand, but also by climatic influences on the production and resorption of plant metabolites. As climate–carbon models incorporate increasingly sophisticated nutrient cycles, these results highlight the need to adequately understand plant physiological responses to climatic variables.     

Combined effects of CO2 enrichment,diurnal light levels and water stress on foliar metabolites of potato plants grown in naturally sunlit controlled environment chambers

Experiments were conducted in outdoor, naturally sunlit, soil–plant–atmosphere research (SPAR) chambers using plants grown in pots. Drought treatments were imposed on potato plants (Solanum tuberosum cv. Kennebec) beginning 10 days after tuber initiation. A total of 23 out of 37 foliar metabolites were affected by drought when measured 11 days after initiating water stress treatments. Compounds that accumulated in response to drought were hexoses, polyols, branched chain amino acids (BCAAs) and aromatic amino acids, such as proline. Conversely, leaf starch, alanine, aspartate and several organic acids involved in respiratory metabolism decreased with drought. Depending upon harvest date, a maximum of 12 and 17 foliar metabolites also responded to either CO₂ enrichment or diurnal treatments, respectively. In addition, about 20% of the measured metabolites in potato leaflets were simultaneously affected by drought, CO₂ enrichment and diurnal factors combined. This group contained BCAAs, hexoses, leaf starch and malate. Polyols and proline accumulated in response to water stress but did not vary diurnally. Water stress also amplified diurnal variations of hexoses and starch in comparison to control samples. Consequently, specific drought responsive metabolites in potato leaflets were dramatically affected by daily changes of photosynthetic carbon metabolism.     

Redox regulation of free amino acid levels in Arabidopsis thaliana

Abiotic stresses induce oxidative stress, which modifies the level of several metabolites including amino acids. The redox control of free amino acid profile was monitored in wild‐type and ascorbate or glutathione deficient mutant Arabidopsis thaliana plants before and after hydroponic treatment with various redox agents. Both mutations and treatments modified the size and redox state of the ascorbate (AsA) and/or glutathione (GSH) pools. The total free amino acid content was increased by AsA, GSH and H₂O₂ in all three genotypes and a very large (threefold) increase was observed in the GSH‐deficient pad2‐1 mutant after GSH treatment compared with the untreated wild‐type plants. Addition of GSH reduced the ratio of amino acids belonging to the glutamate family on a large scale and increased the relative amount of non‐proteinogenic amino acids. The latter change was because of the large increase in the content of alpha‐aminoadipate, an inhibitor of glutamatic acid (Glu) transport. Most of the treatments increased the proline (Pro) content, which effect was due to the activation of genes involved in Pro synthesis. Although all studied redox compounds influenced the amount of free amino acids and a mostly positive, very close (r > 0.9) correlation exists between these parameters, a special regulatory role of GSH could be presumed due to its more powerful effect. This may originate from the thiol/disulphide conversion or (de)glutathionylation of enzymes participating in the amino acid metabolism.     

The role of amino acid metabolism during abiotic stress release

Plant responses to abiotic stress include various modifications in amino acid metabolism. By using a hydroponic culture system, we systematically investigate modification in amino acid profiles and the proteome of Arabidopsis thaliana leaves during initial recovery from low water potential or high salinity. Both treatments elicited oxidative stress leading to a biphasic stress response during recovery. Degradation of highly abundant proteins such as subunits of photosystems and ribosomes contributed to an accumulation of free amino acids. Catabolic pathways for several low abundant amino acids were induced indicating their usage as an alternative respiratory substrate to compensate for the decreased photosynthesis. Our results demonstrate that rapid detoxification of potentially detrimental amino acids such as Lys is a priority during the initial stress recovery period. The content of Pro, which acts as a compatible osmolyte during stress, was adjusted by balancing its synthesis and catabolism both of which were induced both during and after stress treatments. The production of amino acid derived secondary metabolites was up‐regulated specifically during the recovery period, and our dataset also indicates increased synthesis rates of the precursor amino acids. Overall, our results support a tight relationship between amino acid metabolism and stress responses.     

Studies on Brevibacterium linens metabolism in fermentor

Summary Growth and metabolism of Brevibacterium linens were studied in a fermentor regulated for fixed levels of pH (7.5 to 8.5), temperature (20–30° C) and dissolved oxygen (40%–60% of air saturated medium). The curves of disappearance of l-lactate and amino acids were invariable, indicating that phenylalanine, tyrosine, arginine, proline, glutamic acid and histidine are growth-limiting nutrients. Ornithine appeared at the beginning of cultures when oxygen consumption was low. Ammonia was produced, but large quantities were observed only when amino acid concentrations were higher than that of the carbon source. When the latter was low, the ammonia produced was consumed before a number of amino acids as an easily assimilable nitrogen source. Whether alkali or acid was consumed to maintain constant pH depended on the pH of the medium and on maximal growth rates.     

Plant-mediated effects of water-deficit stress on the performance of the jujube lace bug,Monosteira alticarinata Ghauri (Hemiptera: Tingidae) on Jujube tree

Drought phenomenon as a consequence of climate change may become a major limiting factor for agricultural systems, and its impacts on plant-insect interactions are only partially understood. Addressing this issue, life-history responses of the jujube lace bug were evaluated on jujube tree under three water treatments (control, moderate and severe drought stress). Performance responses of the jujube lace bug were evaluated under controlled conditions using arenas consisting of jujube leaves inside Petri dishes. Plant biochemical measurements showed that leaf chlorophyll a, b, and total chlorophyll content, were higher in moderate water stress treatment than other treatments while Total Soluble Solids, total carbohydrate, and proline concentrations were increased in drought stress. Also, the lowest amount of carotenoid, phosphorous, sodium content was measured in severe drought stress treatment. The life history responses of Monosteira alticarinata showed that mean oviposition period and life span were statistically higher on moderate water stress than other treatments. The lowest intrinsic rate of increase (r), the finite rate of increase (λ), the net reproductive rate (R₀), gross rate reproduction (GRR), total fecundity, and the highest doubling time (DT) were measured on severely drought stress treatment. An increase on the performance of the lace bug can be due to an increase on the chlorophyll, nitrogen, protein and free amino acids in moderate water stressed plants. However in severe stress it do worse. Overall, this study revealed that sap feeders benefit from moderate water stress.     

Protein and free amino acids in field-grown cowpea seeds as affected by water stress at various growth stages

Summary This study was undertaken to evaluate water stress effects during vegetative, flowering, and podfilling stages of cowpea plants (Vigna unguiculata L.) grown under natural field conditions in southern California on seed yield and protein and free amino acid content of the cowpea seeds. The lowest concentration of N was found in the seeds of the control treatment plants while the seed yield from these treatments was the highest as compared with the N concentration and yield of seeds from plants subjected to water stress during flowering and podfilling stages. The concentration of N in the seeds was inversely related to the seed dry weight yield. Protein arginine,-threonine,-serine,-cystine,-valine,-methionine, and-isoleucine were significantly affected by water stress at the three growth stages. There was no consistent pattern in the effect of water stress on the individual amino acids. The sum of protein amino acids in the cowpea seeds was not significantly influenced by the various treatments since some of the protein amino acids increased and others decreased producing an averaging effect on the figures comprising the sums of the amino acids. Water stress during the flowering and pod-filling stages increased the free amino acid pool, and at the same time, inhibited incorporation of the amino acids into the protein chain-thus lowering the protein amino acid fraction simultaneously. With the exception of methionine plus cystine, the essential amino acids in the seeds were present at concentrations equal to or greater than recommended by the World Health Organization and FAO. It is of particular importance to note that the concentration of lysine in the cowpeas was substantially higher than that found in wheat grain. It is also important to note that the amount of essential amino acids per gram of protein was not measurably affected by the water stress treatments during any of the growth stages.     

Vegetative growth, aging- and light-induced conidiation ofTrichoderma viride cultivated on different carbon sources

The growth and conidiation of the agedTrichoderma viride culture grown in the dark, and after an induction by a light pulse. was examined in the presence of selected mono-, di(tri)saccharides, amino acids and alcohols as sole carbon sources. Hexoses and disaccharides, but not pentoses and amino acids, promoted proportionally both growth and conidiation induced by aging or light. All compounds but pentoses promoted the conidiation in aged cultures and photoconidiation in a close correlation. Ethanol, glycerol and ethylene glycol supported both growth and conidiation but these processes were not supported equally. Conidia formation with hexoses and amino acids as sole carbon sources seems to be a function of growth promotion, rather than of growth restriction (starvation, stress, aging). With glucose as sole carbon source the conidiation was not triggered by nutrient limitation, nor by the accumulation of waste metabolites. The aging-induced conidiation can be considered to be triggered by the genetic program of the microorganism rather than by its nutrient status.     

A molecular approach to drought‐induced reduction in leaf CO2 exchange in drought‐resistant Quercus ilex

Drought‐induced reduction of leaf gas exchange entails a complex regulation of the plant leaf metabolism. We used a combined molecular and physiological approach to understand leaf photosynthetic and respiratory responses of 2‐year‐old Quercus ilex seedlings to drought. Mild drought stress resulted in glucose accumulation while net photosynthetic CO₂ uptake (P_n) remained unchanged, suggesting a role of glucose in stress signaling and/or osmoregulation. Simple sugars and sugar alcohols increased throughout moderate‐to‐very severe drought stress conditions, in parallel to a progressive decline in P_n and the quantum efficiency of photosystem II; by contrast, minor changes occurred in respiration rates until drought stress was very severe. At very severe drought stress, 2‐oxoglutarate dehydrogenase complex gene expression significantly decreased, and the abundance of most amino acids dramatically increased, especially that of proline and γ‐aminobutyric acid (GABA) suggesting enhanced protection against oxidative damage and a reorganization of the tricarboxylic cycle acid cycle via the GABA shunt. Altogether, our results point to Q. ilex drought tolerance being linked to signaling and osmoregulation by hexoses during early stages of drought stress, and enhanced protection against oxidative damage by polyols and amino acids under severe drought stress.     

亚麻响应盐、碱胁迫的生理特征

利用中性盐NaCl、Na₂SO₄和碱性盐NaHCO₃、Na₂CO₃混合模拟不同强度的盐、碱胁迫条件, 对亚麻(Linum usitatissimum)进行14天胁迫处理, 测定其地上部分和根生长速率、光合特征、离子平衡及有机渗透调节物质积累, 以探讨亚麻对盐、碱两种胁迫的生理响应特点。研究表明: 亚麻生长对盐、碱胁迫的响应存在差异, 在相同盐浓度下, 碱胁迫对亚麻的伤害大于盐胁迫。碱胁迫使地上部分中Na⁺浓度急剧增高, 造成叶绿体破坏、光合色素含量下降, 光合能力及碳同化能力也急剧下降。亚麻中Na⁺含量随着胁迫强度的增加而升高, 而K⁺含量呈下降趋势, 碱胁迫下的变化明显大于盐胁迫。因此, 碱胁迫导致Na⁺过度积累可能是碱胁迫对植物伤害大于盐胁迫的最主要原因。碱胁迫下Ca²⁺和Mg²⁺在根中下降明显, 可见高pH值阻碍根对Ca²⁺和Mg²⁺的吸收。Fe²⁺和Zn²⁺对渗透调节的影响不大, 因为它们的离子含量较低。盐胁迫促进阴离子(Cl^-、H₂PO₄^-和SO₄^2-)的积累来平衡大量涌入的Na⁺, 但是碱胁迫明显减少无机阴离子含量, 可能造成严重营养胁迫(如P和S不足)。亚麻在盐胁迫下积累大量可溶性糖来平衡大量的Na⁺, 但碱胁迫下积累大量有机酸来维持细胞内离子平衡和pH值稳定, 碱胁迫大量积累的有机酸也可能被分泌到根外调节根外的pH值, 这说明亚麻对两种不同胁迫的响应方式不同。研究证明高pH值会直接影响植物根系的生长发育, 影响植物矿质元素的吸收, 阻碍离子稳态重建, 有机酸代谢是亚麻碱胁迫下的关键适应机制。     

Eco-physiological responses of linseed (Linum usitatissimum) to salt and alkali stresses

AimsEffects of salt and alkali stresses (NaCl-Na₂SO₄ and NaHCO₃-Na₂CO₃) were compared on growth, photosynthesis characters, ionic balance and osmotic adjustment of linseed (Linum usitatissimum), to elucidate the mechanisms of salt and alkali stress (high pH value) damage to plants, and their physiological adaptive mechanisms to the stresses. MethodsThe experiment was carried out in an artificial greenhouse. Plants grew at approximately 700 mmol·m^-2·s^-1 photosynthetic photon flux density (PPFD) in greenhouse under photoperiod of 15 h in light and 9 h in dark. In each plastic pot (17 cm diameter) which contained 2.5 kg of washed sand, 20 linseed seeds were sown. The seedlings were exposed to stresses lasting 14 days after 2 months.Important findingsThe inhibitory effects of alkali stress on linseed growth were more remarkable than those of salt stress, indicating that alkali and salt represent two distinct forms of stress. The alkali stress increased the Na⁺ content in shoots, damaged the photosynthetic system, and highly reduced the net photosynthetic rate and C assimilation capacity. Under salinity stress, the Na⁺ content increased, the K⁺ content decreased with increasing stress. Greater changes were observed under alkali than under salt stress. Alkali stress caused the massive influx of Na⁺, which probably explained that the harmful of alkali stress on plants was stronger than that of salt stress. Under alkali stress, Ca²⁺ and Mg²⁺ decreased in roots, showing that high pH value around roots hindered the absorption of them. Fe²⁺ and Zn²⁺ had little effects on the osmotic adjustment, mainly because of they had a low ion content. Under salt stress, anion increased in order to balance the sharp increase of Na⁺. However, alkali stress made severe deficit of negative charge, broke the intracellular ionic balance and pH homeostasis, and caused a series of strain response. Our results showed that linseed enhanced the synthesis of soluble sugars to balance massive influx of Na⁺ under salt stress, but linseed enhanced the synthesis of organic acids to compensate for the shortage of inorganic anions, which might be a key pathway for the pH adjustment. In conclusion, the alkali stress (high pH value) clearly inhibited the growth, element absorption, ion homeostasis reconstruction of plants. Organic acid concentration is possibly a key adaptive factor for linseed to maintain intracellular ion balance and regulate high pH value under alkali stress.     

盐碱胁迫下湖南稷子苗期根系分泌物代谢组学

盐碱胁迫下植物根系分泌物包含丰富生化信息并具有重要生态作用。为了探讨耐盐碱牧草湖南稷子(Echinochloa frumentacea) 在盐碱胁迫下根系分泌物组成,揭示其在盐碱胁迫下的生理及生态作用,以湖南稷子为试验对象,在人工气候室开展水培试验,并在苗期分别进行中性盐(NaCl+Na₂SO₄ 100 mmol/L)、碱性盐(NaCl+NaHCO₃ 100 mmol/L)和碱(Na₂CO₃+NaHCO₃ 50 mmol/L)处理。在处理3 d后,利用液质联用仪(LC-MS/MS)检测对照组和处理组根系分泌物的化合物成分。结果表明,盐碱胁迫下湖南稷子根系分泌物共有334种化合物。依据正交偏最小二乘法判别分析(OPLS、|DA),重要值(VIP)得分及t检验的P值, 发现对照比SaSo100(碱性盐处理 100 mmol/L),对照比Soda50(碱处理50 mmol/L)和对照比Salt100(中性盐处理100 mmol/L)分别有22、15和21个差异根系分泌物。其中碱性盐和碱处理下根系分泌物组成相近,包括脂质、酚酸,生物碱,苯酞类,氨基糖,萜类,醌类,氨基酸及其衍生物;中性盐处理下有脂质、酚酸,生物碱,苯酞类,萜类。京都基因与基因组百科全书注释及富集发现,盐碱胁迫下根系分泌物不仅含有三羧酸循环代谢产生的碳水化合物、核苷酸,氨基酸,脂肪酸,类脂和维生素等物质,而且与瓦博格效应、膜运输,信号传导以及遗传信息处理等途径有关。研究表明,湖南稷子通过根系分泌物渗出,调节自身代谢物浓度,加强或改变碳同化、呼吸作用、信号传导等提高对盐碱胁迫的适应性。     

Increased synthesis of α‐tocopherol,paramylon and tyrosine by Euglena gracilis under conditions of high biomass production

Aims: To analyse the production of different metabolites by dark‐grown Euglena gracilis under conditions found to render high cell growth. Methods and Results: The combination of glutamate (5 g l^?1), malate (2 g l^?1) and ethanol (10 ml l^?1) (GM + EtOH); glutamate (7·15 g l^?1) and ethanol (10 ml l^?1); or malate (8·16 g l^?1), glucose (10·6 g l^?1) and NH₄Cl (1·8 g l^?1) as carbon and nitrogen sources, promoted an increase of 5·6, 3·7 and 2·6‐fold, respectively, in biomass concentration in comparison with glutamate and malate (GM). In turn, the production of α‐tocopherol after 120 h identified by LC‐MS was 3·7 ± 0·2, 2·4 ± 0·1 and 2 ± 0·1 mg [g dry weight (DW)]^?1, respectively, while in the control medium (GM) it was 0·72 ± 0·1 mg (g DW)^?1. For paramylon synthesis, the addition of EtOH or glucose induced a higher production. Amino acids were assayed by RP‐HPLC; Tyr a tocopherol precursor and Ala an amino acid with antioxidant activity were the amino acids synthesized at higher concentration. Conclusions: Dark‐grown E. gracilis Z is a suitable source for the generation of the biotechnologically relevant metabolites tyrosine, α‐tocopherol and paramylon. Significance and Impact of the Study: By combining different carbon and nitrogen sources and inducing a tolerable stress to the cell by adding ethanol, it was possible to increase the production of biomass, paramylon, α‐tocopherol and some amino acids. The concentrations of α‐tocopherol achieved in this study are higher than others reported previously for Euglena, plant and algal systems. This work helps to understand the effect of different carbon sources on the synthesis of bio‐molecules by E. gracilis and can be used as a basis for future works to improve the production of different metabolites of biotechnological importance by this organism.     

Water Stress Induced Changes in the Amounts of Some Photosynthetic Assimilation Products and Respiratory Metabolites of Sunflower Leaves

Sunflower leaves, water-stressed under controlled conditions,contained greater amounts of amino acids as their water potentialdecreased, with glycine, serine, and glutamate increasing morethan alanine and aspartate. Proline accumulated only at severestress. Low O₂ concentration altered the amounts of amino acids,principally decreasing the amount of glycine and increasingserine. The changes in total pool size are related to previousresults on the accumulation of 14C and the specific activityof products. Photorespiration was large under water stress,where leaves accumulated carbon in glycine of low specific activity,and in 21% O₂, where both total amount and specific activityof glycine was greater than in 1.5% O₂. This suggests that thereare two pools of glycine, one controlled by O₂ and closely relatedto photosynthesis, the other non-photosynthetic and affectedby water stress. The organic acids suocinate, citrate, and fumarate increasedat small leaf-water potentials. Sucrose decreased in amountwith stress and was absent at the most severe stress; therewas less glucose and fructose. The amount of carbon lost fromsugars was similar to the amount accumulated in amino acidstogether with the carbon lost in respiration. It is concluded that stress decreased the flux of carbon fromphotosynthesis for the synthesis of amino acids and sugars butmore carbon from stored materials, principally sucrose, wasused in the production of organic acids and amino acids.