Energy state of spring and winter wheat during cold hardening. Soluble sugars and adenine nucleotides

Marked increases were found in the content of total soluble sugars, reducing sugars and ATP in winter wheat ( Triticum aestivum L. cv. Frederick) during cold hardening. The changes in soluble sugars and ATP of spring wheat ( T. aestivum L. cv. Glenlea) grown under similar conditions were less pronounced. The increase in ATP content during hardening of winter wheat was not associated with significant changes in the content of ADP or AMP. The adenylate energy charge did not change during hardening in either cultivar, but it was higher in the winter cultivar under both growth conditions. This difference could be related to the cold hardiness capacity of winter wheat.     

Effect of low temperature and calcium on survival and membrane properties of isolated winter wheat cells

Isolated cells obtained by enzymic digestion of young primary leaves of cold-hardened, dark-grown Kharkov winter wheat (Triticum aestivum L.) were exposed to various low temperature stresses. The initial uptake of ⁸⁶Rb was generally decreased by increasing concentrations of Ca²⁺, but after longer periods of incubation, the inhibiting effect of high Ca²⁺ levels diminished. Viability of isolated cells suspended in water declined rapidly when ice encased at −1°C, while in the presence of 10 millimolar Ca²⁺ viability declined only gradually over a 5-week period. Ice encasement markedly reduced ⁸⁶Rb uptake prior to a significant decline in cell viability or increased ion efflux. Cell damage increased progressively when the icing temperature was reduced from −1 to −2 and −3°C, but the presence of Ca²⁺ in the suspending medium reduced injury. Cell viability and ion uptake were reduced to a greater extent following slow cooling than after rapid cooling to subfreezing temperatures ranging from −10 to −30°C. The results from this study support the view that an early change in cellular properties due to prolonged ice encasement at −1°C involves the ion transport system, whereas cooling to lower subfreezing temperatures for only a few hours results in more general membrane damage, including loss of semipermeability of the plasma membrane.     

Changes in lipid composition of winter wheat leaves under low temperature stress: effect of molybdenum supply

Molybdenum appliedin vivo to winter wheat (Triticum aestivum L., cv. Sadovo 1) grown on acid soil had cryoprotective effect. In Mo-treated plants the amount of digalactosyl diacylglycerol in the leaves increased during cold acclimation and that of monogalactosyl diacylglycerol remained relatively unchanged. An opposite tendency was observed in untreated plants. The most pronounced effect of freezing temperature on lipid changes was observed with respect to the different rate of phospholipid degradation in both kinds of plants. The amount of phospholipids at ?5°C in Mo-treated plants increased while in untreated ones it significantly decreased. In general, the changes observed in Mo-treated plants corresponded to those shown for frost resistant plants.     

Dynamics of energy charge and adenine nucleotides during uncoupling of catabolism and anabolism in Penicillium ochrochloron

Filamentous fungi are able to spill energy when exposed to energy excess by uncoupling catabolism from anabolism, e.g. via overflow metabolism. In current study we tested the hypothesis that overflow metabolism is regulated via the energetic status of the hyphae (i.e. energy charge, ATP concentration). This hypothesis was studied in Penicillium ochrochloron during the steady state of glucose- or ammonium-limited chemostat cultures as well as during three transient states ((i) glucose pulse to a glucose-limited chemostat, (ii) shift from glucose-limited to ammonium-limited conditions in a chemostat, and (iii) ammonium exhaustion in batch culture). Organic acids were excreted under all conditions, even during exponential growth in batch culture as well as under glucose-limited conditions in a chemostat. Partial uncoupling of catabolism and anabolism via overflow metabolism was thus constitutively present. Under all tested conditions, overflow metabolism was independent of the energy charge or the ATP concentration of the hyphae. There was a reciprocal correlation between glucose uptake rate and intracellular adenine nucleotide content. During all transients states a rapid decrease in energy charge and the concentrations of nucleotides was observed shortly after a change in glycolytic flux (“ATP paradoxon”). A possible connection between the change in adenine nucleotide concentrations and the purine salvage pathway is discussed.     

Membrane properties of isolated winter wheat cells in relation to icing stress

Isolated cell preparations of winter wheat (Triticum aestivum L.) were utilized to examine the effect of ice encasement at −1°C and exposure to ethanol on metabolic and biochemical properties of cells. Following icing and ethanol treatments, passive efflux of amino acids increased gradually with duration of exposure to the stress, and closely paralleled the decline in viability of cells. In contrast, uptake of ⁸⁶Rb declined much more rapidly than viability following exposure to icing or ethanol. Electron spin resonance spectroscopy studies revealed no significant change in molecular ordering within the cell membranes following icing or exposure to ethanol, whereas a small but significant increase in order was detected in the noniced controls. O₂ consumption by isolated cells declined only gradually due to icing and ethanol treatments, and remained relatively high even when cell viability was severely reduced. These results indicate that the plasma membrane is a primary site of injury during ice encasement and that damage to the ion transport system is the earliest manifestation of this injury.     

Comparative physiological and proteomic response to abrupt low temperature stress between two winter wheat cultivars differing in low temperature tolerance

Abrupt temperature reduction in winter wheat at either autumn seedling stage prior to vernalisation or early spring crown stage can cause severe crop damage and reduce production. Many studies have reported the physiological and molecular mechanisms underlying cold acclimation in winter wheat by comparing it with spring wheat. However, processes associated with abrupt temperature reduction in autumn seedling stage prior to vernalisation in winter wheat are less understood. In this study, physiological and molecular responses of winter wheat seedlings to abrupt low temperature (LT) stress were characterised in the relatively LT‐tolerant winter wheat cultivar Shixin 828 by comparing it with the relatively LT‐sensitive cultivar Shiluan 02‐1 using a combination of physiological, proteomics and biochemical approaches. Shixin 828 was tolerant to abrupt LT stress, while Shiluan 02‐1 exhibited high levels of reactive oxygen species (ROS) and leaf cell death. Significant increases in relative abundance of antioxidant‐related proteins were found in Shixin 828 leaves, which correlate with observed higher antioxidant enzyme activity in Shixin 828 compared to Shiluan 02‐1. Proteomics analysis also indicated that carbohydrate metabolism‐related proteins were more abundant in Shiluan 02‐1, correlating with observed accumulation of soluble sugars in Shiluan 02‐1 leaves. Amino acid analysis revealed a strong response to LT stress in wheat leaves. A negative effect of exogenous sucrose on LT tolerance was also found. This study indicates that high ROS scavenging capacity and high abundance of photosynthesis‐related proteins might play a role in winter wheat response to abrupt LT stress. In contrast, excess accumulation of soluble sugars might be disadvantageous for LT tolerance in the wheat cultivar Shiluan 02‐1.     

Salicylic acid-induced antioxidant protection against low temperature in cold-hardy winter wheat

“Dongnongdongmai 1” is a cultivated winter wheat which can endure cold temperature as low as ?30 °C with a reviving rate of 85 %. We aimed to explore the involvement of antioxidant protection system in salicylic acid (SA)-enhanced cold resistance of winter wheat. Seedlings were prayed with 0.1 mM SA at three-leaf stage, followed by cold acclimation at tillering stage (4 °C for 5 days) prior to cold treatment at 4, 0, ?10 or ?20 °C for 2 days. Under low temperature, the relative electrical conductivity (REC) of rhizomes and H₂O₂ content in rhizomes were lower compared with leaves, while in the reactive oxygen species (ROS) removal system, only the POD activity was higher. Foliar spray with SA significantly inhibited the cold-increased REC of rhizomes at ?20 °C and REC of leaves at ?10 and ?20 °C. In addition, application of SA prior to ?10 or ?20 °C treatment suppressed the increase in H₂O₂ content both in rhizomes and leaves. SA enhanced the activities of SOD, POD, and CAT in wheat following low-temperature treatment, especially at ?10 and ?20 °C. In addition, spray with SA resulted in 1.1-to-4.9-fold enhanced activities of the key enzymes in AsA–GSH cycle, including APX, DHAR, and MDHAR. Our results suggested that SA could improve the resistance of winter wheat against extreme low temperature by enhancing the activities of antioxidases to eliminate ROS and maintain the redox homeostasis. In addition, the less damage to rhizomes in comparison with leaves may be attributed to enhanced POD activity.     

Changes in intramitochondrial adenine nucleotides in blowfly flight-muscle mitochondria

1. With freshly isolated blowfly mitochondria 38% of the intramitochondrial adenine nucleotide was present as AMP. 2. On incubation with oxidizable substrates the AMP and ADP concentrations fell and that of ATP rose; with pyruvate together with proline the ATP concentration reached its maximum value at 6min; with glycerol phosphate the phosphorylation of endogenous nucleotide was more rapid. 3. Addition of the uncoupling agent carbonyl cyanide phenylhydrazone caused a rapid fall of ATP and a parallel rise in ADP, then ADP was converted into AMP. 4. This was in contrast with rat liver mitochondria endogenous AMP concentrations, which were always lower than those of blowfly mitochondria and changed little under different metabolic conditions. 5. Evidence is presented that adenylate kinase (EC 2.7.4.3) has a dual distribution in blowfly mitochondria, a part being located in the matrix space and a part in the space between the outer and inner mitochondrial membranes, as in liver and other mitochondria. 6. The possible regulatory role of changing AMP concentrations in the mitochondrial matrix was investigated. Partially purified pyruvate carboxylase (EC 6.4.1.1) and citrate synthase (EC 4.1.3.7) were inhibited 30% by 2mm-AMP, whereas pyruvate dehydrogenase (EC 1.2.4.1) was unaffected. 7. AMP activated the NAD(+)-linked isocitrate dehydrogenase (EC 1.1.1.41) activity of blowfly mitochondria in the absence of ADP, but in the presence of ADP, AMP caused inhibition. 8. It is suggested that AMP may exert a controlling effect on the oxidative activity of blowfly mitochondria.     

Adenylic nucleotides and energy charge during the embryonic development of Bufo arenarum

The contents of ATP, ADP and AMP were determined by HPLC and adenylic energy charge (AEC) was estimated during different stages of the embryonic development of Bufo arenarum up to the tailbud stage. All the developmental stages studied showed a high ATP content (about 1.04-1.48 nmol/emb.). The concentration of ADP was low (0.025-0.041 nmol/emb.) but rose slightly at the neural tube stage. AMP was undetectable before the tailbud stage. AEC values were almost constant (about 0.987-0.992) throughout the period studied. Only a fall at the tailbud stage could be detected which can be related to this more advanced cellular differentiation stage.     

Changes in hybridizable RNA in winter wheat embryos during germination and vernalization

DNA-RNA hybridization-competition experiments were performedto analyze RNA heterogeneity in wheat embryos during germinationand vernalization. A significant difference was found betweenRNA populations of 24-hr and 3-day germinated embryos, whileminor differences were detected between 3- and 5-day germinatedembryo RNAs and between 20- and 40-day cold-treated embryo RNAs.RNA populations in 30-day cold-treated embryos were significantlydifferent from those in 50-day ones. The RNA species presentin 50-day cold-treated embryos were not similar to those in3- and 5-day germinated embryos. A great portion of the RNAspecies in 3-day germinated embryos was found in 30-day cold-treatedembryos. These results suggested that some new RNA species aresynthesized in wheat embryos during 30 to 50 days of cold treatment. ¹ Present address: Department of Biochemistry, Faculty of PharmaceuticalSciences, Higashi Nippon Gakuen University, Ishikari-Tobetsu,Hokkaido 061–02, Japan. (Received February 21, 1977; )     

Charges of nicotinamide adenine nucleotides and adenylate energy charge as regulatory parameters of the metabolism in Escherichia coli.

Methods for measurements of catabolic reduction charge (defined as NADH/(NADH+NAD+)) and anabolic reduction charge (defined as NADPH/(NADPH + NADP+)) are described using [14C]nicotinamide labeling of Escherichia coli cultures. Together with these parameters the adenylate energy charge (ATP + 1/2ADP)/(ATP + ADP + AMP) was measured using labeling with [2-3H]adenine. These three charges were found under different exponential growth conditions to have values independent of the growth conditions: catabolic reduction charge, 0.05; anabolic reduction charge, 0.45; and adenylate energy charge, 0.9. The charges were examined during interruption of growth primarily affecting catabolism, respiration, or anabolism, leading to changes of the charges. The changes of charges are evaluated as a possible regulation of the metabolic rates utilizing or producing the nucleotides by their respective charges.     

Adenine nucleotides and energy charge evolution during the induction of flowering in spinach leaves

Changes in adenine nucleotides pool size levels have been investigated in spinach leaves (Spinacia oleracea. L. cv. Nobel) in order to characterize the transition from the vegetative to the reproductive development. The transient changes reported in this study are the earliest responses observed to date in leaves during photoperiodic induction. These results are discussed in relation to Prigogine's theory of systems far from equilibrium.Abbreviations AN adenine nucleotide(s) - MIT mimicked inductive treatment (inductive treatment on already induced plants)     

Catabolism of adenine nucleotides in suspension-cultured plant cells

Profiles of the catabolism of adenine nucleotides in cultured plant cells were investigated. Adenine nucleotides, prelabelled by incubation of suspension-cultured Catharantus roseus cells with [8-14C]adenosine, were catabolized rapidly and most of the radioactivity appeared in 14CO2. Allantoin and allantoic acid, intermediates of the oxidative catabolic pathway of purines, were temporarily labelled. When the cells, prelabelled with [8-14C]adenosine, were incubated with high concentrations of adenosine, the rate of catabolism of adenine nucleotides increased. The results suggest that the relative rate of catabolism of adenine nucleotides is strongly dependent on the concentration of adenine nucleotides in the cells. Studies using allopurinol, coformycin and tiazofurin, inhibitors of enzymes involved in purine metabolism, suggest that participation of AMP deaminase and xanthine oxidoreductase in the catabolism of adenine nucleotides in plant cells. AMP deaminase was found in extracts from C. roseus cells and its activity increased significantly in the presence of ATP. In contrast, no adenosine deaminase or adenine deaminase activity was detected. Qualitative differences in the catabolic activity of AMP were observed between suspension-cultured cells from different species of plants.     

Changes in fatty acid composition of winter wheat during frost hardening

Twelve day old winter wheat seedlings (cvs Kharkov, frost hardy and Champlein, less hardy) accumulated linolenic acid at the expense of linoleic acid during controlled hardening. The change was most pronounced in the roots, where it was not specific to the phospholipid fraction. It was less marked in the leaves, but occurred there mainly in the phospholipids. The lack of differences between fatty acid profiles of the two cultivars rules out the explanation of varietal differences in frost hardiness in winter wheat on the basis of major changes in fatty acid unsaturation.     

灌浆期高温和水分逆境对冬小麦籽粒蛋白质和淀粉含量的影响

灌浆期高温和水分逆境是影响小麦籽粒产量和品质的关键气候因子。以扬麦9号、徐州26和豫麦34三个小麦品种为材料,利用人工气候室模拟灌浆期高温和水分胁迫环境,研究了花后高温及温度和水分互作对小麦籽粒蛋白质和淀粉形成的影响。结果表明,高温显著提高了小麦籽粒蛋白质含量及清蛋白、球蛋白和醇溶蛋白含量,但降低了谷蛋白含量,导致麦谷蛋白／醇溶蛋白比值降低。高温显著降低了籽粒总淀粉和支链淀粉含量及支／直比。籽粒蛋白质和淀粉及其组分形成所需的适宜昼夜温差随小麦品质类型而异,但温度水平对籽粒蛋白质和淀粉的影响较温差大。在高温和水分逆境下,温度对籽粒蛋白质和淀粉含量的影响较水分逆境大,且存在显著的互作效应。小麦籽粒蛋白质含量均表现为干旱〉对照〉渍水,以高温干旱最高,适温渍水最低;淀粉含量为对照〉干旱〉渍水,以适温对照最高,而高温渍水最低。高温和水分逆境显著提高了籽粒醇溶蛋白含量而降低了谷蛋白含量及支链淀粉含量,使蛋白质谷／醇比和淀粉支／直比降低,以高温渍水对籽粒蛋白质和淀粉组分的影响最为显著。不同品种之间,高蛋白小麦籽粒蛋白质和组分的形成受高温和水分逆境的影响更大,而低蛋白品种籽粒淀粉形成显著受温度和水分逆境的调节。分析表明,在高温和水分逆境下籽粒蛋白质含量与清蛋白和醇溶蛋白显著正相关,籽粒淀粉含量与谷蛋白、支链淀粉含量及支／直比显著正相关。     

Changes in the photosynthetic efficiency of winter wheat in response to abiotic stress

The assessment of heat and drought tolerance is of primary importance in breeding programmes designed to improve heat and drought tolerance in cereals. Three winter wheat varieties grown in controlled growth chambers were exposed to heat (H) and drought (D) stress singly and in combination (H+D). The combined effects of H and D stress were much more severe than those of individual treatments for both physiological and yield parameters during grain filling. The chlorophyll content, effective quantum yield of PSII, net assimilation rate, transpiration, stomatal conductance and intercellular CO₂ concentration were greatly reduced by H, D and their interaction. Grain yield decreased to a greater extent (48.3%) in Plainsman V, averaged over the stress treatments, than in Mv Magma (67.8%) and Fatima 2 (53.7%). The least decline was found in grain number, except in Plainsman V. Mv Magma tolerated heat stress better than Fatima 2. In terms of photosynthetic activity, Plainsman V showed better drought tolerance than Mv Magma. The results showed that changes in physiological properties during stress treatment are not always associated with changes in yield parameters, so a combination of methods may be needed to give a more precise picture of the stress tolerance of wheat varieties.