Changes in adenine nucleotides and energy charge in isolated winter wheat cells during low temperature stress

Adenylate energy charge (AEC) and adenine nucleotide levels of isolated winter wheat (Triticum aestivum L. cv Kharkov 22 MC) cells exposed to various low temperature stresses were determined. During ice encasement at −1°C, nucleotide levels decreased gradually in approximate relation to a decline in cell viability. AEC values remained high even after 5 weeks of icing when cell viability was severely reduced. When isolated cell suspensions were exposed to various cooling and freezing regimes ranging from −10 to −30°C, cell damage was dependent on the minimum temperature imposed and the duration of exposure to the freezing stress. The levels of all three adenine nucleotides declined with increasing severity of the imposed stress, but AEC values remained high even at −30°C when nearly all of the cells were killed. The addition of 10 millimolar Ca²⁺ to cell suspensions enhanced survival during low temperature stresses, but did not influence nucleotide levels other than through its effect on cell viability. These results indicate that impairment of the ion transport system during the early stages of ice encasement prior to a detectable decline in cell viability cannot be attributed to changes in the adenylate energy charge system of the cell.     

低温条件下拔节期小麦叶片内源激素含量和抗氧化酶活性的变化

以扬麦16和徐麦30为试验材料,利用人工气候室模拟低温逆境,研究拔节期-3 ℃和-5 ℃低温胁迫对小麦植株受冻率、叶片内源激素含量和抗氧化酶活性的影响.结果表明： 随着处理温度的降低、胁迫时间的延长,小麦植株冻害等级与冻害指数增加,-5 ℃处理72 h两品种五级冻害率均为100%.低温处理结束当天,小麦叶片中内源激素脱落酸（ABA）、玉米素核苷（ZR）含量、抗氧化酶超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）活性随胁迫程度加重呈先升高后降低的趋势;处理结束后3 d,ABA、ZR含量及抗氧化酶活性较处理结束当天升高;至处理结束后6 d,与自然生长的对照处理接近.低温胁迫叶片中赤霉素（GA₃）含量下降,处理结束后3和6 d,扬麦16叶片中GA₃含量呈上升趋势,徐麦30则表现为先升高后下降.-5 ℃ 72 h重度胁迫处理叶片中ABA、ZR、GA₃含量和SOD、POD、CAT活性均较对照显著下降.相关分析表明,较高的ABA、ZR含量、SOD、POD、CAT活性以及较低的GA₃含量可减缓低温胁迫对小麦植株的伤害.     

低温胁迫下喜旱莲子草幼苗膜脂过氧化及保护酶活性的变化

分别在2℃和8℃低温胁迫下,对入侵种喜旱莲子草幼苗抗氧化酶活性进行了研究。与常温28℃的结果相比较,MDA含量、细胞膜透性在8℃条件下变化不明显,2℃处理36h后均显著上升。2℃时SOD、CAT和POD活性在较短时间内(12h)显著升高,随后逐渐下降,CAT活性在48h后受到低温胁迫的严重抑制。8℃处理条件下3种保护酶活性在48h内均有显著升高,但升幅明显小于2℃处理的升幅。8℃低温处理不是抑制而是促进了3种保护酶活性的提高。复温结果显示,8℃低温胁迫的喜旱莲子草幼苗和对照无生长差异,2℃低温胁迫下幼苗表现出形态特征差异。     

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.     

Ultrastructure and lipid composition of etioplasts in developing dark-grown wheat leaves

Changes in the ultrastructure and lipid composition of etioplasts have been evaluated in three regions from the base to the tip of 8-day-old darkgrown wheat leaves and in the upper-2/3 region of etiolated leaves of different ages. In developing darkgrown tissues, the main morphological changes that etioplasts undergo consist of an increase in the amount of thylakoïds which, in the most mature etioplasts, align in parallel arrays. Concomitantly, galactolipids and sulfolipid form an increasing proportion of the total lipids. Trans-3-hexadecenoic acid was not detectable in phosphatidylglycerol (PG) of etioplasts showing appressed thylakoïds isolated from 5-day-old leaves, but was present in significant amounts in etioplasts in the basal part of 8-day-ols leaves in which membrane appression was barely visible. The proportions of this acid increase as etioplasts develop, reaching 25% of the PG fatty acids (1.2% of the total fatty acids) in the most differentiated etioplasts. In wheat etioplasts, it appears that trans-3-hexadecenoic acid may accumulate in considerable amounts in darkgrown tissues and that its accumulation is not directly involved in membrane appression.Abbreviations AP phosphatidic acid - DGDG digalactosyldiacylglycerol - MGDG monogalactosyldiacylglycerol - PC phosphatidylcholine - PE phosphatidylethanolamine - PG phosphatidylglycerol - PI phosphatidylinositol - PS phosphatidylserine - SL sulfolipid     

Interacting effects of CO2 concentration, temperature and nitrogen supply on the photosynthesis and composition of winter wheat leaves

Winter wheat (Triticum aestivum L., cv. Mercia) was grown at two different atmospheric CO₂ concentrations (350 and 700 μmol mol⁻¹), two temperatures [ambient temperature (i.e. tracking the open air) and ambient +4°C] and two rates of nitrogen supply (equivalent to 489 kg ha⁻¹ and 87 kg ha⁻¹). Leaves grown at 700 μmol mol⁻¹ CO₂ had slightly greater photosynthetic capacity (10% mean increase over the experiment) than those grown at ambient CO₂ concentration, but there were no differences in carboxylation efficiency or apparent quantum yield. The amounts of chlorophyll, soluble protein and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) per unit leaf area did not change with long-term exposure to elevated CO₂ concentration. Thus winter wheat, grown under simulated field conditions, for which total biomass was large compared to normal field production, did not experience loss of components of the photosynthetic system or loss of photosynthetic competence with elevated CO₂ concentration. However, nitrogen supply and temperature had large effects on photosynthetic characteristics but did not interact with elevated CO₂ concentration. Nitrogen deficiency resulted in decreases in the contents of protein, including Rubisco, and chlorophyll, and decreased photosynthetic capacity and carboxylation efficiency. An increase in temperature also reduced these components and shortened the effective life of the leaves, reducing the duration of high photosynthetic capacity.     

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.     

Plant stress-related uncoupling protein CSP 310 caused lipid peroxidation in winter wheat mitochondria under chilling stress

The effect of CSP 310 on lipid peroxidation in winter wheat mitochondria was studied by the measurement of primary lipid peroxidation products - dienic conjugates. It was found that some concentrations of CSP 310 caused lipid peroxidation in isolated winter wheat mitochondria in all systems investigated at different concentrations during chilling stress.     

Phosphatidylethanolamine in wheat and barley leaves under water stress

Phosphatidylethanolamine could not be detected in the leaves of less drought-tolerant varieties of wheat (S-308) and barley (BG-25) when the plants wer     

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.     

Characterization of Plasma membrane of winter wheat (TritiCum aestivum L.) Seedlings and Changes in ATP hydrolysing activity under low temparature stress

Plasma membranes were isolated from young seedlings of cold hardened and non-hardened winter wheat using aqueous polymer two phase system consisting of dextran-polyethyleneglycol. Electron microscopic analysis and activities of various marker enzymes revealed that the upper phase was enriched in plasma membrane and free from contamination by other membranes. The fraction thus obtained possessed Mg ATP hydrolysing activity with an optimum pH 6.5 and the activity was stimulated by K⁺. The activity was highly sensitive to orthovanadate, diethylstilbesterol, dicyclohexylcarbodimide, insensitive to azide and nitrate. It was suggested that the ATP hydrolysis was catalysed by H⁺ ATPase. Mg ATP hydrolysing activity in cold hardened seedlings was 38% higher than that from non-hardened seedlings.     

Different mechanisms of the biochemical adaptation of mycelial fungi to temperature stress: Changes in the lipid composition

A comparative study was conducted concerning the effect of temperature stress on the lipid composition of representatives of the subkingdomsEomycota andNeomycota. Changes in the composition of lipid acyl chains (such as saturation and desaturation, isomerization, and changes in the length of fatty acid carbon chains), in the phospholipid composition, and in the contents of sterols and other neutral lipids were revealed. Hyperthermia resulted in (i) an increase in the phosphatidylcholine level, (ii) a decrease in the phosphatidylethanolamine level, (iii) a rise in the content of reserve lipids (triacylglycerols), and (iv) a decline in the free fatty acid level in the neutral lipids. An inverse pattern occurred under hypothermic conditions. The peculiarities in the patterns of the temperature adaptation-related changes in the lipid bilayer composition are considered in terms of the systematic position of the fungi.     

外源一氧化氮对低温胁迫下枇杷叶片AsA-GSH循环的影响

采用0.2、0.5、1.0和1.5 mmol·L^-1的硝普钠(SNP)处理3年生‘早钟6号’枇杷幼苗,以喷清水为对照(CK),于-3 ℃低温胁迫处理6 h后在25 ℃下培养4 d,测定恢复0、1和4 d时叶片非酶抗氧化物质和抗氧化酶活性的变化.结果表明：与CK相比,经SNP处理的枇杷叶片过氧化氢(H₂O₂)含量降低,还原型谷胱甘肽(GSH)和抗坏血酸(AsA)含量及抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)、脱氢抗坏血酸还原酶(DHAR)和单脱氢抗坏血酸还原酶(MDAR)活性升高.低温处理后恢复4 d时,经0.5 mmol·L^-1 SNP处理的枇杷叶片H₂O₂含量下降至CK的75.3%,而GSH和AsA含量分别比CK增加了29.12%和23.40%,APX、GR、DHAR和MDAR活性分别比CK增加了50.0%、44.4%、49.53%和62.68%.适当的外源NO处理可提高枇杷叶片的抗氧化系统活性,减轻细胞在低温胁迫下的损伤,其中以0.5 mmol·L^-1的SNP处理效果较理想.     

Phospholipid changes in wheat and barley leaves under water stress

Total phospholipid content of leaves of wheat and barley increased and phospholipid components changed under water stress. Notable among these were the absence of phosphatidyl serine in barley varieties, decrease in phosphatidyl glycerol content in a less drought-tolerant variety of wheat (S-308) and barley (BG-25), and appearance of phosphatidic acid in both crops. The phospholipid content and its components did not return to normal upon release of the stress by subsequent irrigations. Such observations are indicative of water stress effected alterations in membranes.     

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.