缓慢干旱下春小麦叶片质膜脂肪酸组成、H+-ATPase及5′-AMPase活力的变化

 研究了田间缓慢干旱胁迫下,抗旱性不同的两个小麦(Triticum aestivum)品种的生长状况、质膜极性脂脂肪酸组成以及质膜关键酶活力的变化。在小麦生长发育的早期,干旱胁迫使其叶片质膜极性脂脂肪酸不饱和度下降、质膜微囊消耗O2的速率升高、膜蛋白含量降低、H+-ATPase (EC 3.6.1.35)活力下降、5'-AMPase (EC 3.1.3.5)活力大幅度升高;在小麦发育的后期,随着干旱的持续,小麦叶片质膜的极性脂脂肪酸不饱和度不变或升高、质膜微囊消耗O2的速率降低、膜蛋白含量与H+-ATPase活力升高、5'-AMPase活力下降。以上结果表明,小麦在发育的早期阶段对干旱较敏感,其细胞质膜流动性降低、细胞中能荷贮备降低;而在后期,则又表现出对干旱的适应。这些结果将有助于阐明自然干旱条件下植物的抗旱机制。     

渗透胁迫对小麦根质膜膜脂物理状态的影响

以两相法纯化的小麦（TriticumsativumL．）根质膜微囊为材料,研究了渗透胁迫下质膜物理状态的变化。结果表明,随着介质蔗糖浓度增加,质膜光散射值降低,二苯己三烯（DPH）荧光偏振值升高,MC540荧光强度增强,并且DPH长寿命组分的荧光寿命和平均寿命都缩短,暗示渗透胁迫使质膜微囊收缩变小,降低了质膜流动性和表面电荷密度,并且表明质膜的疏水性减弱。进一步实验发现,质膜内源色氨酸长寿命组分的寿命缩短,质膜H －AT－Pase活力降低,暗示膜蛋白的构象和功能发生了改变。结果表明,质膜膜脂物理状态改变可能是植物感受渗透胁迫的原初响应。     

干旱对棉花根和下胚轴质膜脂肪酸组分及其相关酶活性的影响

棉花根和下胚轴质膜脂肪酸主要由棕榈酸、硬脂酸、亚油酸和亚麻酸组成。干旱胁迫后,ＰＭ脂肪酸饱和度增加,不饱和脂肪到和不饱和指数降低。其中棕榈酸含量上升和亚麻酸含量下降较大,膜透性增高。质膜Ｈ＾＋ＡＴＰａｓｅ和Ｃａ＾２－ＡＴＰａｓｅ活力降低,脂氧合酶活性增强。     

不同生境下木麻黄脯氨酸含量和质膜ATPase活性的研究

采集生长于恶劣环境和中生环境的普通木麻黄(Casuarina)小枝,超速离心提取粗质膜制剂后,用两相系统法纯化得到质膜微囊,研究不同生境下木麻黄的质膜ATPase活性,并测定木麻黄小枝的游离脯氨酸含量。实验结果表明:同一生境中的木麻黄ATPase活性相对一致,而同一树种木麻黄不同生境下质膜微囊H~+-ATPase、Ca~(2+)-ATPase、K~+-ATPase活性有显著差异,表现出以渗透胁迫为主的恶劣环境下的木麻黄质膜微囊ATPase活性和木麻黄细胞内游离脯氨酸明显高于中生环境下生长的木麻黄。说明普通木麻黄在干旱和盐胁迫下能调整生理生化过程来提高其质膜ATPase活性和增加细胞内脯氨酸含量提高渗透调节能力以保证其在恶劣环境的正常生长。     

在耐热性诱导中豌豆叶片过氧化氢和水杨酸含量与质膜ATP酶活性的变化及相互关系

在高温锻炼(37℃,2h)过程中,豌豆(Pisum sativum L.)叶片过氧化氢(H_2O_2)和游离态水杨酸(SA)含量与质膜ATP酶(H~ -ATPase)活性都有一个高峰,H_2O_2的迸发早于游离态SA的积累,而质膜H~ -ATPase活性高峰的出现则迟于SA高峰;活性氧清除剂、抗氧化剂、质膜NADPH氧化酶抑制剂和H_2O_2的淬灭剂预处理均可有效地阻止高温下H_2O_2和SA的积累以及质膜H~ -ATPase活性的增加。根据以上结果推测,H_2O_2、质膜H~ -ATPase和SA均参与耐热性诱导相关的信号传递,前者作用于SA的上游,而后者在SA下游起作用。     

干旱胁迫下La3+在小麦幼苗叶片受伤害中的作用研究

低浓度La3+叶喷小麦整株幼苗后,随着干旱处理时间的延长,除对叶片相对含水量没有影响外,却对叶细胞膜相对透性、丙二醛产生量、O-·2和H2O2的产生速率均有影响,其变化趋势与不加La3+叶喷的正常和干旱处理的小麦整株叶片相比较,变化曲线上升部分却下降,下降部分却略有升高.表明La3+可抑制干旱胁迫后引起的小麦幼苗叶片细胞膜相对透性增加,降低干旱胁迫引起的膜脂过氧化产物MDA含量,抑制O-·2产生速率,减少叶片中H2O2积累,阻止Fe2+的继续生成,限制了由Fe2+催化的Haber-Weiss和Fenton反应中底物O-·2和生成·OH的生成速率,使小麦叶片中·OH产生量相对减少,降低了叶细胞膜脂过氧化水平,减轻膜伤害,起到了保护、防止膜免受干旱胁迫引起的伤害.     

干旱胁迫对转LeFAD7基因番茄光合作用的影响

以‘天津白果’番茄品种的野生株系(WT)和转正义叶绿体ω-3脂肪酸去饱和酶基因(LeFAD7)株系T( )-12为试材,通过盆栽实验测定了它们在PEG-6000模拟干旱胁迫下的叶片类囊体膜脂脂肪酸组成,以及干旱胁迫后的光合参数、叶绿素荧光参数,以探讨类囊体膜脂LeFAD7基因在干旱胁迫下对番茄叶片光合作用的影响。结果表明:与WT株系相比,转正义基因番茄植株类囊体膜脂中亚麻酸(18∶3)含量升高12.67%,亚油酸(18∶2)含量下降26.98%,导致膜脂脂肪酸不饱和度升高;10%、20%和30%PEG干旱胁迫下,转正义基因番茄植株的实际光化学效率(ΦPSⅡ)、净光合速率(Pn)和叶绿素a(Chl a)含量分别比相应对照下降0.36%~5.11%、19.97%~28.13%、13.27%~23.66%,降幅均小于WT株系,且Pn和Chl a在转基因和野生株间大多存在显著差异。可见,LeFAD7基因能增加番茄叶片膜脂脂肪酸不饱和度,从而减轻干旱胁迫对细胞膜的伤害性,保持干旱逆境下光合系统结构和功能的相对稳定,维持较高的光合速率。     

生物膜组分对膜功能和膜相变的调控——Ⅳ.膜脂组分对水稻根端氧化酶活力和透性的影响

水稻根端的呼吸强度,线粒体氧化酶活力和膜脂脂肪酸不饱和度均随低温处理时间的延长而持续下降,抗冷品种下降速率比不抗冷品种慢。质膜透性随低温处理时间的延长而持续上升,抗冷品种的上升速率比不抗冷品种慢。因之,低温下根端质膜透性、呼吸强度、线粒体氧化酶活力和膜脂脂肪酸不饱和度的变化与品种抗冷性有关。低温下根端质膜透性与呼吸强度和膜脂脂肪酸不饱和度间呈负相关关系。但是,在添加氰化物或不饱和脂肪酸的试验中证明,低温下质膜透性与呼吸强度间、质膜透性与膜脂脂肪酸不饱和度间却呈正相关。说明低温下质膜透性与呼吸强度和膜脂脂肪酸不饱和度无直接的内在联系。低温下根端膜脂脂肪酸不饱和度与呼吸强度和线粒体氧化酶活力伺呈正相关,经添加不饱和脂肪酸的试验证明,脂肪酸不饱和度与根端呼吸强度和线粒体氧化酶活力间仍呈正相关,说明两者间确有内在联系。     

NO对小麦叶片干旱诱导膜脂过氧化的调节效应

分析了一氧化氮(NO)对小麦(Triticum aestivum L．)叶片干旱诱导膜脂过氧化的影响。结果表明：不同浓度NO均能使干旱胁迫下小麦叶片的相对含水量先降后升,而：MDA含量先升后降,O2-释放速率下降,SOD活性升高,POD活性降低;同时,NO可以提高脯氨酸含量。这些结果表明NO提高了小麦叶片的抗氧化能力,降低了干旱胁迫诱导的膜脂过氧化损伤。     

土壤干旱对冬小麦旗叶乙烯释放、多胺积累和细胞质膜的影响

土壤水分亏缺时,小麦旗叶的乙烯释放量增加,三种多胺（精胺、亚精胺、腐胺）的含量在小麦灌浆初期显著增加,而在小麦灌浆末期除腐胺外均降低。外施５００ｍｇ／Ｌ的亚精胺溶液能减弱小麦叶片的膜脂质过氧化程度。土壤水分亏缺时膜透性增大,膜脂过氧化物丙二醇含量升高,导致不饱和脂肪酸指数下降,亚麻酸含量下降。严重土壤水分胁迫下,过氧化物酶的活性升高。这些结果表明土壤干旱损害小麦旗叶质膜,促其衰老。研究表明,随着受旱程度的提高,小麦旗叶遭受的损伤逐渐加重,植株对土壤干旱的适应性亦逐渐降低。     

Does triacylglycerol (TAG) serve a photoprotective function in plant leaves? An examination of leaf lipids under shading and drought

Plant survival in many ecosystems requires tolerance of large radiation loads, unreliable water supply and suboptimal soil fertility. We hypothesized that increased production of neutral lipids (triacylglycerols, TAGs) in plant leaves is a mechanism for dissipating excess radiation energy. In a greenhouse experiment, we combined drought and shade treatments and examined responses among four species differing in life form, habitat, and drought‐ and shade‐tolerance. We also present a lipid extraction protocol suitable for sclerophyllous leaves of native Australian trees (e.g. Acacia, Eucalyptus). Fluorescence measurements indicated that plants exposed to full sunlight experienced mild photoinhibition during our experiment. Accumulation of TAGs did not follow photosynthetic capacity, but instead, TAG concentration increased with non‐photochemical quenching. This suggests that plants under oxidative stress may increase biosynthesis of TAGs. Moderate drought stress resulted in a 60% reduction in TAG concentration in wheat (Triticum aestivum). Shading had no effect on TAGs, but increased concentrations of polar lipids in leaves; for example, acclimation to shade in Austrodanthonia spp., a native Australian grass, resulted in a 60% increase in associated polar lipids and higher foliar chlorophyll concentrations. Shading also reduced the digalactosyldiacylglycerol:monogalactosyldiacylglycerol (DGDG:MGDG) ratio in leaves, with a corresponding increase in the degree of unsaturation and thus fluidity of thylakoid membranes of chloroplasts. Our results suggest that prevention of photodamage may be coordinated with accumulation of TAGs, although further research is required to determine if TAGs serve a photoprotective function in plant leaves.     

Effect of drought stress on lipid metabolism in the leaves of Arabidopsis thaliana (ecotype Columbia)

BACKGROUND AND AIMS: Cell membranes are major targets of environmental stresses. Lipids are important membrane components, and changes in their composition may help to maintain membrane integrity and preserve cell compartmentation under water stress conditions. The aim of this work was to investigate the effects of water stress on membrane lipid composition and other aspects of lipid metabolism in the leaves of the model plant, Arabidopsis thaliana. METHODS: Arabidopsis thaliana (ecotype Columbia) plants were submitted to progressive drought stress by withholding irrigation. Studies were carried out in plants with hydration levels ranging from slight to very severe water deficit. Enzymatic activities hydrolysing MGDG, DGDG and PC were measured. Expression of several genes essential to lipid metabolism, such as genes coding for enzymes involved in lipid biosynthesis (MGDG synthase, DGDG synthase) and degradation (phospholipases D, lipoxygenase, patatin-like lipolytic-acylhydrolase), was studied. KEY RESULTS: In response to drought, total leaf lipid contents decreased progressively. However, for leaf relative water content as low as 47.5 %, total fatty acids still represented 61 % of control contents. Lipid content of extremely dehydrated leaves rapidly increased after rehydration. The time-course of the decrease in leaf lipid contents correlated well with the increase in lipolytic activities of leaf extracts and with the expression of genes involved in lipid degradation. Despite a decrease in total lipid content, lipid class distribution remained relatively stable until the stress became very severe. CONCLUSIONS: Arabidopsis leaf membranes appeared to be very resistant to water deficit, as shown by their capacity to maintain their polar lipid contents and the stability of their lipid composition under severe water loss conditions. Moreover, arabidopsis displayed several characteristics indicative of a so far unknown adaptation capacity to drought-stress at the cellular level, such as an increase in the DGDG : MGDG ratio and fatty acid unsaturation.     

Sub cellular Fractionation of the Longitudinal Smooth Muscle/Myenteric Plexus of Dog Ileum: Dissociation of the Distribution of Two Plasma Membrane Marker Enzymes

The distribution of plasma membrane markers, the sodium pump [evaluated as ouabain-sensitive, potassium-stimulated p-nitrophenyl phosphatase (K+-pNPPase)], [3H]saxitoxin binding, and 5'-AMPase, was studied in the subcellular fractions prepared from the homogenates of the longitudinal smooth muscle/myenteric plexus of dog ileum. The K+-pNPPase activity and [3H]-saxitoxin binding were found to be predominantly associated with the synaptosomal fraction as indicated by the high level of these activities in the crude synaptosomal fraction and by the copurification of K+-pNPPase and [3H]saxitoxin binding, but not 5'-AMPase, with several synaptosomal markers during the fractionation of the crude synaptosomal fraction on density gradients. In contrast to the K+-pNPPase activity and [3H]saxitoxin binding, the 5'-AMPase activity was found to be concentrated in the microsomal pellet. Further fractionation of microsomes on density gradient resulted in copurification of 5'-AMPase but not K+-pNPPase or [3H]saxitoxin binding, with other smooth muscle plasma membrane-bound enzymes, such as high-affinity Ca2+-ATPase, Mg2+-ATPase, and Ca2+-ATPase. It was concluded that in the longitudinal smooth muscle/myenteric plexus, the sodium pump activity is present in higher density in the neuronal plasma membranes whereas 5'-AMPase activity is concentrated in the smooth muscle plasma membranes.     

不饱和脂肪酸在逆境胁迫中的作用

生物膜是将细胞与环境分开的第一道屏障,是环境胁迫造成损伤的主要位点.脂肪酸是生物膜的主要组成成分,不饱和脂肪酸在决定生物膜的生理特性中具有重要作用,增加脂肪酸的不饱和程度能增加膜脂的流动性.近年来,很多研究发现,生物通过脂肪酸脱饱和维持膜的流动性来适应外界环境变化.本文主要从不饱和脂肪酸在环境温度胁迫、盐胁迫、氧化胁迫、酸碱胁迫、干旱胁迫、乙醇胁迫及铝胁迫中的作用研究进展进行了综述.     

Omega-3 fatty acid desaturase (FAD3, FAD7, FAD8) gene expression and linolenic acid content in cowpea leaves submitted to drought and after rehydration

Membrane polyunsaturated fatty acids (PUFA) and particularly linolenic acid (18:3, LA) are known to be implicated in plant tolerance to low temperature. Their role in resistance to drought is much less investigated. In this work, three full-length cDNAs corresponding to omega-3 fatty acid desaturases: fad3 (endoplasmic reticulum), fad7 and fad8 (chloroplastic) were isolated from Vigna unguiculata leaves. Two cowpea cultivars, one drought-tolerant, EPACE-1, and one drought-susceptible, 1183, were compared in terms of fad isoform gene expression and leaf LA contents in plants submitted to water stress followed by rehydration. In EPACE-1, LA content in the main leaf polar lipids increased in response to mild water deficit. Severe water deficits induced a decrease in MGDG LA content while those of PC and DGDG continued to increase. Variations in FAD gene expression, matched those in LA contents. In 1183, LA contents decreased in all lipid classes in response to water stress, as did FAD3 and FAD8 gene expression levels. Rehydration after a moderate water stress induced stimulation mostly in FAD3 gene expression in both cvs. LA contents were equivalent to control levels in EPACE-1. In 1183, they were back to control levels in PC shortly after rehydration but remained low in galactolipids. These results suggested that omega-3 FAD activities were involved in the increase in leaf membrane unsaturation, in the drought tolerant plants whereas the sensitive plants lost PUFAs in response to the treatment. The significance of this discrepancy between the two cvs. in terms of adaptation to drought is discussed.     

Differences in the susceptibility of plant membrane lipids to peroxidation

Peroxidation of three membrane lipid preparations from plants was initiated using Fe-EDTA and ascorbate and quantified as the production of aldehydes and loss of esterified fatty acids. Using liposomes prepared from commercial soybean asolecithin, the degree of peroxidation was shown to be dependent on: the free radical dose, which was varied by the ascorbate concentration; the presence of tocopherol in the liposome; the configuration, of the liposome, multilamellar or unilamellar; and time after initiation. There were dramatic interactions among these factors which led to the conclusion that in comparing the susceptibility of different membrane preparations it is essential to examine the kinetics of the peroxidation reactions. The composition of the liposome was a major determinant of the degree of peroxidation and of the type of degradative reactions initiated by the oxygen free radicals. A fresh polar lipid extract from Typha pollen had very similar fatty acid composition to the soybean asolecithin, but was more resistant to peroxidation as shown by less aldehyde production and increased retention of unsaturated fatty acids after treatment. Similarly, microsomal membranes from the crowns of non-acclimated and cold acclimated winter wheat (Triticum aestivum L.) seedlings had a much higher linolenic acid content than soybean asolecithin but was much more resistant to peroxidation. In the winter wheat microsomes, the loss of esterified fatty acids was not selective for the unsaturated fatty acids; consequently, even with 40% degradation, the degree of unsaturation in the membrane did not decrease. These different reaction mechanisms which occur in plant membranes may explain why measurements of fatty acid unsaturation fail to detect peroxidative reactions during processes such as senescence, aging and environmental stress.     

Effects of moderately enhanced levels of ozone on the acyl lipid composition and dynamical properties of plasma membranes isolated from garden pea (Pisum sativum)

Plasma membranes were isolated from leaves of 16-day-old garden pea, Pisum sativum L., that had been grown in the absence or presence of 65 nl l⁻¹ ozone for 4 days prior to membrane isolation. Plasma membranes from ozone-fumigated plants contained significantly more acyl lipids per protein than those from leaves of plants grown in filtered air on a molar/weight ratio. The ratio between the major acyl lipids, phosphatidylethanolamine (PE) and phosphatidylcholine (PC), also increased due to the ozone fumigation, while the fatty acid unsaturation level was unaltered in total plasma membrane acyl lipids, as well as in PC and PE. The amount of free sterols per protein was unaltered, but the percentage of campesterol increased, concomitant with a decrease in stigmasterol. The dynamical properties of the isolated plasma membranes were assessed using Laurdan fluorescence spectroscopy, which monitors water penetration and mobility at the hydrophilic-hydrophobic interface of the membrane. At 0°C, the molecular mobility was slightly lower in plasma membranes from ozone-fumigated plants than in plasma membranes from control plants, possibly reflecting the increased PE/PC, campesterol/stigmasterol and lipid/protein ratios, and suggesting that ozone-fumigated pea plants may be more susceptible to freezing injuries.     

Very Long-Chain Fatty Acids in Composition of Plant Membrane Lipids

Literary data on very long-chain fatty acids (VLCFAs) that are present in polar lipids of the plant cell membranes are discussed. Large amounts of VLCFA are found in polar lipids of some cellular organelles as well as in nonextractable lipids from diverse plant objects, where the influence of surface lipids on the relative content of these FAs is excluded. In some plants, the VLCFA fraction in membrane lipids increases under several kinds of stress. Amounts and diversity of VLCFAs are lower in flowering plants as compared with the representatives of more ancient taxons—gymnosperms, ferns, and marine algae. Presence of VLCFAs in the composition of annular lipids of the cell membranes is assumed. Biosynthesis of VLCFAs, enzymes involved in the process, and encoding genes are discussed.     

干旱条件下冬小麦幼苗根细胞膜脂组成的变化

干旱条件下小麦幼苗根膜脂总脂肪酸含量、磷脂含量及总脂肪酸双键指数均下降,而游离甾醇含量却明显增加,结果导致游离甾醇／磷脂比率上升。用薄层层析法测得小麦根细胞磷脂主要由磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)、磷脂酰肌醇(PI)及磷脂酸(PA)组成。干旱降低了各种磷脂的含量,但不改变其相对配比。文中讨论了膜脂代谢变化与植物抗旱性的关系。