草莓果实成熟过程中细胞Ca2+-ATPase活性的变化

‘春星’草莓果实成熟时,总糖和花青苷含量增加,呼吸速率也显著升高;同时,细胞溶质Ca2+-ATPase活性和微粒体膜的Ca2+-ATPase总活性变化具有相似的特点,即先升高,至粉红期达到高峰,全红期又下降,在微粒体膜中以质膜Ca2+-ATPase占的比例最高。抑制质膜Ca2+-ATPase活性的Na3VO4能促进草莓果实花青苷积累、降低可溶性总糖含量,但对呼吸速率的影响则因草莓果实成熟度不同而异。     

灵武长枣果实质膜和液泡膜H~+-ATPase和H~+-PPase活性与果实糖分积累

以不同发育时期灵武长枣(Ziziphus jujuba cv.Lingwuchangzao)的果实为材料,通过测定与分析果肉组织中细胞质膜、液泡膜H+-ATPase和H+-PPase活性、果实糖分含量变化,研究了灵武长枣果实质膜、液泡膜H+-ATPase和H+-PPase活性与糖积累特性的关系。结果表明:(1)果实第二次快速生长期之前主要积累葡萄糖和果糖,之后果实迅速积累蔗糖,葡萄糖和果糖含量则逐渐下降,成熟期果实主要积累蔗糖。(2)在果实发育的缓慢生长期S1,质膜H+-ATPase活性最低;第一次快速生长期,质膜H+-ATPase活性最高;缓慢生长期S2,其活性降低;第二次快速生长期,质膜H+-ATPase活性升至次高;完熟期,质膜H+-ATPase活性下降幅度较大。(3)在果实发育过程中,液泡膜H+-ATPase和H+-PPase活性的变化趋势相似。缓慢生长期S1,液泡膜H+-ATPase和H+-PPase活性较低;从缓慢生长期S1至第一次快速生长期缓慢下降至最低;从第一次快速生长期开始,液泡膜H+-ATPase和H+-PPase活性呈现为逐渐增高的变化趋势;除第二次快速生长期以外,液泡膜H+-PPase活性始终高于H+-ATPase。由此推测,质膜H+-ATPase和液泡膜H+-ATPase、H+-PPase对灵武长枣果实糖分的跨膜次级转运起到重要的调控作用。     

NaCl胁迫对盐芥质膜和液泡膜ATPase活性的影响

以盐生植物盐芥和中生植物拟南芥幼苗为材料,研究了盐胁迫对它们叶片和根质膜、液泡膜H+-ATPase、Ca2+-ATPases和K+-ATPase活性以及H+-ATPase、Na+/H+ 逆向转运蛋白表达的影响.结果显示:在NaCl胁迫下,盐芥叶片和根质膜的H+-ATPase活性分别比对照显著升高41%～212%和35%～53%,液泡膜的H+-ATPase分别显著升高281%～373%和4%～38%,而拟南芥却比相应对照都显著降低;相同盐浓度胁迫下,盐芥叶片的H+-ATPase活性比根部高4～8倍,盐芥根也远高于拟南芥.在NaCl胁迫下,盐芥叶片和根的液泡膜H+-ATPase蛋白质β亚基含量变化与其酶活性变化趋势一致,质膜Na+/H+ 逆向转运蛋白的表达量与Na+含量变化趋势一致.盐胁迫下盐芥根中Ca2+-ATPases和K+-ATPase活性的增加与根中Ca2+和K+含量呈显著正相关.研究发现,在盐胁迫条件下,盐芥能有效增强H+-ATPase蛋白和Na+/H+逆向转运蛋白表达,显著提高其根系与叶片质膜和液泡膜的H+-ATPase、Ca2+-ATPase和K+-ATPase活性,维持细胞质中较高的Ca2+和K+水平,从而缓解盐胁迫的伤害,增强耐盐性.     

草莓果实成熟过程中细胞Ca2+-ATPase活性的变化

‘春星’草莓果实成熟时,总糖和花青苷含量增加,呼吸速率也显著升高;同时,细胞溶质Ca2 -ATPase活性和微粒体膜的Ca2 ．ATPase总活性变化具有相似的特点,即先升高,至粉红期达到高峰,全红期又下降,在微粒体膜中以质膜Ca2 -ATPase占的比例最高。抑制质膜Ca2 -ATPase活性的Na3VO4能促进草莓果实花青苷积累、降低可溶性总糖含量,但对呼吸速率的影响则因草莓果实成熟度不同而异。     

陕西红富士苹果矿质营养DRIS标准研究

通过对60个红富士苹果园叶片中N、P、K、Ca、Mg、Fe等6种元素含量的测定和统计分析表明,苹果产量和品质是叶片N、P、K、Ca、Mg、Fe等元素含量综合作用的结果;通过主因素分析,各因素对产量的影响顺序为产量(Y1)x3(K)>x2(P)>x1(N)>x4(Mg)>x5(Ca)>x6(Fe);对可溶性固形物浓度(Y2)的影响顺序为x4(Mg)>x3(K)>x6(Fe)>x1(N)>x5(Ca)>x2(P);对硬度(Y3)的影响顺序为x1(N)>x6(Fe)>x3(K)>x4(Mg)>x2(P)>x5(Ca);对>80%果面红色果占有率(Y4)的影响顺序为x3(K)>x2(P)>x1(N)>x4(Mg)>x5(Ca)>x6(Fe);对低产组(产量<22 500 kg/hm2)大量元素需肥强度影响最大的为K,占该组的53.4%,对中量元素需肥强度影响最大的为Ca和Mg,均占该组的40%;对高产组(产量≥22 500 kg/hm2)DRIS重要参数的表达式为N/K、P/N、K/P,Ca/Mg、Fe/Ca、Fe/Mg,N/K、Ca/K、Ca/N,K/P、Fe/P、Fe/K,Ca/K、Mg/K、Ca/Mg,根据试验结果制定出红富士苹果园N、P、K、Ca、Mg、Fe的DRIS适宜指数范围分别为-8.355～23.011、-13.784～22.078、-12.772～9.544、-2.114～11.532、-2.503～8.902、-4.617～5.291;各园的需肥顺序用图示法和指数法判断,其结果一致.     

脱硫废弃物对碱胁迫下水稻叶片钙分布、Ca2+-ATPase活性及抗氧化特征的影响

为了探讨脱硫废弃物提高水稻抗盐碱的作用机制,采用盆栽法,研究脱硫废弃物对碱胁迫下水稻幼苗叶片总钙含量、Ca2+分布、细胞膜Ca2+-ATPase活性及活性氧含量等的变化.结果表明:对照处理的细胞中钙颗粒零星分布于细胞壁和叶绿体中,添加脱硫废弃物和CaSO4处理的细胞质膜、细胞间隙、细胞壁和液泡中有大量的钙颗粒分布;随着脱硫废弃物和CaSO4添加量的增加,叶片总钙含量增加,质膜和液泡膜Ca2+-ATPase活性呈上升趋势,质膜透性、MDA含量和活性氧O2-产生速率呈下降趋势,SOD、POD等保护酶活性升高.添加脱硫废弃物在一定程度上能够减缓碱胁迫对水稻造成的细胞伤害,起主要作用的物质可能是其主要成分CaSO4.     

氮和钙交互作用对番茄氮素吸收的影响

以沈阳草甸土为栽培基质,采用二次D-饱和最优设计,进行番茄N、Ca两因素效应的盆栽试验,结果表明,在本试验条件下,番茄茎、叶、果实中氮素含量随生育期的进展而下降,氮肥对番茄果实中NO3^--N含量影响呈直线正效应,但各处理番茄果实中NO3^--N含量均不超过可生食标准．适量施用钙肥有利于控制果实中NO3^--N含量,氮肥对番茄植株地上部分和果实中N含量影响均符合报酬递减律．氮肥和钙肥对番茄N素积累符合报酬递减律;番茄从开花期到果实膨大期属N素最大效应期．番茄叶片硝酸还原酶活性受氮素影响较大．N、Ca配合施用,有利于提高番茄叶片硝酸还原酶活性,钙素可降低硝酸还原酶活性,土壤中NO3^--N含量和电导率EC值均随氮肥施用增加而提高。     

脱硫废弃物对碱胁迫下水稻叶片钙分布、Ca2+-ATPase活性及抗氧化特征的影响

为了探讨脱硫废弃物提高水稻抗盐碱的作用机制,采用盆栽法,研究脱硫废弃物对碱胁迫下水稻幼苗叶片总钙含量、Ca2+分布、细胞膜Ca2+-ATPase活性及活性氧含量等的变化.结果表明： 对照处理的细胞中钙颗粒零星分布于细胞壁和叶绿体中,添加脱硫废弃物和CaSO4处理的细胞质膜、细胞间隙、细胞壁和液泡中有大量的钙颗粒分布;随着脱硫废弃物和CaSO4添加量的增加,叶片总钙含量增加,质膜和液泡膜Ca2+-ATPase活性呈上升趋势,质膜透性、MDA含量和活性氧O2〖SX(B-*3〗-〖〗·〖SX)〗产生速率呈下降趋势,SOD、POD等保护酶活性升高.添加脱硫废弃物在一定程度上能够减缓碱胁迫对水稻造成的细胞伤害,起主要作用的物质可能是其主要成分CaSO4.     

压力超负荷性心肌肥厚大鼠心肌细胞核钙转运的改变

通过腹主动脉缩窄(abdominalaorticcoarctation ,AAC)心肌肥厚大鼠模型制备、差速离心提纯心肌细胞核、酶学方法测定Ca2 +-ATPase活性、45Ca2 +同位素法测定核钙摄取和荧光分光光度计测定细胞核内自由钙浓度 ,初步揭示压力超负荷心肌肥厚大鼠心肌细胞核钙转导异常的环节。结果发现 :心肌细胞核上存在具有[Ca2 +]和ATP依赖性的高亲和力Ca2 +-ATPase ,以[Ca2 +]依赖的方式摄取45Ca2 +,并呈先升高后降低趋势。AAC术后4周大鼠心肌显著肥厚 ,伴有明显的血流动力学异常 ,与对照组比较 ,AAC大鼠心肌细胞核Ca2 +-ATPase活性减少51.93 %(p<0.001) ,但核45Ca2 +摄入量(核外[Ca2 +]浓度为800 -1600nmol/L时)和核内[Ca2 +](核外[Ca2 +]浓度为0 -1000nmol/L时)均明显增加(p<0.05) ;正常组离体心肌细胞核Ca2 +摄取受PKA刺激(p<0.05) ,而被PKC抑制剂和CaM抑制剂显著抑制(p<0.05) ,AAC大鼠心肌细胞核Ca2 +摄取仅受CaM抑制剂抑制(p<0.01) ,而PKA和PKC抑制剂对其无明显影响(p>0.05)。结论为心肌肥厚时 ,心肌细胞核Ca2 +转运系统及其磷酸化调节可能发生改变。     

叶面喷施钙镁肥对‘妃子笑''荔枝果肉苹果酸积累的影响

为探讨叶面喷施钙镁肥对‘妃子笑''荔枝果肉苹果酸积累的影响,该文对‘妃子笑''荔枝树冠作喷布0.3%氯化钙(Ca)、0.3%氯化镁(Mg)及其二者混合(Ca+Mg)等水溶液处理,以树冠喷布清水为对照(CK),测定不同生长时期果肉水溶性钙和镁、苹果酸等含量及苹果酸代谢相关酶活性的动态变化,并作多元线性相关分析。结果表明:(1)苹果酸含量呈“L”型变化,Mg、Ca和Ca+Mg处理在果实发育前期促进苹果酸积累,Ca处理在后期促进苹果酸积累。(2)果肉水溶性钙含量总体呈上升趋势,水溶性镁含量大致呈“M”的动态变化趋势。(3)CK和Ca处理的苹果酸含量与NADP-ME活性、Ca+Mg处理的苹果酸含量与PEPC和NAD-MDH活性等均呈正相关,CK的苹果酸含量与PEPC活性、MS活性呈负相关。(4)水溶性钙抑制NAD-MDH、NADP-ME等活性,水溶性镁抑制NAD-MDH、MS等活性。综上认为,钙、镁叶面营养通过改变水溶性钙、镁等含量和苹果酸代谢途径不同关键酶活性而影响果肉苹果酸积累,其中Ca处理可能通过积累更多的苹果酸而抑制果肉呼吸作用,进而使果肉减少糖分损失,在生产中可作施肥技术应用。该研究结果为我国荔枝实际生产提供一定的理论参考和技术支持。     

ca2 +对苹果果实细胞膜透性 保护酶活性和保护物质含量的影响

CaCl2处理“金冠”苹果果肉圆片,明显地增加Ca2+含量,降低K+外渗。不同浓度Ca2+程度不同地提高圆片超氧化物歧化酶、过氧化氢酶活性和谷胱甘肽、抗坏血酸含量,降低H2O2含量和相对电导率。本文认为：Ca2+能提高保护酶活性和保护物质含量,有利于保护细胞膜结构。     

Effect of diet of Varying Protein concentrations on the Activity of Erythrocyte Membrane Ca2+Mg2+ ATPase in Dogs

Alterations in protein diet have been reported to result in alterations in calcium homeostasis in the body. Ca2+Mg2+ATPase is an ubiquitous enzyme important in calcium homeostasis in the body. The effect of varying protein diet on the activities of Ca2+ pump across cell membranes is however yet to be fully elucidated. In this study, the activity of erythrocyte membrane calcium pump in response to varying protein concentration in diet was therefore studied in the dog. The study was carried out in 24 dogs, randomly divided into 4 groups. The groups were fed with diets containing 30%, 26%, 16% and 0% proteins (high, medium, low and zero) for six weeks respectively. Blood samples were collected from each animal to determine packed cell volumes, hematocrit, blood urea, electrolyte studies and erythrocyte ghost membrane studies. The effects of Ca2+ and ATP on the activity of Ca2+Mg2+ ATPase were determined in the isolated ghost membrane. The result of the study shows that there was a protein diet dependent increase in the activity of Ca2+Mg2+ ATPase in the presence and absence of ATP in all the groups with the highest activity recorded in the high protein diet group and the lowest activity observed in the zero protein group. There was also a protein diet dependent increase in the protein concentration of the membranes in all groups observed with the highest protein concentration recorded in the high protein diet group and the lowest activity observed in the zero protein group. There was a significant decrease in K+ concentration (P <0.05) and a significant increase in urea concentration of animals fed with high protein diet (P <0.05). There was also a significant increase (P <0.05) in HCO3- concentration in the animals fed with medium protein diet and no significant difference in the PCV and heamatocrit values in all groups. This study has shown that high protein diets increase the activity of the Ca2+Mg2+ ATPase in the presence and absence of ATP. Keywords; Protein diet, Membrane proteins, Ca2+Mg2+ ATPase activity, Calcium.     

The modification of the unidirectional calcium fluxes of sarcoplasmic reticulum vesicles by monovlent cation ionophroes

Calcium uptake by rabbit skeletal muscle sarcoplasmic reticulum vesicles in phosphate-containing media exhibits time-dependent changes that arise from changing rates of calcium influx and efflux. The monovalent cation ionophore gramicidin, added before the start of the calcium uptake reaction, delayed the spontaneous calcium release that normally occurred after approx. 6 min in such reactions; the rate of calcium efflux was inhibited while calcium influx was little affected. Under these conditions, Ca2+-activated ATPase activity could remain unaltered. Gramicidin stimulated calcium uptake irrespective of the presence of a K+ gradient across the vesicle membrane. Valinomycin stimulated calcium uptake in a manner similar to that for gramicidin even in an NaCl-containing medium lacking potassium. Thus, dissipation of a transmembrane K+ gradient is unlikely to account for the effects of these ionophores on the spontaneous changes in calcium flux rates. Addition of gramicidin to partially calcium-filled vesicles inhibited the phase of spontaneous calcium reuptake because both calcium influx and efflux wre inhibited. Addition of gramicidin to partially calcium-filled vesicles in the presence of a water-soluble protein, such as bovine serum albumin, creatine kinase or pyruvate kinase, markedly stimulated calcium uptake. This stimulatory effect was due primarily to inhibition of calcium efflux, calcium influx being minimally influenced by the ionophore. After cleavage of the 100,000 dalton ATPase to 50,000 dalton fragments, which was not associated with changes in Ca2+-activated ATPase activity or initial calcium uptake rate, gramicidin increased rather than decreased calcium content when added to vesicles after the initial maximum in calcium content. Thus, the ability of monovalent cation ionophores to block calcium efflux from calcium-filled vesicles may reflect their interaction with a portion of the Ca2+-activated ATPase protein.     

Regulation of steady state filling in sarcoplasmic reticulum. Roles of back-inhibition, leakage, and slippage of the calcium pump

Calcium filling of sarcoplasmic reticulum vesicles in the steady state is greatly increased by precipitation of lumenal calcium with oxalate. We find that low concentrations (1 mM) of Pi also allow greater loading by forming a soluble complex with lumenal calcium, an effect that is likely to be of physiological relevance. Furthermore, ADP scavenging by ATP regenerating systems favors calcium loading by preventing reversal of the pump. We also find that uncoupling of ATPase and transport activities is another factor limiting calcium loading. In fact, calcium uptake and ATP utilization occur with a molar ratio of 2:1 in the transient state following addition of ATP but decrease to much lower values in the steady state. Even in the absence of the highly conductive channel which is present only in "heavy" vesicles, "light" vesicles display calcium leakage which is inhibited by medium Ca2+ in the concentration range of ATPase activation and is likely related to an ATPase channel which is involved in calcium transport. It is apparent that, under conditions of ATPase turnover and in the presence of high lumenal Ca2+ and ADP, slippage of calcium through this channel produces true uncoupling of catalytic and transport activities. Coupling is improved by complexation of lumenal Ca2+ and by ATP regeneration and is influenced by the solvent characteristics of the reaction medium. The synergistic effects of lumenal Ca2+ and ADP, and the role of alternate pathways for phosphoenzyme cleavage, are clarified by steady state analysis of a multiple step reaction mechanism. It is concluded that the ideal (2:1) stoichiometric coupling of transport and ATPase activities is not insured by an obligatory pathway of catalysis (as predicted by all reaction schemes published so far); rather, coupling is influenced by the concentrations of ligands and their effects on second order reactions and the consequent distribution of intermediate states.     

Ionic and substrate requirements of the high affinity calcium pumping ATPase in endoplasmic reticulum of pancreas

1. Calcium transport and ATPase activities were determined in microsomal vesicles from pancreatic tissue enriched in endoplasmic reticulum membranes. 2. Calcium transport and ATPase share the following properties: (i) magnesium was required with a K0.5 of 0.7 mM and maximal pumping ATPase activity at 5 mM Mg-ATP; (ii) at saturating magnesium concentrations, calcium increased ATP splitting activity up to three times with an apparent K0.5 close to 0.3 microM calcium; (iii) potassium stimulated the high calcium affinity Mg2+-dependent ATPase and calcium transport. 3.The properties of the calcium pumping system fulfil the cationic and substrate requirements from a physiological point of view.     

Effects of Mg2+ on calcium accumulation by two fractions of sarcoplasmic reticulum from rabbit skeletal muscle

Calcium accumulation by two fractions of sarcoplasmic reticulum presumably derived from longitudinal tubules (light vesicles) and terminal cisternae (heavy vesicles) was examined radiochemically in the presence of various free Mg2+ concentrations. Both fractions of sarcoplasmic reticulum exhibited a Mg2+-dependent increase in phosphate-supported calcium uptake velocity, though half-maximal velocity in heavy vesicles occurred at a much higher free Mg2+ concentration than that in light vesicles (i.e., approx. 0.90 mM vs. approx. 0.02 mM Mg2+). Calcium uptake velocity in light vesicles correlated with Ca2+-dependent ATPase activity, suggesting that Mg2+ stimulated the calcium pump. Calcium uptake velocity in heavy vesicles did not correlate with Ca2+-dependent ATPase activity, although a Mg2+-dependent increase in calcium influx was observed. Thus, Mg2+ may increase the coupling of ATP hydrolysis to calcium transport in heavy vesicles. Analyses of calcium sequestration (in the absence of phosphate) showed a similar trend in that elevation of Mg2+ from 0.07 to 5 mM stimulated calcium sequestration in heavy vesicles much more than in light vesicles. This difference between the two fractions of sarcoplasmic reticulum was not explained by phosphoenzyme (EP) level or distribution. Analyses of calcium uptake, Ca2+-dependent ATPase activity, and unidirectional calcium flux in the presence of approx. 0.4 mM Mg2+ suggested that ruthenium red (0.5 microM) can also increase the coupling of ATP hydrolysis to calcium transport in heavy vesicles, with no effect in light vesicles. These functional differences between light and heavy vesicles suggest that calcium transport in terminal cisternae is regulated differently from that in longitudinal tubules.     

GM1对肌质网Ca~(2+)-ATPase活性及膜流动性的影响

外源性GM1对肌质网Ca2+-ATPase的水解及转运活性都有明显的抑制作用.在GM1浓度为0～8nmol/mg蛋白质范围内抑制作用具有浓度依赖性.当GM1浓度达到8nmol/mg蛋白质时,酶活性受到最大抑制,此时水解活性降低51%,转运活性降低49%.荧光偏振测定结果表明:GM1参入后,肌质网膜流动性降低.     

Subcellular effects of some anesthetic agents on rat myocardium.

The effects of ether, chloroform, and halothane on calcium accumulation and ATPase activity of rat heart microsomes and mitochondria as well as on myofibrillar ATPase activity were investigated. Chloroform and halothane depressed microsomal and mitochondrial calcium uptake and binding in a parallel fashion. Ether decreased microsomal calcium binding and mitochondrial calcium uptake to varying degrees, while mitochondrial calcium binding was slightly enhanced. Whereas ether had no effect, chloroform depressed microsomal and mitochondrial total APTase activities and halothane decreased microsomsl ATPase and slightly stimulated mitochondrial total ATPase activities. Halothane was found to depress myofibrillar Mg2+-ATPase and ether was capable of decreasing myofibrillar Ca2+-ATPase. Chloroform was seen to inhibit both myofibrillar enzymes. These results suggest that the cardiodepressant actions of volatile anesthetic agents may be due to alterations in the calcium accumulating abilities of microsomal and mitochondrial membranes while direct myofibrillar effects may contribute to the depression seen with relatively higher concentrations of anesthetics.     

Inhibition of calcium ATPase by phencyclidine in rat brain

Phencyclidine (PCP) is a potent psychotomimetic drug of abuse and has profound effect on the functioning of the central nervous system (CNS). Many of the CNS functions are known to be mediated by calcium (Ca2+). In the present study we have investigated the effects of PCP on Ca2+ ATPase activity in rat brain both in vitro and in vivo. For in vitro studies, synaptic membrane fractions prepared from normal rat brain were incubated with PCP at different concentrations (25-100 M) before the addition of substrate. For n vivo studies, rats were treated with a single moderate dose of PCP (10 mg/kg, IP) and animals were sacrificed at 1,2, 6 and 12 h after treatment. Ca2+ ATPase activity in synaptic membrane fractions was assayed by estimation of inorganic phosphate. PCP inhibited the Ca2+ ATPase in vitro in a concentration dependent manner with significant effect at 50 and 100 M. A significant time-dependent reduction of the Ca2+ ATPase activity was evident in vivo. As early as 2 h after the treatment of rats with PCP the ATPase activity was significantly reduced. The reduction of Ca2+ ATPase observed even at 12 h after treatment suggesting a prolonged presence of the drug in the brain tissue. Further, kinetic studies in vitro indicated PCP to be a competitive inhibitor of Ca2+ ATPase with respect to the substrate, ATP. The present findings indicate that PCP inhibits synaptic membrane Ca2+ ATPase thus altering cellular Ca2+ homeostasis in CNS which may partially explain the pharmacological effects of the drug and/or its neurotoxicity.     

壳聚糖涂膜对机械伤苹果抗性生理特征的影响

为了提高苹果采后抗机械损伤能力,减少贮藏损失,以红富士苹果为材料,通过人工模拟机械伤试验,研究壳聚糖涂膜对损伤红富士苹果常温贮藏条件下果肉褐变、相关酶活性及抗性相关物质的影响,探索壳聚糖涂膜在果蔬防腐保鲜上的应用。结果显示:壳聚糖涂膜处理科显著减少红富士苹果果实机械伤口的扩张,提高机械伤果实的总酚含量,降低PPO活性,从而有效抑制机械伤苹果贮藏期间的果肉褐变的发生。同时,壳聚糖涂膜处理可提高机械伤苹果的POD和PAL活性,延缓酚类物质含量的下降,促进体内木质素的合成。研究表明,壳聚糖涂膜处理能够有效防止机械伤苹果贮藏期间的酶促褐变,减少果肉组织中有害物质的积累,促进愈伤组织的形成,从而增强了机械伤苹果的抗性,有效延缓了果实衰老。