钒酸钠和氯化锂对盐藻质膜排钠系统的影响

杜氏盐藻(Dunaliella salina)有着极强的耐盐碱性.盐藻在低pH值条件下,主要依靠质膜上的Na+/H+逆向转运蛋白实现Na+的外排.在pH值较高的环境中,由于Na+/H+逆向转运蛋白不能够很好的发挥效能,所以盐藻质膜上很可能存在着不依赖于跨膜质子电化学势梯度的排钠机制.通过将盐藻培养液的pH值稳定在7.1和8.9,比较Na+/H+逆向转运蛋白的特异性抑制剂LiCl,以及P型ATPase的特异性抑制剂钒酸钠对盐藻生长的影响,推测盐藻质膜上存在着由不依赖于跨膜质子电化学势梯度的P型ATPase所介导的排钠机制.     

高盐胁迫对坛紫菜(Pyropia haitanensis)叶状体光合作用的影响

坛紫菜(Pyropia haitanensis)具有极强的耐高盐胁迫能力,但其耐盐机理尚不明确。检测了不同高盐（100、110）胁迫处理不同时间（0,4,8,24 h）时坛紫菜Z-61藻体的最大光化学效率（Fv/Fm）和净光合作用,并选取高盐胁迫4 h后的藻体提取RNA,采用第二代高通量测序技术分析正常盐度（对照组,30）与高盐胁迫处理的坛紫菜叶状体转录组数据,探究坛紫菜响应高盐胁迫过程中的光合生理机制。结果发现:盐度100对坛紫菜藻体Fv/Fm无显著影响;而盐度110下,藻体Fv/Fm逐渐下降至0,但转移至对照海水后其仍能恢复到初始水平,可将110称之为“亚致死”（sub-lethal）盐度;当盐度增加至120时,藻体Fv/Fm急剧下降至0,且转移到对照海水中不能恢复,即120是致死的盐度。转录组数据显示,高盐胁迫下的转录本与对照组有很大差异,光合作用相关基因在高盐胁迫下显著上调表达,包括多条碳酸酐酶基因和天线蛋白基因,并且在盐度100胁迫下其上调趋势更为明显。以上结果说明:坛紫菜在耐受盐度100条件下可以通过积极提高光合作用相关基因的表达,维持光合活性,为藻体生长提供物质和能量;而在亚致死盐度110条件下,坛紫菜光合活性逐渐下降,光合基因表达受到抑制,活性氧的产生减少,避免藻体细胞遭受过氧化损伤。这说明坛紫菜可以通过积极响应调节光合系统来应答高盐胁迫。     

盐胁迫下5种木本植物体内Na~+、K~+含量变化及其与抗盐性的关系研究

对NaHCO3胁迫下5种木本植物苗木Na+、K+的吸收和积累及其与木本植物耐盐性的关系进行了研究.结果表明:随盐碱胁迫强度的增加,5个树种苗木地上部分Na+含量、地上部分Na+/K+总体上逐渐增加,地上部分K+含量逐渐降低.地上部分Na+含量依次为枸杞樟子松丁香南蛇藤白榆;Na+/K+含量依次为樟子松南蛇藤丁香枸杞白榆.研究表明,地上部分Na+含量可以作为评价不同树种抗盐性的有效指标,但对于非盐生植物与盐生植物应该区别对待,即非盐生植物地上部分Na+含量与抗盐性负相关,盐生植物地上部分Na+含量与抗盐性正相关.地上部分Na+/K+与抗盐性没有普遍联系,不能作为不同生活型木本植物抗盐性评价标准.     

植物液泡膜Na~+/H~+逆向转运蛋白的研究与应用

液泡膜Na~+/H~+逆向转运蛋白是植物体内广泛存在的一种跨膜转运蛋白,负责植物体内Na~+、H~+的交换。本文对Na~+/H~+逆向转运蛋白的克隆、分子结构、功能及应用等方面展开论述,旨在为研究者系统的理解Na~+/H~+逆向转运蛋白的研究进展,为调控该蛋白的表达来改进植物的生长发育及抗盐胁迫能力等方面的应用提供参考。     

不同盐处理对海滨木槿种子离子含量及抗氧化酶活性的影响

以不同浓度NaCl、KCl溶液及两者等摩尔比混合溶液培养海滨木槿种子,测定种皮与胚中离子含量及胚中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)活性.结果发现:盐胁迫下海滨木槿的种皮有阻隔Na+、K+进入种子内部的作用,种皮及胚对Na+的吸收比例高于对K+的吸收,但胚中有较高的K+含量.种皮积累Na+,种胚积累K+维持较低Na+/K+以适应高盐毒害.同时,不同盐处理下胚中的抗氧化酶活性均呈现先升高后降低的趋势,即低盐下活性升高启动保护机制,高盐下其活性受到抑制,但SOD、CAT、POD能协调一致发挥作用.     

转海蓬子Na+/H+逆向运输蛋白基因番茄的耐盐性研究

以转海蓬子Na /H 逆向运输蛋白基因(Nhap)的番茄(Lycopersicon esculentum Mill.)阳性植株和对照植株为材料,测定叶片相对含水量、质膜透性、K /Na 比值和叶绿素含量4项耐盐指标.结果表明:在盐胁迫条件下,转基因植株的相对含水量、 K /Na 比值和叶绿素含量明显高于对照植株,而叶片质膜透性明显低于对照植株,说明Nhap基因已经在番茄转化植株体内表达,提高了转基因植株的耐盐性.     

坛紫菜耐低盐品系的选育及经济性状的比较

对人工选育的褐绿色、翠绿色、红棕色坛紫菜F1、F2、F3代叶状体在低盐(盐度为15、10)培养条件下的生长和品质进行了研究.结果显示:3种色泽坛紫菜在盐度为15、10的条件下培养20 d,均无成熟、腐烂现象发生,藻体长度增长、鲜重增长和总藻胆蛋白含量均高于野生型坛紫菜(对照组).红棕色F1代叶状体与褐绿色、翠绿色藻体及对照组相比,具有明显的耐低盐能力;通过体细胞克隆和单性生殖获得的红棕色F2、F3代叶状体,在盐度为15、10的条件下,长度增长和鲜重增长较F1代叶状体稳定;红棕色F2代叶状体,在盐度为5的条件下培养20 d,藻体色泽鲜艳,无成熟、腐烂现象发生,生长良好;3～4 cm长的红棕色F3代叶状体,在盐度为5的低盐环境中培养20 d后,平均长度为(46.66±8.68)cm,比对照组长11.89 cm,总藻胆蛋白质量比(干品)达(76.52±3.80)mg*g-1.由此可见,在盐度为10左右生长良好、性状稳定的耐低盐品系,可为坛紫菜向河口区或低盐水域扩大栽培提供有用的种质.     

低盐胁迫对黄鳍棘鲷幼鱼存活率、鳃ATP酶和肝脏抗氧化酶的影响

为了解低盐胁迫对黄鳍棘鲷(Acanthopagrus latus)养殖所产生的影响,采用快速或缓慢2种梯度淡化方法,研究盐度由20淡化至10,5甚至淡水后对黄鳍棘鲷幼鱼的存活率、鳃ATP酶和肝脏抗氧化酶的影响.结果表明:1)除了淡水组的黄鳍棘鲷全部死亡之外,其他低盐组中的成活率为100%;2)在响应不同盐度的低盐胁迫时,不同酶的敏感性有所不同,其中超氧化物歧化酶(SOD)仅对淡水胁迫敏感,Na+/K+-ATP酶和Ca2+/Mg2+-ATP酶对盐度10的低盐胁迫不敏感,过氧化氢酶(CAT)对所有的实验组都敏感,盐度胁迫强度由强到弱依次为淡水、盐度5和盐度10;3)不同的淡化速度影响着Na+/K+-ATP酶和CAT活力恢复到正常水平所需的时间,其中快速淡化所需时间要比缓慢淡化长.本研究结果表明黄鳍棘鲷幼鱼可以较好适应低盐度养殖环境,但淡化速率会对黄鳍棘鲷幼鱼产生一定的应激影响,并且在淡水中应激非常明显.     

外源NO供体硝普钠(SNP)显著缓解NaCl对小麦幼根的生长和质子分泌抑制

笔者研究了NO供体硝普钠(SNP)对NaCl胁迫下小麦(鲁麦15)幼根生长、质子分泌及离子含量的影响.结果表明:75 mmol·L-1的NaCl显著抑制小麦幼根的生长、质子分泌速率,增加Na+/K+比和MDA含量,而同时添加0.08 mmol·L-1SNP后,能显著缓解NaCl胁迫对小麦幼根生长的抑制作用,提高幼根质子分泌速率,降低Na+/K+比和MDA含量.这些结果表明NO可能通过增加根系质子分泌速率,维持低的Na+/K+ 比和膜脂过氧化水平,从而缓解NaCl引起的生长抑制.     

变水植物直叶灰藓渗透调节性研究

用不同浓度的PEG-6000溶液处理直叶灰藓,研究渗透胁迫处理对其渗透调节的影响.结果表明,不同强度的渗透胁迫处理引起直叶灰藓可溶性蛋白质含量先增后减,而其脯氨酸、游离氨基酸和可溶性糖含量随着渗透胁迫的加强呈增加趋势.直叶灰藓Na+含量大于K+含量,Na+/K+为4.50;在渗透胁迫处理过程中Na+积累显著大于K+,Na+/K+达到7.74.说明Na+是直叶灰藓主要的细胞液渗透调节剂.     

转AlNHX1基因大豆后代遗传稳定性及耐盐性分析

为了获得耐盐性有所提高的转AlNHX1基因大豆后代材料,以已获得的转AlNHX1基因的6个株系中的3个株系后代为研究对象,通过分别对这3个株系转基因大豆各后代进行PCR分子检测并结合耐盐性鉴定,以分析外源基因在转基因大豆中遗传稳定性和耐盐性.结果表明:AlNHX1基因在转基因植株的后代中遗传;选取3个株系中部分阳性植株做耐盐性检测,结果表明:转基因大豆耐盐性状均好于野生型大豆.在150mmol/L NaCl胁迫下,转基因大豆叶片中维持了相对较高的K+/Na+比值,相对含水量较野生型提高了9%,而渗透势降低了39%,表明转基因大豆具有较好的吸水和保水能力;在盐胁迫下,超氧化物歧化酶(SOD)与过氧化物酶(POD)活性较野生型大豆分别提高了45%与69%.综上,通过耐盐筛选获得的转AlNHX1基因大豆具有较强的耐盐性.     

Molecular characterization of GmNHX2, a Na＋/H＋ antiporter gene homolog from soybean,and its heterolosous expression to improve salt tolerance in Arabidopsis

Na＋/H＋ antiporters have been well documented to enhance plant salt tolerance by regulating cellular ion homeostasis. Here, a putative Na＋/H＋ antiporter gene homolog GmNHX2 from soybean was cloned and predicted to encode a protein of 534 amino acids with 10 putative transmembrane domains. GmNHX2 was expressed in all soybean plant tissues but enriched in roots and its expression was induced by NaCI and polyethylene glycol （PEG） treatments. GmNHX2 exhibits greater sequence similarity with LeNHX2 and AtNHX6 than that of AtNHX1 and AtSOS1. Although phylogenetic analysis clustered GmNHX2 with organellar （tonoplast and vesicles） antiporters, the GmNHX2-EGFP （enhanced green fluorescent protein） fusion protein was possibly localized in the plasma membrane or organelle membrane of transgenic plant cells, Furthermore, transgenic Arabidopsis plants expressing GmNHX2 were more tolerant to high NaCl concentrations during germination and seedling stages when compared with wild-type plants. These results suggest that GmNHX2 is a membrane Na＋/H＋ antiporter and may function to regulate ion homeostasis under salt stress.     

OsNHX1基因转化84K杨的研究

采用农杆菌介导的方法开展了水稻Na /H 逆向转运蛋白基因OsNHX1转化84K杨的研究.建立了84K杨的叶外植体高频再生系统,经诱导不定芽及诱导生根阶段卡那霉素连续筛选,获得了大量卡那霉素抗性转化植株.PCR检测和Southern杂交检测表明,OsNHX1基因已成功整合到84K杨基因组.耐盐实验表明,转基因植株能在200 mmol NaCl条件下正常生长.     

Red cell sodium fluxes catalysed by the sodium pump in the absence of K+ and ADP

In the absence of extracellular Na+ or K+, the sodium pump catalyses an ouabain-sensitive "uncoupled" Na+ efflux1-4. With red cell ghosts Glynn and Karlish5 showed that this Na+ efflux is accompanied by ATP hydrolysis and that extracellular sodium ions, at low concentrations, inhibit this efflux as well as the associated ATP hydrolysis. At higher concentrations, extracellular sodium ions restore the hydrolysis of ATP3,6 but it is not known whether there is an associated increase in Na+ efflux and, perhaps, an influx. To answer this question we have used inside-out red cell membrane vesicles which are specially suitable for controlling the composition of the medium at the two membrane surfaces while measuring 22Na+ fluxes in both directions. We report here that the sodium pump can operate in a mode in which influx and efflux of sodium are associated with ATP hydrolysis. This mode is different from the Na-Na exchange described by Garrahan and Glynn7, and Glynn and Hoffman8, which requires ADP as well as ATP9 and is probably associated with ADP-ATP exchage rather than ATP hydrolysis10,11.     

Beta-adrenoceptor agonists increase membrane K+ conductance in cardiac Purkinje fibres

Hormonal modulation of the ionic conductance of cell membranes is a topic of considerable current interest; it has a major role, for example, in the improved performance of the vertebrate heart elicited by sympathetic nerve stimulation or by circulating catecholamines, an effect involving enhanced calcium influx. beta-Agonist catecholamines also abbreviate the action potential of cardiac Purkinje fibres, and increase the resting potential in a variety of cells, including cardiac cells, a hyperpolarization usually attributed to stimulation of the electrogenic Na+/K+ pump. We show here that nanomolar concentrations of beta-catecholamines cause hyperpolarization of cardiac Purkinje fibres, not by increasing Na+/K+ pump current, but by increasing resting membrane K+ conductance. The hyperpolarization and shortening of the action potential should increase availability of Na+ channels and reduce the refractory period, effects tending to safeguard impulse propagation through the ventricular conducting system despite the increased heart rate caused by beta-catecholamine action on the sinus node pacemaker.     

Vanadate inhibits uncoupled Ca efflux but not Na--Ca exchange in squid axons.

Nerve cells can maintain a very low intracellular calcium concentration ([Ca2+]i) against large Ca2+ electrochemical gradients (see ref. 1 for review). The properties of the calcium efflux from these cells depend on [Ca2+]i (ref. 2), and within the physiological range, most Ca efflux depends on ATP (which stimulates with high affinity) and is insensitive to Na1, Na0 and Ca0 (uncoupled Ca efflux). When the [Ca2+]i is well above the physiological range, Ca efflux becomes only partially dependent on ATP (acting now with low affinity), is inhibited by Nai and is stimulated by Na0 and Ca0 (Na--Ca exchange). Orthovanadate, a powerful inhibitor of the (Na+ + K+)ATPase and the Na pump, also inhibits the Ca-stimulated ATPase activity, which is the enzymatic basis for the uncoupled Ca pump, in human red cells. The experiments reported here show that in squid axons the ATP-dependent uncoupled Ca efflux can be fully and reversibly inhibited by vanadate, whereas concentrations of vanadate 10 times higher have no effect on the Na--Ca exchange. This is another indication that the uncoupled Ca efflux represents an ATP-driven Ca pump, and supports the suggestion that the uncoupled Ca efflux and Na--Ca exchange are mediated by different mechanisms.     

用于坛紫菜光合效率的DIVING-PAM参数优化研究

采用DIVING-PAM叶绿素荧光测定仪比较了坛紫菜（Porphyra haitanensis)藻体在不同光合有效辐射强度（photosynthetically active radiation,PAR）、暗处理时间和不同测量时间间隔下光合效率值的变化。结果表明,为了在最短的测量时间内获得准确的坛紫菜藻体叶绿素荧光指标测量结果,叶绿素荧光测定仪的最佳参数应设定为:PAR 100 μmol·m-2·s-1,最佳测量时间间隔30 s,最适暗处理时间15min。同时发现坛紫菜藻体光合效率的日变化在每天的10:30~18:30时间段内最为稳定,可以作为测量叶绿素荧光指标的最适时间段。