Na+/H+逆向转运蛋白和植物耐盐性

Na⁺H⁺逆向转运蛋白对植物耐盐起着重要作用 ,它利用质膜H⁺ATPase或液泡膜H⁺ATPase及Ppiase泵H+产生的驱动力把Na⁺排出细胞或在液泡中区隔化以消除Na⁺的毒害。主要讨论植物中Na⁺H⁺逆向转运蛋白研究在分子水平的最新进展.     

拟南芥根皮层细胞质膜内向K+通道电生理特性分析

利用膜片钳技术对模式植物拟南芥根皮层细胞原生质体的内向跨膜钾电流进行了全细胞记录 ,并对内向K⁺通道的特性进行了分析 .结果表明 ,拟南芥根细胞质膜上的内向K⁺通道由超极化膜电位所激活 ;该通道具有较高的K^+/Na⁺选择性 ,可被TEA⁺和Ba^{2 +}等K+通道阻断剂所抑制 ,而且对胞内自由Ca^{2 +}浓度变化不敏感 .这为进一步利用模式植物拟南芥进行植物K⁺吸收机制以及植物抗盐机制的研究奠定了基础 .     

吸收与分配规律的初步研究

选择苗期耐盐性较强的水稻(Oryza sativa)品种（株系）‘AB52’、‘02402’和‘02435’及敏感品种‘日本晴’, 在网室周转箱内,设置5 000和8 000 mg•L^－1 NaCl两种盐处理,以清水为对照, 研究盐胁迫下苗期水稻植株不同部位Na⁺和K⁺的吸收和分配与品种耐盐性的关系。结果表明,盐胁迫下,株高、绿叶干重和绿叶面积下降,绿叶中的水分含量降低,但茎鞘中的水分含量有所上升。5 000 mg•L^－1 NaCl胁迫处理10 d,耐盐品种所受的生长影响和叶片伤害程度低于敏感品种,但8 000 mg•L^－1 NaCl胁迫处理下品种间差异变小。盐胁迫下,水稻植株吸收 Na⁺和置换出K⁺,但不同器官部位中Na⁺和K⁺的区域化分布特征明显,各部位的Na⁺含量由低到高依次为绿叶、根、茎鞘和枯叶。下部老叶能优先积累较多Na⁺而枯黄;绿叶吸收Na⁺ 相对较少,维持较低的Na⁺水平,同时保持较高且稳定的K⁺含量;植株茎鞘通过选择性吸收大量Na⁺ 和置换出一部分K⁺到叶片中,保持绿叶较稳定的K⁺含量和相对较低的Na⁺含量,维持较高的K⁺ /Na⁺比,从而使植株少受盐害。敏感品种‘日本晴’在盐胁迫下绿叶中的Na⁺含量相对较高,且 5 000 mg●L^－1 NaCl胁迫下绿叶Na⁺含量已接近高值,与在8 000 mg●L^－1 NaCl胁迫下差异不大, 而耐盐品种绿叶吸收较少的Na⁺。另一方面,耐盐品种茎鞘的含K⁺相对较高,在盐胁迫下能吸收容纳较多的Na⁺,而绿叶中K⁺/Na⁺比较高。可以认为,绿叶的K⁺/Na⁺比可作为一个衡量耐盐性的相对指标。     

盐胁迫下不同基因型冬小麦渗透及离子的毒害效应

以4种不同基因型冬小麦为试验材料,利用分根法研究了盐胁迫对小麦的渗透胁迫和离子毒害的效应。结果表明,在盐胁迫下,小麦既受渗透胁迫,也受盐离子胁迫。渗透胁迫效应比较快,大约在处理后1-2d内发生;离子毒害效应比较缓慢,大约需3-4d时间。在一半盐胁迫(200mmol/L NaCl)和一半非盐胁迫的分根条件下,小麦没有明显的渗透胁迫效应,小麦植株地上部Na⁺ 累积到毒性水平之前盐处理对小麦生长无抑制效应。小麦具有将Na⁺ 从盐胁迫一侧转移非盐一侧的能力,说明小麦吸收的Na⁺ 有一部分可以从地上部回流到根系中,回流率可达76%-89%。无水分胁迫(不加入PEG)的回流率大于水分胁迫(加入PEG)的回流率。不同基因型小麦在盐分吸收累积和回流,及渗透和离子胁迫的速度和程度等方面具有明显差异。NR 9405和小偃6号的Na⁺ 累积速度要少于陕229和RB 6;NR 9405根系排Na⁺ 能力强于陕229和RB 6。因此,NR 9405和小偃6号的耐盐性高于陕229和RB 6。     

大鼠前脂细胞质膜Na+/H+交换同时参与细胞的增殖和分化

以原代培养的大鼠前脂细胞为模型 ,以 2′ ,7′ bis ( 2 carboxyethyl) 5 ( 6 ) carboxyfluorescein (BCECF)作为检测胞内pH(pHi)的荧光探针 ,测定不同生长因子刺激下胞内pH的变化 ,证明大鼠肾周前脂细胞质膜存在Na⁺/H⁺交换活性 ,胎牛血清(FCS)能快速激活Na⁺/H⁺交换 ,导致pHi升高 (约 0 .2pH单位 ) ,并引起DNA合成 .Ethyl isopropyl amiloride (EIPA)抑制Na⁺/H⁺交换与DNA合成 .在无血清条件下 ,胰岛素不刺激DNA合成但引起细胞分化 ,表现为胞内脂滴积累和 3 磷酸 甘油脱氢酶(G₃ PDH酶 )活性增强 ,同时激活Na⁺/H⁺交换活性导致pHi升高 ;EIPA既抑制胰岛素对Na⁺/H⁺交换的激活 ,也抑制G₃ PDH酶活性增强 .结果证明 :Na⁺/H⁺交换的激活不仅与大鼠前脂细胞增殖相关 ,同时也是细胞分化的早期事件 .     

不同季节荷斯坦牛3个微卫星座位与GSH-Px、SOD和Na+/K+-ATP酶活性及日产奶量的关系

选择分别与谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)和Na⁺/K⁺-ATP酶基因紧密连锁的3个微卫星座位BMS2258、SOD1和BM723, 采用PCR及非变性聚丙烯酰胺凝胶电泳分析其在130头荷斯坦牛中的遗传变异, 计算了3个微卫星座位的多态信息含量、有效等位基因数和遗传杂合度, 并利用最小二乘法拟合线性模型初步探索了它们与荷斯坦牛夏、秋季GSH-Px、SOD、Na⁺/K⁺-ATP酶活性及日产奶量的关系。结果表明, 3个微卫星座位与其紧密连锁基因的酶活性及日产奶量均存在显著相关(P<0.05)。BMS2258座位182 bp/164 bp的GSH-Px活性和日产奶量对应的最小二乘均值较高; SOD1座位148 bp/146 bp对应的SOD活性的最小二乘均值较高, 148 bp/148 bp对应的日产奶量最小二乘均值较高; BMS2258座位161 bp/111 bp对应的Na⁺/K⁺-ATP酶活性和日产奶量的最小二乘均值较高, 它们是各自座位上的最有利基因型。     

耐盐和盐敏感型小麦品种对NaCl胁迫的生理响应及耐盐性差异

为探究小麦品种类型对盐胁迫的生理响应及敏感性差异,以耐盐型品种济麦22和盐敏感型品种河农6425为材料,对幼苗期植株施以不同浓度的NaCl胁迫处理,比较分析了两个品种幼苗在胁迫条件下的生长、生理生化和叶绿体荧光特征等方面的差异。结果表明,NaCl胁迫对幼苗地上和地下部分的生长表现浓度依赖性的抑制效应,对耐盐型品种的抑制程度较小。在生理响应方面,幼苗体内的Na⁺含量随NaCl 浓度的增加而上升,K⁺和Ca²⁺含量则表现相反的变化趋势,耐盐型品种幼苗在胁迫处理前、后均具有较高的K⁺/Na⁺比和Ca²⁺/Na⁺比;NaCl胁迫导致幼苗的光系统Ⅱ受损,表现在Fv/Fm、Fv/Fo、qP 和YⅡ 等叶绿体荧光参数数值下降,降幅在耐盐性品种上相对较小。在氧化胁迫和抗氧化系统方面,NaCl胁迫导致幼苗体内活性氧水平的上升和过氧化物酶（POD）、过氧化氢酶（CAT）等抗氧化酶的响应;POD活性在处理后的0-12 d范围内呈先下降后上升的趋势,CAT活性则呈先上升后下降的趋势。耐盐品种的POD活性在胁迫早期受抑制时间较短,随后的响应更迅速且上升幅度更高;耐盐品种的CAT活性上升幅度更高,且在胁迫后期对高浓度NaCl和长时间胁迫导致的酶活性抑制的耐受性更强。耐盐品种抗氧化酶的这一响应特征与其较低的活性氧上升幅度一致,也与其较低水平的代表膜损伤程度的丙二醛（MDA）积累一致。耐盐型品种根部的MDA积累经200 mmol/L NaCl处理1 d后达到峰值,而盐敏感品种根部的MDA积累经150 mmol/L NaCl处理1 d后即达到峰值。以上研究结果表明,耐盐型小麦品种济麦22可分别通过其较强的K⁺/Na⁺、Ca²⁺/Na⁺调节能力和抗氧化酶体系缓解盐胁迫所导致的渗透胁迫和活性氧伤害,从而表现出耐盐的特征。     

川楝素对K+、Ca2+通道活动及细胞内Ca2+浓度的调控

川楝素是我国学者从驱蛔中药中分离、鉴定的一个三萜化合物，已证明具选择地影响神经递质释放，有效地对抗肉毒中毒，促进细胞分化、凋亡，抑制肿瘤增殖，抑制昆虫发育和取食，影响K⁺、Ca²⁺通道活动等多种生物效应. 综述了证明川楝素抑制多种K⁺通道，选择地易化L型Ca²⁺通道和进而升高胞内Ca⁺浓度的研究资料，并对川楝素产生这些生物效应的机制进行了讨论.     

(Na+/K+)-ATPase研究概况

本文概述(Na⁺/K⁺)-ATPase的一般分子性质。介绍神经元和脂肪细胞中两种不同分子形式(Na⁺/K⁺)-ATPase的分离鉴定和功能性质,以及(Na⁺/K⁺)-ATPase主要功能亚基一级序列和高级结构研究所取得的一些进展。     

抗VacA+CagA+幽门螺杆菌IgY的抗感染作用研究*

为研究抗VacA⁺CagA⁺幽门螺杆菌(Hp)IgY的抗感染作用,以VacA⁺CagA+Hp为抗原免疫蛋鸡,聚乙二醇法和水稀释法从鸡卵黄中提取抗-VacA⁺CagA⁺Hp-IgY,酶联免疫吸附实验(ELISA)测定IgY抗体效价。建立胃腔感染VacA⁺CagAHp的昆明系小鼠模型,观察抗-VacA⁺CagA⁺     

Enhancement of the salt tolerance of Triticum turgidum L. by the Kna1 locus transferred from the Triticum aestivum L. chromosome 4D by homoeologous recombination

Durum wheat, Triticum turgidum L. (2n= 4x=28, genome formula AABB) is inferior to bread wheat, T. aestivum L. (2n=6x=42, genome formula AABBDD), in the ability to exclude Na⁺ under salt strees, in the ratio of the accumulated K⁺ to Na⁺ in the leaves under salt stress, and in tolerance of salt stress. Previous work showed that chromosome 4D has a major effect on Na⁺ and K⁺ accumulation in the leaves of bread wheat. The 4D chromosome was recombined with chromosome 4B in the genetic background of durum wheat. The recombinants showed that Na⁺ exclusion and enhanced K⁺/Na⁺ ratio in the shoots were controlled by a single locus, Kna1, in the long arm of chromosome 4D. The recombinant families were grown in the field under non-saline conditions and two levels of salinity to determine whether Kna1 confers salt tolerance. Under salt stress, the Kna1 families had higher K⁺/Na⁺ ratios in the flag leaves and higher yields of grain and biomass than the Kna1 ^- families and the parental cultivars. Kna1 is, therefore, one of the factors responsible for the higher salt tolerance of bread wheat relative to durum wheat. The present work provides conceptual evidence that tolerance of salt stress can be transferred between species in the tribe Triticeae.     

长苞香蒲对人工盐碱湿地Na+和K+的吸收与转运特征

为揭示长苞香蒲(Typha domingensis)对盐生湿地生态系统中Na⁺和K⁺的吸收与转运特征,探讨长苞香蒲对盐生湿地的生态修复效果,该研究采用人工模拟盐生湿地的方法,设置CK(对照)、T1(浇灌100 mmol·L^-1盐水)、T2(浇灌200 mmol·L^-1盐水)及T3(浇灌300 mmol·L^-1盐水)4种不同盐浓度的人工湿地生态系统,并分别于5月5日(开始盐胁迫处理,S0)、5月30日(S1)、6月30日(S2)和7月30日(S3)测量其株高和干重、植株地上与地下部分Na⁺和K⁺的含量以及底泥和水体中Na⁺和K⁺的含量以分析长苞香蒲对盐碱湿地的脱盐作用。结果表明：(1)各处理的长苞香蒲的株高和干重随着处理时间的延长呈增加趋势,但与CK相比,各处理生长量随盐浓度升高出现下降趋势。(2)高浓度盐处理(T3)使长苞香蒲的地上部分和地下部分的Na⁺分别增加了2.5...     

RETRACTED ARTICLE: Analysis of the physiological mechanism of salt-tolerant transgenic rice carrying a vacuolar Na+/H+ antiporter gene from Suaeda salsa

Salt stress is one of the most serious factors limiting the productivity of agricultural crops. Increasing evidence has demonstrated that vacuolar Na⁺/H⁺ antiporters play a crucial role in plant salt tolerance. In the present study, we expressed the Suaeda salsa vacuolar Na⁺/H⁺ antiporter SsNHX1 in transgenic rice to investigate whether this can increase the salt tolerance of rice, and to study how overexpression of this gene affected other salt-tolerant mechanisms. It was found that transgenic rice plants showed markedly enhanced tolerance to salt stress and to water deprivation compared with non-transgenic controls upon salt stress imposition under outdoor conditions. Measurements of ion levels indicated that K⁺, Ca²⁺ and Mg²⁺ contents were all higher in transgenic plants than in non-transformed controls. Furthermore, shoot V-ATPase hydrolytic activity was dramatically increased in transgenics compared to that of non-transformed controls under salt stress conditions. Physiological analysis also showed that the photosynthetic activity of the transformed plants was higher whereas the same plants had reduced reactive oxygen species generation. In addition, the soluble sugar content increased in the transgenics compared with that in non-transgenics. These results imply that up-regulation of a vacuolar Na⁺/H⁺ antiporter gene in transgenic rice might cause pleiotropic up-regulation of other salt-resistance-related mechanisms to improve salt tolerance.Fengyun Zhao and Zenglan Wang contributed equally to this work.     

Salt tolerance in rice in vitro: Implication of accumulation of Na+, K+ and proline

The implication of accumulation of both inorganic (Na⁺, K⁺) and organic (proline) solutes were evaluated in unadapted and NaCl-adapted callus of a salt-sensitive (Basmati 370) and a salt-tolerant (SR-26B) cultivar of rice (Oryza sativa L.) after a NaCl shock. Accumulation of Na⁺,K⁺ and/or proline in callus was co relatable and the relative presence of these components in tissues after shock treatment was found to be important factors to support differential regrowth capacities of the shock treated calluses. Presence or retention of K⁺ in rice callus was a key factor for salt tolerance as it was observed to be positively correlated with growth in both the varieties. The results indicated that K⁺ was the first candidate to counteract the negative water potential of outside milieu, while proline was probably the last metabolic device that rice calluses opted for when exposed to salt stress.     

Effect of increased alkalinity on Na+ and K+ contents, lipid peroxidation and antioxidative enzymes in two populations of Populus cathayana

We compared two populations of Populus cathayana Rehder, originating from altitudes 2 840 m and 1 450 m, to determine whether trees from different altitudes exhibit different tolerance to alkalinity. The tree cuttings were exposed to nutrient solutions with pH 7.9, 8.8, 9.8 and 10.4 and the salt concentration 200 mM. Na⁺ and K⁺ contents, and Na⁺/K⁺ ratios in leaves and roots were greatly affected by pH values. At pH 10.4, the Na⁺/K⁺ ratios in both leaves and roots sharply dropped in the higher altitude population but were maintained at higher levels in the lower altitude population. The patterns of pH-induced changes in contents of malondialdehyde and free proline, and antioxidative enzyme activities indicated that the higher altitude population exhibits greater tolerance to alkalinity stress than does the lower altitude population.     

Na+/H+ antiport activity in tonoplast vesicles isolated from sunflower roots induced by NaCl stress

Na⁺ transport across the tonoplast and its accumulation in the vacuoles is of crucial importance for plant adaptation to salinity. Mild and severe salt stress increased both ATP- and PP_i-dependent H⁺ transport in tonoplast vesicles from sunflower seedling roots, suggesting the possibility that a Na⁺/H⁺ antiport system could be operating in such vesicles under salt conditions (E. Ballesteros et al. 1996. Physiol. Plant. 97: 259–268). During a mild salt stress, Na⁺ was mainly accumulated in the roots. Under a more severe salt treatment, Na⁺ was equally distributed in shoots and roots. In contrast to what was observed with Na⁺, all the salt treatments reduced the shoot K⁺ content. Dissipation by Na⁺ of the H⁺ gradient generated by the tonoplast H⁺-ATPase, monitored as fluorescence quenching of acridine orange, was used to measure Na⁺/H⁺ exchange across tonoplast-enriched vesicles isolated by sucrose gradient centrifugation from sunflower (Helianthus annuus L.) roots treated for 3 days with different NaCl regimes. Salt treatments induced a Na⁺/H⁺ exchange activity, which displayed saturation kinetics for Na⁺ added to the assay medium. This activity was partially inhibited by 125 μM amiloride, a competitive inhibitor of Na⁺/H⁺ antiports. No Na⁺/H⁺ exchange was detected in vesicles from control roots. The activity was specific for Na⁺. since K⁺ added to the assay medium slightly dissipated H⁺ gradients and displayed non-saturating kinetics for all salt treatments. Apparent K_m for Na⁺/H⁺ exchange in tonoplast vesicles from 150 mM NaCl-treated roots was lower than that of 75 mM NaCl-treated roots, V_max remaining unchanged. The results suggest that the existence of a specific Na⁺/H⁺ exchange activity in tonoplast-enriched vesicle fractions, induced by salt stress, could represent an adaptative response in sunflower plants, moderately tolerant to salinity.     

QTL Analysis of Na+ and K+ Concentrations in Roots and Shoots under Different Levels of NaCl Stress in Rice (Oryza sativa L.)

The key to plant survival under NaCl salt stress is maintaining a low Na⁺ level or Na⁺/K⁺ ratio in the cells. A population of recombinant inbred lines (RILs, F_2∶9) derived from a cross between the salt-tolerant japonica rice variety Jiucaiqing and the salt-sensitive indica variety IR26, was used to determine Na⁺ and K⁺ concentrations in the roots and shoots under three different NaCl stress conditions (0, 100 and 120 mM NaCl). A total of nine additive QTLs were identified by QTL Cartographer program using single-environment phenotypic values, whereas eight additive QTLs were identified by QTL IciMapping program. Among these additive QTLs, five were identified by both programs. Epistatic QTLs and QTL-by-environment interactions were detected by QTLNetwork program in the joint analyses of multi-environment phenotypic values, and one additive QTL and nine epistatic QTLs were identified. There were three epistatic QTLs identified for Na⁺ in roots (RNC), three additive QTLs and two epistatic QTLs identified for Na⁺ in shoots (SNC), four additive QTLs identified for K⁺ in roots (RKC), four additive QTLs and three epistatic QTLs identified for K⁺ in shoots (SKC) and one additive QTL and one epistatic QTL for salt tolerance rating (STR). The phenotypic variation explained by each additive, epistatic QTL and QTL×environment interaction ranged from 8.5 to 18.9%, 0.5 to 5.3% and 0.7 to 7.5%, respectively. By comparing the chromosomal positions of these additive QTLs with those previously identified, five additive QTLs, qSNC9, qSKC1, qSKC9, qRKC4 and qSTR7, might represent novel salt tolerance loci. The identification of salt tolerance in selected RILs showed that a major QTL qSNC11 played a significant role in rice salt tolerance, and could be used to improve salt tolerance of commercial rice varieties with marker-assisted selection (MAS) approach.