小麦幼苗拒Na+部位的拒Na+机理

采用日立Z 80 0 0原子吸收分光光度计测Na 、K 含量 ,采用不连续蔗糖梯度离心分离质膜和液泡膜微囊。递增盐度和盐冲击处理下 ,耐盐品种德抗 96 1的SK ,Na(吸收 ) 值和SK ,Na(运输 ) 值均明显大于盐敏感品种鲁麦 15 ;德抗 96 1根部和鲁麦 15根茎结合部Na 含量均呈递增趋势 ,表现出累积效应 ;德抗 96 1根细胞质膜微囊和液泡膜微囊H ATP酶活性均明显大于鲁麦15 ,鲁麦 15根茎结合部液泡膜微囊H ATP酶活性大于德抗 96 1,在同一品种的植株里 ,盐冲击的根和根茎结合部细胞质膜微囊和液泡膜微囊H ATP酶活性均小于递增盐度的酶活性。小麦拒Na 部位细胞质膜和液泡膜H ATP酶活性与其耐盐性强弱成正相关     

DNA超甲基化在小麦耐盐胁迫中的作用

运用高效液相色谱技术测定小麦(Triticum aestivumL.)耐盐品种‘德抗961’和盐敏感品种‘豫麦34’盐胁迫后叶片和根DNA中5-甲基胞嘧啶百分含量的变化,结果表明,经150 mmol/L NaCl处理6 d后,‘德抗961’叶片和根DNA中的5-甲基胞嘧啶的百分含量显著下降,但经150 mmol/L NaCl处理10 d后,耐盐品种‘德抗961’叶片和根DNA中的5-甲基胞嘧啶的百分含量都比盐敏感品种‘豫麦34’的高。由此推测DNA超甲基化可能是植物耐盐机制的一部分。     

盐诱导的依赖叶黄素循环的热耗散提高了小麦的耐盐性

以小麦抗盐品种‘DK961’和盐敏感品种‘LM15’为材料,探讨盐胁迫条件下叶黄素循环与膜脂过氧化的关系。结果表明：200mmol·L-1NaCl处理后二者地上部分鲜重、含水量、K＋含量显著下降,Na＋含量、Na＋/K＋比、丙二醛含量显著升高,膜透性显著增大,‘LM15’的变化幅度均明显大于‘DK961’,而‘LM15’脱环氧化状态（A＋Z）/（A＋Z＋V）的增加明显小于‘DK961’。这表明盐胁迫下‘DK961’通过增加依赖叶黄素循环的热耗散减轻了膜脂过氧化,提高其耐盐性。     

NaCl胁迫下棉花体内 Na~+ 、K~+分布与耐盐性

采用盐化土壤方法 ,选择苗期耐盐性较强的陆地棉品种枝棉 3号和中棉所 1 9及耐盐性较弱的品种泗棉 2号和苏棉 1 2号 ,研究了盐胁迫下棉苗体内 Na+、K+的运输和分配与耐盐性的关系。结果表明 ,耐盐品种根系具有一定的截留 Na+作用。棉花地上部盐分器官水平上的区域化分布特征明显 :2 0 0 mmol/L Na Cl胁迫下 ,枝棉 3号叶片中的 Na+含量显著低于泗棉 2号 ,茎及叶柄中的 Na+含量显著高于泗棉 2号 ;棉株地上部茎、叶柄、叶片中的 Na+含量分别由下而上逐渐减小 ,相同节位的茎、叶柄中的 Na+含量大于叶片 ,枝棉 3号更显著。1 0 0 mmol/L和 1 50 mmol/L Na Cl胁迫下 ,枝棉 3号和中棉所 1 9K+/Na+显著高于泗棉 2号和苏棉 1 2号。Na+在茎和叶柄中滞留和积累 ,根中的 K+向地上部选择性运输 ,以维持叶片中较高的 K+/Na+,是棉花耐盐性的一个重要特点     

NaCl胁迫对阿月浑子各器官Na+、K+吸收的影响

在温室盆栽条件下,用NaCl处理新疆长果阿月浑子与美国品种Kerman,处理浓度为50、150、250和500 mmol·L-1,盐处理后5、10、20 d分别取叶、茎和根,测定各器官中Na+、K+含量及Na+/K+变化规律.研究结果表明,随着土壤盐浓度的增加,长果阿月浑子和Kerman的叶、茎和根中Na+离子含量在盐处理5、10、20 d后每个处理都表现出升高趋势;而两个品种的叶、根和茎中的K+含量在盐处理后其变化规律都不稳定.长果阿月浑子叶片、茎和根系中Na+/K+的比值在盐处理5 d和10 d都升高.Kerman的叶和根系中Na+/K+比值在处理5、10、20 d后升高;茎中Na+/K+处理10 d后升高,但20 d后没有明显变化.实验结果表明NaCl胁迫后,长果阿月浑子的叶片中Na+含量增加幅度均大于Kerman,Kerman根和茎中的Na+含量增加幅度大于长果阿月浑子;NaCl胁迫5 d和10 d后,长果阿月浑子叶片中的Na+/K+的比值高于Kerman,Kerman茎和根部中的Na+/K+的比值高于长果阿月浑子,表明了长果阿月浑子和Kerman都具有一定的耐盐性,但Kerman的耐盐性强于长果阿月浑子.由于叶片和根中Na+和Na+/K+比值的变化很稳定,可以确定为阿月浑子主要抗盐指标,茎中Na+/K+的变化规律较稳定,可以作为抗盐性参考指标.     

盐胁迫下水稻苗期Na+和K+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-1NaCl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-1NaCl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-1NaCl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-1NaCl胁迫下绿叶Na+含量已接近高值,与在8 000 mg·L-1NaCl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.     

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.     

NaCl胁迫对4种豆科树种幼苗生长和K~+、Na~+含量的影响

以合欢、刺槐、国槐和皂荚4种豆科树种盆栽实生幼苗为试验材料,研究了NaCl胁迫下4个树种幼苗的生长、耐盐临界浓度和Na+、K+含量的变化,并对其耐盐性进行了比较.结果表明:NaCl胁迫抑制了4个树种幼苗的生长,苗木的干物质积累量减小、根冠比增大,尤其对合欢和皂荚的影响较大;以相对干质量降至对照组50%时的NaCl浓度作为生长临界NaCl浓度(C50)指标,4个树种的耐盐强弱顺序为:刺槐(5.0‰)>国槐(4.5‰)>皂荚(3.9‰)>合欢(3.0‰);随NaCl浓度的增加,各树种幼苗根、茎、叶中Na+含量逐渐增加,K+含量先增加后减小(合欢根除外),而K+/Na+差异较大.相同浓度NaCl胁迫下,幼苗器官的Na+分布为根>茎>叶,K+因树种和NaCl浓度不同而各异,以叶片中较多,K+/Na+为叶>茎>根.NaCl胁迫下,刺槐的K+含量和K+/Na+较高,地上部分Na+含量较低,幼苗干物质量大,耐盐性较强;而合欢的K+/Na+较小,高浓度NaCl胁迫下地上部分的Na+含量较高,幼苗干物质量小,耐盐性较差.苗木地上部分对K+的积累和根部对Na+的滞留是影响豆科树种耐盐性能的主要因素.     

NaCl处理对盐地碱蓬开花及Na~+、K~+含量的影响

为探讨盐地碱蓬(Suaeda salsa)开花与盐的关系,研究了1(对照)、200和400 mmol·L-1 NaCl处理对盐地碱蓬不同分枝和单株开花数目、叶片和茎及花器官中的Na+、K+含量及Na+/K+比的影响。结果表明:与对照相比,200 mmol·L-1 NaCl处理下盐地碱蓬分枝及单株开花数目增加最显著,单株开花数目增加了69.90%,花器官中的Na+含量、Na+/K+比分别增加了1.41倍和1.77倍,而一级叶片的Na+含量、Na+/K+比分别增加了3.96倍和4.96倍,茎中Na+含量、Na+/K+比分别增加了7.00倍和12.39倍。400 mmol·L-1 NaCl处理下,单株开花数目增加了19.00%,花器官中的Na+含量、Na+/K+比分别增加了2.09倍和3.21倍,而一级叶片的Na+含量、Na+/K+比分别增加了4.28倍和6.50倍,茎中Na+含量、Na+/K+比分别增加了7.65和15.40倍。这些结果表明,NaCl处理下真盐生植物盐地碱蓬可能通过把Na+区域化到茎叶而动员其中的K+到花器官中维持花器官中合适的Na+/K+比而促进开花。     

NaCl胁迫对醋栗番茄、樱桃番茄和番茄幼苗生长、叶片气体交换和离子平衡的影响

以醋栗番茄( Solanum pimpinellifolium Linn.)、樱桃番茄品种‘秦皇贵妃红’( S. lycopersicum var. cerasiforme‘Qinhuangguifeihong’)和番茄品种‘浙粉202’(S. lycopersicum‘Zhefen 202’)幼苗为材料,研究了0(对照)、100、200 mmol·L-1 NaCl胁迫对其生长、叶片气体交换参数和离子平衡的影响。结果表明：在100和200 mmol·L-1 NaCl胁迫下,‘秦皇贵妃红’和‘浙粉202’幼苗单株总干质量的降幅较大,醋栗番茄的降幅较小。 NaCl胁迫明显增加醋栗番茄幼苗的根冠比,但不同胁迫条件下‘秦皇贵妃红’和‘浙粉202’幼苗的根冠比差异不显著。与对照相比,在100 mmol·L-1 NaCl胁迫下,醋栗番茄幼苗叶片的净光合速率( Pn)、胞间CO2浓度( Ci)和蒸腾速率( Tr)的降幅明显低于‘秦皇贵妃红’和‘浙粉202’,而醋栗番茄幼苗叶片气孔导度(Gs)的降幅明显高于后二者;在200 mmol·L-1 NaCl胁迫下,三者叶片Pn、Gs、Ci和Tr值的降幅接近。在100和200 mmol·L-1 NaCl胁迫下,醋栗番茄、‘秦皇贵妃红’和‘浙粉202’幼苗叶片的水分利用效率和气孔限制值均较各自对照显著升高,其中‘秦皇贵妃红’的增幅最大。在100和200 mmol·L-1 NaCl胁迫下,醋栗番茄、‘秦皇贵妃红’和‘浙粉202’幼苗根、茎和叶中Na+含量均较各自对照显著升高,而K+含量和K+/Na+比总体上较各自对照显著降低。与对照相比,经不同浓度NaCl处理后醋栗番茄幼苗根、茎和叶的Na+含量增幅以及K+含量降幅在供试3种植物中均最小,而其不同部位的K+/ Na+比总体上较高。上述研究结果表明：醋栗番茄的耐盐性较强,‘秦皇贵妃红’次之,‘浙粉202’较弱。 NaCl胁迫显著抑制‘秦皇贵妃红’和‘浙粉202’幼苗根的生长,但显著促进醋栗番茄幼苗根的生长,使其维持较强的耐盐性,且NaCl胁迫下醋栗番茄对Na+的吸收和运输减少,以维持体内的离子平衡及较强的光合作用。     

The effect of Na+ and K+ on the thermal denaturation of Na+ and + K+-dependent ATPase.

To increase our understanding of the physical nature of the Na+ and K+ forms of the Na+ + K+-dependent ATPase, thermal-denaturation studies were conducted in different types of ionic media. Thermal-denaturation measurements were performed by measuring the regeneration of ATPase activity after slow pulse exposure to elevated temperatures. Two types of experiments were performed. First, the dependence of the thermal-denaturation rate on Na+ and K+ concentrations was examined. It was found that both cations stabilized the pump protein. Also, K+ was a more effective stabilizer of the native state than was Na+. Secondly, a set of thermodynamic parameters was obtained by measuring the temperature-dependence of the thermal-denaturation rate under three ionic conditions: 60 mM-K+, 150 mM-Na+ and no Na+ or K+. It was found that ion-mediated stabilization of the pump protein was accompanied by substantial increases in activation enthalpy and entropy, the net effect being a less-pronounced increase in activation free energy.     

CD34+-derived CD11c+ + + BDCA-1+ + CD123+ + DC: expansion of a phenotypically undescribed myeloid DC1 population for use in adoptive immunotherapy

BACKGROUND: DC are commonly defined as HLA-DR+/Lin- cells that can be CD11c+ + + CD123+/ -, termed DC1/myeloid DC that induce a Th1 response, or CD11c- CD123+ + +, termed DC2/lymphoid DC that induce a Th2 response. However, significant heterogeneity within DC preparations is apparent and supports the existence of several distinct DC subpopulations. This study aimed to expand and characterize CD34+ DC for use in immunotherapy. METHODS: CD34+ cells were seeded at 1 x 10(5)/mL and expanded for 14 days in RPMI + 10% autologous plasma supplemented with GM-CSF, IL-4, Flt-3L and SCF. Maturation was induced with TNF-alpha and PGE2 for 2 days. DC were analyzed morphologically, phenotypically with a panel of MAb to lineage and DC markers, and functionally in MLR, T-cell assays and T-cell cytokine secretion by ELISA. RESULTS: Significant cellular expansion was observed: 60+/-5 x 10(6) DC from 1 x 10(6) CD34+ cells (n=28). Phenotypically DC were characterized as HLA-DR+ +, CD11c+ + +, CD80+ +, CD83+, CD86+ +, CD123+ +, CD15+ +, CD33+ +, BDCA-1+ +, CD4+ and Lin-. DC displayed potent allostimulatory capacity and efficient presentation of KLH and tetanus toxin. DC-primed T cells secreted IFN-gamma (Th1); however, no detectable IL-4 (Th2) was noted. DISCUSSION: We present features of CD34+ DC that have not been previously described. The CD34+ DC generated represent a population of myeloid DC functioning as DC1 but phenotypically expressing markers characteristic of both DC1 and DC2. This novel DC population is capable of inducing naive T-cell responses and can be expanded to clinically useful numbers. CD34+-derived DC represent attractive candidates for use in adoptive T-cell immunotherapy.     

Hydrolysis of adenylyl imidodiphosphate in the presence of Na+ + Mg+ by (Na+ + K+)-activated ATPase

(1) Contrary to what has usually been assumed, (Na⁺ + K⁺)-ATPase slowly hydrolyses AdoPP[NH]P in the presence of Na⁺ + Mg²⁺ to ADP-NH₂ and P_i. The activity is ouabain-sensitive and is not detected in the absence of either Mg²⁺ or Na²⁺. The specific activity of the Na⁺ + Mg²⁺ dependent AdoPP[NH]P hydrolysis at 37°C and pH 7.0 is 4% of that for ATP under identical conditions and only 0.07% of that for ATP in the presence of K⁺. The activity is not stimulated by K⁺, nor can K⁺ replace Na⁺ in its stimulatory action. This suggests that phosphorylation is rate-limiting. Stimulation by Na⁺ is positively cooperative with a Hill coefficient of 2.4; half-maximal stimulation occurs at 5–9 mM. The K_m value for AdoPP[NH]P is 17 μM. At 0°C and 21°C the specific activity is 2 and 14%, respectively, of that at 37°C. AMP, ADP and AdoPP[CH₂]P are not detectably hydrolysed by (Na⁺ + K⁺)-ATPase in the presence of Na⁺ + Mg²⁺. (2) In addition, AdoPP[NH]P undergoes spontaneous, non-enzymatic hydrolysis at pH 7.0 with rate constants at 0, 21 and 37°C of 0.0006, 0.006 and 0.07 h^?1, respectively. This effect is small compared to the effect of enzymatic hydrolysis under comparable conditions. Mg²⁺ present in excess of AdoPP[NH]P reduces the rate constant of the spontaneous hydrolysis to 0.005 h^?1 at 37°C, indicating that the MgAdoPP[NH]P complex is virtually stable to spontaneous hydrolysis, as is also the case for its enzymatic hydrolysis. (3) A practical consequence of these findings is that AdoPP[NH]P binding studies in the presence of Na⁺ + Mg²⁺ with enzyme concentrations in the mg/ml range are not possible at temperatures above 0°C. On the other hand, determination of affinity in the (Na⁺ + K⁺)-ATPase reaction by competition with ATP at low protein concentrations (μg/ml range) remains possible without significant hydrolysis of AdoPP[NH]P even at 37°C.     

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 Hp-IgY对小鼠胃腔感染VacA CagA Hp的防治效果。ELISA法测定IgY效价均为1∶20,480;抗-VacA CagA Hp-IgY防治小鼠胃腔感染VacA CagAHp效果较理想,IgY高、中剂量组效果优于阳性对照组(P<0.05);低剂量组效果等同于阳性对照组(P>0.05)。抗-VacA CagA Hp-IgY较好的体内抗感染作用,提示该IgY有望成为较理想的治疗VacA CagA Hp感染的生物制剂。     

Different cation sensitivities and binding site domains of Na+-Ca2+-K+ and Na+-Ca2+ exchangers

We examined inhibitory effects of external multivalent cations Ni(2+), Co(2+), Cd(2+), La(3+), Mg(2+), and Mn(2+) on reverse-mode exchange of the K(+)-dependent Na(+)/Ca(2+) exchanger NCKX2 and the K(+)-independent exchanger NCX1 expressed in CCL-39 cells by measuring the rate of Ca(2+) uptake with radioisotope tracer and electrophysiological techniques. The apparent affinities for block of Ca(2+) uptake by multivalent cations was higher in NCKX2 than NCX1, and the rank order of inhibitory potencies among these cations was different. Additional experiments also showed that external Li(+) stimulated reverse-mode exchange by NCX1, but not NCKX2 in the presence of 5 mM K(+). Thus, both exchangers exhibited differential sensitivities to not only K(+) but also many other external cations. We attempted to locate the putative binding sites within the alpha motifs for multivalent cations by site-directed mutagenesis experiments. The cation affinities of NCKX2 were altered by mutations of amino acid residues in the alpha-1 motif, but not by mutations in the alpha-2 motif. These results contrast with those for NCX1 where mutations in both alpha-1 and alpha-2 motifs have been shown previously to affect cation affinities. Susceptibility tests with sulfhydryl alkylating agents suggested that the alpha-1 and alpha-2 motifs are situated extracellularly and intracellularly, respectively, in both exchangers. A topological model is proposed in which the extracellular-facing alpha-1 motif forms an external cation binding site that includes key residues N203, G207C, and I209 in NCKX2, while both alpha-1 and alpha-2 motifs together form the binding sites in NCX1.     

Lyt phenotype of cytotoxic T cells: shift from Lyt-1+2+3+ to a mixed population of Lyt-1+2+3+ and Lyt-1-2+3+ during in vitro culture

The Lyt phenotype of cytotoxic T cells generated in the primary H-2 response was investigated kinetically. The cytotoxicity generated in the early stage of culture was abolished by treatment with alpha Lyt-1,2,3, and complement (C), whereas that generated in the late stage was only partially eliminated by alpha Lyt-1, but was abolished by alpha Lyt-2, 3, and C. This suggested late expansion of the Lyt-1-2+3+ population. Lack of Lyt-1 antigen was confirmed with cells that were depleted of Lyt-1+ from primary culture and then stimulated in the secondary response by elimination of cytotoxicity and by direct Lyt typing. Results indicated that the response of proliferative and cytotoxic T cells of the Lyt-1+2+3+ phenotype in the early stage of culture was followed by activation of Lyt-1-2+3+ T cells. Cytotoxic T cells in the late stage were shown to be a mixture of Lyt-1+2+3+ and Lyt-1-2+3+ cells. This was confirmed with cytotoxic T cells from secondary culture and uncloned long-term T cell lines.