外源CaCl2对NaCl胁迫下酸枣幼苗氮代谢的影响

为了研究CaCl₂对NaCl胁迫下酸枣幼苗根、茎、叶的氮代谢影响,探索钙缓解幼苗NaCl胁迫的作用途径。该研究以酸枣幼苗为试验材料,检测不同浓度CaCl₂(0、5、10、20 mmol/L)对NaCl(150 mmol/L)胁迫下幼苗叶片H₂O₂、O^-·₂含量,根、茎、叶中硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)活性及游离氨基酸、可溶性蛋白、硝态氮含量的影响,并采用主成分分析法筛选出评价CaCl₂缓解NaCl胁迫效应的生理指标。结果表明：与NaCl胁迫相比,盐胁迫幼苗叶片的H2O₂、O^-·₂积累量在5、10 mmol/L CaCl₂处理下显著减少;GOGAT活性在5、10 mmol/L CaCl₂处理下的植株根和茎内以及各浓度 CaCl₂处理的叶内均显著升高, GS、NR活性在10、20 mmol/L CaCl₂处理的根内和10 mmol/L CaCl₂处理的茎内以及5、10、20 mmol/L CaCl₂处理的叶内均显著升高;可溶性蛋白含量在5、10、20 mmol/L CaCl₂处理的根、茎、叶内均显著升高,游离氨基酸含量在10、20 mmol/L CaCl₂处理的根和茎内以及10 mmol/L CaCl₂处理的叶内均显著升高,硝态氮含量在10 mmol/L CaCl₂处理的根和茎内以及5、10、20 mmol/L CaCl₂处理的叶内均显著升高。研究发现,150 mmol/L NaCl胁迫对酸枣幼苗造成明显过氧化伤害,抑制了体内氮代谢;外源CaCl₂可通过促进幼苗根和茎内GS/GOGAT循环对NH₄⁺的同化作用,提高叶片NR活性,加快硝态氮的转化速率,从而增强幼苗对NaCl胁迫的适应性,并以10 mmol/L CaCl₂处理缓解效果最佳;游离氨基酸、GOGAT、NR可以作为CaCl₂缓解幼苗NaCl胁迫伤害的评价指标。     

葡萄糖、根浸出液对丛枝菌根真菌吸收不同外源氮产生精氨酸的影响

以大葱(Allium fistulosum)为宿主植物, 接种丛枝菌根(Arbuscular mycorrhizal, AM)真菌Glomus intraradices, 采用三室隔离盆栽培养系统, 在菌丝室施加浓度为4 mmol/L的不同形态外源氮、1%葡萄糖及根浸出液, 通过测定根外菌丝(Extraradical mycelium, ERM)和菌根中精氨酸的含量, 探究葡萄糖、根浸出液对AM真菌吸收不同形式外源氮产生精氨酸的影响。结果表明, 不同外源氮对ERM中精氨酸含量的影响为尿素>Gln>NH₄NO₃>Arg/Gly>NH₄Cl>KNO₃, 对菌根中精氨酸含量的影响为Arg>Gln>尿素>NH₄NO₃>Gly>NH₄Cl>KNO₃; 施加葡萄糖和根浸出液在不同程度上提高ERM干重和菌丝室孢子数量, 但使ERM和菌根中的精氨酸含量降低。说明AM真菌吸收同化不同外源氮产生精氨酸的能力不同, 葡萄糖和根浸出液降低AM真菌吸收同化外源氮产精氨酸的能力。     

外源H2O2对盐胁迫下小白菜种子萌发和幼苗生理特性的影响

以小白菜"甜脆青"为试材,研究不同浓度（5、10、25、50和100 mmol/L）过氧化氢（H₂O₂）浸种处理对100 mmol/L NaCl胁迫下小白菜（Brassica chinensis L.）种子萌发、幼苗生长及生理特性的影响。结果表明：100 mmol/L NaCl胁迫明显抑制小白菜种子的萌发状况和幼苗生长,发芽势、发芽指数、活力指数及幼苗根和芽长度和鲜重均明显降低,根和芽中CAT的活性及K⁺含量明显受到抑制,渗透调节物质、活性氧和MDA含量显著增加。不同浓度H₂O₂浸种处理提高了NaCl胁迫下小白菜种子发芽势、发芽指数和活力指数,促进小白菜根和芽的生长,增强了NaCl胁迫下根和芽中SOD、CAT和APX的活性及K⁺含量,降低O₂^-·产生速率及H₂O₂和MDA含量,进一步促进脯氨酸和可溶性糖含量的增加,降低体内Na⁺含量。其中以10 mmol/L H₂O₂处理缓解盐胁迫效果最好,明显缓解NaCl胁迫对小白菜种子萌发和幼苗生长的抑制。     

青霉素对草莓组培苗硝酸还原酶活性的影响

本实验利用植物组织培养的方法,在基本培养基中添加不同浓度的青霉紊对草莓进行根的诱导,以测定植株的 NR 活性。结果表明:实验剂量范围内,其 NR 活性随青霉素浓度增加呈正相关,同时发现草莓体内 NR 主要存在于地上部分。本实验设两个处理:MS 青霉素50mg/1;MS 青霉素100mg/1;对照 MS,各80瓶,在无菌条件下,分别取草莓组培丛芽接种于上述培养基中70天后进行 NR 活性的测定:取草莓组培植株0.5克,置三角瓶中,各瓶中分别加入5ml 0.1M/L 硝酸缓冲液,5ml 0.2M/L     

无机磷影响叠氮钠对ATP合成酶合成活性的抑制

利用ADP和放射性磷直接合成ATP的方法,研究了无机磷(P_i)和叠氮钠对猪心线粒体ATP合成酶（F₁F_O-ATPase）ATP合成活性的影响.结果发现无机磷除作为合成ATP的底物参与F₁F_O-ATPase的合成反应外,还对F₁F_O-ATPase的合成活性呈现抑制作用,在1 mmol/L ADP存在时,随着P_i浓度由0.01～10 mmol/L增加,抑制合成作用越来越强.与叠氮钠在低浓度时(小于1 mmol/L)只抑制ATP水解,不影响ATP合成的观点不同.实验结果显示0.1 mmol/L叠氮钠表观激活F₁F_O-ATPase的ATP合成活性,且激活程度与反应体系中所加P_i的浓度呈负相关.当固定P_i浓度（0.1 mmol/L）后,随着叠氮钠浓度的增加表观激活程度也在变化,叠氮钠与磷浓度相等时表观激活程度最大,直至叠氮钠浓度接近0.5 mmol/L时,开始呈现表观抑制现象,叠氮钠浓度高于1 mmol/L之后,就出现解偶联现象.     

施氮量对小麦叶片硝酸还原酶活性、一氧化氮含量和气体交换的影响

研究了不同施氮量对冬小麦分蘖到抽穗期叶片硝酸还原酶(NR)活性、一氧化氮(NO)含量、气体交换参数和籽粒产量的影响.结果表明:叶片光合速率（P_n）、蒸腾速率（T_r）、瞬时水分利用效率（IWUE）和产量均随施氮量的增加呈先升高后降低的趋势,在180 kg·hm^-2氮处理时达到最高.随施氮量的增加,叶片NR活性提高; 在分蘖期和拔节期,叶片NR活性与NO含量呈显著线性相关（R²≥0.68,n=15）,NO含量和气孔导度（G_s）呈显著正二次相关（R²≥0.43,n=15）;低氮处理下,NR活性较低使叶片NO含量维持在较低水平,促进气孔开放,高氮处理下,NR活性较高使叶片NO含量增加,诱导气孔关闭;在抽穗期叶片NR活性和NO含量无显著相关关系,虽然NO含量和G_s也呈显著正二次相关（R²≥0.36,n=15）,但不能通过施氮提高NR活性来影响叶片NO含量,进而调节叶片气孔行为.合理施氮使小麦叶片NO含量维持在较低水平,可提高叶片G_s、T_r和IWUE,增强作物抗旱能力,促进光合作用,提高小麦产量.     

胞浆型磷脂酶A2介导弱氧化修饰低密度脂蛋白诱导的人脐静脉内皮细胞凋亡

探讨弱氧化修饰低密度脂蛋白(MM-LDL)能否诱导人脐静脉内皮细胞(HUVECs)凋亡以及胞浆型磷脂酶A₂(cPLA₂)在此过程中的作用.MTT法测定细胞存活率;相差显微镜、荧光显微镜和流式细胞仪检测细胞凋亡;³H-花生四烯酸(³H-AA)预标法测定PLA₂活性;蛋白质印迹检测cPLA₂磷酸化;激光共聚焦显微镜检测单个细胞内钙离子浓度的变化.结果表明,MM-LDL(100～300 mg/L)作用后的HUVECs呈现凋亡典型的形态特征,凋亡率随MM-LDL浓度的增加而上升.MM-LDL能引起胞内钙离子浓度增加,cPLA₂的活化及磷酸化.15 μmol/L AACOCF₃和5 mmol/L EGTA在抑制cPLA₂活性的同时,部分抑制MM-LDL诱导的HUVECs凋亡.加入外源性AA(50 μmol/L)能逆转AACOCF₃引起的凋亡抑制.结果提示,cPLA₂参与了MM-LDL诱导HUVECs凋亡的信号传递.     

AM真菌对盐胁迫下番茄幼苗生理特征及AVP1表达的影响

采用温室盆栽试验研究不同NaCl浓度（0、50 和85 mmol/L）持续胁迫接种摩西球囊霉和地表球囊霉 2种AM真菌对加工番茄耐盐性的影响。结果显示:（1）在0 mmol/L NaCl处理条件下,2种菌的番茄菌根化苗的根系活力、叶片中可溶性糖、可溶性蛋白、根系脯氨酸含量以及超氧化物歧化酶和过氧化物酶活性均高于非菌根植株,且丙二醛含量低于非菌根植株,但差异不显著。（2）在50、85 mmol/L NaCl浓度胁迫下,接种2种菌根真菌可显著提高番茄植株根系活力,促进叶片中可溶性糖、可溶性蛋白及根系脯氨酸含量的积累,显著提高叶片中与抗逆相关的超氧化物歧化酶和过氧化物酶的活性,减少丙二醛在根系中的积累;随着NaCl浓度的增加,效果更为明显。（3）RT-PCR分析显示,AM真菌和盐胁迫共同调控H⁺转运无机焦磷酸酶H⁺- PPase的表达,随NaCl浓度的增加,AVP1基因表达量下降,但菌根化番茄植株的AVP1基因表达量显著高于非菌根植株。研究表明,接种AM真菌后,菌根化植株可通过显著促进幼苗体内渗透调节物质积累和抗氧化酶活性的提高,有效降低体内膜脂过氧化水平,同时过量表达AVP1基因增加了番茄植株中离子向液泡膜的转运,从而缓解盐胁迫对植株的伤害,增强番茄幼苗对盐胁迫的耐性。     

外源H2O2对盐碱胁迫下苹果矮化砧木幼苗生理特性的影响

以2年生苹果矮化砧木M9 T337为试材,采用盆栽试验法,设置浇灌清水(CK)和盐碱胁迫(0.1 mol/L NaCl+NaHCO₃溶液)+ 喷施5种浓度的H₂O₂ ［0(T₁)、0.2 mmol/L(T₂)、0.4 mmol/L(T₃)、0.6 mmol/L(T₄)、0.8 mmol/L(T₅)］ 处理,测定各处理叶片叶绿素含量、光合气体交换参数、渗透调节物质含量、抗氧化酶活性和细胞膜透性,并利用相关性与主成分分析进行综合评价,以探讨外源过氧化氢(H₂O₂)增强其盐碱耐性的生理机制。结果表明：(1)随着盐碱胁迫(T₁)的时间延长,M9 T337幼苗叶片叶绿素a(Chl a)含量、叶绿素b (Chl b)含量、叶绿素总量(Chl t)、净光合速率(P_n)、气孔导度(G_s)、蒸腾速率(T_r)、可溶性蛋白(SP)含量均呈逐渐下降趋势;胞间CO₂浓度(C_i)、可溶性总糖(TSS)含量、脯氨酸(Pro)含量、过氧化氢酶(CAT)活性、抗坏血酸过氧化物酶(APX)活性、相对电导率(REC)、丙二醛(MDA)含量均呈上升趋势;超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性均呈先升后降趋势。(2)与CK相比,盐碱胁迫+外源H₂O₂(T₂- T₅)处理后M9 T337幼苗叶片各指标均呈现不同幅度变化,且存在明显浓度效应,并以T₃(0.4 mmol/L H₂O₂)处理叶片的Chl a、Chl b、Chl t、SP和G_s降幅最小,C_i、REC、MDA升幅最小,TSS、Pro、APX升幅最大。(3)M9 T337幼苗叶片P_n与T_r、G_s、Chl a、Chl b、Chl t、SP、SOD、POD呈显著正相关,与C_i、MDA、CAT、APX、REC呈显著负相关。(4)综合评价表明,各处理对M9 T337幼苗叶片生理特性的效应依次为：CK>T₃>T₄>T₂>T₅>T₁。研究发现,叶面喷施适宜浓度H₂O₂可有效改善盐碱胁迫下M9 T337幼苗光合能力,显著提高抗氧化酶活性和渗透调节物质的含量,降低细胞膜透性,从而达到缓解盐碱胁迫的作用,并以0.4 mmol/L H₂O₂处理效果最佳。     

增铵营养对小麦光合作用及硝酸还原酶和谷氨酰胺合成酶的影响

采用水培试验研究不同形态氮营养(NH4^+／NO3^-分别为0／100、50／50和100／0)对小麦光合作用及氮代谢关键酶活性的影响．结果表明,增铵营养较单—NO₃^-营养显著提高叶片叶绿素含量、净光合速率及可溶性糖含量,叶、根中可溶性蛋白质含量和叶片硝酸还原酶活性。而对谷铵酰胺合成酶活性影响较小．与单—NO₃^-营养相比。增氨营养下叶片较高的可溶性糖含量与净光合速率的提高相关。而维持较高的叶片和根系可溶性糖／可溶性蛋白质比例有利于氮同化和生长．因此,增铵营养下提高了叶片净光合速率、可溶性糖含量和硝酸还原酶活性。维持较高叶片和根系可溶性糖／蛋白质比例。从而促进小麦生长．     

Seedling Nitrate Reductase Activity and Nitrate Partitioning in Maize (Zea mays L.) Strains Divergently Selected For Post-anthesis Leaf-Lamina Nitrate Reductase Activity

Two experiments were conducted to evaluate the effects of phenotypicrecurrent selection for high and low post-anthesis leaf-laminain vivo NRA on nitrate uptake, nitrate partitioning and in vitroNRA of seedling roots and leaves. In Experiment 1, intact plantsof cycle 0, 4, and 6 of the high and low NRA strains were grownon NH₄-N for 11 d, then exposed to 1.0 mol m^–3 KNO₃, andcultures sampled at 6 h and 28 h (induction and post-inductionperiods). Nitrate uptake, tissue nitrate concentration and invitro NRA were determined. The pattern of response to selectionin seedling leaf NRA was similar to that observed for in vivoNRA of field grown plants. Leaf NRA increased between 6 h and28 h. Root NRA was not affected by selection or sampling time.Treatments differed in total fresh weight but not in reductionor uptake of nitrate per unit weight, indicating a lack of correspondencebetween NRA and reduction and supporting the idea that concomitantreduction by NR is not obligatorily linked to nitrate influxin the intact plant. In Experiment 2, dark-grown plants of cycle 0, and 6 of thehigh and low NRA strains were cultured without N, detopped onday 6, transferred the following day to 0-75 mol m^–3 KNO₃and sampled at 6 h and 28 h. In contrast to Experiment 1, selectionpopulations differed in nitrate reduction and root NRA, whichby 28 h reached higher average levels than root NRA of intactplants. Translocation and reduction were inversely related amongstrains within each sampling time. The high level of translocationin detopped plants of the low NRA strain was difficult to reconcilewith its low leaf NRA level of Experiment 1. It is suggestedthat nitrate transport in detopped roots is altered relativeto the intact system in a way which permits greater NRA inductionand nitrate reduction. The results indicate that nitrate partitioningby detopped root systems should be interpreted with caution. Key words: Zea, nitrate reductase activity, nitrate uptake, nitrate reduction, nitrate partitioning, selection     

Induction and Turnover of Nitrate Reductase in Zea mays (Influence of Light)

Both light and NO3- are necessary for the appearance of nitrate reductase (NR) activity (NRA) in photosynthetic tissues. To define the light effect more precisely, we examined the response to light/dark transitions on NRA, NR protein (NRP), and NR mRNA in 6-d-old maize (Zea mays cv W64A x W182E) seedlings that had been grown in a light/dark regime for 5 d and then induced with 5 mM KNO3 for 24 h. The decay of NRA and NR mRNA in the shoot was immediate, but there were only minor changes in NRP during the initial 4 h in the dark. In root tissues, in contrast, there was a 4-h delay in the loss of NRA, NRP, and NR mRNA after transfer to the dark. When the seedlings were returned to light after a 2-h interval in the dark, shoot NRA reached 92% of the initial levels within 30 min of illumination. These results indicate that in the shoots (a) NR message production requires light and (b) the NRP that appears with light treatment and that is active is inactivated in the dark. The NRP can be reactivated when the light is turned on after short periods of darkness (2 h). Root tissues, on the other hand, probably respond to the supply of photosynthetically produced metabolites rather than to immediate products of the light reactions of photosynthesis.     

Time-course of Nitrate Uptake and Nitrate Reductase Activity in Nitrogen-depleted Dwarf Bean

The rate of nitrate uptake by N-depleted French dwarf bean (Phaseolus vulgaris L. cv. Witte Krombek) increased steadily during the first 6 h after addition of NO₃ -After this initial phase the rale remained constant for many hours. Detached root systems showed the same time-course of uptake as roots of intact plants. In vivo nitrate reductase activity (NRA) was assayed with or without exogenous NO₃^- in the incubation medium and the result ing activities were denoted potential and actual level, respectively. In roots the difference between actual and potential NRA disappeared within 15 min after addition of nitrate, and NRA increased for about 15 h. Both potential and actual NRA were initially very low. In leaves, however, potential NRA was initially very high and was not affected by ambient nitrate (0.1–5 mol m^-3) for about 10 h. Actual and potential leaf NRA became equal after the same period of time. In the course of nitrate nutrition, the two nitrate reductase activities in leaves were differentially inhibited by cycloheximide (3.6 mmol m^-3) and tungstate (1 mol m^-3). We suggest that initial potential NRA reflects the activity of pre-existing enzyme, whereas actual NRA depends on enzyme assembly during NO₃^- supply. Apparent induction of nitrate uptake and most (85%) of the actual in vivo NRA occurred in the root system during the first 6 h of nitrate utilization by dwarf bean.     

小麦叶内硝酸还原的代谢库

随营养液中No_3~-浓度升高,叶片内No_3~-总量、代谢库大小(NIPS)及硝酸还原酶(NR)活性均升高,其中MPS与NK活性呈同步变化;No_3~-浓度达2.0mmol/L时,两者趋于稳值;若再增加NO_3~-浓度,则被吸收的NO_3~-积累于液泡中,而代谢库中NO_3~-含量(MPS)与NO_3~-总量之比有一定程度降低。低氮(NO_3~-浓度为1.0 mmol/L)情况下,反应液中无NO_3~-时,叶片内NR活性品种间有差异,但在50 mmol/L NO_3~-反应液中则品种间无差异;NK活性高的品种鲁麦8号及品种321叶内有大的NO_3~-代谢库,反应液中NO_3~-对NR活性刺激程度低,代谢库NO_3~-含量与叶NO_3~-总量之比高,而叶组织长时间反应过程中其NR活性衰减速率低。     

Nitrate Reductase Activity in Maize (Zea mays L.) Leaves: II. Regulation by Nitrate Flux at Low Leaf Water Potential

Experiments were conducted to determine whether the nitrate flux to the leaves or the nitrate content of the leaves regulated the nitrate reductase activity (NRA) in leaves of intact maize (Zea mays L.) seedlings having low water potentials (ψ_w) when other environmental and endogenous factors were constant. In seedlings that were desiccated slowly, the nitrate flux, leaf nitrate content, and NRA decreased as ψ_w decreased. The decrease in nitrate flux was caused by a decrease in both the rate of transpiration and the rate of nitrate delivery to the transpiration stream. Upon rewatering, the recovery in NRA was correlated with the nitrate flux but not the leaf nitrate content.     

Effects of Root Temperature and Form of Nitrogen Nutrition on Nitrate Reductase Activity in Oilseed Rape (Brassica napus L.)

The effect of root temperature and form of inorganic nitrogensupply on in vitro nitrate reductase activity (NRA) was studiedin oilseed rape (Brassica napus L. cv. bien venu). Plants weregrown initially in flowing nutrient solution containing 10 µMNH₄NO₃ and then supplied with either nitrate or ammonium for15 d at root temperatures of 3, 7, 11 or 17 °C. Shoot temperatureregime was similar for all plants; 20/15 °C, day/night.Root NRA was highest when roots were grown at 3 and 7 °C.In laminae and petioles NRA was highest when roots were 11 or17 °C. The plants supplied with ammonium had much lowerlevels of NRA in roots after 5 d than the plants supplied onlywith nitrate. NRA in the laminae of plants supplied with ammoniumwas low relative to that in plants supplied with nitrate onlywhen root temperature was 11 or 17 °C. Values of the apparent activation energy (E_a) of NR, calculatedfrom the Arrhenius equation, in laminae and petioles were differentfrom roots suggesting difference in enzyme conformation. Evidencethat the temperature at which roots were growing affected E_awas equivocal. Oilseed rape, Brassica napus L., activation energy, ammonium, Arrhenius equation, nitrate, root temperature, nitrate reductase     

Nitrate Reductase Activity in Maize (Zea mays L.) Leaves: I. Regulation by Nitrate Flux

The roles that leaf nitrate content and nitrate flux play in regulating the levels of nitrate reductase activity (NRA) were investigated in 8- to 14-day old maize (Zea mays L.) plants containing high nitrate levels while other environmental and endogenous factors were constant. The nitrate flux of intact plants was measured from the product of the transpiration rate and the concentration of nitrate in the xylem. NRA decreased when the seedlings were deprived of nitrate. The nitrate flux and the leaf nitrate content also decreased. When nitrate was resupplied to the roots, all three parameters increased.     

Triadimenol Increases Nitrate Levels and Nitrate Reductase Activity in Canola Leaves

Nitrate reductase activity in the first true leaves of canola(Brassica napus L.) seedlings grown in one-quarter strengthHoagland's solution from seeds pretreated with triadimenol (0.3or 30 g (a.i.) kg^–1 of seed) was higher than controlsduring the growth period of 15 to 25 d after planting. Triadimenolalso increased chlorophyll levels, the increase being more pronouncedat its lower concentration. The treatment also increased theweight and nitrate content of the leaves. When seedlings weregrown in nutrient solution containing 1 to 20 mM nitrate, theincrease in nitrate reductase activity by triadimenol was higherat lower rather than at higher nitrate concentrations. The nitratelevels and Kjeldahl nitrogen in the triadimenol-treated leaveswas higher than the controls at concentrations of added nitrateabove 2 mM. Addition of nitrate to plants grown in ammonium,increased nitrate reductase activity more in plants grown fromtriadimenol-treated seeds than controls. However, addition of10µM triadimenol for 24 h to ammonium-grown plants hadlittle effect on enzyme activity, both in the absence as wellas the presence of nitrate. This study demonstrates that triadimenolincreases nitrate reductase activity and nitrate accumulationin the leaves and at least part of the increased enzyme activityis independent of nitrate accumulation. Key words: Triazoles, nitrate content, nitrate reductase activity