硝酸还原酶抑制剂钨酸钠对油菜硝态氮积累的影响

采用溶液培养方法,选取硝酸盐积累差异明显的两个油菜品种（低硝态氮积累品种‘红油3号’和高硝态氮积累品种‘中双6号’,研究苗期根系硝酸还原酶（NR）活性被抑制以后两个油菜品种叶片、叶柄和根系中NR活性和硝态氮含量的变化。结果表明：1.0mmol.L-1的NR活性抑制剂Na2WO4对两个油菜品种的根系NR活性抑制效果最佳;根系NR活性被抑制以后,两个油菜品种的根系NR活性、硝态氮吸收速率均显著下降,而硝态氮含量却显著上升;且Na2WO4对‘中双6号’硝态氮吸收的抑制程度强于其对‘红油3号’的抑制。叶片和叶柄的NR活性变化不显著,但叶柄硝态氮含量显著下降,叶片硝态氮含量稳定,且这一趋势在低积累品种‘红油3号’中表现得更为明显。     

菠菜叶片中硝态氮还原与叶柄中硝态氮累积的关系

测定了不同生长期在不同施氮水平下3个菠菜品种各器官的硝态氮含量、叶片的硝酸还原酶活性、叶片细胞硝态氮的贮存库和代谢库大小.结果表明:叶柄中硝态氮含量远高于其它器官,其含量与叶片内源/外源硝酸还原酶活性的比值呈负相关;叶片细胞中硝态氮代谢库的大小与叶柄中硝态氮含量之间没有确定的关系.     

不同覆盖材料对夏秋苹果根区土壤硝酸盐代谢的影响

以3年生新红星苹果树为试验材料,在春季将稻草苫、农用地毯、透明塑料膜和园艺地布覆盖地表,于夏秋季调查根区土壤硝化-反硝化作用、硝酸还原酶(NR)和亚硝酸还原酶(NiR)活性以及铵态氮、硝态氮、亚硝态氮含量和植株生长的变化.结果表明: 4种覆盖处理均降低了夏季土壤硝化强度和夏秋之交的土壤NiR活性,提高了秋季土壤铵态氮含量以及夏秋之交的土壤反硝化强度、NR活性和铵态氮含量,降低了夏秋季土壤硝化强度、反硝化强度和NR活性的变异系数;稻草苫提高了夏季和秋季土壤反硝化强度与硝态氮含量,降低了夏季土壤NR和NiR活性;在4种处理中,稻草苫覆盖的土壤硝化与反硝化强度及NR活性在整个夏秋季的变异系数最低;农用地毯降低了夏季土壤反硝化强度,提高了夏季土壤NR和NiR活性、夏秋之交土壤硝态氮含量和秋季土壤反硝化强度;透明塑料膜降低了夏季土壤硝态氮含量,提高了夏季土壤亚硝态氮含量、夏秋之交土壤硝态氮含量以及秋季土壤硝化强度和NiR活性;园艺地布提高了夏季土壤反硝化强度、夏秋之交和秋季土壤的硝化强度以及秋季土壤硝态氮含量.4种覆盖处理均促进了植株生长,其中稻草苫和园艺地布促进新梢和干径增粗的效果更显著;4种覆盖处理对夏秋季土壤硝酸盐代谢的影响不同,但对土壤硝酸盐代谢与转化都具有稳定作用,其中稻草苫的稳定效果最好.     

外源亚精胺对盐胁迫下黄瓜植株氮化合物含量和硝酸还原酶活性的影响

研究了外源亚精胺(Spd)在营养液栽培中,对盐胁迫下耐盐性不同的两品种黄瓜幼苗体内硝态氮、铵态氮、脯氨酸(Pro)含量和硝酸还原酶(NR)活性的影响。结果表明,外源Spd显著减小了盐胁迫引起的铵态氮、Pro含量的升高幅度和NR活性、硝态氮含量的降低幅度,且对盐敏感型黄瓜品种影响幅度较大。表明Spd可明显减缓盐胁迫对黄瓜幼苗氮素营养代谢的影响。     

外源亚精胺对高温胁迫下黄瓜幼苗氮素代谢的影响

以较为耐热的黄瓜品种‘津春4号’为试材,在人工气候箱中,采用石英砂培加营养液浇灌的栽培方式,研究了外源亚精胺( Spd)对高温胁迫(42℃)下黄瓜幼苗氮素代谢的影响.结果表明:短期高温胁迫处理,尤其是4h内,植株硝态氮含量降低而铵态氮含量升高;外源Spd预处理使幼苗体内硝态氮和铵态氮含量升高且硝酸还原酶(NR)活性增强.较长期高温胁迫处理下,幼苗根系中硝态氮含量升高但向地上部运输受阻,根系NR钝化,根系和叶片中铵态氮含量均显著升高;高温胁迫下喷施Spd,除进一步促进根系吸收硝态氮且向地上部运输外,根系和叶片NR活性亦有所升高,从较长期的效果看,外源Spd还具有防止铵态氮过度积累、促进幼苗体内氮素代谢趋于正常的作用.     

缺氮和复氮对菘蓝幼苗生长及氮代谢的影响

对基质育苗后水培的菘蓝进行缺氮与复氮处理,分析其生长情况及氮代谢产物含量的变化,探讨缺氮和复氮对菘蓝幼苗生长及氮代谢的影响,以提高菘蓝产量和品质以及栽培过程中的氮素利用效率。结果显示:(1)正常供氮条件下,菘蓝幼苗的叶绿素含量、谷氨酰胺合成酶(GS)活性、硝态氮含量、靛玉红含量为最高,而其株高、主根直径、根的鲜重与干重、叶的鲜重与干重、根系活力均最小。(2)缺氮处理增加了菘蓝幼苗的主根直径和根干重,提高其根系活力和硝酸还原酶(NR)活性,促进游离氨基酸在叶中的积累;但降低了GS的活性,也降低了叶中硝态氮、可溶性蛋白、靛玉红及根中游离氨基酸的含量;缺氮对叶中靛蓝的含量无明显影响。(3)复氮处理增加了菘蓝幼苗的株高、主根长、根鲜重、叶鲜重、叶干重,提高了其根系活力,降低了NR和GS的活性;与对照相比,复氮降低了叶中硝态氮含量,提高了叶中可溶性蛋白、靛蓝及根中游离氨基酸的含量,但对叶中游离氨基酸和靛玉红含量影响较小。研究表明,缺氮后再复氮有利于菘蓝幼苗叶的生长,同时有利于增加其叶内靛蓝含量,从而提高其产量和品质。     

叶施NO对NaCl胁迫下红砂幼苗叶片和根系中氮代谢酶及营养物质的影响

以当年生红砂(Reaumuria soongorica)幼苗为材料,采用盆栽实验,考察叶面喷施不同浓度(0、0.01、0.10、0.25、0.50、1.00 mmol·L^-1)NO供体硝普钠 (SNP) 对NaCl(300 mmol·L^-1)胁迫下红砂根、叶中可溶性蛋白、游离氨基酸和硝态氮含量,以及谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、硝酸还原酶(NR)活性的影响,并采用主成分分析和隶属函数法筛选NO对NaCl胁迫缓解效应的氮代谢指标和最佳NO浓度,以探讨外源NO对NaCl 胁迫下红砂缓解效应的氮代谢响应机制。结果表明：(1)在300 mmol·L^-1 NaCl胁迫处理下,红砂幼苗根、叶中可溶性蛋白、硝态氮含量以及GS、GOGAT、NR活性均比对照显著下降。(2)外源NO能显著提高盐胁迫下红砂叶、根中GS、GOGAT、NR活性和硝态氮含量,增加根中可溶性蛋白和游离氨基酸含量。(3)NR和GOGAT活性可用于评价NO对NaCl胁迫下红砂幼苗的缓解作用,外源NO(SNP)对红砂幼苗在NaCl胁迫下的缓解效果强弱表现为0.25 mmol·L^-1> 0.50 mmol·L^-1> 0.10 mmol·L^-1> 1.00 mmol·L^-1> 0.01 mmol·L^-1。研究发现,300 mmol·L^-1 NaCl胁迫显著抑制了红砂幼苗氮代谢,外源NO(SNP)有助于提高盐胁迫下红砂NR活性,加快硝态氮转化为铵态氮,促进红砂叶片和根中GS/GOGAT对转化物的同化,从而增强红砂幼苗的耐盐性,并以0.25 mmol·L^-1SNP处理时缓解作用最佳;NR和GOGAT活性可作为NO缓解盐胁迫的评价指标。     

不同氮素形态配比对甘薯前期氮代谢的影响及其生理机制

以3种不同类型的甘薯(Ipomoea batatas (L.) Lam.)为实验材料,根据氮素的3种形态设置5个配比处理(N1～N5),分别在栽秧后15、25和35 d取样测定甘薯不同器官的氮含量、功能叶氮代谢酶活性变化以及酶调控基因表达情况。结果显示:在同一生育期,N4和N5处理铵态氮和硝态氮配施下植株氮素的积累量明显高于其它处理;在甘薯生长发育前期,叶片含氮量先降低后上升,茎、须根和膨大根以及全株含氮量均呈上升趋势; N4处理能够显著提高硝酸还原酶(NR)、谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH)活性; N3处理能够明显提高谷氨酸合成酶(GOGAT)的活性; NR活性随肥料中硝态氮比例的增加而提高;增加肥料配比中硝态氮比例可使调控NR活性的基因上调表达,N4和N5处理可使GS调控基因上调表达,但抑制GOGAT调控基因的表达。酰胺态氮在前期对氮代谢相关酶调控基因无显著影响。研究结果表明,在甘薯生长发育前期,硝态氮和铵态氮配施能够显著提高氮素积累量、代谢酶活性和调控基因表达量,铵态氮∶硝态氮∶酰胺态氮=1∶2∶0的配施方案为本实验条件下的最佳配施组合。     

氮素形态对樱桃番茄果实发育中氮代谢的影响

以樱桃番茄为材料,采用基质 营养液共培养的方法,研究了全硝态氮(NO₃^-）、铵态氮和硝态氮配施（75%NO₃^-∶25%NH₄⁺）及全铵态氮（NH₄⁺）营养对樱桃番茄果实氮代谢及硝酸还原酶（NR）和谷氨酰胺合成酶（GS）基因表达的影响.结果表明： 铵态氮和硝态氮配施处理下樱桃番茄的单果质量比全硝态氮处理略有增加,且果实中NH₄⁺、总氨基酸、氮含量和氮素累积量均显著高于全硝态氮处理;全硝态氮及铵态氮和硝态氮配施处理下果实NR活性及其基因表达没有明显差异,但都显著高于全铵态氮处理;铵态氮和硝态氮配施处理下果实GS活性都高于全硝态氮处理.不同形态氮素及配施处理下,同工酶GS1（胞质型GS）和GS2（叶绿体型GS）的表达与GS的活性不一致,说明氮素对GS活性的影响主要发生在转录后水平.     

氮肥形态对冬小麦根际土壤氮素生理群活性及无机氮含量的影响

采用大田盆栽方法研究了硝态氮肥、铵态氮肥、酰胺态氮肥3种氮肥形态对冬小麦品种豫麦50生育中后期(拔节期、开花期、花后14 d、花后28 d)根际土壤氮转化相关微生物活性、酶活性和根际土壤NH+4离子、NO-3离子含量的影响。结果表明:随着生育期的推进,除脲酶外,氨化细菌、硝化细菌、亚硝化细菌、反硝化细菌和蛋白酶活性变化的均为"倒V"型变化特征,以花后14 d活性最强;而脲酶活性在拔节期最强,并且其活性远大于其它微生物及酶。氮肥形态对根际土壤氮素生理群及无机氮的影响不同。酰胺态氮肥促进了根际氨化细菌、反硝化细菌、脲酶、蛋白酶的活性,而硝化细菌、亚硝化细菌在硝态氮肥条件下活性较强。除拔节期外,土壤中NH+4离子在铵态氮肥处理下含量较高,NO-3离子在酰氨态氮肥处理下含量较高。因此,酰胺态氮能够促进小麦根际土壤有机氮的分解,硝态氮肥可以促进土壤中氨的转化,以利于小麦根系的吸收与利用。氮肥形态主要是通过影响土壤中氮素生理类群及酶的活性,从而影响土壤中无机氮的含量。     

外源脯氨酸对盐胁迫下甜瓜幼苗硝酸还原的影响

采用营养液水培方法,以＂雪美＂品种甜瓜（Cucumis melo L.）为材料,研究了外源脯氨酸（Proline）对盐胁迫下甜瓜幼苗叶片和根系硝酸还原的影响。结果表明：（1）盐胁迫提高了甜瓜幼苗叶片和根系内铵态氮（NH4＋-N）和可溶性蛋白含量;降低了硝态氮（NO-3-N）含量和硝酸还原酶（nitrate reductase,NR）活性。（2）外源施用脯氨酸明显地提高了盐胁迫下甜瓜幼苗叶片和根系内NO-3-N和可溶性蛋白含量;降低了盐胁迫下甜瓜幼苗叶片和根系内NH＋4-N含量;增强了盐胁迫下甜瓜幼苗体内NR活性。研究结果表明,外源脯氨酸可以通过调节甜瓜幼苗体内硝酸还原酶活性和氮化合物含量来缓解盐胁迫对甜瓜幼苗植株的伤害。     

不同氮效率水稻生育后期根表和根际土壤硝化特征

通过田间试验研究了不同氮效率粳稻品种4007(氮高效)和Elio(氮低效)生育后期在N0(0 kgN hm-2)、N180(180 kgN hm-2)和N300(300 kgN hm-2)水平下根表、根际和土体土壤pH值、铵态氮(NH+4-N)和硝态氮(NO-3-N)含量、硝化强度和氨氧化细菌(AOB)数量.结果表明无论是齐穗期、灌浆期还是成熟期,根表土壤pH值均显著低于根际和土体土壤.土壤pH值范围在5.95至6.84之间变化.土壤NH+4-N含量随水稻生长显著下降,且随施氮量增加而显著增加.根表土壤NH+4-N有明显亏缺区,且随距水稻根表距离增加,NH+4-N含量逐渐升高.土壤NO-3-N含量随水稻生长显著增加,施氮处理均显著高于不施氮处理,但N180和N300处理差异不显著.NO-3-N含量表现为根际>土体>根表.水稻根表和根际土壤硝化强度随水稻生长显著下降,而土体土壤硝化强度随时间延长小幅增加.施氮显著提高4007水稻根表土壤在齐穗和收获期硝化强度以及Elio在齐穗期根际硝化强度,但在施氮处理N180和N300中无显著差异.在整个采样期间,土壤硝化强度均表现为根际>根表>土体.水稻根表和根际AOB数量随水稻生长而显著降低,而土体土壤AOB数量无显著变化.例如,根表土壤AOB数量在齐穗期、灌浆期和收获期分别为16.7×105、8.77×105个g-1 dry soil和8.01×105个g-1 dry soil.根表和根际土壤AOB数量无显著差异,但二者显著高于土体土壤AOB数量.就两个氮效率水稻品种而言,土壤pH值基本无差异.4007土壤NH+4-N含量均显著高于Elio.在齐穗期水稻根表、根际和土体土壤NO-3-N含量在N180水平下均表现为Elio显著高于4007.而在灌浆期和收获期,水稻根表、根际和土体土壤则表现为4007显著高于Elio.在所有采样期,两个水稻品种土体土壤硝化强度和AOB数量在3个施氮量下均无显著差异.Elio根表和根际土壤硝化强度和AOB数量在水稻灌浆期之前一直显著高于4007,而在灌浆期之后则显著低于4007,且最终产量和氮素利用率(NUE)显著低于4007,这可能是由于4007灌浆期后硝化作用强,根际产生的NO-3-N含量高,从而4007根吸收NO-3-N的量也高造成的.因此水稻灌浆期和收获期根表和根际硝化作用以及AOB与水稻高产及氮素高效利用密切相关.     

不同氮效率水稻全生育期内对增硝营养的响应及其生理机制

通过添加硝化抑制剂（二氰胺,DCD）来控制硝化作用的水培试验方法,研究了氮高效水稻品种南光和氮低效水稻品种ELIO的籽粒产量对增硝营养（NH4＋∶NO3-比例为100∶0和75∶25）的响应,同时从产量构成、不同生育时期水稻生长、氮素吸收和同化4个方面研究了造成其产量差异的生理机制。结果表明：增NO3-营养可以显著促进氮高效水稻品种南光的生长,从而使其籽粒产量水平提高21%,而对氮低效水稻品种ELIO的籽粒产量没有显著影响。进一步分析表明：在增NO3-营养条件下,南光的穗粒数增加了25%,结实率增加了16%,而氮低效水稻品种ELIO的结实率和穗粒数在两种营养条件下没有显著变化;增NO3-营养可以促进南光对氮素的吸收,使其在苗期、分蘖盛期、齐穗期和成熟期对氮素的吸收量平均增加了36%,进而促进了其生长,干物质积累量在四个生育时期平均增加了30%;南光叶片硝酸还原酶和根系谷氨酰胺合成酶的活力在增硝营养条件下分别增加了100%和95%,说明增硝营养促进了南光对NH4＋和NO3-的同化利用。与氮低效水稻品种（ELIO）相比,氮高效水稻品种（南光）对增硝营养表现出较强的生理响应。     

Influence of Different Concentrations of NO-3 and NH+4 on the Activity of Glutamine Synthetase and Other Relevant Enzymes of Nitrogen Metabolism in Wheat Roots

Influence of different concentrations of NO3 and NH+ on the activity of glutamine synthetase (GS), asparagine synthetase (AS), glutamate dehydrogenase (GDH), nitrate reductase (NR) and the changes of GS-mRNA in wheat roots have been studied with enzymes activity assay and Northern blot. The results showed that the higher GS activity was found in roots of wheat when NH+4-N was the sole nitrogen source than when NO3-N was the sole nitrogen source. GS-mRNA of Northern blot was simillar to GS activity. 3 mmol/L NO3- promoted the activity of AS. The change of AS was independent of the change of GS. GDH activity was not been detected, and change in regulation of NR activity was not found.     

Nitrate reductase activity of two leafy vegetables as affected by nickel and different nitrogen forms

The author studied the effect of different nickel concentrations (0, 0.4, 40 and 80 μM Ni) on the nitrate reductase (NR) activity of New Zealand spinach (Tetragonia expansa Murr.) and lettuce (Lactuca sativa L. cv. Justyna) plants supplied with different nitrogen forms (NO₃ ⁻–N, NH₄ ⁺–N, NH₄NO₃). A low concentration of Ni (0.4 μM) did not cause statistically significant changes of the nitrate reductase activity in lettuce plants supplied with nitrate nitrogen (NO₃ ⁻–N) or mixed (NH₄NO₃) nitrogen form, but in New Zealand spinach leaves the enzyme activity decreased and increased, respectively. The introduction of 0.4 μM Ni in the medium containing ammonium ions as a sole source of nitrogen resulted in significantly increased NR activity in lettuce roots, and did not cause statistically significant changes of the enzyme activity in New Zealand spinach plants. At a high nickel level (Ni 40 or 80 μM), a significant decrease in the NR activity was observed in New Zealand spinach plants treated with nitrate or mixed nitrogen form, but it was much more marked in leaves than in roots. An exception was lack of significant changes of the enzyme activity in spinach leaves when plants were treated with 40 μM Ni and supplied with mixed nitrogen form, which resulted in the stronger reduction of the enzyme activity in roots than in leaves. The statistically significant drop in the NR activity was recorded in the aboveground parts of nickel-stressed lettuce plants supplied with NO₃ ⁻–N or NH₄NO₃. At the same time, there were no statistically significant changes recorded in lettuce roots, except for the drop of the enzyme activity in the roots of NO₃ ⁻-fed plants grown in the nutrient solution containing 80 μM Ni. An addition of high nickel doses to the nutrient solution contained ammonium nitrogen (NH₄ ⁺–N) did not affect the NR activity in New Zealand spinach plants and caused a high increase of this enzyme in lettuce organs, especially in roots. It should be stressed that, independently of nickel dose in New Zealand spinach plants supplied with ammonium form, NR activity in roots was dramatically higher than that in leaves. Moreover, in New Zealand spinach plants treated with NH₄ ⁺–N the enzyme activity in roots was even higher than in those supplied with NO₃ ⁻–N.     

Nitrate-independent expression of plant nitrate reductase in Lotus japonicus root nodules

Nitrate-independent nitrate reductase (NR) activity is generally found in legume root nodules. Therefore, the effects of nitrate on plant NR activity and mRNA were investigated in the root nodules of Lotus japonicus (L. japonicus). Both NR activity and mRNA levels in roots and root nodules were up-regulated by the addition of nitrate. In the absence of nitrate, NR activity and mRNA were detected in root nodules but not in roots. Southern blotting analysis indicates that NR is encoded by a single gene in L. japonicus. No nitrate was detected in the root nodules or roots of plants grown in the absence of nitrate, while its accumulation was observed in plants supplied with exogenous nitrate. These results indicate that inducible-type NR can be expressed in root nodules in the absence of nitrate. The activation state of the nitrate-independent activity of NR was as high as that of NR activity induced by nitrate. NR mRNA expressed independently of nitrate in root nodules without nitrate was localized in the infected regions of the root nodules. Thus, the expression could be related to the specific structure and environment of root nodules.     

Rapid modulation of nitrate reductase in pea roots

The regulatory properties of nitrate reductase (NR; EC 1.6.6.1) in root extracts from hydroponically grown pea (Pisum sativum L. cv. Kleine Rheinländerin) plants were examined and compared with known properties of NR from spinach and pea leaves. Nitrate-reductase activity (NRA) extracted from pea roots decreased slowly when plants were kept in the dark, or when illuminated plants were detopped, with a half-time of about 4 h (= slow modulation in vivo). In contrast, the half-time for the dark-inactivation of NR from pea leaves was only 10 min. However, when root tip segments were transferred from aerobic to anaerobic conditions or vice versa, changes in NRA were as rapid as in leaves (= rapid modulation in vivo). Nitrate-reductase activity was low when extracted from roots kept in solutions flushed with air or pure oxygen, and high in nitrogen. Okadaic acid, a specific inhibitor of type-1 and type-2A protein phosphatases, totally prevented the in vivo activation by anaerobiosis of NR, indicating that rapid activation of root NR involved protein dephosphorylation. Under aerobic conditions, the low NRA in roots was also rapidly increased by incubating the roots with either uncouplers or mannose. Under these conditions, and also under anaerobiosis, ATP levels in roots were much lower than in aerated control roots. Thus, whenever ATP levels in roots were artificially decreased, NRA increased rapidly. The highly active NR extracted from anaerobic roots could be partially inactivated in vitro by preincubation of desalted root extracts with MgATP (2 mM), with a half-time of about 20 min. It was reactivated by subsequently incubating the extracts with excess AMP (2 mM). Thus, pea root NR shares many of the previously described properties of NR from spinach leaves, suggesting that the root enzyme, like the leaf enzyme, can be rapidly modulated, probably by reversible protein phosphorylation/ dephosphorylation.     

Foliar and fine root nitrate reductase activity in seedlings of four forest tree species in relation to nitrogen availability

Summary Seedlings of red maple, white pine, pitch pine and red pine were fertilized with nutrient solutions containing 4 levels of nitrate or ammonium additions. These levels corresponded to approximately 0.5, 1, 2 and 4 times normal availability of nitrogen in northeastern forests. Nitrate reductase (NR) activity was assayed in roots and leaves. Red maples treated with nitrate showed much higher leaf activities and higher ratios of leaf NR activity to root NR activity than any other species. Ammonium additions to red maple and white pine appeared to inhibit NR activity in leaves. With high nitrate additions, NR activity was induced in roots and leaves of pine species, but activity in roots remained much higher than in leaves.     

Nitrate regulation of nitrate uptake and nitrate reductase expression in barley grown at different nitrate:ammonium ratios at constant relative nitrogen addition rate

Barley (Hordeum vulgare L. cv. Golf) was cultured using the relative addition rate technique, where nitrogen is added in a fixed relation to the nitrogen already bound in biomass. The relative rate of total nitrogen addition was 0.09 day^?1 (growth limiting by 35%), while the nitrate addition was varied by means of different nitrate: ammonium ratios. In 3- to 4-week-old plants, these ratios of nitrate to ammonium supported nitrate fluxes ranging from 0 to 22 μmol g^?1 root dry weight h^?1, whereas the total N flux was 21.8 ± 0.25 μmol g^?1 root dry weight h^?1 for all treatments. The external nitrate concentrations varied between 0.18 and 1.5 μM. The relative growth rate, root to total biomass dry weight ratios, as well as Kjeldahl nitrogen in roots and shoots were unaffected by the nitrate:ammonium ratio. Tissue nitrate concentration in roots were comparable in all treatments. Shoot nitrate concentration increased with increasing nitrate supply, indicating increased translocation of nitrate to the shoot. The apparent V_max for net nitrate uptake increased with increased nitrate fluxes. Uptake activity was recorded also after growth at zero nitrate addition. This activity may have been induced by the small, but detectable, nitrate concentration in the medium under these conditions. In contrast, nitrate reductase (NR) activity in roots was unaffected by different nitrate fluxes, whereas NR activity in the shoot increased with increased nitrate supply. NR-mRNA was detected in roots from all cultures and showed no significant response to the nitrate flux, corroborating the data for NR activity. The data show that an extremely low amount of nitrate is required to elicit expression of NR and uptake activity. However, the uptake system and root NR respond differentially to increased nitrate flux at constant total N nutrition. It appears that root NR expression under these conditions is additionally controlled by factors related to the total N flux or the internal N status of the root and/or plant. The method used in this study may facilitate separation of nitrate-specific responses from the nutritional effect of nitrate.