生境因子作用下NO_3~-/NH_4~+吸收及硝酸还原酶活性变化

土壤中的氮素因土壤类型和季节变化产生异质性。在长期的进化过程中,植物适应各自的氮营养生境,形成了对NO3-/NH4+吸收的分子机制。饱和高亲和传输系统(HATS)中,植物在不同的转录基因控制下吸收NO3-/NH4+,表现出对两种氮源的偏选性。这种偏选性主要取决于植物种的特性,但是NO3-/NH4+的吸收受光照、介质N强度、pH值、外源氨基酸和温度等生境因子的影响,同时植物的营养生境也因NO3-/NH4+的吸收被深刻改造。硝酸还原酶(NR)在氮同化过程中作用于NO3-还原阶段,其活性受各种生境因子的制约,影响植物对NO3-吸收利用。     

增硝营养对不同基因型水稻苗期吸铵和生长的影响

随着水稻节水栽培技术越来越得到广泛的应用与推广以及水稻在淹水条件下根际氧化圈的存在,水稻的硝酸盐营养作用受到更大的关注。利用水培方法研究了4种具代表性的水稻基因型(常规籼稻、常规粳稻、杂交籼稻、杂交粳稻)在苗期(2 8d)的铵(NH 4 )吸收动力学特性以及硝(NO-3 )对NH 4 吸收动力学特征和叶片谷氨酰胺合成酶活性(GSA)的影响。结果表明:增NO-3 营养可以增加水稻对氮素的吸收,提高氮素利用率,进而促进水稻生长;不同基因型之间NH 4 吸收速率的差异为:杂交籼稻>常规籼稻>杂交粳稻>常规粳稻;NO-3 的存在促进了水稻对NH 4 的吸收,增加水稻吸收NH 4 的Vmax值(4个品种平均增加31 5 % ) ,而对其Km 值影响不大(4个品种平均增加4 2 6 % ) ,说明NO-3 对NH 4 吸收的影响主要在于影响NH 4 载体的运转速率而非吸收位点与NH 4 之间的亲和性;增NO-3 营养可以增加叶片谷氨酰胺合成酶(GS)的活性,提高水稻同化NH 4 的能力     

利用gs1.1和gs1.2突变体研究外源蔗糖对高铵胁迫拟南芥碳氮代谢的影响

以拟南芥野生型Col-0、谷氨酰胺合成酶敲除突变体gs1.1和gs1.2为实验材料,采取土培试验,比较正常培养液(4 mmol/L NH4⁺)培养(CK)、正常培养液(4 mmol/L NH4⁺)下外源添加5%蔗糖(T1)、高NH4⁺胁迫(20 mmol/L)(T2)以及高NH4⁺胁迫(20 mmol/L)下外源添加5%蔗糖(T3)对拟南芥各株系各生理指标的影响;通过测定地上部分的鲜重、叶绿素、游离NH4⁺、可溶性糖、可溶性蛋白、谷氨酰胺合成酶(GS)、谷氨酸脱氢酶(GDH)、矿质元素含量等指标,研究外源蔗糖对NH4⁺胁迫拟南芥碳氮代谢的影响。结果表明,高NH4⁺胁迫下,拟南芥生长受到严重的抑制,鲜重、GS、GDH酶活性降低,游离NH4⁺含量、叶绿素含量、可溶性糖和可溶性蛋白含量增加,植株的N、P、K、Ca的含量增加,Mg、Fe的含量减少,其中gs1.1和gs1.2在高NH4⁺处理下受到的抑制比Col-0更为显著。外源添加5%蔗糖显著缓解了高NH4⁺毒害,提高了可溶性糖和可溶性蛋白含量,提高了GS和GDH的活性,降低了叶绿素和游离NH4⁺的含量,提高了植株体内的N、P、K、Ca,Mg的含量,降低了植株Fe的含量,其中,外源蔗糖对gs1.1和gs1.2高NH4⁺毒害的缓解更为显著。     

不同氮素形态比例条件下接种AMF对玉米氮同化关键酶的影响

以珍珠岩为基质,通过供应3种不同的NH4+∶NO3ˉ比例营养液,研究了接种丛枝菌根真菌对玉米氮同化关键酶活性的影响。结果看出,与不接种的玉米植株相比,接种Glomus intraradices和Glomus mosseae分别在NH4+∶NO3ˉ=3∶1和NH4+∶NO3ˉ=1∶3形态下提高了植物叶片的硝酸还原酶活性;接种AMF对叶片谷氨酰胺合成酶活性（GS）影响不大,但在NH4+∶NO3ˉ=3∶1形态下接种3种AMF处理均显著提高了根系GS活性,相对提高了铵态氮在地下部的同化比例。在铵态氮比例较高时,接种AMF的促生效应较好,且AMF提高根系GS活性作用较大。表明丛枝菌根真菌在促进宿主植物对铵态氮的利用作用较大。     

氮素诱导的甜菜gln2的克隆及序列分析

本文探索了在氮素诱导下,甜菜gln2序列变化情况及氮素对gln2的调控表达情况。根据已知甜菜谷氨酰胺合成酶基因(gln2,登录号为AY026353)设计特异引物,利用RT-PCR方法克隆甜菜质体型谷氨酰胺合成酶的cDNA(GS2 cDNA)片段和甜菜质体型谷氨酰胺合成酶基因组DNA(GS2 DNA)序列,并进行生物信息学分析。获得甜菜GS2 cDNA片段序列,并首次得到甜菜GS2 DNA序列,并将GS2DNA登录到了NCBI网站的GenBank(登录号为EU558132)。生物信息学分析结果表明,GS2 DNA长度为6 144bp,含有13个外显子和12个内含子;氮素诱导的GS2 cDNA序列长度为1 296bp,与gln2序列相似性达99.92%,可编码431个氨基酸残基;其氨基酸序列与其它植物的质体型谷氨酰胺合成酶(GS2)有很高的同源性,与菠菜GS2的同源性最高,属于混合型蛋白,具有Gln-synt保守功能域,N端为beta-Grasp domain,C端为catalytic domain。采用半定量PCR分析该基因在氮素处理下的诱导表达情况,结果表明,NO3-N∶NH4+-N=80∶20和NO3-N∶NH4+-N=50∶50的处理对GS2基因表达的促进效果最好,NO3-N∶NH4+-N=0∶100对GS基因诱导作用最差。研究甜菜GS基因的结构功能及表达调控对甜菜实现高同化氨及增产增糖有重要意义。     

外源H2S对盐胁迫下红砂幼苗叶片和根系氮代谢的影响

H₂S作为新型气体信号分子在调控植物生长发育和抗逆境胁迫中发挥着重要作用。为探究外源H₂S对盐胁迫下红砂(Reaumuria soongorica)氮代谢的影响机制及最佳叶施浓度,以当年生红砂幼苗为材料,采用盆栽试验,以1/2 Hoagland浇灌为对照(CK)考察在300 mmol·L^-1NaCl胁迫(CK300)下,叶面喷施不同浓度(0、0.010、0.025、0.050、0.100、0.250、0.500、1 mmol·L^-1)H₂S供体硫氢化钠(NaHS)对红砂叶片和根系中硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合成酶(GOGAT)活性以及硝态氮、可溶性蛋白和游离氨基酸含量的影响。结果表明,盐胁迫下,红砂幼苗根、叶中可溶性蛋白、游离氨基酸、硝态氮含量以及NR、GOGAT、GS活性均较对照(CK)显著下降。经不同浓度外源H₂S处理后,红砂根、叶中可溶性蛋白含量与单独盐胁迫对照(CK300)相比显著降低,NR、GOGAT、GS活性和硝态氮、游...     

外源ABA提高甘蓝型油菜抗镉胁迫能力和氮素生理利用效率

  【目的】  植物脱落酸 (ABA) 可调节植物生长、种子休眠和叶片衰老,还可对非生物胁迫作出反应。比较添加外源ABA前后镉 (Cd) 胁迫下甘蓝型油菜的生理反应,可深化对脱落酸提高作物抗性和氮素生理利用效率作用机理的认识。  【方法】  供试甘蓝型油菜品种为814号,试验设3个处理:正常培养 (CK)、10 μmol/L CdCl₂ (Cd) 和5 μmol/L ABA + 10 μmol/L CdCl₂ (A + Cd)。在处理4天后,观察甘蓝型油菜表型,测定脯氨酸及丙二醛 (MDA) 含量、Cd含量、硝酸盐和铵盐含量以及硝酸还原酶 (NR)、谷氨酰胺合成酶 (GS) 活性等相关指标。  【结果】  与Cd处理相比,A + Cd处理甘蓝型油菜新叶SPAD值提高了1.1倍,根部脯氨酸含量增加了29.0%,MDA 含量降低了48.3%。外源ABA的添加会抑制甘蓝型油菜对Cd的吸收,降低甘蓝型油菜地上部和地下部Cd含量,减小Cd对甘蓝型油菜的毒害作用,影响甘蓝型油菜对NO₃–的吸收,降低油菜体内的总氮含量。与Cd处理相比,A + Cd处理甘蓝型油菜根部NR活性显著提高82.8%,更多的NH₄+分配到了地上部,地上部GS活性升高了32.1%,增强了甘蓝型油菜体内的氮同化和代谢过程,氮素生理利用效率显著提高了17.2%。  【结论】  外源ABA的添加可降低Cd的吸收量,提高氮代谢相关酶活性,促进脯氨酸的合成和降低丙二醛的生成,进而增强甘蓝型油菜对Cd的抗性,增强甘蓝型油菜体内氮同化和代谢的过程,并提高甘蓝型油菜的氮素生理利用效率。     

外源蔗糖对高NH4+胁迫下拟南芥碳氮代谢的影响

以拟南芥野生型、amt1.1和amt1.3为实验材料，采取土培的方法，以正常培养液（4mmol/L NH4+）培养，在20mmol/L NH4+的胁迫下，通过在培养液中添加0%（T1）蔗糖、5%（T2）蔗糖，测定地上部分的鲜重，叶绿素，游离NH4+，可溶性糖，可溶性蛋白，谷氨酰胺合成酶（GS）、谷氨酸脱氢酶（GDH），矿质元素含量等指标，研究外源蔗糖对NH4+胁迫拟南芥碳氮代谢的影响。结果表明，T1处理下，拟南芥生长受到严重的抑制，鲜重，GS，GDH酶活性降低，游离NH4+含量，叶绿素含量，可溶性糖和可溶性蛋白含量增加，植株的N、P、K、Ca的含量增加，Mg、Fe的含量减少。其中col-0在T1处理下受到的抑制比amt1.1和amt1.3更为显著。与T1处理相比较，T2处理增加了拟南芥植株的鲜重，显著提高了可溶性糖和可溶性蛋白含量，提高了GS和GDH的活性；降低了叶绿素和游离NH4+的含量，提高了植株体内的N、P、K、Ca，Mg的含量，降低了植株Fe的含量，其中，外源蔗糖对col-0高NH4+毒害的缓解更为显著。     

水稻根际的硝化作用与水稻的硝态氮营养

由于淹水条件下土壤硝化作用被强烈抑制 ,人们对水稻氮素 (N)营养的研究主要侧重在铵(NH 4 )营养而忽略了对硝 (NO-3 )营养的研究。但值得注意的是 ,水稻根系能分泌氧气 (O2 ) ,这些O2 能被硝化微生物利用 ,从而将NH 4 氧化成NO-3 ,在根表形成的NO-3 立即被水稻吸收。但通常情况下从水稻土中采集的土样中较难测到NO-3 或数量极微。事实上 ,即便是完全淹水 ,水稻根系也是处于铵、硝混合营养中。本文首先论述了水稻根际通过硝化作用产生NO-3 的过程 ,然后从吸收速率和根系生物量两方面提出了NO-3 对水稻NH 4 吸收和同化的促进机理 ,并比较了NO-3 对侧根生长发育的局部刺激作用和系统抑制作用 ,其中对于NO3 -对侧根生长发育的局部刺激作用是由于NO3 -的营养作用 (NO-3 对植物体内糖类、氨基酸和内源激素的影响 )还是信号物质作用进行了详细阐述 ,最后提出了今后在水稻硝酸盐营养方面的研究方向。     

植物氮素利用途径中硝酸盐转运基因的研究进展

氮素是植物生长发育必不可少的大量元素之一,土壤中的硝酸盐是植物获取氮素的主要来源。植物对硝酸盐的吸收与利用是通过一个精密的信号调控网络来实现的,其中硝酸盐转运蛋白在植物体内硝酸盐的运输和分配过程中发挥着重要的作用。通过对氮素利用途径中不同硝酸盐转运基因在硝酸盐的吸收、转运、同化和再利用进行功能鉴定,可以更好地解析硝酸盐在植物体内的吸收机制,从而找到提高植物氮素利用效率的关键环节。因此,综述了植物硝酸盐转运蛋白对土壤中硝酸盐的响应和信号的传递;硝酸盐转运蛋白在植株体内参与硝酸盐的转运、储存和再利用的功能以及硝酸盐在植物育种中的应用,并从对硝酸盐转运基因的单碱基编辑、关键结构域的改造和基因功能鉴定等方面进行展望。综述了有利于揭示硝酸盐转运基因的功能,拓宽植物吸收转运硝酸盐的分子机制认识,为提高植物氮素利用效率、培育氮高效利用农作物品种提供理论支撑。     

增铵营养对低温胁迫下棉花幼苗氮代谢的影响

【目的】探明增铵营养提高棉花幼苗抗低温胁迫能力的机制。【方法】以棉花新陆早13号为供试品种,在人工气候室内模拟不同温度处理（15℃和25℃）,研究了不同铵硝态氮配比（NH4+-N/NO3--N分别为0/100、25/75、50/50、75/25、100/0）对低温（15℃）胁迫下棉花苗期生长、氮素吸收量及氮代谢相关酶活性的影响。【结果】常温条件（25℃）下,较单一铵、硝营养,铵硝混合营养显著提高棉苗各器官的生物量,地上部和根系干物质量在NH4+-N/NO3--N比为50/50处理时最大,单一铵营养处理时最小;对棉苗生物量的影响效果表现出铵硝混合营养处理优于单一铵、硝营养处理。低温胁迫（15℃）后棉苗各器官生物量减小,且差异显著。常温和低温条件下,随着营养液中NH4+-N比例增加,棉苗全氮含量逐渐递增,氮素吸收量先升后降;棉苗根系、茎秆及叶柄内硝态氮含量呈明显降低趋势;棉花幼苗叶片NR活性明显减小,相反,GS和GOGAT活性则极显著提高。常温处理下棉苗各器官的氮素累积量显著高于低温胁迫处理,低温抑制了棉苗对硝态氮的吸收,降低NR、GS和GOGAT活性。【结论】低温胁迫下,增铵营养可显著提高氮素养分含量,促进棉苗生长,同时通过提高GS、GOGAT等氮代谢相关酶活性,维持氮代谢平衡,增强棉花幼苗对低温的抗性。     

Ammonium assimilation in rice based on the occurrence of 15N and inhibition of glutamine synthetase activity

Assimilation of ammonium (NH₄) into free amino acids and total reduced nitrogen (N) was monitored in both roots and shoots of two‐week old rice seedlings supplied with 5 mM 99% (¹⁵NH₄)₂SO₄ in aerated hydroponic culture with or without a 2 h preincubation with 1 mM methionine sulfoximine (MSX), an inhibitor of glutamine synthetase (GS) activity. ¹⁵NH₄ was not assimilated into amino acids when the GS/GOGAT (glutamate synthase) cycle was inhibited by MSX. Inhibition of glutamine synthetase (GS) activity in roots with MSX increased both the amount of NH₄ and the abundance of ¹⁵N labeled NH₄. In contrast, the amount of Gln and Glu, and their proportions as ¹⁵N, decreased in roots when GS activity was inhibited. This research confirms the importance of GS/GOGAT in NH₄ assimilation in rice roots.

¹⁵N‐labeled studies indicate that NH₄ ions incorporated by roots of rice are transformed primarily into glutamine (Gln) and glutamic acid (Glu) before being converted to other amino acids through transamination (15). The formation of amino acids such as aspartic acid (Asp) and alanine (Ala) directly from free NH₄ in roots also has been reported (4,15). Translocation of free NH₄ to plant shoots, based on the concentration of free NH₄ in xylem exudate, has been reported in tomato (13), although NH₄ in shoots primarily originates from nitrate reduction in the shoot. Photorespiration also can contribute to the accumulation of NH₄ in leaves (7).

The GS/GOGAT cycle appears to be primarily responsible for the assimilation of exogenously supplied NH₄ and NH₄ derived from nitrate reduction in leaves, as well as NH₄ derived from photorespiration (2,3,6,8). Genetic evidence cited to support this conclusion includes the lethal effect of photorespiratory conditions on plant mutants deficient in chloroplast‐localized GS and GOGAT activities (2,3,9), and the rapid accumulation of free NH₄ in GS‐deficient mutants under photorespiratory conditions (2,3,5).

The present study was initiated to quantify the in vivo amino acid synthesis in rice roots and shoots by analysis of ¹⁵N labeling, and should provide a more complete understanding of this important system for NH₄ utilization.     

氮素形态, 光合作用, 光呼吸

Under high light conditions, ammonium nutrition has a negative effect on plant growth. This suggests that the adverse effects of ammonium nutrition on plant growth may be related to carbon gain, photosynthesis, and photorespiration. However, there is no consistent evidence of a specific mechanism that could explain the plant growth reduction under ammonium supply. It is generally accepted that during the light reaction, a surplus of nicotinamide adenine dinucleotide hydrogen phosphate （NADPH） is produced, which is not completely used during the assimilation of CO2, Nitrate reduc- tion in the leaf represents an additional sink for NADPH that is not available to ammonium-grown plants. Nitrate and ammonium nutrition may use different pathways for NADPH consumption, which leads to differences in photosynthesis and photorespiration. The morphological （i.e., cell size, mesophyll thickness, and chloroplast volume） and enzymic （i.e., ribulose-1,5-bisphosphate carboxylase/oxygenase （Rubisco）, phosphoenolpyruvate carboxylase （PEPCase）, and glutamine synthetase/glutamate synthetase （GS/GOGAT）） differences that develop when plants are treated with either nitrate or ammonium nitrogen forms are related to photosynthesis and photorespiration. The differences in photorespiration rate for plants treated with nitrate or ammonium are related to the conversion of citrate to 2-oxoglutarate （2-OG） and photorespiratory CO2 refixation.     

不同铵硝比例对水稻铵吸收代谢基因表达的影响

以水稻南光为材料,研究了不同铵硝摩尔比例处理时,水稻NH4^＋吸收代谢基因的表达情况.档结果表明：（1）应用荧光定量PCR方法,可以精确检测水稻氮素吸收代谢基因在不同铵硝处理间的表达量变化;（2）从各基因的表达量上来看,吸收基因中以OsAMT1.1表达量最高,编码GS的基因中以OsGln1.1表达量最高,编码GOGAT的基因中,OsGlu表达量最高;（3）总体来说,不同铵硝处理对NH4^＋吸收代谢基因的表达有显著影响,吸收基因对不同铵硝比例的反应要比代谢基因更敏感;（4）氮吸收基因中OsAMT4.1显著受到NO3的抑制,NH4^＋的诱导.而OsAMT1.1,OsAMT1.2,OsAMT1.3和OsNRT2在铵硝摩尔比例由100：0变为50：50过程中,受到NO3^-的显著抑制,在铵硝摩尔比例由50：50变为0：100过程中OsAMT1.2和OsAMT1.3受到NO3^-的显著诱导,OsAMT1.1和OsNRT2变化不显著;（5）编码GS的基因OsGln1.1表达受NO3诱导,受NH4^＋抑制.OsGln2在铵硝摩尔比例由100：0变为50：50过程中,受到NO3^-增加的诱导,同时,OsGln1.2受到NO3增加的显著抑制作用,铵硝达到50：50以后,NO3^-比例的增加对OsGln1.2和OsGln2的表达没有显著影响;（6）编码GOGAT的基因OsGlt1和OsGlu在不同铵硝摩尔比例中的变化趋势一致：铵硝比例由100：0变为50：50过程中没有显著变化,铵硝比例50：50基础上表达量受到NO3^-比例增加的显著抑制,而OsGlt2的表达受NO3^-的显著抑制,NH;的显著诱导.     

Nitrogen metabolism is related to improved water‐use efficiency of nodulated alfalfa grown with sewage sludge under drought

Leguminous plants grown in sewage sludge–amended soils can acquire nitrogen by assimilation of nitrate and ammonium from the soil solution or from atmospheric‐dinitrogen (N₂) fixation through association with N₂‐fixing bacteria. We proposed that operation of both metabolic processes could contribute to alleviate the impact of drought in sludge‐treated plants. A greenhouse experiment was conducted to evaluate the involvement of nodule metabolism in the use efficiency of water and N in sludge‐treated plants. Treatments comprised (1) plants inoculated with rhizobia and amended with sewage sludge; (2) plants inoculated with rhizobia without any amendment; and (3) noninoculated plants supplied with ammonium nitrate, each under well‐watered and drought conditions. Under drought, sludge‐treated plants had increased plant growth and higher photosynthetic and water‐use efficiencies than untreated plants. Drought stimulated nitrate reductase and GS/GOGAT activities but did not affect the activities of phosphoenolpyruvate carboxylase and malate dehydrogenase or the leghemoglobin concentration. The results suggest that under drought conditions, both N₂ fixation and nitrate assimilation in nodules of sludge‐treated plants contributed to improve plant N supply and to increase the drought tolerance of alfalfa.     

干湿交替灌溉与氮肥形态耦合对水稻光合特性及氮素利用的影响

  【目的】  探讨干湿交替灌溉与氮肥形态对水稻光合特性及氮肥利用的影响。  【方法】  以徐稻3号为材料,在防雨棚内按处理数量构建9 m × 1.5 m × 0.4 m水泥池,用于2因素3水平完全区组试验。因素1为灌溉方式:浅水层灌溉 (0 kPa,CK)、轻度干湿交替灌溉 (?20 kPa)、重度干湿交替灌溉 (?40 kPa)。因素2为氮素形态:100%NH₄+-N (NH)、50%NH₄+-N+50%NO₃–-N (1/2NH+1/2NN)、100%NO₃–-N (NN)。在水稻分蘖盛期、幼穗分化始期、抽穗期和成熟期取植株样品,测定水稻根系氮代谢酶活性、叶片光合荧光特性及植株各部位氮素含量。  【结果】  在相同氮肥形态下,轻度干湿交替灌溉根系硝酸还原酶 (NR)、谷氨酰胺合成酶 (GS)、谷氨酸合成酶 (GOGAT)、谷氨酸脱氢酶 (GDH) 活性与浅水对照相比分别增加6.4%～80.4%、8.1%～85.9%、5.1%～61.8%与13.4%～94.0%;叶片光合速率及最大光化学效率得到提升;水稻产量、光合氮素利用率及氮肥农学效率明显提高,重度干湿交替灌溉则抑制根系NR、GS、GOGAT及GDH活性,降低叶片光合速率及最大光化学效率,最终导致水稻产量、光合氮素利用率及氮肥农学效率显著降低 (P < 0.05)。在浅水对照下,NH处理可改善根系氮代谢酶活性,提高叶片光合速率及最大光化学效率,有利于水稻产量、光合氮素利用率及氮肥农学效率的提升。干湿交替灌溉下,铵硝混合处理提高了根系氮代谢酶活性,增加了叶片光合速率及最大光化学效率,提高了水稻产量、光合氮素利用率及氮肥农学效率。相关分析表明,根系GS、GOGAT及GDH活性及叶片光合速率、最大光化学效率与氮素农学效率呈显著 (P < 0.05) 或极显著 (P < 0.01) 的正相关关系,而非光化学猝灭系数则与氮肥吸收利用率呈显著的负相关关系 (P < 0.05)。  【结论】  水稻生长期一直保持浅水层时,供应100%铵态氮可以充分发挥水肥的耦合效应,促进根系氮代谢酶活性,提高叶片的光合速率及最大光化学效率,有利于水稻的高产及氮高效利用。轻度干湿交替灌溉则以施用50%铵态氮和50%硝态氮混合氮肥最佳。     

Potential Nitrate Leaching Under Common Landscaping Plants

Two Elsholtzia haichowensis S. populations, copper-tolerant (TLS) and non-tolerant (HA) ones were studied in hydroponic experiment for the nitrogen assimilation and plant growth under excess Cu conditions. The results demonstrated that there were surely the differences in nitrogen assimilation and plant growth between the two populations. Excess Cu caused evident decreases in the shoot and root biomass and root/shoot biomass ratio in HA population while no significant changes happened in TLS population. In addition, in HA population, excess Cu also induced apparent declines in activities of nitrate reductase (NR, EC 1.6.6.1) and glutamine synthetase (GS, EC 6.3.1.2) in the leaves and roots as well as the contents of nitrate, ammonium and amino acids in the roots. In TLS population, excess Cu did not significantly affect the NR activities in the leaves and roots and the nitrate content in the roots, and apparently elevated the root ammonium and amino acids contents, although it also clearly reduced the GS activities in the leaves and roots. Besides, with the addition of Cu in the culture solution, the Cu contents in the leaves and roots of the two populations markedly increased. But this increase was significantly lower in TLS population than that in HA population; the fact might be partly responsible for the relative stabilization of nitrogen assimilation in TLS population compared to that in HA population.     

Enzymes of Nitrogen Metabolism and Proteases Activity in Hairy Roots of Clover and Carrots,with and without Arbuscular Mycorrhizal Fungi

Hairy roots can be used for metabolic studies and also as a substrate for arbuscular mycorrhizal fungi (AMF). However, little is known about the behavior of these roots when infected with AMF. The metabolism of hairy roots grown with or without AMF, on two culture media, was investigated. An increase in protease activity was observed when the roots were cultured on medium with high nitrogen (N)-content. Glutamate synthase (GOGAT) and glutamate dehydrogenase (GDH) activities increased along the development in both carrot and clover hairy roots. With the advancement of root senescence (at 132 d) a greater degradation of root tissue occurred, which was characterized by an increase in catabolic enzymes activity (proteases and GDH deamination). The results indicate that the glutamine synthetase/GOGAT pathway, characteristic of plant tissues, ceases to act in synchronism and is replaced by the GDH/GOGAT system under conditions of stress or senescence when excess ammonium is present.