植物吸收转运无机氮的生理及分子机制

氮是植物生长必需的营养元素。植物从土壤中吸收的氮素主要是NO3-和NH4 +等无机氮源。植物吸收NO3-和NH4+的系统均有高亲和转运系统(high-affinity transport system, HATS)和低亲和转运系统(low-affinity transport system, LATS)之分。近10多年的研究已对这些转运系统的分子基础有了较好的理解, 本文着重对近年来植物吸收无机氮分子机制的研究进展进行了综述。     

高等植物硝酸盐转运蛋白的功能及其调控机制

硝酸盐是植物从土壤中吸收的重要无机氮素形态。植物为适应含有不同浓度NO3-的土壤环境,进化出了高亲和硝酸盐转运系统(HATS)和低亲和硝酸盐转运系统(LATS),两个基因家族NRT1和NRT2家族分别参与了LATS和HATS的NO3-的吸收和转运。近年来,随着分子生物学技术和植物基因组学的快速发展,研究人员克隆出了大量参与硝酸盐吸收和转运的基因,并对这些基因的功能进行了深入研究,逐渐形成了复杂的硝酸盐调控网络。综述了植物中硝酸盐转运蛋白基因的克隆、表达及调控,并对进一步的研究作了展望,这些结果对于理解植物硝酸盐吸收的调控机制具有重要作用。     

植物吸收利用铵态氮和硝态氮的分子调控

本文介绍了植物吸收和利用NH4^＋、NO3^-的转运系统和相关基因的表达调控特征,以及与氮还原、同化相关的酶及其调控基因的发现和功能的研究进展。     

拟南芥无机氮素转运蛋白及其磷酸化调控研究进展

氮元素是植物必需的营养元素之一, 氮素供需失衡会严重影响植物的生长发育。无机氮(硝酸根NO₃^-和铵根NH₄⁺)是植物体内氮素的主要来源, 对其有效吸收和利用依赖于多种类型转运蛋白的协同作用。其中, 部分无机氮素转运蛋白的活性受到可逆磷酸化作用的精准调控。该文将对模式植物拟南芥(Arabidopsis thaliana)中硝酸根和铵根转运蛋白的分类、结构、定位和功能特点等进行总结, 并重点对可逆磷酸化调控转运蛋白的分子机制加以阐述。     

八种氨基酸对水稻硝酸盐吸收的影响

目的:在较低NO3-浓度下,研究不同氨基酸对水稻硝酸盐吸收的调控作用.方法:使用非侵害性硝酸盐检测系统测定了谷氨酸、天门冬氨酸、精氨酸、赖氨酸、脯氨酸、苯丙氨酸、缬氨酸和丝氨酸8种氨基酸对水稻硝酸盐吸收的动力学影响.结果:加入谷氨酸、天门冬氨酸后对水稻硝酸盐的吸收有短暂的促进作用,硝酸盐净吸收率分别上升258±123%和217±34%;另外,加入赖氨酸和精氨酸以后发现水稻硝酸盐的吸收率降低-63±34%和-64±27%,反应延迟时间为30min以上,而其他4种氨基酸并没有产生明显的影响.结论:氨基酸不是直接调控水稻高亲和力NO3-转运系统,而可能是对NO3-同化过程进行调控.     

原始森林土壤NH4+/NO3-生境特征与某些针叶树种的适应性

在陆地生态系统中,生存地段的土壤养分环境构成了植物的“营养生境”。植物在长期进化过程中往往产生对原生营养生境的生态适应,其中对NH4 和NO3-两种无机氮源的吸收、利用特性便可能是这种适应的一个重要方面。由于硝化抑制(限制)或微生物对NO3-的强烈吸收、固持作用,酸性、弱酸性的原始森林土壤中NH4 含量大都远高于NO3-,从而形成了以NH4 占绝对优势的“氮营养生境”。很多针叶树种(尤其是演替晚期阶段占优势者)对其长期所处的NH4 优势生境产生了充分适应,以致对非还原态氮(NO3-)的吸收、利用能力严重下降。这些针叶树往往表现出典型的“喜铵性”,而在NO3-优势环境中则会引起氮代谢失调和生长下降。从氮同化酶、高耐铵性、根对NH4 和NO3-的相对吸收能力及NO3-吸收的反馈控制、养分关系与养分平衡、根部碳流失、光合作用及耐荫性等多方面阐述了喜铵针叶树适应的生理生化机制。这种生态适应可能是顶极森林群落维持长期稳定的重要机制之一,而采伐干扰后NO3-明显增加的立地条件则可能会导致喜铵的“原优势针叶树种”更新困难。在温带退化森林生态系统恢复与重建过程中,顶极针叶树种对NH4 营养生境的固有适应性是必须充分考虑的问题。     

粉绿狐尾藻和凤眼莲对不同形态氮吸收动力学研究

采用改进常规耗竭法,研究了粉绿狐尾藻(Myriophyllum aquaticum)、凤眼莲(Eichhornia crassipes)在抑菌和非抑菌两种处理中对NO3-、NH4+吸收的动力学特征。结果表明:两种植物对NO3-和NH4+的最大吸收速率(Imax)和亲和力(1/Km)有显著差异,凤眼莲对NO3-、N H4+的吸收速率显著高于狐尾藻,说明凤眼莲更适宜用于污染水体养分的去除;不同植物对NO3-、NH4+表现出吸收偏好性,凤眼莲对NO3-有较高的Imax值和亲和力,而狐尾藻对NH4+有较高的Imax值和亲和力;抑菌处理能显著增加凤眼莲对NH4+的吸收速率,NO3-浓度高于1.00 mmol/L时,抑菌处理能显著减小凤眼莲对NO3-的吸收速率,狐尾藻对NO3-、N H4+的吸收未受到抑菌处理的显著影响。       

水源涵养林不同植被类型土壤NH_4~+-N和NO_3~--N分布特征

以辽宁东部山地水源涵养林为对象,选择槭树-蒙古栎林、山杨林、白桦-山杨林和落叶松人工林等4种植被类型,测定其土壤NH4+-N、NO3--N、pH值、容重、有机碳和全氮等理化指标,分析了植被类型、土壤层次与土壤无机氮分布特征之间的关系。结果表明:4种植被类型土壤NH4+-N、NO3--N在土壤表层(0~5 cm)含量最高,由表层向下逐渐降低;土壤总无机氮含量大小为落叶松人工林(27.46 mg·kg-1)山杨林(21.76 mg·kg-1)槭树-蒙古栎林(19.09 mg·kg-1)白桦-山杨林(17.88 mg·kg-1);阔叶林中NH4+-N是土壤无机氮的主要存在形式,而落叶松人工林土壤中NO3--N所占比例较高;水源涵养林土壤NH4+-N、NO3--N均与土壤有机质、土壤含水量呈极显著正相关(P0.01)。总体而言,植被类型对土壤无机氮分布有较大影响,研究结果可为辽东山区水源涵养林植被类型的选择和结构调控提供参考。     

南黄海表层沉积物中氮的潜在生态学功能

首次探讨了南黄海表层沉积物中不同形态可转化氮与该海域浮游植物、浮游动物的丰度、生物量以及初级生产力的关系 ,研究了沉积物中可转化氮在海洋生物生长、繁衍中的潜在生态学功能。结果表明 ,不同粒度的沉积物中各形态氮的生态学功能有较大差异 ,一般细粒度沉积物中可转化各形态氮与浮游植物、底栖生物有较密切的关系 ,而中、粗颗粒沉积物中的可转化氮主要与浮游动物有关 ;在南黄海不同粒度的表层沉积物的 4项可转化氮中 ,强氧化剂可浸取态氮 (SOEF- N )、强碱可浸取态氮(SAEF- N)与浮游植物的生长繁殖、提升海域的生产力有密切关系 ;两种无机形式的氮 (NH4 - N和 NO3- N)和叶绿素 a、浮游植物细胞总量以及初级生产力具有正相关关系 ,说明表层沉积物中的可转化态 NH4 - N和 NO3- N对于促进浮游植物的生长以及提高初级生产力具有非常重要的作用 ,是浮游植物可以直接吸收利用的氮的两种主要形式 ,其中 NO3- N的作用要稍大一些 ,且粒度越细 ,影响越大 ;NH4 - N和 NO3- N对于促进浮游动物以及底栖动物生长繁殖的作用则不明显 ,因其不能直接被它们吸收利用 ,而与通过食物链的传输等一系列中间环节有关 ;且在南黄海表层沉积物中 ,氮无论以什么形态存在 ,只有转化为无机形式的 NH 4后 ,其生态学功能才易被显     

退化草地恢复过程中土壤氮素状况以及与植被地上绿色生物量形成关系的研究

 研究了在不同放牧率下形成的不同退化阶段的草地各形态氮素（全氮、硝态氮、铵态氮、无机氮和微生物氮）的变化情况,同时也研究了植被地上绿色生物量与各形态氮素季节变化的同步性关系。土壤全氮含量相对稳定,随草地植被状况和植物生长时期变化不大,说明土壤总氮库有相当的弹性。土壤硝态氮（NO－3-N）、铵态氮（NH+4-N）、无机氮（IN）和微生物氮（Micro-N）季节变化明显。土壤Micro-N和NO-3-N含量随植物生长逐渐降低,到植物枯黄期含量又回复到较高的水平;土壤NH+4-N含量随植物生长有逐渐升高的趋势;IN则随着植物的生长出现低-高-低-高的特点,且与植被地上绿色生物量呈显著负相关（R=-0.247, p<0.01）。在放牧条件下草原植物优先利用NO－3-N,NO－3-N与植被地上绿色生物量有显著的负相关性,是形成草原植被地上绿色生物量的有效性氮素。Micro-N能解释土壤IN 22.3%的变异（R2=0.223, p<0.01）,Micro-N是土壤无机氮的重要来源。土壤NH+4-N与Micro-N呈显著负相关（R=-0.222, p<0.01）,说明土壤微生物对土壤NH+4-N有偏好吸收。总体上,不同形态的氮素在各土壤层次间差异显著,随土壤层次的加深含量逐步降低。连续放牧11年恢复两年后,各氮素组分对放牧压力消除的响应并不一致。土壤全氮含量与停止放牧前相比变化差异不显著;而Micro-N对放牧压力消失的响应在不同处理下整个生长季的结果比较一致,即以前过度和中度放牧处理的Micro-N含量较高,无牧和轻牧含量较低;IN、NH+4-N和NO－3-N变化比较复杂,在不同放牧恢复处理上结果并不一致。总的来看,以前中度和过度放牧的IN、NH+4-N和NO－3-N含量较高,存在潜在损失的可能。经过两年的恢复,植被地上绿色生物量（8月）过牧处理与无牧处理差异不显著。     

Foliar application of nitrate or ammonium as sole nitrogen supply in Ricinus communis. II. The flows of cations, chloride and abscisic acid

Following a precultivation with pedospheric nitrogen nutrition, Ricinus plants were supplied with nitrogen solely by spraying nitrate or ammonium solution onto the leaves during the experimental period. The chemical composition of tissues, xylem and phloem exudates was determined and on the basis of the previously determined nitrogen flows (Peuke et al., New Phytologist (1998), 138 , 657–687) the flows of potassium, sodium, magnesium, calcium, chloride and ABA were modelled. These data, which permit quantification of net-uptake, transport in xylem and phloem, and utilization in shoot and root, were compared with results obtained in plants with pedospherically-supplied nitrate or ammonium and data in the literature. Although the overall effects on the chemical composition of supplying ammonium to the leaves were not as pronounced as in pedospherically supplied plants, there were some typical responses of plants fed with ammonium (ammonium syndrome). In particular, in ammonium-sprayed plants uptake and transport of magnesium decreased and chloride uptake was increased compared with nitrate-sprayed plants. Furthermore, acropetal ABA transport in the xylem in ammonium-sprayed Ricinus was threefold higher than in nitrate-sprayed plants. Additionally, concentrations of anions were more or less increased in tissues, particularly in the roots, and transport fluids. The overall signal from ammonium-sprayed leaves without a direct effect of ammonium ions on uptake and transport systems in the root is discussed.     

Gene structure and expression of the high-affinity nitrate transport system in rice roots

Rice has a preference for uptake of ammonium over nitrate and can use ammonium-N efficiently. Consequently, transporters mediating ammonium uptake have been extensively studied, but nitrate transporters have been largely ignored. Recently,some reports have shown that rice also has high capacity to acquire nitrate from growth medium, so understanding the nitrate transport system in rice roots is very important for improving N use efficiency in rice. The present study identified four putative NRT2 and two putative NAR2 genes that encode components of the high-affinity nitrate transport system (HATS) in the rice (Oryza sativa L. subsp, japonica cv. Nipponbare) genome. OsNRT2.1 and OsNRT2.2 share an identical coding region sequence, and their deduced proteins are closely related to those from monocotyledonous plants. The two NAR2 proteins are closely related to those from mono-cotyledonous plants as well. However, OsNRT2.3 and OsNRT2.4 are more closely related to Arabidopsis NRT2 proteins. Relative quantitative reverse tranecdption-polymerase chain reaction analysis showed that all of the six genes were rapidly upregulated and then downregulated in the roots of N-starved rice plants after they were re-supplied with 0.2 mM nitrate, but the response to nitrate differed among gene members.The results from phylogenetic tree, gene structure and expression analysis implied the divergent roles for the individual members of the rice NRT2 and NAR2 families. High-affinity nitrate influx rates associated with nitrate induction in rice roots were investigated and were found to be regulated by external pH. Compared with the nitrate influx rates at pH 6.5, alkaline pH (pH 8.0) inhibited nitrate Influx, and acidic pH (pH 5.0) enhanced the nitrate influx In I h nitrate induced roots, but did not significantly affect that in 4 to 8 h nitrate induced roots.     

Molecular and physiological interactions of urea and nitrate uptake in plants

While nitrate acquisition has been extensively studied, less information is available on transport systems of urea. Furthermore, the reciprocal influence of the two sources has not been clarified, so far. In this review, we will discuss recent developments on plant response to urea and nitrate nutrition. Experimental evidence suggests that, when urea and nitrate are available in the external solution, the induction of the uptake systems of each nitrogen (N) source is limited, while plant growth and N utilization is promoted. This physiological behavior might reflect cooperation among acquisition processes, where the activation of different N assimilatory pathways (cytosolic and plastidic pathways), allow a better control on the nutrient uptake. Based on physiological and molecular evidence, plants might increase (N) metabolism promoting a more efficient assimilation of taken-up nitrogen. The beneficial effect of urea and nitrate nutrition might contribute to develop new agronomical approaches to increase the (N) use efficiency in crops.     

A two-component high-affinity nitrate uptake system in barley

The analysis of genome databases for many different plants has identified a group of genes that are related to one part of a two-component nitrate transport system found in algae. Earlier work using mutants and heterologous expression has shown that a high-affinity nitrate transport system from the unicellular green algae, Chlamydomonas reinhardtii required two gene products for function. One gene encoded a typical carrier-type structure with 12 putative trans-membrane (TM) domains and the other gene, nar2 encoded a much smaller protein that had only one TM domain. As both gene families occur in plants we investigated whether this transport model has more general relevance among plants. The screening for nitrate transporter activity was greatly helped by a novel assay using (15)N-enriched nitrate uptake into Xenopus oocytes expressing the proteins. This assay enables many oocytes to be rapidly screened for nitrate transport activity. The functional activity of a barley nitrate transporter, HvNRT2.1, in oocytes required co-injection of a second mRNA. Although three very closely related nar2-like genes were cloned from barley, only one of these was able to give functional nitrate transport when co-injected into oocytes. The nitrate transport performed by this two-gene system was inhibited at more acidic external pH and by acidification of the cytoplasm. This specific requirement for two-gene products to give nitrate transport function has important implications for attempts to genetically manipulate this fundamental process in plants.     

Interaction of sulfur and nitrogen nutrition in tobacco (Nicotiana tabacum) plants: significance of nitrogen source and root nitrate reductase

Abstract: The significance of root nitrate reductase for sulfur assimilation was studied in tobacco (Nicotiana tabacum) plants. For this purpose, uptake, assimilation, and long-distance transport of sulfur were compared between wild-type tobacco and transformants lacking root nitrate reductase, cultivated either with nitrate or with ammonium nitrate. A recently developed empirical model of plant internal nitrogen cycling was adapted to sulfur and applied to characterise whole plant sulfur relations in wild-type tobacco and the transformant. Both transformation and nitrogen nutrition strongly affected sulfur pools and sulfur fluxes. Transformation decreased the rate of sulfate uptake in nitrate-grown plants and root sulfate and total sulfur contents in root biomass, irrespective of N nutrition. Nevertheless, glutathione levels were enhanced in the roots of transformed plants. This may be a consequence of enhanced APR activity in the leaves that also resulted in enhanced organic sulfur content in the leaves of the tranformants. The lack of nitrate reductase in the roots in the transformants caused regulatory changes in sulfur metabolism that resembled those observed under nitrogen deficiency. Nitrate nutrition reduced total sulfur content and all the major fractions analysed in the leaves, but not in the roots, compared to ammonium nitrate supply. The enhanced organic sulfur and glutathione levels in ammonium nitrate-fed plants corresponded well to elevated APR activity. But foliar sulfate contents also increased due to decreased re-allocation of sulfate into the phloem of ammonium nitrate-fed plants. Further studies will elucidate whether this decrease is achieved by downregulation of a specific sulfate transporter in vascular tissues.     

Nitrogen acquisition by roots: physiological and developmental mechanisms ensuring plant adaptation to a fluctuating resource

Background

Nitrogen (N) is one of the key mineral nutrients for plants and its availability has a major impact on their growth and development. Most often N resources are limiting and plants have evolved various strategies to modulate their root uptake capacity to compensate for both spatial and temporal changes in N availability in soil. The main N sources for terrestrial plants in soils of temperate regions are in decreasing order of abundance, nitrate, ammonium and amino acids. N uptake systems combine, for these different N forms, high- and low-affinity transporters belonging to multige families. Expression and activity of most uptake systems are regulated locally by the concentration of their substrate, and by a systemic feedback control exerted by whole-plant signals of N status, giving rise to a complex combinatory network. Besides modulation of the capacity of transport systems, plants are also able to modulate their growth and development to maintain N homeostasis. In particular, root system architecture is highly plastic and its changes can greatly impact N acquisition from soil.

Scope

In this review, we aim at detailing recent advances in the identification of molecular mechanisms responsible for physiological and developmental responses of root N acquisition to changes in N availability. These mechanisms are now unravelled at an increasing rate, especially in the model plant Arabidopsis thaliana L.. Within the past decade, most root membrane transport proteins that determine N acquisition have been identified. More recently, molecular regulators in nitrate or ammonium sensing and signalling have been isolated, revealing common regulatory genes for transport system and root development, as well as a strong connection between N and hormone signalling pathways.

Conclusion

Deciphering the complexity of the regulatory networks that control N uptake, metabolism and plant development will help understanding adaptation of plants to sub-optimal N availability and fluctuating environments. It will also provide solutions for addressing the major issues of pollution and economical costs related to N fertilizer use that threaten agricultural and ecological sustainability.     

根系氮吸收过程及其主要调节因子

氮（N）是植物根系吸收最多的矿质元素之一．全球变化将使土壤中N的有效性发生改变,影响陆地生态系统碳分配格局与过程．研究根系N吸收及其调控对预测生态系统结构和功能具有重要理论意义．由于土壤中存在多种形态的N源,长期的生物进化和环境适应导致植物根系对不同形态N的吸收部位、机理及调控有较大差别．因此,植物长期生长在以某一形态N源为主的土壤上就形成了不同的N吸收机制和策略．本文简述了近年来在植物根系N吸收和调控方面的最新研究进展,重点评述了不同形态N在土壤中的生物有效性,根系N吸收部位,N在木质部中的装载和运输,不同形态N（NO3^-、NH4^＋和有机氮）的吸收机制,以及根系N吸收的自身信号调控和环境因子对根系N吸收的影响．在此基础上,提出了目前根系N吸收研究中存在的几个问题．