植物中硝酸还原酶和亚硝酸还原酶的作用

植物通过硝酸盐同化途径以硝酸盐和氨的形式吸收氮元素。硝酸盐的同化是一个受到严格控制的过程,其中两个先后参加反应的酶——硝酸还原酶(NR)和亚硝酸还原酶(NiR)对初级氮的同化起主要调控。在高等植物中,NR和NiR基因的转录及转录后加工受到各种内在和外在因素的影响,翻译后调控是消除亚硝酸盐积累的重要机制。随着分子生物学技术的发展,可以更容易地通过突变体和转基因方式来研究NR和NiR基因的调控。     

植物抗寒基因表达调控研究进展

低温胁迫是植物生长过程中的主要非生物胁迫因子之一,也是影响农作物生产的主要因素之一。研究表明,在植物体内存在着一个复杂的对低温胁迫信号感知及传导的网络系统,该系统中大量的相关基因已有报道,这些基因不仅仅涉及植物激素的应答,还涉及到植物基因的转录调控及转录后的修饰与调控等各个方面。该文就近年来国内外有关植物抗寒基因表达调控的研究进展进行综述。     

C3和C4植物的氮素利用机制

提高植物的氮素利用效率(NUE)不仅有利于保障全球粮食安全, 也是实现农业可持续发展的重要途径。近半个世纪以来, 植物氮素利用机理研究已取得重要进展, 但NUE的调控机制仍不明确, NUE的提高仍然十分有限。高等植物集光合碳素同化和氮素同化于一体, 只有碳氮代谢相互协调, 才能维持植物体内的碳氮平衡, 保证植物正常生长发育。由于C₃和C₄植物的光合氮素利用率(PNUE)存在差异, 对氮素的利用效率也会存在差异。为了更有效地提高作物的NUE, 须更全面地了解C₃和C₄植物对氮素吸收、转运、同化和信号转导等关键因子的功能和调控机制。此外, 面对大气CO₂浓度增高和全球气候变暖条件下的植物碳氮同化及其机理的研究也不容忽视。该文综述了C₃和C₄植物氮素利用关键因素的差异及其调控机制, 并对提高C₃禾本科作物氮素利用效率的遗传改良途径进行了展望。     

植物氮代谢硝酸还原酶水平调控机制的研究进展

氮代谢是植株体内最基本的物质代谢之一,硝酸还原酶是植物氮代谢的关键酶。主要对植物氮代谢在硝酸还原酶水平上调控的研究新进展,尤其是其合成／降解及活性调控机制进行了较为系统的综述。硝酸还原酶合成的调控主要发生在转录水平和翻译水平上,硝酸还原酶降解的调控主要发生在翻译后水平上,同时NO3^-及光在硝酸还原酶转录水平调控上的作用重大,硝酸还原酶编码基因转录的mRNA的稳定性强弱影响植物的氮代谢,而影响mRNA稳定性的因素很多,机理复杂;磷酸化／去磷酸化在硝酸还原酶活性调控中占举足轻重的地位,研究也比较深入。钝化蛋白也能够影响硝酸还原酶活性,许多小分子物质对硝酸还原酶活性有影响。     

硝酸盐调控豆科植物与根瘤菌共生固氮的机制研究

氮是植物生长发育所需的大量营养元素之一。硝态氮不仅可以被植物直接吸收利用,还可以作为重要的信号分子调控植物对氮素的响应、吸收、代谢相关基因的表达,从而影响植物的生长和发育。豆科植物可以通过与根瘤菌共生互作来获得生长所需的氮,但共生固氮是一个耗费植物能量的过程。当土壤中存在高浓度的氮素时,氮作为信号分子会影响共生固氮基因的功能从而抑制共生固氮过程。目前的研究表明,硝酸盐通过局部和系统的调控方式抑制共生固氮过程;结瘤自主调控(Autoregulation of nodulation,AON)和NLPs(NIN-like proteins)转录因子在硝酸盐抑制豆科植物根瘤形成中有着重要的作用。本文结合最近的研究进展,重点讨论NLPs转录因子和AON途径在硝酸盐抑制共生固氮过程的作用。     

水培硝态氮浓度对冬小麦幼苗氮代谢的影响

以Hoagland营养液为培养基质,以冬小麦为试材,动态测定高(含NO3--N15mmol·L-1)、中(含NO3--N7.5mmol·L-1)、低(含NO3--N2.5mmol·L-1)三种氮水平处理条件下硝态氮的吸收和累积、硝酸还原酶活性、铵态氮含量、小麦吸氮量及根系活力,分析不同供氮水平对冬小麦硝态氮吸收、还原、转运的影响,探讨不同供氮条件下,植物地上、地下部分硝态氮代谢的变化。结果表明:水培条件下,营养液NO3-的消耗量、pH变化、植株全氮以及根系活力均能较好地反映不同氮水平对植株硝态氮代谢的影响;高氮条件下植物体内NO3-进一步同化较中氮弱,冬小麦植株积累了较多的NO3-,而非过多的吸收营养液中的NO3-。不同氮浓度处理下,NO3-的供应与植株NRA间无相关关系,根系与地上部的变化曲线不同;NO3-供应浓度高时,植物地上部是主要同化部位;低浓度时根部是主要同化部位。虽然NO3-是一种安全的氮源,但供应过高则抑制体内硝态氮进一步同化,而供应过低,植物吸收NO3-量不足、根系活力下降,不利于小麦幼苗氮素营养。     

C_3和C_4植物的氮素利用机制

提高植物的氮素利用效率(NUE)不仅有利于保障全球粮食安全,也是实现农业可持续发展的重要途径。近半个世纪以来,植物氮素利用机理研究已取得重要进展,但NUE的调控机制仍不明确, NUE的提高仍然十分有限。高等植物集光合碳素同化和氮素同化于一体,只有碳氮代谢相互协调,才能维持植物体内的碳氮平衡,保证植物正常生长发育。由于C_3和C_4植物的光合氮素利用率(PNUE)存在差异,对氮素的利用效率也会存在差异。为了更有效地提高作物的NUE,须更全面地了解C_3和C_4植物对氮素吸收、转运、同化和信号转导等关键因子的功能和调控机制。此外,面对大气CO_2浓度增高和全球气候变暖条件下的植物碳氮同化及其机理的研究也不容忽视。该文综述了C_3和C_4植物氮素利用关键因素的差异及其调控机制,并对提高C_3禾本科作物氮素利用效率的遗传改良途径进行了展望。     

可变剪切在植物成花转换中的作用

精确调控成花转换,确保植物在适宜环境下开花,对于植物的成功繁殖和物种繁衍至关重要。开花由多种分子机制在转录、转录后和蛋白质水平进行调控。可变剪切(AS)是一种普遍的转录后水平调控过程,可从单个基因产生多个转录本,从而丰富转录组和蛋白质组的多样性。大量研究表明,可变剪切在成花转换过程中发挥重要作用。根据发育和环境条件, AS能够影响mRNA的稳定性和/或蛋白亚型的功能,从而调控开花相关基因的功能转录本和/或功能蛋白水平。揭示成花相关pre-mRNA的AS作用将进一步增进人们对开花相关基因功能以及整个成花转换调控网络的认识。该文归纳了涉及成花转换的AS研究进展,并针对各个调控途径进行总结,以期为进一步研究植物AS和成花转换调控机制提供参考。     

植物冷驯化相关基因研究进展

摘要 冷驯化是与提高植物抗冷性有关的生物化学及生理学过程, 主要包括寒驯化(cool acclimation)和冻驯化(freezing acdimation)。在冷驯化过程中, 植物体内许多基因在转录水平上的表达受到影响, 已经克隆了大量的相关基因,它们组成复杂的分子调控网络。目前研究表明不依赖ABA的低温信号转导途径是植物冷驯化机制的重要组成部分, 其中CBF/DREB1是该调控过程的关键转录因子, 与植物通过冷驯化而提高冰冻耐受能力密切相关。进一步利用转基因技术, 可有效地改善作物的耐冷性状。     

真盐生植物盐角草对不同氮形态的响应

盐角草(Salicornia europaea L.)是一种喜盐植物,其最佳生长的实现需要200—400 mmol/L NaCl。为了解盐渍环境下盐角草氮素吸收利用特点,在水培添加200或400 mmol/L NaCl情况下,从生长指标,光合参数,根系体积和活力,硝酸还原酶与谷氨酰胺合成酶活力,蛋白、总氮、硝态氮及铵态氮含量等方面检测了硝态氮、铵态氮和尿素3种氮形态对盐角草生长的影响。研究发现以氮摩尔浓度(mmol/L)计,在0.1—400 mmol/L浓度范围进行测试,盐角草在0.1 mmol/L低氮条件下仍能维持生长,同时,抑制盐角草生长的氮浓度域值较高,其中铵态氮、尿素和硝态氮分别为50、50及400 mmol/L。研究结果表明盐角草吸收利用氮素的能力强,对氮素的浓度耐受范围宽,3种氮形态都可作为氮源满足其生长需要,但有效促进生长的效果存在差异,总体顺序从高到低依次为硝态氮、铵态氮和尿素。研究为揭示盐生植物氮吸收利用特点提供了基础数据,对提高盐生植物生产力,指导沿海滩涂生态建设具有一定的指导意义。     

草甸草原土壤与植物系统自然丰度氮稳定同位素的年际变化及其指示作用

氮是陆地生态系统生产力的首要限制性养分,利用自然丰度δ¹⁵N(¹⁵N/¹⁴N)可以有效指示生态系统氮循环过程。本试验研究了内蒙古草甸草原土壤与植物系统自然丰度δ¹⁵N、土壤净氮矿化潜势的年际变化。结果表明: 2017—2020年,土壤NO₃^--N含量(9.83～14.79 mg·kg^-1)均显著高于NH₄⁺-N含量(3.92～5.00 mg·kg^-1);土壤NH₄⁺的δ¹⁵N值(13.3‰～18.3‰)显著高于NO₃^-的δ¹⁵N值(3.76‰～6.14‰),土壤NO₃^-的δ¹⁵N值与土壤NO₃^-含量呈显著负相关;干旱年NH₄⁺的δ¹⁵N值相对较高,降水较高或较低年NO₃^-的δ¹⁵N值显著降低。干旱年土壤净氮矿化速率、净氨化速率显著高于湿润年,而土壤硝化速率与年降水量无显著相关性。植物δ¹⁵N值与土壤δ¹⁵N值无显著相关性,但与植物N含量呈显著负相关;豆科植物与非豆科植物δ¹⁵N值、N含量均呈显著正相关,在一定程度上表明豆科植物对非豆科植物的N吸收具有促进作用。研究结果可为草原土壤-植物系统氮循环过程及其对降水变化的响应提供数据支撑。     

Photosynthate costs associated with the utilization of different nitrogen–forms: influence on the carbon balance of plants and shoot–root biomass partitioning

The photosynthate costs of processes (amino acid and protein synthesis and turnover, and pH regulation) associated with the utilization of nitrate (NO₃⁻), ammonium (NH₄⁺) or glutamine (Gln) for plant growth were estimated. Based on these estimates, the effects of these forms of nitrogen (N) on the carbon balance of plants and on shoot–root biomass allocation were evaluated. The results indicated that NO₃⁻ as an N source for plant growth is not substantially more expensive to utilize than either NH₄⁺ or Gln, particularly in the long term when costs due to protein turnover dominate the total costs of N utilization. It is also suggested that the photosynthate use in processes associated with N assimilation has little impact on the carbon balance of plants, and hence on shoot–root biomass allocation.     

Varying the ratio of 15N-labelled ammonium and nitrate-N supplied to creeping bent: effects on nitrogen absorption and assimilation, and plant growth

The relative rates of ammonium and nitrate-N uptake and assimilation by creeping bent ( Agrostis stolonifera ), were investigated for plants grown in soil and supplied with three different ratios of ammonium and nitrate-N. Following two preliminary defoliations, plants were supplied with the equivalent of 150 kg N ha⁻¹, given as ¹⁵N-(differentially) labelled NH₄⁺ and NO₃⁻-N in three different ratios (20:80, 50:50 and 80:20), followed by sequential destructive harvests of shoots and roots at four points during a 35-d regrowth period. Maximum use of labelled nitrogen and 'exhaustion' of soil mineral nitrogen reserves occurred much earlier when plants were supplied with half or more of their nitrogen as ammonium, than occurred when they were supplied predominately with nitrate-N. The lack of consistency in the patterns of ammonium and nitrate-N absorption, however, implied that the plants had no specific preference for either nitrogen form. Supplying plants with different combinations of ammonium and nitrate produced distinctive differences in plant morphology. In the high nitrate treatment, plants preferentially partitioned resources into shoot and stolon formation, whereas in the high ammonium treatment, resources were preferentially partitioned into root production. These changes in plant morphology might be adaptations to aid species survival in environments associated with a predominance of either nitrogen form.     

Sink control of nitrogen uptake and assimilation during regrowth after-cutting of ryegrass (Lolium perenne L.)

Abstract. The ¹⁵N isotope was used to compare the uptake and the assimilation of NH₄⁺ and NO₃ nitrogen in ryegrass ( Lolium perenne L.) during regrowth after cutting. Uptake of nitrate-N, expressed per plant, was at all times greater than ammonium-N uptake and assimilation decreased in roots and stubble while its assimilation was maintained at a high level in leaves. It has been suggested that ammonium assimilation is directly related to the availability of carbohydrates in the sink organ (leaves) resulting from their remobilization from the source organs (roots and stubble). Nitrate reduction decreased in all organs, while the uptake of NO₃ was still high. After this first period of regrowth, nitrogen assimilation both from nitrate and ammonium increased in all the plants. Nitrate reduction capacity (expressed in μg NO₃-N reduced per g D.W. per d) is 7.5 and 22.5 times greater in leaves than in stubble and roots, respectively. Therefore, nitrogen assimilation in stubble and particularly in roots was mainly dependent on ammonium nitrogen.     

Assimilation of nitrate and ammonium by the Scots pine (Pinus sylvestris) seedling under conditions of high nitrogen supply

Seedlings of Scots pine ( Pinus sylvestris L.) were grown on perlite for 21 days under controlled conditions. Apart from the water control, KNO₃ (15 m M ), (NH₄)₂SO₄ (7.5 m M ), and NH₄NO₃ (15 m M ) were offered to study the effects of a high nitrogen supply on nitrogen assimilation. In some experiments 1.3 m M potassium was added to the basic ammonium solutions. In labelling studies nitrate and ammonium were 2.3 atom%¹⁵N-enriched. It was found that over the 21-day period approximately three times more ammonium-N was taken up than nitrate-N. However, nitrate and ammonium, applied simultaneously, were taken up to the same extent as if they were applied separately (additivity). The presence of K⁺ in the medium did not affect N-uptake. Among the soluble N-containing compounds nitrate, ammonium and 8 amino acids were quantified. It was found that assimilation of nitrate can cope with the uptake of NO⁻₃ under all circumstances. Neither free nitrate nor ammonium or amino acids accumulated to an extent exceeding the values of water-grown seedlings. On the other hand, in case of high ammonium supply considerably more nitrogen was taken up than could be incorporated into nonsoluble N-containing substance ('protein'). The remaining nitrogen was found to accumulate in intermediary storage pools (free NH₄⁺, glutamine, asparagine, arginine). Part of this accumulated N could be incorporated into protein when potassium was offered in the nutrient solution. It is concluded that potassium is a requirement for a high rate of protein synthesis not only in crop plants but also in conifers.     

抑制剂和猪粪对尿素氮在稻田土壤中转化的影响

为了阐明稻田土壤中尿素在配施抑制剂和猪粪的情况下不同形态氮的响应特征,探究不同管理措施下稻田土壤氮素保持和供给能力。本研究采用¹⁵N标记尿素进行盆栽试验,设置不施肥(CK)、猪粪(M)、尿素(N)、猪粪+尿素(NM)、尿素+抑制剂(NI)和尿素+抑制剂+猪粪(NIM)6个处理。抑制剂选用脲酶抑制剂(PPD+NBPT)和硝化抑制剂(DMPP)组合,测定返青期、分蘖期和成熟期土壤氮库的分配、尿素氮在氮库中的保存及水稻吸氮状况。结果表明: 施用猪粪显著提高了土壤铵态氮、固定态铵含量和微生物生物量氮,提高了分蘖期尿素氮在各氮库中的贮存,显著增加了水稻产量。与N处理相比,添加抑制剂促进了NH₄⁺的矿物固定和微生物对尿素氮的固持;与NM处理相比,施用抑制剂增加了黏土矿物对¹⁵NH₄⁺的固定。通径分析表明,施用猪粪能促进水稻吸收肥料氮,增加水稻产量;添加抑制剂可通过铵的矿物固定将更多的肥料氮暂时储存;NIM能将更多的氮贮存在微生物生物量氮中,至作物生长后期,铵的释放和微生物周转矿化可为水稻提供更多的有效氮源。在我国北方稻田,NM和NIM处理是推荐的施肥方式。     

ASSAY OPTIMIZATION AND REGULATION OF UREASE ACTIVITY IN TWO MARINE DIATOMS

An in vitro urease enzyme assay was developed for the marine diatoms Thalassiosira pseudonana Hasle et Heimdal (clone 3H) and T. weissflogii (Grunow) Fryxell et Hasle (clone Actin). This assay involves the colorimetric measurement of ammonium following the hydrolysis of urea in crude cell homogenates and it is the first assay to account for the rate of nitrogen assimilation in both species grown on urea as the sole nitrogen source. Urease activity was found to be present regardless of nitrogen source, although activities showed distinctly different patterns depending on the species examined and form of nitrogen supplied. Under nitrogen-replete conditions, urease activity in T. pseudonana was present constitutively when grown on NH₄⁺ and upregulated when grown on NO₃⁻ or urea. In nitrogen-replete T. weissflogii , urease activity was present at high constitutive levels regardless of the nitrogen source and showed no upregulation. Nitrogen starvation did not upregulate activity in either species.     

Response of nitrogen metabolism to boron toxicity in tomato plants

Boron (B) toxicity has become important in areas close to the Mediterranean Sea where intensive agriculture has been developed. The objective of this research was to study the effects of B toxicity (0.5 m m and 2.0 m m B) on nitrogen (N) assimilation of two tomato cultivars that are often used in these areas. Leaf biomass, relative leaf growth rate (RGR_L), concentration of B, nitrate (NO₃⁻), ammonium (NH₄⁺), organic N, amino acids and soluble proteins, as well as nitrate reductase (NR), nitrite reductase (NiR), glutamine synthase (GS), glutamate synthetase (GOGAT) and glutamate dehydrogenase (GDH) activities were analysed in leaves. Boron toxicity significantly decreased leaf biomass, RGR_L, organic N, soluble proteins, and NR and NiR activities. The lowest NO₃⁻ and NH₄⁺ concentration in leaves was recorded when plants were supplied with 2.0 m m B in the root medium. Total B, amino acids, activities of GS, GOGAT and GDH increased under B toxicity. Data from the present study prove that B toxicity causes inhibition of NO₃⁻ reduction and increases NH₄⁺ assimilation in tomato plants.     

Functional characterization of an ammonium transporter gene from Lotus japonicus

NH₄⁺ is the main product of symbiotic nitrogen fixation and the external concentration of combined nitrogen plays a key regulatory role in all the different step of plant-rhizobia interaction. We report the cloning and characterization of the first member of the ammonium transporter family, LjAMT1;1 from a leguminous plant, Lotus japonicus. Sequence analysis reveals a close relationship to plant transporters of the AMT1 family. The wild type and two mutated versions of LjAMT1;1 were expressed and functionally characterized in yeast. LjAMT1;1 is transcribed in roots, leaves and nodules of L. japonicus plants grown under low nitrogen conditions, consistent with a role in uptake of NH₄⁺ by the plant cells.     

较长时间内15N标记的氨态氮和硝态氮在小嵩草草甸中的运移规律

运用15N稳定性同位素技术,对15N标记的硝酸盐和铵盐在输入小嵩草(Kobresia pygaea C.B.Clarke)草甸11～13个月后的运移规律进行了研究.在经历11～13个月后,进入无机氮库中的15N很少,一般不超过所输入氮素的l%,而较多的1 5N为土壤有机质、土壤微生物和植物所固持.NO3--15N和NH4 -1 5N在小嵩草草甸中的运移规律差异很大.在11、12和13个月后,NO3--15N的总恢复率分别为92.83%、92.64%和79.96%;而NH4 -15N的则分别为49.6%、63.33%和66.22%.两者的差异在土壤有机质、土壤微生物和植物等库之间的分配中更加明显.输入NO3--15N时在11、12个月后植物所固持的15N最多,而土壤微生物和土壤有机质所固持的15N比较接近,而在13个月后,土壤有机质和植物所固持的15N接近,而土壤微生物所固持的15N下降许多;当输入NH4 -15N,土壤有机质所固持的1 5N比植物和土壤微生物所固持的都多,而且植物所固持的15N比较稳定,而土壤微生物所固持的15N则有较大变化.这表明在较长的时间内嵩草草甸对NO3-和NH4 的固持能力是不一样的.