Intraspecific differences in nitrogen assimilating enzymes in spinach under contrasting forms of nitrogen supply 1

Intraspecific differences in the activities of nitrate reductase (NR), glutamine synthetase (GS), NADH dependent glutamate synthase (NADH‐GOGAT), and glutamate dehydrogenase (GDH) under contrasting forms of nitrogen (N) supply were studied in tissues of three spinach (Spinacia oleracea L.) cultivars. The varieties (Viroflay, Butterflay, and Giant) were smooth, curly and semicurly leaved, respectively. The plants were grown in nutrient solutions containing NO₃ as the sole source of N (100:0) and NO₃ plus NH₄ (80:20). Giant, the NH₄ tolerance of which had been evaluated in growth and on the basis of nutrient uptake, had much higher GS and GDH activities in the roots and higher NR and NADH‐GOGAT levels in the leaves of plants grown on NO₃ and NH₄ than that grown on NO₃ alone. On the level of N assimilating enzymes of Butterflay, mixed N nutrition caused an increase of GDH and NADH‐GOGAT in leaves and roots and at the same time a decrease of GS in the roots and NR in the leaves. An inverse relationship between GS and GDH activities was detected in the leaves and foots of Virofiay grown with both N sources. Finally, Viroflay gave the highest levels of GDH irrespective of the NO₃:NH₄ assayed, whereas the leaves of Giant were GDH deficient in comparison with the other cultivars. In addition, the GS activity approached zero in the roots of spinach cultivars characterized by hardly any NH₄ tolerance, whereas in those of Giant it increased remarkably with the supply of NO₃ plus NH₄.     

Determining potential glutamine synthetase and glutamate dehydrogenase activity in soil

Ammonium (NH₄⁺), an important nitrogen (N) source for microorganisms, is assimilated via two major pathways. One route is catalyzed by glutamate dehydrogenase (GDH), while the other mechanism involves two enzymes, glutamine synthetase (GS) and glutamate synthase (GOGAT). The GS/GOGAT enzyme system requires more energy to operate, but has a much higher affinity for NH₄⁺ than GDH. We describe procedures to determine potential GS and GDH activity in soil samples. GS and GDH are intracellular enzymes. We used chloroform fumigation to make cell membranes permeable for substrates and products of the enzymes. Fumigation for 4 h increased GS activity almost ten-fold compared to the unfumigated control. Under optimized assay conditions, GS activity increased linearly for at least 80 min, indicating that the substrates were not limiting. In contrast to what was found for GS activity, direct addition of substrates to the soil to assay GDH activity did not result in a linear increase in GDH activity over time. A linear response for 3 h, however, resulted when the soil samples were first extracted with buffer solution and the reagents were added after centrifugation. The differences between the assays explain why fumigation for 3 d prior to the assay increased GDH activity by only 60%. In a microcosm study with glucose and NH₄⁺ addition, the activity of the two enzymes depended on the carbon (C) to N ratio of the amendment. With increasing C to N ratios from 5 to 120, GS activity doubled, while C to N ratios higher than 120 did not further increase GS activity. In contrast, GDH activity decreased by 13% with increasing C to N ratios from 5 to 200. The GDH to GS activity ratio in soil may therefore yield valuable information about the availability of N relative to C at a specific time.     

Do reactive oxygen species (ROS) induced by NaCl contribute to ammonium accumulation in Spartina alterniflora?

Growth, activity of antioxidant enzymes viz. glutathione reductase (GR), superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and guaiacol peroxidase (GPX), and some metabolic processes related to ammonium metabolism were investigated in a salt‐tolerant Spatina alterniflora. In comparison to 0 mM–NaCl treatment, growth of S. alterniflora plant increased significantly at 200 mM NaCl, but was highly inhibited at 500 mM NaCl. Ammonium concentration in the leaves and roots increased 2.1–3.4 times when plants were treated with 500 mM NaCl. Under 200 mM NaCl, antioxidant‐enzyme activities increased, however, at 500 mM the antioxidant system was unable to compensate reactive oxygen species induced by NaCl. At this high level of salinity, ammonium production through nitrate reductase (NR) was inhibited, but no significant changes in the activities of glutamine synthetase (GS) or glutamate dehydrogenase (GDH) were found. We conclude that the accumulation of ammonium under high salt stress was not due to inhibition of the assimilatory activities of GS or GDH. Ammonia accumulation under high salinity may result from amino acid and protein catabolism activated by reactive oxygen species (ROS) and/or a lack of carbon skeletons to incorporate ammonium into organic molecules due to a decrease in photosynthetic activity in salt‐stressed plants.     

Growth response and nitrogen metabolism of sunflower (Helianthus annuus L.) to vermicompost and biogas slurry under salinity stress

Sunflower was grown under saline media with or without vermicompost amendment and biogas slurry, the organic fertilizers. A randomized complete block design with five replications was used. Forty-five pots were divided in three sets comprising of 15 pots each. Out of 15 pots of each sets, five pots of each were subjected to different levels of saline water irrigation i.e. electrical conductivity (EC): 0.5, 4.8 8.6 dS/m). Amendments of vermicompost and biogas slurry have shown some reduction of sodium induced inhibitory effects. Analyses of fresh and dry weight of leaves, ions, amino acid, protein and nitrogen metabolism enzymes in leaves etc., have been undertaken with reference to above-mentioned treatments. Organic manure amendments improved growth yield, nitrate and protein content and decreased the amount of sodium (Na⁺), chloride (Cl^?), ammonium and total amino acid under saline or non-saline condition. Activities of nitrogen (N)-assimilating enzymes i.e. nitrate reducatse (NR, EC 1.6.6.1), nitrite reductase (NiR, EC 1.6.6.4), glutamine synthetase (GS, EC 6.3.1.2) and glutamate synthase (NADH-GOGAT, EC 1.4.1.14) were enhanced to some extent irrespective to non saline or saline condition. Under salinity NADH-glutamate dehydrogenase (GDH, EC 1.4.1.2) activity was stimulated concomitantly with the increasing ammonium contents and proteolysis activity in the leaves and organic manure did not show a significant difference as compared to their respective control. With respect to salt stress, among the two above-mentioned organic manure, vermicompost showed better result in the entire studied parameter as compared to the biogas slurry.     

Effects of supplemental nitrogen on nitrogen‐assimilation enzymes,free amino nitrogen,soluble sugars,and crude protein of rice

Abstract

An upland rice variety IAC‐47 was grown in a greenhouse to determine the effect of foliar nitrogen (N) supplementation during grain development on the activity of the N assimilation enzymes, nitrate reductase (NR) and glutamine synthetase (GS), on free amino‐N content and leaf soluble sugars, and on grain crude protein content. At 10 and 20 days after anthesis (DAA), the leaves were fertilized with a liquid fertilizer containing 32% N as 12.8% urea, 9.6% ammonium (NH₄), and 9.6% nitrate (NO₃) in increasing rates corresponding to 0,20+20, 40+40, and 60+60 kg N ha^‐1. Leaves were collected twice (at 12 DAA and 14 DAA for GS activity, sugar and amino‐N content, and at 11 and 13 DAA for NRA) after each application of leaf N. The late foliar application of N increased significantly grain crude protein without a corresponding decrease in grain weight. The NR activity (NRA) increased after the foliar application of N. In the flag leaf, 60+60 kg N ha^‐1 (21 DAA) resulted in higher NRA (20x over the control), while GS activity was smaller than the control. At 22 DAA there was an increase in GS activity in the flag leaf at 20+20 N level. However, the GS activity decreased as applied N levels increased. Also at the 20+20 level, there were increases in free amino‐N in the flag leaf and second leaf at the final harvest. Throughout the experiment, plants at the 60+60 N level had the lowest levels of soluble sugars. Increases in crude protein were highest at 40+40 N level (27.9%), followed by 60+60 (18.7%).     

Effect of Amino Acid Fertilization on Nitrate Assimilation of Leafy Radish and Soil Chemical Properties in High Nitrate Soil

Abstract

The objective of the present work was to investigate the corresponding uptake and assimilation of nitrate (NO₃ ^?) in leafy radish and effects on soil chemical properties by foliar application of amino acid fertilizer (AAF). The activity of the enzymes related to the process of NO₃ ^? reduction (NR: nitrate reductase; NiR: nitrite reductase; GS: glutamine synthetase) and the content of NO₃ ^?, total N, and the end products of this process (amino acids and proteins) were analyzed. The soils were sampled twice, and some chemical properties were analyzed. The results of this study showed that application of AAF increased biomass production, and nitrogen (N) utilization in the treatment of a low rate of AAF increased 55% over the control. In addition, the activities of the enzymes were affected differently depending on applied rate of AAF. Furthermore, the NO₃ ^? content was reduced 4–24%, and total N content was increased 14–32% by AAF treatments. Finally, application of AAF improved uptake efficiency of N from soil and prevented N loss by leaching.     

不同氮素水平对甜菜硝酸还原酶和亚硝酸还原酶活性的影响

以当前甜菜（Beta Vulgaris L.）生产主栽品种KWS0143为试材,设立4个氮素水平（N 0、60、120、180 kg/hm2）,研究硝态氮肥对甜菜硝酸还原酶（nitrate reductase, NR）和亚硝酸还原酶（nitrite reductase, NiR）活性以及光合速率与叶绿素总含量的影响,探讨了氮素水平与NR及NiRA之间的关系。结果表明:在甜菜生育期间,光合速率呈单峰曲线变化,NR、NiR活性及叶绿素总含量基本呈双峰曲线变化,叶绿素变化曲线的高峰期早于NR活性,NR活性早于NiR活性。氮肥用量在N 120 kg/hm2时显著提高了甜菜光合速率、叶绿素总含量、NR活性和NiR活性。相比于N 120 kg/hm2,N 180 kg/hm2时光合速率及叶绿素总含量没有明显变化,NR与NiR活性则有一定的提高。本试验中,甜菜产量随氮水平的增加而提高;含糖率则相反,随氮水平的增加而降低,N 120 kg/hm2水平下产糖量最高。     

Nitrogen Metabolism in Tomato Plants Under Cadmium Stress

Abstract

Young seedlings of tomato (Lycopersicon esculentum Mill.) grown on nitric medium and treated with different cadmium (Cd) concentrations (0–50 µM) were used. Results obtained show that Cd remains predominantly located in the roots, which then seem to play the role of trap‐organs. Increasing Cd concentration in the medium leads particularly to a decrease in NO₃ ^? accumulation, together with decrease in the activities of nitrate reductase (NR), nitrite reductase (NiR), and glutamine synthetase (GS). On the other hand, stimulations were observed for ammonium accumulation, anabolitic activity of glutamate dehydrogenase (GDH) and protease activity. Contrarily, decreases were obtained for soluble protein contents and catabolitic activity of GDH. These results were discussed in relation to the hypothesis attributing to the mitochondrial enzyme NADH‐dependent GDH, an important role in the plant defense processes, in maintaining the glutamate pool in conditions where the need for NH₄ ⁺ detoxification is increased by heavy metals.     

INFLUENCE OF HIGH TEMPERATURE AND UREA FERTILIZATION WITH N-(N-BUTYL) THIOPHOSPHORIC TRIAMIDE AND DICYANDIAMINDE ON COTTON GROWTH AND PHYSIOLOGY

The objective of this growth chamber study was to evaluate the effect of adding N-(n-butyl) thiophosphoric triamide (NBPT) and dicyandiaminde (DCD) to urea fertilizer, on the physiology and growth of cotton (Gossypiumhirsutum L.) under normal and high temperatures. Treatments consisted of two day temperature regimes, 30°C and 38°C, and five nitrogen fertilization applications: unfertilized control, 125 kg ha^?1 of urea, 93 kg ha^?1 of urea, 93 kg ha^?1 urea + NBPT, and 93 kg ha^?1 urea + NBPT + DCD. The addition of NBPT to urea fertilizer had positive effects on leaf chlorophyll, leaf area, dry matter, nitrogen (N) uptake, and N use efficiency. The absence of a significant interaction effect indicated that N fertilization was not influenced by temperature. Deficiency of N significantly decreased leaf chlorophyll, increased glutathione reductase, decreased protein and increased leaf nitrate reductase. Physiological changes under high temperature included increased plant N uptake, glutamine synthetase, leaf chlorophyll, protein content, plant height and leaf area were due to high N uptake and utilization.     

Nitrate leaching,nitrous oxide emission and N use efficiency of aerobic rice under different N application strategy

A field study was conducted in the sub-humid tropical region of India to examine the effect of different nitrogen (N) management strategies on nitrate leaching, nitrous oxide (N₂O) emission and N use efficiency in aerobic rice. Treatments were: control (no N), 120 kg N ha^?1 applied as prilled urea (PU) in conventional method, 120 kg N ha^?1 applied as neem coated urea (NCU) in conventional method, N applied as PU on the basis of leaf colour chart (LCC) reading, N applied as NCU on the basis of LCC reading, and 120 kg N ha^?1 applied as PU and farm yard manure (FYM) in 1:1 ratio. Results showed that 3.4–16.1 kg NO₃-N ha^?1 was leached below 45 cm depth and 0.61–1.12 kg N₂O-N ha^?1 was emitted from aerobic rice during the growing season. NCU when applied conventionally reduced nitrate-nitrogen (NO₃-N) leaching and N₂O emission by 18.6% and 21.4%, respectively However when applied on the basis of LCC reading NCU reduced NO₃-N leaching by 39.8% as compared to PU applied in conventional method. NCU when applied on the basis of LCC reading synchronized N supply with demand and reduced N loss, which resulted in higher yield and N use efficiency.     

Effect of Deficit Irrigation and Dairy Manure on Winter Wheat Yield,Soil Physical Health,and Nitrate Leaching

ABSTRACT

The effect of deficit irrigation (DI) on wheat crop yield, soil physical parameters and on nitrate nitrogen movement in soil profile was evaluated under application of dairy manure and nitrogen fertilizer. Two levels of DI were taken as I_0.6 (60% FC) and I_0.8 (80% FC) along with two dairy manure levels (20 and 25 Mg ha^?1) and three nitrogen levels (80, 100, and 120 kg ha^?1). The grain yield was high under I_0.8 than I_0.6, whereas the irrigation level has no significant effect on soil organic carbon contents. Dairy manure, irrigation, and nitrogen indicated strong interaction with each other for all yield-related parameters during both years of study, however, results for 2nd year were highly positive. Soil nitrate nitrogen movement was significantly affected under I_0.8 with high rate of dairy manure (25 Mg ha^?1) and nitrogen fertilizer (120 kg ha^?1). Results concluded that combined application of dairy manure (25 Mg ha^?1) and nitrogen fertilizer (120 kg ha^?1) under DI level I_0.8 resulted in high grain yield. To overcome water scarce conditions, further experiments can be designed by addition of various organic matters in different combination that enhances the yield and soil health.     

Fate of fertilizer nitrogen applied to grassland. I. Field leaching results

Results are presented from a 3 year investigation into nitrate leaching from isolated 0.4 ha grassland plots fertilized with 250, 500 and 900 kg N ha^?1 a^?1. Cumulative nitrate leaching over the 3 years was equivalent to 1.5%, 5.4% and 16.7% of the fertilizer applied at 250, 500 and 900 kg N ha^?1 rates respectively. Over a whole drainage season, mean nitrate leachate concentrations at 250 kg N ha^?1 did not exceed 4 mgl^?1, although maximum values of 13.3 mgl^?1 were observed. In contrast, at 900 kg N ha^?1, the mean nitrate leachate concentration in two of the years exceeded 90 mgl^?1. Mineral nitrogen balances constructed for the 1979 growing season indicated that leaching at 250 kg N ha^?1 was low because net mineralization of soil organic nitrogen was small, and crop nitrogen uptake almost balanced fertilizer application. Although the pattern of nitrate leaching suggested that by-passing occurred in the movement of water down the soil profile, it was not possible to confirm this using simulation models of leaching. Possible reasons for this, including the occurrence of rapid water flow down gravitationally drained macropores, are discussed.     

Fate of fertilizer nitrogen.

Results are presented from a three year lysimeter investigation, employing single (¹⁵NH₄NO₃) and double (¹⁵NH₄¹⁵NO₃) labelled ammonium nitrate to study the uptake of soil and fertilizer nitrogen by cut ryegrass at 250, 500 and 900 kg N ha^?1 a^?1. Average annual recoveries of nitrogen were equivalent to 99,76 and 50% of the nitrogen added at 250, 500 and 900 kg N ha^?1, respectively. At 250 kg N ha^?1 the difference between the overall nitrogen recovery and the fertilizer recovery was almost entirely attributable to pool substitution resulting from mineralization/immobilization turnover (MIT). At 900 kg N ha^?1 both the low overall recovery of nitrogen and the low fertilizer recovery reflected the large excess of available nitrogen over crop requirements. No evidence of ‘priming’ was obtained. Analysis of the results from single and double labelled lysimeters using simultaneous equations indicated that at 250 kg N ha^?1,～70% of the nitrogen in the crop was derived from the ammonium pool. At 500 kg N ha^?1 this dropped to 64%, while at 900 kg N ha^?1 the figure was 59%. There was a suggestion that at the lower application rates, preferential uptake of ammonium was occurring but that as N supply exceeded crop requirements, nitrate was the major N source. Despite the preferential exploitation of the ammonium pool, at 250 and 500 kg N ha^?1 pool substitution resulting from MIT resulted in lower recoveries of fertilizer ammonium compared with fertilizer nitrate.     

镉对超富集植物圆锥南芥氮素代谢的影响研究

采用土培方法,研究了不同Cd添加(01、02、04、08、01、60和240 mg kg-1)对超富集植物圆锥南芥叶片中氮素含量以及氮代谢关键酶活性的影响。结果表明,随着Cd添加浓度的增加,圆锥南芥的生物量、叶片数、直径长以及Cd含量均呈增加趋势。Cd添加浓度为240 mg kg-1时,生物量增加了137%,叶片数增加了1.02倍,直径较对照增加了130%,叶片中Cd含量达到451 mg kg-1。NO3--N和NH4+-N呈先降低后升高的变化趋势,在Cd浓度为240 mg kg-1时,达最大值。在Cd处理浓度为102、04、08、0 mg kg-1时,NO3--N和NH4+-N含量间无显著性差异(p>0.05)。在Cd添加浓度为240 mg kg-1时,硝酸还原酶(NR)活性只相当于对照的67%,与对照间的差异显著。在Cd处理浓度为10~160 mg kg-1时,NR活性与对照间无显著性差异(p>0.05)。谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)活性以及可溶性蛋白质含量在Cd处理浓度为10~240 mg kg-1时,与对照间无显著性差异(p>0.05)。谷氨酸脱氢酶(GDH)活性呈先升高后降低的变化趋势。游离脯氨酸含量在Cd浓度为102、0、408、01、602、40 mg kg-1时,分别为对照的0.67倍1、.29倍1、.54倍、2.38倍、2.54倍、1.14倍。     

Impact of nitrogen supply on glucosinolate content and phenolic acids in cauliflower varieties

Application of nitrogen (N) is a common practice used to achieve profitable yields in horticultural crops and N application can be used as a tool to manipulate the enhancement of phytochemicals in vegetable crops to address consumer-oriented quality production. Our previous findings recommended 90?kg?ha^?1 for certain types of cauliflower varieties without compromising yields. Thus, this study was aimed to investigate the effect of N application on glucosinolates and phenolic acids, at harvest, in varieties ‘Largardo’, ‘Eskimo’ and ‘CF-744’ grown in the field. N was applied as ammonium nitrate (NH₄NO₃) at concentrations from 0, 60, 90, 120, 150 to 180?kg?ha^?1. Variety ‘CF-744’ was more sensitive to N supply and at 180?kg?ha^?1 N it showed the highest accumulation of glucosinolates (sinigrin, glucoiberin, progoitrin, 4-methoxyglucobrassicin) at harvest. However, 90?kg?ha^?1 N supply demonstrated the highest accumulation of majority of the glucosinolates in varieties ‘Largardo’ and ‘Eskimo’. Different varieties responded differently to N supply and glucosinolate levels in cauliflowers. Also, different varieties responded differently to N supply and antioxidant property. In all three varieties, the N supply at 120?kg?ha^?1 showed the highest accumulation of protocatechoic acid, 4-hydroxy benzoic acid, ρ-coumaric acid and caffeic acid.     

A lysimeter study of the fate of fertilizer nitrogen in spring barley crops grown on shallow soil overlying Chalk: crop uptake and leaching losses

The objective of this work was to determine the fate of fertilizer nitrogen (labelled with nitrogen-15) applied to an undisturbed shallow soil overlying Chalk contained in 10 lysimeters (80 cm diameter, 135 cm deep). Measurements are reported of the nitrogen uptake by four spring barley crops and the rate and extent of leaching of nitrate beyond the roots. The crops were fertilized with 0, 80 or 120 kg N ha^?1 in each of four years, but only the first application in 1977 was labelled with nitrogen ?15. Rainfall and irrigation approximated to the long-term average, but in two treatments dry or wet spring conditions were imposed for the 10 weeks after sowing the first crop in 1977. The dry matter and grain yields of the spring barley crops varied from year to year in the ranges 8.7–14.0 t ha^?1 and 3.5–6.1 t ha^?1 respectively. The total nitrogen harvested in the crop approximated to the amount of nitrogen applied in each year with an apparent recovery of fertilizer in the range 38–76%. The recovery of nitrogen derived from fertilizer (labelled with nitrogen-15) was 46–54% in the first crop and after 2 years rapidly declined to below 1%. The total amount of nitrogen-15 labelled fertilizer recovered in four barley crops was 49–57% of that applied. Mean annual nitrate concentrations in water draining from the base of the lysimeters were in the range 11.8–26.7 mg N 1^?1 and did not differ significantly between nitrogen fertilizer treatments (0, 80 and 120 kg N ha^?1 a^?1). In all treatments nitrate concentrations varied considerably within each growing season, with a cycle of peaks and troughs. Annual losses of nitrate were in the range 39–128 kg N ha^?1, and the mean annual losses over the 4 years varied between lysimeters from 65 to 83 kg N ha^?1. Nitrogen-15 labelled nitrate was detected in the first drainage water collected in autumn following its spring application, 5 months earlier. Recovery of fertilizer-derived nitrogen in drainage water was greatest during the winter following the second barley crop, and was 3.4–3.7% of the nitrogen-15 applied. Over the 4 years of the experiment 6.3–6.6% of labelled fertilizer was accounted for in drainage water, representing 2–3% of the total nitrogen lost by leaching.     

Nitrate leaching loss under annual and perennial pastures with and without lime on a duplex (texture contrast) soil in humid southeastern Australia

Mineral N accumulates in autumn under pastures in southeastern Australia and is at risk of leaching as nitrate during winter. Nitrate leaching loss and soil mineral N concentrations were measured under pastures grazed by sheep on a duplex (texture contrast) soil in southern New South Wales from 1994 to 1996. Legume (Trifolium subterraneum)‐based pastures contained either annual grass (Lolium rigidum) or perennial grasses (Phalaris aquatica and Dactylis glomerata), and had a control (soil pH 4.1 in 0.01 m CaCl₂) or lime treatment (pH 5.5). One of the four replicates was monitored for surface runoff and subsurface flow (the top of the B horizon), and solution NO₃^– concentrations. The soil contained more mineral N in autumn (64–133 kg N ha^?1 to 120 cm) than in spring (51–96 kg N ha^?1), with NO₃^– comprising 70–77%. No NO₃^– leached in 1994 (475 mm rainfall). In 1995 (697 mm rainfall) and 1996 (666 mm rainfall), the solution at 20 cm depth and subsurface flow contained 20–50 mg N l^?1 as NO₃^– initially but < 1 mg N l^?1 by spring. Nitrate‐N concentrations at 120 cm ranged between 2 and 22 mg N l^?1 during winter. Losses of NO₃^– were small in surface runoff (0–2 kg N ha^?1 year^?1). In 1995, 9–19 kg N ha^?1 was lost in subsurface flow. Deep drainage losses were 3–12 kg N ha^?1 in 1995 and 4–10 kg N ha^?1 in 1996, with the most loss occurring under limed annual pasture. Averaged over 3 years, N losses were 9 and 15 kg N ha^?1 year^?1 under control and limed annual pastures, respectively, and 6 and 8 kg N ha^?1 year^?1 under control and limed perennial pastures. Nitrate losses in the wet year of 1995 were 22, 33, 13 and 19 kg N ha^?1 under the four respective pastures. The increased loss of N caused by liming was of a similar amount to the decreased N loss by maintaining perennial pasture as distinct from an annual pasture.     

叶面喷施尿素对葡萄氮代谢相关基因表达的影响

本研究利用VitisEST数据库中EST序列片段结合RT-PCR方法克隆了与氮代谢相关的硝酸盐还原酶（NR）,亚硝酸还原酶（NiR）,谷氨酰胺合成酶（GS）,谷氨酸脱氢酶（GDH）和天冬酰胺合成酶（AS）的基因,并对它们进行亚细胞定位分析。同时,采用半定量RT-PCR和荧光定量PCR法研究葡萄叶面喷施不同浓度尿素后5个基因的表达变化。结果表明,5个基因在葡萄幼叶中的表达水平显著高于老叶,说明叶面喷施尿素以喷布到幼叶对5个基因的表达水平影响显著;5个基因在不同时间段的表达水平差异不一致,但表达水平均高于对照,以0.3%和0.5%浓度的尿素对其影响明显;适当提高尿素水平既能提高氮代谢基因的表达水平,又能提高果实大小等果实指标,使其达到同步增加。喷施尿素6 h后,GS的表达量上升幅度明显高于其它基因,而NR在喷施尿素后48 h内一直保持着比较高的表达水平;NiR、AS表达量的变化趋势分别与NR、GS相一致,并且NR﹥NiR,GS﹥AS。     

INTERACTIVE EFFECTS OF SALINITY AND N ON PEPPER (CAPSICUM ANNUUM L.) YIELD,WATER USE EFFICIENCY AND ROOT ZONE AND DRAINAGE SALINITY

The aim of this study was to determine the salt tolerance of pepper (Capsicum annuum L.) under greenhouse conditions and to examine the interactive effects of salinity and nitrogen (N) fertilizer levels on yield. The present study shows the effects of optimal and suboptimal N fertilizer levels (270 kg ha^?1 and 135 kg ha^?1) in combination with five different irrigation waters of varying electrical conductivity (EC) (EC_iw = 0.25, 1.0, 1.5, 2.0, 4.0, and 6.0 dS m^?1) and three replicates per treatment. At optimal N level, yield decreased when the irrigation water salinity was above EC_iw 2 dS m^?1. At the suboptimal N level, a significant decrease in yield occurred only above EC_iw 4 dS m^?1. At high salinity levels the salinity stress was dominant with respect to yield and response was similar for both N levels. Based on the results it can also be concluded that under saline conditions (higher than threshold salinity for a given crop) there is a lesser need for N fertilization relative to the optimal levels established in the absence of other significant stresses.