Assimilation of [N]Nitrate and [N]Nitrite in Leaves of Five Plant Species under Light and Dark Conditions

Light dependency of nitrate and nitrite assimilation to reduced-N in leaves remains a controversial issue in the literature. With the objective of resolving this controversy, the light requirement for nitrate and nitrite assimilation was investigated in several plant species. Dark and light assimilation of [¹⁵N]nitrate and [¹⁵N]nitrite to ammonium and amino-N was determined with leaves of wheat, corn, soybean, sunflower, and tobacco. In dark aerobic conditions, assimilation of [¹⁵N]nitrate as a percentage of the light rate was 16 to 34% for wheat, 9 to 16% for tobacco, 26% for corn, 35 to 76% for soybean, and 55 to 63% for sunflower. In dark aerobic conditions, assimilation of [¹⁵N]nitrite as a percentage of the light rate was 11% for wheat, 7% for tobacco, 13% for corn, 28 to 36% for soybeans, and 12% for sunflower. It is concluded that variation among plant species in the light requirement for nitrate and nitrite assimilation explains some of the contradictory results in the literature, but additional explanations must be sought to fully resolve the controversy.

In dark anaerobic conditions, the assimilation of [¹⁵N]nitrate to ammonium and amino-N in leaves of wheat, corn, and soybean was 43 to 58% of the dark aerobic rate while dark anaerobic assimilation of [¹⁵N]nitrite for the same species was 31 to 41% of the dark aerobic rate. In contrast, accumulation of nitrite in leaves of the same species in the dark was 2.5-to 20-fold higher under anaerobic than aerobic conditions. Therefore, dark assimilation of nitrite cannot alone account for the absence of nitrite accumulation in the in vivo nitrate reductase assay under aerobic conditions. Oxygen apparently inhibits nitrate reduction in the dark even in leaves of plant species that exhibit a relatively high dark rate of [¹⁵N]nitrite assimilation.

     

Enhanced N-Transfer from a Soybean to Maize by Vesicular Arbuscular Mycorrhizal (VAM) Fungi

Using a split-root technique, roots of soybean plants were divided between two pots. In one of the two pots, two maize plants were grown and half of those pots were inoculated with the vesicular arbuscular mycorrhizal (VAM) fungus, Glomus fasciculatus. Fifty-two days after planting, ¹⁵N-labeled ammonium sulfate was applied to the pots which contained only soybean roots. Forty-eight hours after application, significantly higher values for atom per cent ¹⁵N excess were found in roots and leaves of VAM-infected maize plants as compared with the non-VAM-infected maize plants. Results indicated that VAM fungi did enhance N transfer from one plant to another.     

Nitrate Reduction and Partitioning of Nitrogen in Komatsuna (Brassica campestris L. var. rapa) Plants: Compartmental Analysis in Combination with 15N Tracer Experiments

Komatsuna (Brassica campestris L. var. rapa cv. Misugi) is aleafy vegetable that readily accumulates nitrate in its tissues.Plants grown hydroponically with 2 mM nitrate in a greenhousewere fed ¹⁵N-labeled nitrate for 2 h, followed with nonlabelednitrate for 8 h. At intervals of 2 h, the plants were sampledand analyzed for the distribution of 15N in the nitrate, aminoacids, and proteins in the tissues of roots, petioles plus midribs,and leaves. Nitrate reduction and nitrogen fluxes were examinedusing a compartmental analysis with 19 compartments and 28 transferrate constants. Nitrate existed in the three types of tissues as a large storagepool and a small metabolic pool. Nitrogen reduction was observedin these tissues, but mainly in the leaf tissue. Nitrate uptakeand reduction rates were smaller in the dark than in light,and particularly nitrate reduction in the shoot was less inthe dark. The rate of protein synthesis was much greater inthe light. The simulation, using compartment models and ¹⁵Ndistribution data, may be useful for estimating actual ratesof nitrogen transport and metabolism in the whole plant system. (Received October 15, 1986; Accepted March 26, 1987)     

Preferential Assimilation of Ammonium Ions from Ammonium Nitrate Solutions by Apple Seedlings

Apple seedlings, Pyrus malus L., were grown in complete nutrient solutions containing nitrate, ammonium, or ammonium plus nitrate as the nitrogen source. Uptake of nitrogen was calculated from depletion measurements of the nutrient solutions and by using ¹⁵N labelled nitrate and ammonium salts. If the plants received nitrogen as ammonium only or as nitrate only, the amounts of nitrogen taken up were similar. However, if the seedlings were supplied with ammonium nitrate, the amount of nitrate-nitrogen assimilated was only half that of ammonium. Nevertheless, if ammonium and nitrate were supplied to a plant with a split-root system, with each root half receiving a different ion, the uptakes were similar. The possibility of independent inhibition by ammonium of both nitrate uptake and reduction in the roots is discussed.     

Absorption and Re-distribution of Nitrogen during Growth and Development of Field Bean, Vicia faba

The absorption of nitrate by field bean plants at different times of development was investigated to determine the distribution and subsequent utilization of N up to the maturity of the beans. Nodulated plants were grown in sand culture and pulse-labelled with ¹⁵NO₃ for 4 short periods during development. Whole plants were sampled at intervals until maturity and analysed for total nitrogen and ¹⁵N. There was a rapid increase in the bean dry weight and total N after fruit set and, at maturity, the beans contained about three times as much N as had ever been in the rest of the plant. Nitrate absorption continued steadily throughout plant development but nodule N fixation made a progressively greater contribution to the total N in the plant. Over a quarter of the total ¹⁵N absorbed was found in the roots immediately after each treatment and little of this was subsequently translocated upwards in spite of the large increase in shoot N taking place. Above ground the most powerful “sink” for recently absorbed and redistributed N was initially the youngest vegetative tissue. However after fruit set competition developed between the growing tip and the developing beans until at the later stages the beans became the more powerful. Quantitatively redistribution made only a small contribution to bean N.     

Amino acids as a nitrogen source for tomato seedlings: The use of dual-labeled (C, N) glycine to test for direct uptake by tomato seedlings

Direct uptake of organic nitrogen (ON) compounds, rather than inorganic N, by plant roots has been hypothesized to constitute a significant pathway for plant nutrition. The aim of this study was to test whether tomatoes (Solanum lycopersicum cv. Huying932) can take up ON directly from the soil by using ¹⁵NH₄Cl, K¹⁵NO₃, 1, 2-¹³C₂¹⁵N-glycine labeling techniques. The ¹³C and ¹⁵N in the plants increased significantly indicating that a portion of the glycine-N was taken up in the form of intact amino acids by the tomatoes within 48 h after injection into the soil. Regression analysis of excess ¹³C against excess ¹⁵N showed that approximately 21% of the supplied glycine-N was taken up intact by the tomatoes. Atom% excesses of ¹⁵N and ¹³C in the roots were higher than in any shoots. Results also indicated rapid turnover of amino acids (e.g., glycine) by soil microorganisms, and the poor competitive ability of tomatoes in absorbing amino acids from the soil solution. This implies that tomatoes can take up ON in an intact form from the soil despite the rapid turnover of organic N usually found under such conditions. Given the influence of climatic change and N pollution, further studies investigating the functional ecological implications of ON in horticultural ecosystems are warranted.     

Nitrate Reduction in Roots and Shoots of Barley (Hordeum vulgare L.) and Corn (Zea mays L.) Seedlings: I. N Study

Nitrate reduction in roots and shoots of 7-day-old barley seedlings, and 9-day-old corn seedlings was investigated. The N-depleted seedlings were transferred for 24 h or 48 h of continuous light to a mixed nitrogen medium containing both nitrate and ammonium. Total nitrate reduction was determined by ¹⁵N incorporation from ¹⁵NO₃⁻, translocation of reduced ¹⁵N from the roots to the shoots was estimated with reduced ¹⁵N from ¹⁵NH₄⁺ assimilation as tracer, and the translocation from the shoots to the roots was measured on plants grown with a split root system. A model was proposed to calculate the nitrate reduction by roots from these data. For both species, the induction phase was characterized by a high contribution of the roots which accounted for 65% of the whole plant nitrate reduction in barley, and for 70% in corn. However, during the second period of the experiment, once this induction process was finished, roots only accounted for 20% of the whole plant nitrate reduction in barley seedlings, and for 27% in corn. This reversal in nitrate reduction localization was due to both increased shoot reduction and decreased root reduction. The pattern of N exchanges between the organs showed that the cycling of reduced N through the plant was important for both species. In particular, the downward transport of reduced N increased while nitrate assimilation in roots decreased. As a result, when induction was achieved, the N feeding of the roots appeared to be highly dependent on translocation from the leaves.     

A novel morphological response of maize (Zea mays) adult roots to heterogeneous nitrate supply revealed by a split‐root experiment

Approximately 35–55% of total nitrogen (N) in maize plants is taken up by the root at the reproductive stage. Little is known about how the root of an adult plant responds to heterogeneous nutrient supply. In this study, root morphological and physiological adaptations to nitrate‐rich and nitrate‐poor patches and corresponding gene expression of ZmNrt2.1 and ZmNrt2.2 of maize seedlings and adult plants were characterized. Local high nitrate (LoHN) supply increased both lateral root length (LRL) and density of the treated nodal roots of adult maize plants, but only increased LRL of the treated primary roots of seedlings. LoHN also increased plant total N acquisition but not N influx rate of the treated roots, when expressed as per unit of root length. Furthermore, LoHN markedly increased specific root length (m g^?1) of the treated roots but significantly inhibited the growth of the lateral roots outside of the nitrate‐rich patches, suggesting a systemic carbon saving strategy within a whole root system. Surprisingly, local low nitrate (LoLN) supply stimulated nodal root growth of adult plants although LoLN inhibited growth of primary roots of seedlings. LoLN inhibited the N influx rate of the treated roots and did not change plant total N content. The gene expression of ZmNrt2.1 and ZmNrt2.2 of the treated roots of seedlings and adult plants was inhibited by LoHN but enhanced by LoLN. In conclusion, maize adult roots responded to nitrate‐rich and nitrate‐poor patches by adaptive morphological alterations and displayed carbon saving strategies in response to heterogeneous nitrate supply.     

Nodule Nitrate Reductase as a Source of Reduced Nitrogen in Soybean, Glycine max

Hydroponically grown soybeans were fed ¹⁵N-enriched NaNO₃ at nine reproductive stages of development. The stem exudates contained excess ¹⁵N in the fully reduced nitrogen fraction. The soybean nodules had high nitrate reductase activity, whereas the roots had no detectable nitrate reductase activity. Based on these results, we concluded that the nodule nitrate reductase system has the potential of contributing significantly to the nitrogen economy of the plant.     

Experimental evidence for diel δ15N‐patterns in different tissues,xylem and phloem saps of castor bean (Ricinus communis L.)

Nitrogen isotope signatures in plants might give insights in the metabolism and allocation of nitrogen. To obtain a deeper understanding of the modifications of the nitrogen isotope signatures, we determined δ¹⁵N in transport saps and in different fractions of leaves, axes and roots during a diel course along the plant axis. The most significant diel variations were observed in xylem and phloem saps where δ¹⁵N was significantly higher during the day compared with during the night. However in xylem saps, this was observed only in the canopy, but not at the hypocotyl positions. In the canopy, δ¹⁵N was correlated fairly well between phloem and xylem saps. These variations in δ¹⁵N in transport saps can be attributed to nitrate reduction in leaves during the photoperiod as well as to ¹⁵N‐enriched glutamine acting as transport form of N. δ¹⁵N of the water soluble fraction of roots and leaves partially affected δ¹⁵N of phloem and xylems saps. δ¹⁵N patterns are likely the result of a complex set of interactions and N‐fluxes between plant organs. Furthermore, the natural nitrogen isotope abundance in plant tissue is not constant during the diel course – a fact that needs to be taken into account when sampling for isotopic studies.     

Estimation of N2-fixation by sugarcane,Saccharum sp., and soybean,Glycine max,grown in soil with15N-labelled organic matter

Summary Two varieties of sugarcane, and nodulated and non-nodulated soybean isolines, were planted in a soil previously mixed with¹⁵N-labelled plant material. 45 days was allowed to elapse before planting, to permit initiation of organic matter mineralization. Plants were grown for 60 days, then harvested, dried, weighed and analysed for total N. Analysis of soil samples pre-incubated in the laboratory was carried out to evaluate ammonium and nitrate from added organic matter. Dry weights of the soybean isolines were similar, but total N was higher for the nodulated line. Both sugarcane varieties showed similar weight and total N. Nitrogen derived from applied organic matter (NdfOM) was higher in non-nodulated soybean than in all other plants. Although there is the possibility of different¹⁵N availabilities between species, nitrogen derived from fixation (Nfix) was calculated based on the¹⁵N enrichment of the non-nodulating soybean. Nfix was 72% for nodulating soybean and ranged from 19 to 39% for different parts of sugarcane plants, despite high levels of available-N. Nitrogen derived from soil was calculated by difference. NdfOM was lower in roots than in upper parts (leaves+stalks) of plants. Use of¹⁵N labelled organic matter seems a useful approach to the longer term measurement of N₂-fixation.IAEA Project BRA/5/009-CENA.     

Uptake of nitrogen by flue-cured tobacco during maturation and senescence

A field experiment with flue-cured tobacco,Nicotiana tabacum L., was conducted to estimate the uptake and partitioning of nitrogen during maturation and senescence. On the 83rd day after transplanting (crop day 83), nitrate which had been leached from the plow layer was replaced with an equivalent amount of¹⁵N-labeled nitrate. Plants were harvested at crop day 83, 90, 96, 106, 113, and 127, and each of 11 plant parts was analyzed for nitrogen derived from the soil (NDS) and from the applied¹⁵N-labeled fertilizer (NDF). Equivalent quantities of NDF and NDS were taken up during the initial week after¹⁵N-fertilizer application; in the subsequent 5 weeks, ten times more NDS than NDF were taken up. It appears likely that the leached nitrate (NDS) accumulated below the hard pan where it became available to plants as their roots penetrated this layer via fractures originating from prior deep chiseling. Of the NDF taken up during the initial week, 20% was partitioned to the root and 42%, 24%, 14% respectively, to the upper, middle and bottom node positions (leaves plus stems). The partitioning reflected the respective growth rates of the tissues. Little change in partitioning was evident during the subsequent 5-week period, indicating that little remobilization of NDF from older to younger tissues occurred. In contrast, some remobilization of NDS was apparent between crop day 96 and 106 when the uptake of both NDF and NDS was negligible. During this period root growth was sustained by the apparent transfer of NDS from the root stump and from the adjacent lower leaf and stem tissues. These responses occurred in tobacco grown under higher nitrogen fertility levels than those usually considered optimal for the growth of flue-cured tobacco, but under conditions which are sometimes encountered. Paper no. 11640 of the Journal series of the North Carolina Agricultural Research Service, Raleigh, NC 27695-7601. Part of a thesis submitted by the senior author in partial fulfilment of the requirements of the Ph.D degree.     

Tobacco plants can use nitrogen taken up before mechanical wounding to synthesize nicotine afterwards

Mechanical wounding stimulates nicotine synthesis in tobacco plants. In the practice of tobacco production, most nitrogen (N) is taken up before removal of the shoot apex, while nicotine is mainly synthesized afterwards. Since N is required for nicotine synthesis, it is interesting to know whether plants can use N taken up before removal of the shoot apex to synthesize nicotine after wounding. To address this question, a hydroponics culture experiment was carried out, in which N was supplied as NH₄NO₃ at two levels (1 mM and 6 mM) in pre-culture, and N was either withdrawn or replaced by ¹⁵N after removing the shoot apex for the next seven days. Removal of the shoot apex caused a marked increase in nicotine concentration in various organs, also when plants grew under low-N conditions and showed symptoms of N deficiency. Increased nicotine accumulation even occurred when N was withdrawn from the growth medium before the apex was removed, indicating that tobacco plants can use N taken up previously to synthesize nicotine after mechanical wounding. The amount of N used for nicotine synthesis accounted for 5–6% of the total N, irrespective of treatment. Although most of the nicotine in intact plants and plants with the apex removed was synthesized de novo, as evidenced by the data when N was replaced by ¹⁵N-labeled NH₄NO₃, a large amount of the N absorbed before the N replacement was incorporated into the newly formed nicotine. The proportion of nicotine-¹⁵N to total nicotine-N was almost the same as that of ¹⁵N to total N in various organs. The results show the utilization of remobilized N taken up before excision of the shoot apex for nicotine synthesis afterwards, and highlight the importance of N cycling within plants, both when grown under N-sufficient and N-deficient conditions.Key words: ¹⁵N-isotope nitrogen, mechanical wounding, nicotine concentration, nicotine synthesis, nitrogen deficiency, removal of the shoot apex, tobacco (Nicotiana tabacum L.)     

Translocation of nitrogen in osmotically stressed wheat seedlings

Wheat (Triticum aestivum L., cv. Drabant) seedlings were grown in a ‘split root’ system where either the whole root system or one root half was subjected to osmotic stress for 24 h, using 200 g polyethylene glycol (PEG, molecular weight 4000) dm^?3 nutrient solution. ¹⁵N-Labelled nitrate was fed to one of the root compartments and total N and ¹⁵N-labelling were measured in plant material and xylem sap. Untreated plants translocated 87% of the N taken up to the shoot, and 10% of this was then retranslocated back to the root. Recalculated on a root nitrogen basis, 36% of the label recovered in the root after 24 h had passed through the shoot. Significant labelling of xylem sap collected from non-labelled roots indicated cycling of organic N through the roots. PEG-treatment of the whole root system caused significant water loss in both roots and shoots. Uptake of nitrate and retranslocation of N to roots were inhibited, whereas cycling of organic nitrogen through the root was still measurable. Treatment of half the root system with PEG had minor effects on shoot water content, but reduced the water content of the treated root part. The total uptake of nitrate by the root system was unaffected, and the effect on the treated root half was comparatively small. Nitrate reductase activity (NRA) declined in PEG-treated roots even if high nitrate uptake rates were maintained. Shoot NRA was unaffected by osmotic stress. The data indicate that the reduction in water content of the root per se has only small effects on nitrate uptake. Major inhibition of nitrate uptake was observed only after treatment of a sufficiently large portion of the root system to given an effect on shoot water content.     

Nitrite reduction and carbohydrate metabolism in plastids purified from roots of Pisum sativum L.

Nitrate reduction in roots and shoots and exchange of reduced N between organs were quantitatively estimated in intact 13-d-old seedlings of two-row barley (Hordeum vulgare L. cv. Daisengold) using the ¹⁵N-incorporation model (A. Gojon et al. (1986) Plant Physiol. 82, 254–260), except that NH ₊ ⁴ was replaced by NO _- ² . N-depleted seedlings were exposed to media containing both nitrate (1.8 mM) and nitrite (0.2 mM) under a light-dark cycle of 12:12 h at 20°C; the media contained different amounts of ¹⁵N labeling. Experiments were started either immediately after the beginning (expt. 1) or immediately prior to the end (expt. 2) of the light period, and plants were sampled subsequently at each light-dark transition throughout 36 h. The plants effectively utilized ¹⁵NO _- ³ and accumulated it as reduced ¹⁵N, predominantly in the shoots. Accumulation of reduced ¹⁵N in both experiments was nearly the same at the end of the experiment but the accumulation pattern in roots and shoots during each 12-h period differed greatly depending on time and the light conditions. In expt. 1, the roots accounted for 31% (light), 58% (dark), and 9% (light) of nitrate reduction by the whole plants, while in expt. 2 the contributions of the root were 82% (dark), 20% (light), and 29% (dark), during each of the three 12-h periods. Xylem transport of nitrate drastically decreased in the dark, but that of reduced N rather increased. The downward translocation of reduced ¹⁵N increased while nitrate reduction in the root decreased, whereas upward translocation decreased while nitrate reduction in the shoot increased. We conclude that the cycling of reduced N through the plant is important for N feeding of each organ, and that the transport system of reduced N by way of xylem and phloem, as well as nitrate reduction by root and shoot, can be modulated in response to the relative magnitude of reduced-N demands by the root and shoot, with the one or the other predominating under different circumstances.Symbols Anl accumulation of reduced ¹⁵N from ¹⁵NO _- ³ in ¹⁴NO _- ³ -fed roots of divided root system - Ar accumulation in root of reduced ¹⁵N from ¹⁵NO _- ³ - As accumulation in shoot of reduced ¹⁵N from ¹⁵NO _- ³ - Rr ¹⁵NO _- ³ reduction in root - Rs ¹⁵NO _- ³ reduction in shoot - Tp translocation to root of shoot-reduced ¹⁵N from ¹⁵NO _- ³ in phloem - Tx translocation to shoot of root-reduced ¹⁵N from ¹⁵NO _- ³ in xylem     

Nitrogen form, availability, and mycorrhizal colonization affect biomass and nitrogen isotope patterns in Pinus sylvestris

Nitrogen (N) isotope patterns are useful for understanding carbon and nitrogen dynamics in mycorrhizal systems but questions remain about how different N forms, fungal symbionts, and N availabilities influence δ¹⁵N signatures. Here, we studied how biomass allocation and δ¹⁵N patterns in Pinus sylvestris L. cultures were affected by nitrogen supply rate (3% per day or 4% per day relative to the nitrogen already present), nitrogen form (ammonium versus nitrate), and mycorrhizal colonization by fungi with a greater (Laccaria laccata) or lesser (Suillus bovinus) ability to assimilate nitrate. Mycorrhizal (fungal) biomass was greater with ammonium than with nitrate nutrition for Suillus cultures but similar for Laccaria cultures. Total biomass was less with nitrate nutrition than with ammonium nutrition for nonmycorrhizal cultures and was less in mycorrhizal cultures than in nonmycorrhizal cultures. The sequestration of available N by mycorrhizal fungi limited plant N supply. This limitation and the higher energetic cost of nitrate reduction than ammonium assimilation appeared to control plant biomass accumulation. Colonization decreased foliar δ¹⁵N by 0.5 to 2.2‰ (nitrate) or 1.7 to 3.5‰ (ammonium) and increased root tip δ¹⁵N by 0 to 1‰ (nitrate) or 0.6 to 2.3‰ (ammonium). Root tip δ¹⁵N and fungal biomass on root tips were positively correlated in ammonium treatments (r ²?=?0.52) but not in nitrate treatments (r ²?=?0.00). Fungal biomass on root tips was enriched in ¹⁵N an estimated 6–8‰ relative to plant biomass in ammonium treatments. At high nitrate availability, Suillus colonization did not reduce plant δ¹⁵N. We conclude that: (1) transfer of ¹⁵N-depleted N from mycorrhizal fungi to plants produces low plant δ¹⁵N signatures and high root tip and fungal δ¹⁵N signatures; (2) limited nitrate reduction in fungi restricted transfer of ¹⁵N-depleted N to plants when nitrate is supplied and may account for many field observations of high plant δ¹⁵N under such conditions; (3) plants could transfer assimilated nitrogen to fungi at high nitrate supply but such transfer was without ¹⁵N fractionation. These factors probably control plant δ¹⁵N patterns across N availability gradients and were here incorporated into analytical equations for interpreting nitrogen isotope patterns in mycorrhizal fungi and plants.     

Fate and effects on yield components of extra applications of nitrogen on spring wheat (Triticum aestivum L.) grown in solution culture

Spring wheat (Triticum aestivum L.) was grown with daily additions of nitrate-N. The relative addition rate of nitrate-N was decreased stepwise, and after 125 days of growth, 58 mg N plant^-1 had been introduced. The fate and effect of an extra addition of nitrate (20 mg N plant^-1) at six different times during the ontogeny (37, 54, 66, 79, 94 and 108 days from sowing) on grain yield and grain protein concentration was investigated. The plants absorbed all or most of the extra nitrate at all stages of development evaluated. Dry matter production of both aerial vegetative parts and grains, but not roots, generally increased as a result of the extra nitrate addition. The increase in grain dry matter was mainly an effect of an increased number of grains per plant. Extra nitrate applications had large effects on grain nitrogen content at all stages, but the effect on main shoot and tiller ears varied depending on the time of application. Early applications, i.e. before anthesis, mainly led to increased yield with unchanged protein concentration whereas late applications also led to increased grain protein concentration. The largest effect on grain nitrogen concentration (25–30% increase) was obtained when the extra nitrate was applied late after sowing, i.e. less than four weeks before final harvest. As the extra dose of nitrate was labelled with ¹⁵N, it was possible to follow the movement of the extra nitrogen addition within the plant. Samples were taken at one and five days after ¹⁵N-addition and at final harvest. There were differences in the movement of ¹⁵N depending on when it was introduced. Generally, net movement of the ¹⁵N-labelled N into the grain increased with age at application until 94 days after sowing when a maximum of 90% of the added ¹⁵N ended up in the grain.Abbreviations RN Relative increase in nitrogen content - RA Relative nitrogen addition rate - RG Relative growth rate - N nitrogen     

Effects of timing and supply of nitrogen on nitrogen remobilization from vegetative organs and redistribution to developing seeds of sunflower

A glasshouse study was made of the distribution of ¹⁵N among vegetative organs of sunflower and its later remobilization and redistribution to seeds, as influenced by the developmental stage at which ¹⁵N was provided, and by the N status of the plants. Plants of Hysun 30 sunflower were grown in sand culture and provided with K¹⁵NO₃ for a 3-day period at: (a) 3 days before the end of floret initiation; (b) 3 days before anthesis; (c) the start of anthesis; (d) full anthesis; and (e) 8 days after full anthesis. The plants were grown on a range of N supply rates, from severely deficient to more than adequate for maximum growth. Nitrogen-15 was distributed to all parts of the plant at the end of the ¹⁵N uptake periods. With the exception of the most N-stressed plants, subsequent remobilization of ¹⁵N from roots, stems and leaves occurred irrespective of the time the ¹⁵N was taken up. However, the percentage redistribution to seeds of ¹⁵N taken up at the end of floret initiation was less than for ¹⁵N taken up at anthesis. Remobilization of ¹⁵N from leaves and roots was higher (70%) for ¹⁵N taken up during and after anthesis than for ¹⁵N taken up at the end of floret initiation (45%), except for plants grown on the lowest N supply. By contrast, remobilization of ¹⁵N from the stem was lower for ¹⁵N taken up after full anthesis (40%) than before or during anthesis (>70%). The proportion of ¹⁵N remobilized from the top third of the stem was less than that from the bottom third, and decreased with increasing plant N status. Nitrogen-15 taken up over the 3-day supply periods during anthesis contributed from 2 to 11% of the total seed N at maturity; the contribution to seeds was greatest for plants grown on the highest N supply. Nitrogen taken up just before and during anthesis contributed most of the N accumulated in mature seeds of plants grown on an adequate N supply, but N taken up between the end of floret initiation and just before anthesis, or after full anthesis seemed to make an equally important contribution to mature seeds as N taken up during anthesis for plants grown on a very low N supply. It was concluded that the development of florets and seeds of sunflower is supported by N taken up by the plant between the end of floret initiation and anthesis, and by N redistributed from vegetative organs. Unless soil N is so low as to impair early growth, split applications of N fertilizer would be best made just before the end of floret initiation (‘star stage’) and just before anthesis.     

Utilization of 15NO3 − by nodulated soybean plants under conditions of root hypoxia

Waterlogging of soils is common in nature. The low availability of oxygen under these conditions leads to hypoxia of the root system impairing the development and productivity of the plant. The presence of nitrate under flooding conditions is regarded as being beneficial towards tolerance to this stress. However, it is not known how nodulated soybean plants, cultivated in the absence of nitrate and therefore not metabolically adapted to this compound, would respond to nitrate under root hypoxia in comparison with non-nodulated plants grown on nitrate. A study was conducted with ¹⁵N labelled nitrate supplied on waterlogging for a period of 48 h using both nodulated and non-nodulated plants of different physiological ages. Enrichment of N was found in roots and leaves with incorporation of the isotope in amino acids, although to a much smaller degree under hypoxia than normoxia. This demonstrates that nitrate is taken up under hypoxic conditions and assimilated into amino acids, although to a much lesser extent than for normoxia. The similar response obtained with nodulated and non-nodulated plants indicates the rapid metabolic adaptation of nodulated plants to the presence of nitrate under hypoxia. Enrichment of N in nodules was very much weaker with a distinct enrichment pattern of amino acids (especially asparagine) suggesting that labelling arose from a tissue source external to the nodule rather than through assimilation in the nodule itself.     

Cross-polarization NMR of N-15 labeled soybeans

Cross-polarization ¹⁵N nmr spectra of ¹⁵N-labeled soybean seeds, pods, and leaves have been obtained at 9.12 MHz both with and without high-speed sample rotation at the magic angle. Spectral resolution is sufficient to permit a determination of the relative concentrations of amide and amine nitrogens, as well as of a few specific amino acid residues of proteins in the solid, intact samples. Utilization by soybean of nitrogen from labeled fertilizer in the presence of dinitrogen fixation can be determined from these spectra. A double-cross polarization ¹³C nmr spectrum of a spinning, ¹⁵N-labeled seed has been obtained in which resonances are observed only from these carbons directly bonded to nitrogens. This technique leads to a qualitative estimate of amino-acid composition of the protein which is complementary to that obtained directly from the ¹⁵N nmr spectrum.