The Effect of Soil Water Regimes on Leaf Water Potential, Growth and Development of Soybeans

The growth and development of soybeans (Glycine max L. cv. Amsoy) was studied at soil matric potentials of ?0.1 to ?1.0 bars. Chlorophyll, photosynthesis, and leaf nitrogen per plant was greatest at ?4 bars leaf water potential. Leaf area, number of internodes, plant height and dry weight of vegetative parts declined as leaf water potential decreased from ?2 to ?19 bars. Nitrogen content and nitrate reductase activity per g fresh weight determined the percentage protein of individual seeds but nitrogen content and nitrate reductase activity per plant determined the amount of total seed protein. The protein synthesized in the seed changed little in amino acid composition with changes in leaf water potential. Leaf water potentials above or below ?4 bars decreased yield, total protein and total lipid but plants produced the largest percentage of individual seed protein at ?19 bars leaf water potential.     

Nitrogen assimilation in opaque and normal sorghum during grain development

Activity of key nitrogen assimilating enzymes was studied in developing grains of high-lysine opaque sorghum P-721 and normal sorghum CSV-5. The higher percentage of protein in opaque sorghum was mainly due to lower starch content since protein per grain was less than in CSV-5. During grain development, albufn and globulin decreased while prolafne and glutelin increased. Prolafne content in CSV-5 was higher than in opaque sorghum. Average nitrate reductase activity in flag and long leaf were similar in both the varieties. The nitrate reductase activity decreased during grain development. Glutamate dehydrogenase activity was higher during early development and lower at later stages in opaque sorghum than in CSV-5. Glutamate oxaloacetate transaminase activity was higher and glutamine synthetase lower in opaque sorghum than in CSV-5 grains during development. Glutamate synthase activity was higher in opaque sorghum up to day 20 and lower thereafter than in CSV-5. It is suggested that reduced activities of glutamine synthetase as well as glutamate synthase in opaque sorghum as compared to CSV-5 during later stages of development may restrict protein accumulation in the former.     

Heterotrophy and nitrate metabolism in a cyanobacteriumPhormidium uncinatum

Nitrate-supported heterotrophic growth ofPhormidium uncinatum was achieved after repeated exposure to glucose in the presence of a photosystem (PS) II inhibitor. Nitrate and glucose utilization as well as activities of their metabolizing enzymes were measured comparatively in photoautotrophic and heterotrophic cells. Nitrate and glucose were taken up together at the ratio of 1:8 (molar basis) and glucose catabolism via glucose-6-phosphate dehydrogenase (G6PDH), and 6-phosphogluconate dehydrogenase (6PGDH) activities transferred desired electrons for nitrate reduction to ammonia through coupled ferredoxin-NADP⁺ reductase (FNR) activity. Ammonia thus generated was assimilated mainly by NADPH-glutamate dehydrogenase (GDH) activity. These data demonstrate an operation of nitrate assimilation in this cyanobacterium under heterotrophic conditions.     

Effect of nodulation on the nitrate assimilation in vegetative soybean plants

Summary Nitrate assimilation in the first trifoliate leaf of vegetative soybean plants (Glycine max L. Merr, cv Hodgson) was studied in relation to nodulation. Nodulated and non-nodulated plants were grown in a nitrate medium (4 mM). As a control nodulated plants were grown in a nutrient medium without combined nitrogen. This study included measurements of the acetylene reduction activity of the whole plant and of thein vitro nitrate reductase, glutamine synthetase and glutamate dehydrogenase activities in the first leaf and of the nitrate concentration. Nitrate accumulation and nitrate reductase activity were depressed in nodulated plants; root growth was decreased in the presence of nitrate. The relationships between nitrate assimilation and nodulation are discussed.     

Effect of decreased irradiance on N and C metabolism in leaves and roots of maize

The effects of decreased irradiance on fresh and dry weight, root respiration, levels of carbohydrates and N-compounds, and extractable activities of enzymes involved in C and N metabolism were evaluated in maize ( Zea mays L. cv. Plauto) seedlings during the 7 days following transfer from 450 to 200 μmol m⁻² s⁻¹ PAR. The fresh weight of roots and stems, the initiation of new leaves, root respiration rate, and the accumulation of dry matter, soluble sugars, starch, malate and amino acids in both leaves and roots were strongly reduced at low irradiance. In contrast, the level of nitrate was increased in leaves and only marginally affected in roots. Leaf phosphoenolpyruvate carboxylase (EC 4.1.1.31) activity started to decrease after 24–34 h, whereas ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39) activity and chlorophyll content were unaffected or only slightly reduced. In both leaves and roots, the adjustment of N metabolism to low irradiance occurred through a relatively rapid (30% after 10 h) and large (60% after 3 days) decrease of nitrate reductase (NR; EC 1.6.6.1) activity, followed by slower and smaller changes in the activity of nitrite reductase (EC 1.7.7.1), glutamine synthetase (EC 6.3.1.2) and NAD-dependent glutamate dehydrogenase (EC 1.4.1.2). We suggest that the preferential decrease of NR activity relative to other N-assimilating enzymes may be important for preventing the accumulation of toxic N-compounds like ammonia in both leaf and root tissues.     

Nitrate reductase activity and protein concentration of two populas clones

Nitrate reductase activity and protein percentage of various tree parts of two Populus clones were determined in relation to nitrate ion activity. Nitrogen was supplied as NH(4)NO(3) in a nutriculture system. Wisconsin-5 had significantly greater nitrate reductase activity than Tristis No. 1. Protein percentages of leaf plastochron index 10 leaves (tenth leaf below first leaf lamina exceeding 20 mm in length), bottom leaves, and roots in relation to nitrate ion activity were not appreciably different between clones. The nitrate reductase activity and protein percentage of Tristis No. 1 apex started to level off at the same nitrate ion activity, about 0.09 mm. In Wisconsin-5 apex protein percentage continued to increase at nitrate ion activities where nitrate reductase activity decreases sharply, suggesting that protein nitrogen was being supplied by ammonium ion. The difference in nitrate reductase activity between clones was probably due to genetically determined ability to synthesize nitrate reductase in response to nitrate ion. The expression of nitrate reductase activity was not an index of nitrogen assimilation ability but may be a useful index of growth potential when nitrate ion does not limit nitrate reductase synthesis.     

Changes in the activities of nitrogen assimilation enzymes ofLolium perenne L. during regrowth after cutting

The activities of key enzymes involved in N assimilation were investigated after defoliation of 6-week-old ryegrass plants grown in water culture conditions. In a first experiment, nitrate reductase, glutamine synthetase and glutamate dehydrogenase activities were measured in roots, stubble and leaves on the day of cutting and at 7-day intervals over the following 5-week period of regrowth. Ammonia assimilation enzymes showed little change whereas the nitrate reductase activity sharply decreased 2 weeks after clipping. In a second experiment, the nitrate reductase activity was measured at 2- or 3-day intervals 1 week before and 3 weeks after clipping.In vivo andin vitro assays both showed an increasing activity in leaves up to 8 days after cutting while root activity decreased. The opposite changes then occurred and both organs recovered their initial nitrate reductase activity levels after 12–14 days of regrowth. These fluctuations in nitrate reductase activity were considered to be related to the capacity for C assimilation and the nitrate availability.     

An Analysis of the Response of Young Barley Seedlings to Time of Application of Nitrogen

The reasons for the sensitivity of young barley seedlings totime of application of nitrogen have been examined. It is shownthat the transfer of nitrogenous reserves from endosperm toembryo which begins at about 36 h from planting proceeds ata faster relative rate than that of dry matter as a whole. Inconsequence embryo and endosperm nitrogen contents become temporarilysimilar some 24–36 h earlier than is the case for dryweights. Addition of nitrate on day 2 does not affect ratesof transfer of endosperm reserves but leads to a significantlyhigher nitrogen content in the embryo of treated plants, particularlyin the shoots. This additional nitrogen is present as nitrateup to around day 5 when reduction of accumulated nitrate commencesin the first leaf in significant amounts. For plants up to 14 days old delay in application of nitrateleads to a lowering of total nitrogen level which is proportionalto the delay in treatment. This is so for all parts of the plantexcept the first leaf for which the evidence indicates thatlevels of total and organic nitrogen and of accumulated nitrateare much lower when treatment is made late. It is argued thatnitrate accumulation by the leaf becomes progressively lessas it reaches full expansion, but irrespective of time of nitrateapplication about 95 per cent of the additional nitrogen presentin the leaf is in organic form Significant increases in organic nitrogen are found from day6 for plants supplied with nitrate up to day 4; for plants suppliedon day 6, or day 8nitrateand nitrate reductase activity in leafextracts are found within 6 h of treatment. Peak levels of nitratereductase activity are found for all treatments around days8–10 when the first leaf is fully expanded and when photosyntheticactivity is maximal. However, late supply of nitrate leads toa lower level of enzyme activity. Nitrate reduction in the rootsystem is undetectably low, and it is concluded that a substantialamount of carbon translocated from leaf to roots is in the formof nitrogenous compounds. The effects of time of application are also found when ammoniumnitrogen is substituted for nitrate indicating that the responseis independent of effects on the nitrate reducing system inthe leaf. Some inhibition of growth, particularly of the roots,is found due to ammonium toxicity. Why plants supplied early with nitrate show superior growthand enhanced photosynthetic activity in the first leaves isexplained in terms of treatment alleviating the restrictiveeffects of declining endosperm reserves. This is only possibleif nitrogen is supplied while the first leaf is expanding andable to accumulateand utilise the available nitrogen. Late supplyis associated with failure to use the nitrogen provided leadingto a lower protein level in the leaf; this can be correlatedwith the smaller size of leaf and the lower rates of carbonfixation occurring there.     

Influence of Age and Light on the Distribution and Development of Nitrate Reductase in Greening Barley Leaves

The relation between leaf age and the induction of nitrate reductase activity by continuous and intermittent light was studied with barley seedlings (Hordeum vulgare L. cv. Club Mariout). In general, nitrate reductase activity declined as the period of growth in darkness was extended beyond 5 days. Maximum activity was found near the leaf tip while activity was lowest in the morphologically youngest tissue near the base of the lamina. Increased activity was observed after continuous illumination of dark-grown seedlings for 24 hours. The increase in activity in response to light was greatly reduced when the dark pretreatment period was extended beyond 8 days. The amount of nitrate reductase activity present in the different sections of the leaf was closely related to the amount of polyribosomes present. The pattern of chlorophyll accumulation closely parallelled that of increases in nitrate reductase activity. The initial lag in the induction of nitrate reductase activity was removed by a 10-minute light treatment 6 hours before placing dark-grown barley seedlings in light. The enzyme was also induced under flashing light with various dark intervals. These induction curves closely resembled those of chlorophyll accumulation under the same conditions. The development of photosynthetic CO₂ fixation follows the same induction pattern in this system. Our results suggest that photosynthetic products may be required for the induction of significant levels of nitrate reductase activity in leaves of dark-grown seedlings, although other light effects may not be discounted.     

Reversible Inactivation of Nitrate Reductase by NADH and the Occurrence of Partially Inactive Enzyme in the Wheat Leaf

Nitrate reductase from wheat (Triticum aestivum L. cv Bindawarra) leaves is inactivated by pretreatment with NADH, in the absence of nitrate, a 50% loss of activity occurring in 30 minutes at 25°C with 10 micromolar NADH. Nitrate (50 micromolar) prevented inactivation by 10 micromolar NADH while cyanide (1 micromolar) markedly enhanced the degree of inactivation.

A rapid reactivation of NADH-inactivated nitrate reductase occurred after treatment with 0.3 millimolar ferricyanide or exposure to light (230 milliwatts per square centimeter) plus 20 micromolar flavin adenine dinucleotide. When excess NADH was removed, the enzyme was also reactivated by autoxidation. Nitrate did not influence the rate of reactivation.

Leaf nitrate reductase, from plants grown for 12 days on 1 millimolar nitrate, isolated in the late photoperiod or dark period, was activated by ferricyanide or light treatment. This suggests that, at these times of the day, the nitrate reductase in the leaves of the low nitrate plants is in a partially inactive state (NADH-inactivated). The nitrate reductase from moisture-stressed plants showed a greater degree of activation after light treatment, and inactive enzyme in them was detected earlier in the photoperiod.

     

A role for young leaves in vernalization of cauliflower: I. Analysis of leaf development during curd induction

Growth analysis techniques are used to test the hypothesis that chilling induces curd (flower) initiation in the cauliflower ( Brassica oleracea Botrytis L. cv. Perfection) through inhibiting leaf growth, thereby increasing the availability of growth factors to the stem apex and enabling differentiation of the curd. Effects of chilling on leaf growth and curd induction are compared in juvenile and mature, vegetative plants. Chilling at 5°C reduced dry matter accumulation in the total leaf complement by ca 60% in juvenile plants and 40% in mature plants, compared to control plants growth at 20°C. Juvenile plants showed slower rates of leaf initiation than mature plants. Leaf initiation was retarded by chilling in both plant types with the most marked effect seen in the juvenile plants. This was consistent with dry matter availability to the stem apex limiting differentiation more severely in juvenile plants than in mature plants. The rate of dry matter accumulation in existing leaves, however, was faster in juvenile plants than in mature plants at 20°C. Plants that were juvenile during chilling produced an average of 43 leaves below the curd whereas those that were mature produced 25.
Dry matter accumulation in younger leaves was more markedly inhibited by chilling than in older leaves. Chilling also reduced the rate at which enlarging leaves became positionally more remote from the stem apex. Possible roles for such leaves in regulating apical development are considered.     

Temperature-dependent growth and emergence of functional leaves: an adaptive mechanism in the seedlings of the western Himalayan plant Podophyllum hexandrum

As an adaptive mechanism, hypocotyl dormancy delays emergence of functional leaf until favorable season of growth in Podophyllum hexandrum, an endangered medicinal plant of the western Himalayas. However, upon exposure of the freshly germinated seedlings to favorable temperature (25°C), functional leaves emerged within 20 days. Therefore, we examined regulation mechanisms of growth and development of this alpine plant by temperature under laboratory conditions. The seedlings were exposed to (1) 25°C (temperature prevailing at the time of maximum vegetative growth), (2) 4°C (mean temperature at the onset of winter in its natural habitat), and (3) 10°C (an intermediate temperature). Slackened growth at 4°C was followed by senescence of aerial parts and quiescence of roots and predetermined leaf primordia. Rapid development of leaf primordia at 25°C was associated with increased starch hydrolysis. This was evident from higher α-amylase activity and reducing sugars. These parameters decreased on sudden exposure to 4°C. In contrast, the roots (perennating organs) showed a slight increase (1.36-fold) in α-amylase activity. Growth and development in seedlings growing at 10°C (temperature less adverse than 4°C) were comparatively faster. The content of reducing sugars and α-amylase activity were also higher in all the seedling parts at 10°C as compared to 4°C. This indicated larger requirements for sugar by the seedlings at 10°C. Irrespective of temperature, maximum changes in nitrate and nitrate reductase occurred during the initial 10 days, i.e., when the readily available form of sugars (reducing sugar) was highest. This indicated that a temperature-dependent availability of carbon, but not temperature itself, was an important regulator of uptake and reduction of nitrogen. IHBT Publication number 508a.     

Carbon Economy and Translocation of 14C in Leaflets of the Seventh Leaf of Tomato During Leaf Expansion

The growth pattern (leaf expansion, carbon metabolite accumulation)and the assimilation and translocation of leaf assimilate inthe leaflets of the seventh leaf of tomato were examined onsix occasions during a period of 35 days beginning 10 days afterthe leaf was first visible. Leaf expansion progressed basipetally within the leaf. The primarypartitioning of the currently-fixed ¹⁴C and the 24 h incorporationof both self-fixed and imported ¹⁴C were closely related tothe degree of leaf expansion. Therefore, leaflets of up to halffinal size within the same leaf, were markedly different inphysiological function. Maximum import of ¹⁴C-leaf assimilate from the six basal leavesinto the seventh leaf was recorded at 15 per cent expansion.The terminal and top-pair of leaflets reached one quarter fullsize first and exported ¹⁴C leaf assimilate to the basal-pairof leaflets and to the rest of the plant when the seventh leaf,as a whole, was only one tenth expanded. The transition fromimport to export in a given leaflet was gradual and overlapping. During the early part of leaflet expansion, more imported ¹⁴C-leafassimilate was used for protein synthesis than for the synthesisof sugars and starch. Meanwhile, the capacity for sucrose synthesisfrom the self-fixed carbon increased significantly precedingthe onset of export. Each leaflet resembles a whole leaf in functional development.     

Nitrate reduction in ramets of a clonal plant Eichhornia crassipes responding to nitrate availability during clonal growth stage

Effect of nitrate availability on nitrate reduction was examined in inter-connected ramets of invasive clonal plant Eichhornia crassipes grown with two nitrate supply regimes during different clonal growth stage. Increase of nitrate availability accelerated nitrate reductase activity (NRA) in parent and offspring ramets of E. crassipes, and there was greatly different pattern in inter-connected ramets during clonal growth stage. Leaf NRA was lower in offspring than that in parent ramets in phase 1, while significantly higher leaf NRA in offspring ramets was detected during phase 2. The results indicated NRA in inter-connected ramets of E. crassipes was highly dependent on nitrate availability and growth stage.     

Nitrate reductase,nitrite reductase and glutamate dehydrogenase levels in roots and leaves of maize seedlings

Total activities of nitrate and nitrite reductases were higher in 4 to 20 day old maize plants in the leaves than in the roots. The ratio of activities found in the leaves and in the roots respectively was much higher in the case of nitrate reductase than in the case of nitrite reductase. On the other hand higher glutamate dehydrogenase activity in the roots than in the leaves clearly indicates that the roots play a more important role in the assimilation of ammonium than in the assimilation of nitrate. When comparing the distribution of seminal and nodal adventitious roots of maize seedlings with the assimilation of inorganic nitrogen on the basis of enzyme levels, it could be deduced that during the first 20 days of seedling growth seminal roots were more involved in the assimilation of nitrate whereas nodal adventitious roots were more active in ammonium assimilation.     

Repression of nitrate reductase in cucumber leaves caused by calcium deficiency

Growth of young cucumber plants was strongly inhibited, whencalcium was removed from the culture solution. The activitiesof nitrate reductase, glutamate dehydrogenase and glutaminesynthetase were investigated after the removal of calcium. Thoughthe activities of glutamine synthetase and glutamate dehydrogenasewere not altered much, nitrate reductase activity, measuredby in vitro and in vivo assays, decreased dramatically. Theloss of nitrate reductase activity coincided with the levelof nitrate in the leaves. When nitrate was supplied to the cucumberswith a nitrate deficiency, the plants induced nitrate reductasetogether with a distinct accumulation of nitrate. However, cucumberstreated for both calcium and nitrate deficiency failed to inducenitrate reductase and to accumulate nitrate on the additionof large amounts of nitrate. Leaf sections that had been treatedfor both calcium and nitrate deficiency could induce nitratereductase when floated on nitrate solution under the light.This indicates that the drastic loss of nitrate reductase causedby the removal of calcium was due mainly to the deficiency ofnitrate as the inducer in leaves. (Received December 19, 1979; )     

Proline accumulation and nitrate reductase activity in contrasting sorghum lines during mid-season drought stress

Six lines of sorghum ( Sorghum bicolor L. Moench) with differing drought resistance (IS 22380, ICSV 213, IS 13441 and SPH 263, resistant and IS 12739 and IS 12744, susceptible) were grown under field conditions in the semi-arid tropics and analysed for proline and nitrate reductase activity (NRA; EC 1.6.6.1) during a mid-season drought. The resistant lines accumulated high levels of proline, while the susceptible lines showed no significant proline accumulation. Most of the proline was accumulated after growth of the plants had ceased. In a separate greenhouse experiment, most of the proline was found in the green rather than the fired portions of leaves. The levels returned to that of irrigated controls within 5 days of rewatering. Proline levels increased as leaf water potential and relative water content fell, and there was no apparent difference among the different sorghum lines with change in plant water status. Susceptible lines accumulated less proline than resistant lines as leaf death occurred at higher water potentials. Proline accumulation may, however, contribute to the immediate recovery of plants from drought. Leaf NRA reached high levels at about 35 days after sowing in both the stressed and irrigated plants, after which it declined. The decline in NRA was more pronounced in the stressed than in the irrigated plants and closely followed changes in the growth rate. Upon rewatering, NRA increased several-fold in all the lines and, in contrast to proline accumulation, genotypic differences in NRA were small, both during stress and upon rewatering. The high sensitivity of NRA to mild drought stress was reflected in the rapid decline of activity with small changes in leaf water potential and relative water content. The results are discussed in the light of a possible role for proline during recovery from drought, and the maintenance of NRA during stress and its recovery upon rewatering.     

Mobilisation of NADPH-glutamate dehydrogenase mRNA in regreening cultures of Euglena gracilis

Exposure of dark-grown resting Euglena gracilis Klebs strain Z Pringsheim to light results in a transient increase in the specific activity of NADPH-glutamate dehydrogenase. NADPH-glutamate dehydrogenase antibody was used to detect NADPH glutamate dehydrogenase resulting from the translation of total polyadenylated RNA and polysomal RNA from Euglena in a cell-free rabbit reticulocyte lysate system. NADPH-glutamate dehydrogenase mRNA was present in cells at all stages of development and present on polysomes from dark-grown and regreening cells but not on polysomes from dark-grown resting cells. These results indicate that the light-induced increase in NADPH-glutamate dehydrogenase in dark-grown resting cells represent an increase in the rate enzyme synthesis resulting from the mobilisation of NADPH-glutamate dehydrogenase mRNA onto polysomes.