Effect of nitrate in the rooting medium on carbohydrate composition of soybean nodules

Nodulated soybean plants (Glycine max [L.] Merr) were grown in sand culture. Carbohydrate composition of nodules, roots, and leaf blades was determined and related to the effects of nitrate in nutrient solution on nodule growth and on nitrogenase activity of nodules.     

Glutamine synthetase isoenzymes of Phaseolus vulgaris L.: subunit composition in developing root nodules and plumules

In the legume Phaseolus vulgaris L., glutamine synthetase (GS) (EC.6.3.1.2.) occurs as three cytosolic polypeptides, , and , and a plastidic polypeptide, . This paper describes the subunit composition of active octameric GS isoenzymes from root nodules and plumules using ionexchange high-performance liquid chromatography followed by two-dimensional denaturing gel electrophoresis and Western immunodetection. Root nodules contained four separable GS activities, three of which were composed mainly of cytosolic , / and GS polypeptides, whereas the fourth activity, consisted of plastidic GS polypeptides. The increase in GS activity during nodulation was due largely to the appearance of -containing isoenzymes, and to a lesser extent on the isoenzyme, whereas the -isoenzyme activity remained approximately constant throughout. Plumule GS from imbibed seeds was found to be composed of separate and isoenzymes, but 2 d after germination, plumule GS consisted of a mixture of , / and isoenzymes. The results from both nodules and plumules indicate that different cytosolic GS polypeptides in P. vulgaris are able to assemble into both homo-octameric and heterooctameric isoenzymes. Moreover, the changes in the patterns of isoenzymes observed during nodule development and plumule growth are interpreted to be caused both by temporal changes in the denovo synthesis of the polypeptides and also by their spatial separation in different cell types.Abbreviations 1D, 2D one-, two-dimensional - GS glutamine synthetase - GS_s GS semibiosynthetic activity - GS_t GS transferase activity - IEX-HPLC ion-exchange high-performance liquid chromatography - kDa kilodaltons - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis     

Drought and shade deplete nonstructural carbohydrate reserves in seedlings of five temperate tree species

Plants that store nonstructural carbohydrates (NSC) may rely on carbon reserves to survive carbon‐limiting stress, assuming that reserves can be mobilized. We asked whether carbon reserves decrease in resource stressed seedlings, and if NSC allocation is related to species' relative stress tolerances. We tested the effects of stress (shade, drought, and defoliation) on NSC in seedlings of five temperate tree species (Acer rubrum Marsh., Betula papyrifera Marsh., Fraxinus americana L., Quercus rubra L., and Quercus velutina Lam.). In a greenhouse experiment, seedlings were subjected to combinations of shade, drought, and defoliation. We harvested seedlings over 32–97 days and measured biomass and NSC concentrations in stems and roots to estimate depletion rates. For all species and treatments, except for defoliation, seedling growth and NSC accumulation ceased. Shade and drought combined caused total NSC decreases in all species. For shade or drought alone, only some species experienced decreases. Starch followed similar patterns as total NSC, but soluble sugars increased under drought for drought‐tolerant species. These results provide evidence that species deplete stored carbon in response to carbon limiting stress and that species differences in NSC response may be important for understanding carbon depletion as a buffer against shade‐ and drought‐induced mortality.     

Carbohydrate and organic acid composition of effective and ineffective root nodules of Phaseolus vulgaris

Dicarboxylic acid transport mutants of Rhizobium species are usually deficient in their ability to fix atmospheric dinitrogen. We report here a study comparing the physiology of root nodules on Phaseolus vulgaris L. cv. Goldie induced by an effective strain of Rhizobium leguminosarum biovar phaseoli and a C₄-dicarboxylic acid utilization mutant. The mutant, while able to form nodules, was ineffective in N₂ fixation. Carbohydrates and organic acids of roots and nodules formed by the 2 strains were monitored at 3-day intervals from 13 to 34 days after inoculation. Both carbohydrates and organic acids accumulated in ineffective nodules in comparison with the effective nodules. The concentration of malic acid was tenfold higher in ineffective nodules than in effective nodules. Other organic acids, i.e., lactate, malonate, ascorbate and gluconate, were also detected. Lactate and ascorbate were the only other organic acids accumulating in ineffective nodules. The most prevalent carbohydrates found in both types of nodules were sucrose, glucose and fructose. Myo-inositol was the only cyclitol detected in both types of nodules. Carbohydrates and organic acids were present in lower concentration in roots than in nodules, except for lactate. These compounds were not consistently detected in higher concentration in roots from plants inoculated with the mutant strain, as was the case in nodules.     

Effect of phosphorus deficiency on growth angle of basal roots in Phaseolus vulgaris

Root architectural plasticity might be an important factor in the acquisition by plants of immobile nutrients such as phosphorus (P). In this study, we examined the effect of P availability on the orientation of basal roots with respect to gravity, and thereby on the growth angle of these roots of common bean (Phaseolus vulgaris L.). In one set of studies the growth angle of basal roots of bean seedlings was measured over time. Sixteen bean genotypes were examined; six showed a decrease in root orientation with respect to gravity in low P media, one increased orientation, and nine showed no difference within 5 d of basal root emergence. Bean taproots also showed decreased root orientation with respect to gravity in low P. Growth angle after 5 d was correlated with field performance of contrasting genotypes in low P tropical soils. Mineral deficiencies other than P did not cause changes in root angle. In a split pouch system that provided high or low P solution to different parts of the root system, the decrease in root angle in low P was found to be a response to global P availability, and not local to the portion of the root system in low P. Effects of P availability on root angle were associated with reduced shoot P concentration, but preceded effects on plant biomass accumulation and leaf area expansion. Results from growth pouches for genotype G 19833 were confirmed using a solid-phase buffered sand-culture system supplying P at three levels. Pea (Pisum sativum), soybean (Glycine max Williams), chickpea (Cicer arietinum), lima bean (Phaseolus lunatus), and lentil (Lens culinaris) were grown with and without P; soybean and pea also showed decreased basal root angles in low P.     

Actin isoforms in non-infected roots and symbiotic root nodules of Phaseolus vulgaris L.

The present report describes an initial characterization of actin from non-infected roots and symbiotic nodules of Phaseolus vulgaris L. cv. Negro damapa. Using anti-actin monoclonal antibodies, a 42-kDa polypeptide was identified in plant extracts. After two-dimensional SDS-PAGE analysis and Western blotting of actin fractions enriched using diethylaminoethyl-resin, the presence of one major isoform of actin in symbiotic nodules and two main isoforms in non-infected roots was revealed. Possible implications of this finding are discussed.Abbreviations MAB(s) monoclonal antibody(ies) - IEF isoelectric focusing We thank Jose Luis Zitlalpopoca for technical assistance. H.P. gratefully acknowledges financial support from CONACYT (Mexico) and the National Institute of Public Health of Mexico. This work was partially supported by grants DGAPA-UNAM IN208489 and CEE. C11 06228-M.     

Fluorescent Pseudomonas influences palisade mesophyll development and spatial root development in Phaseolus vulgaris

Three strains of plant growth promoting fluorescent Pseudomonads (HPR6, RRLJ008 and RRLJ134) were studied for their effect on growth and yield of French bean (Phaseolus vulgaris L.) under field conditions. The effect of these strains on nature of root development and leaf palisade tube length were also examined. The strains induced positive response on growth and physiological parameters resulting in higher yield in P. vulgaris. Strain HPR6 produced the most promising results in thickening of leaf palisade layer, spreading of lateral roots and production of root hairs. The increase in specific leaf weight (SLW), net assimilation rate (NAR) and relative growth rate (RGR) by these strains were 68%, 152% and 167%, respectively. The growth and yield parameters were also significantly improved compared to the uninoculated control. Antibiotic resistant mutant strains demonstrated that these bacteria effectively colonized the rhizosphere of French bean. The results suggest that the strains could be developed for field application on a large scale.     

Effects of gibberellic acid on patterns of carbohydrate distribution and acid invertase activity in Phaseolus vulgaris

The application of gibberellic acid (GA₃,10 μ M ) as a root drench to 16-day-old plants of Phaseolus vulgaris L. cv. Masterpiece stimulated growth of the stem internodes and reduced root growth. GA₃ treatment did not affect the export of ¹⁴C from a primary leaf to which [¹⁴C]-sucrose was applied, but greatly increased upward translocation to the elongation region of the stem at the expense of transport to the hypocotyl and root system. The observed changes in the patterns of growth and [¹⁴C]-labelled assimilate distribution were correlated with an increase in the specific activity of acid invertase in the elongating stem internodes and a decrease in invertase activity in the hypocotyl and root. Sucrose concentration in the elongating internodes fell substantially after treatment with GA₃ while the concentration of hexose sugars increased. We suggest that by stimulating acid invertase synthesis in the elongating internodes, GA₃ acts to establish a more favourable sucrose gradient between these sinks and source leaves. Under source-limiting conditions this, in turn, will lead to a reduced rate of assimilate translocation to competing sinks in the root system.     

Effect of gibberellic acid on growth and carbohydrate metabolism of developing tubers of potato (Solanum tuberosum)

Potato plants (Solanum tuberosum L. cv. Ostara) were grown in aerated water culture in a controlled environment. When the tubers had reached a diameter of 1–3 cm. ¹⁴C-labelled or unlabelled gibberellic acid (GA₃) was applied to the surface of the stolons at points approximately 1 crn from the developing tubers, and treatment continued for 10 days. - Significant quantities of GA₃ moved into tuber tissue within 2–4 days of hormone application. This influx of GA₃ was accompanied by a marked reduction in both the activity of ADPG-pyrophospharylase and the ratio ADPG-pyrophosphorylase/starch phosphorylase and an increase in the activity of UDPG-pyrophosphorylase. Starch phosphorylase activity initially increased slightly but then fell, whereas the activity of starch synthase remained constant throughout the experiment. The soluble sugar composition of the tubers changed qualitatively towards a pattern characteristic of growing stolon tips prior to tuber initiation, but there was no clear evidence of net starch degradation. Changes in the activities of the enzymes were observed prior to noticeable effects of the hormone on tuber growth rate or the development of new stolons at the tuber eyes. - GA₃- treated tubers imported more ¹⁴C from labelled photosynthate than expected on the basis of growth rate. However, the capacity to convert solub#e-¹⁴C to ethaTiol-insoluble-¹⁴C (predominantly starch) was reduced in comparison with non-treated tubers. - The observed changes in carbohydrate composition and enzyme activities indicate that GA₃ induces a drastic change in potato tuber metabolism towards a pattern characteristic for the termination of the storage process.     

The effect of phosphate deficiency on membrane phospholipid composition of bean (Phaseolus vulgaris L.) roots

Plasma membranes were isolated from roots of bean (Phaseolus vulgaris L.) plants cultured on phosphate sufficient or phosphate deficient medium. The phospholipid composition of plasma membranes was analyzed and compared with that of the microsomal fraction. Phosphate deficiency had no influence on lipid/protein ratio in microsomal as well as plasma membrane fraction. In phosphate deficient roots phospholipid content was lower in the plasma membrane, but did not change in the microsomal fraction. Phosphatidylcholine and phosphatidylethanolamine were two major phospholipids in plasmalemma and microsomal membranes (80 % of the total). After two weeks of phosphate starvation a considerable decrease (about 50 %) in phosphatidylcholine and phosphatidylethanolamine in microsomal membranes was observed. The decline in two major phospholipids was accompanied by an increase in phosphatidic acid and lysophosphatidylcholine content. The effect of alterations in plasma membrane phospholipids on membrane function e.g. nitrate uptake is discussed.     

Effects of NaCl on ion relations and carbohydrate status of roots and on osmotic regulation of roots and shoots of Atriplex amnicola

Abstract Atriplex amnicola was grown at 25, 200 or 400 mol m³ NaCl. Root tissues at different stages of development were investigated for concentrations of K⁺, Na⁺ and Mg²⁺, and in some cases for Cl^?. Sugar and starch concentrations were measured for plants grown at 25 or 400 mol m³ NaCl. In the ‘slightly vaeuolated’ root tips, Na⁺ was only 40 mol m^?3 at an external concentration of 400 mol m^?3 NaCl. The concentrations of K⁺ were not affected substantially by external NaCl between 25 mol m^?3 and 400 mol m^?3. The ‘highly vacuolated’ root tissues had substantially higher concentrations of K⁺, Na⁺ and Cl^? in plants grown at 200 and 400 mol m ³ NaCl than in plants grown at 25 mol m^?3 NaCl. Concentrations of Cr and of the sum of the cations in recently expanded tissue were similar to those in the bulk of the roots, consisting mainly of old cells. However, the K⁺: Na⁺ decreased with age; at 400 mol m^?3 external NaCl with a K⁺: Na⁺ of 0.012, the K⁺: Na⁺ in recently expanded 12 mm root tips was as high as 1.6, compared with 0.7 for the bulk of the roots. These ion data were used to estimate cytoplasmic and vacuolar concentrations of K⁺ and Na ⁺. Such calculations indicated that between 25 mol m³ and 400 mol m^?3 external NaCl the concentration of the sum of (Na⁺+K⁺) in the cytoplasm was maintained at about 180–200 mol m^?3 (cell water basis). In contrast, the (Na⁺+ K⁺) concentration in the vacuole was 170 mol m^?3 for plants grown at 25 mol m^?3 NaCl and 420 mol 400 mol m^?3 NaCl. The expanding root (issues exhibited greatly decreased soluble sugars and starch between dusk and dawn. Ai both times, sugar and starch concentrations in these tissues were 2.5–4.0 times greater in plants grown at 400 mol m^?3 NaCl compared with plants grown at 25 mol m^?3 NaCl. In contrast, carbohydrate concentrations in expanded root tissues were very similar at 25 and 400 mol m^?3 and showed little diurnal fluctuation. This paper considers the causes for the slower growth of A. amnicola at 400 than at 25 mol m”³ NaCl, using the data for the roots described here, and those for the shoots presented in the preceding paper (Aslam et al., 1986). There is no support for possible adverse effects by high internal ion concentrations. Instead, there may be deficiencies in supply of organic solutes for osmotic regulation; during part of the night a limited supply of such solutes may well restrict the rate of expansion of cells in plants growing at 400 mol m^?3 NaCl. There is insufficient evidence to decide whether this limitation in the expanding tissues is particularly prominent for the roots or for the shoots.     

Location of two isoenzymes of NADH-dependent glutamate synthase in root nodules of Phaseolus vulgaris L.

The two isoenzymes of NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14), previously identified in root nodules of Phaseolus vulgaris L., have both been shown to be located in root-nodule plastids. The nodule specific NADH-GOGAT II accounts for the majority of the activity in root nodules, and is present almost exclusively in the central tissue of the nodule. However about 20% of NADH-GOGAT I activity is present in the nodule cortex, at about the same specific activity as this isoenzyme is found in the central tissue. Glutamine synthetase (GS; EC 6.3.1.2) occurs predominantly as the polypeptide in the central tissue, whereas in the cortex, the enzyme is represented mainly by the polypeptide. Over 90% of both GS and NADH-GOGAT activities are located in the central tissue of the nodule and GS activity exceeds NADH-GOGAT activity by about twofold in this region. Using the above information, a model for the subcellular location and stoichiometry of nitrogen metabolism in the central tissue of P. vulgaris root nodules is presented.Abbreviations Fd-GOGAT ferredoxin-dependent glutamate synthase - GOGAT glutamate synthase - GS glutamine synthetase - NADH-GOGAT NADH-dependent glutamate synthase - IEX-HPLC ion-exchange high-performance liquid chromatography     

Effect of storage on redgram [Cajanus cajan (L.) Millsp] and greengram [Vigna radiata (L.) Wilczek] varieties with particular reference to carbohydrate composition

The effect of storage on carbohydrate composition of three redgram (TTB-7, BRG-1 and ICP-8863) and three greengram (Pusa Bisaki, KDM-1 and China Moong) varieties during storage was studied. The total free sugars, starch and dietary fibre were evaluated both in control and in samples stored for six months. Decrease in percentage of (a) free non-reducing sugars, (b) starch and (c) pentose sugar of dietary fibre was observed during storage. Concomitantly insect infested samples showed increase in reducing sugar, dietary fibre and also hexose sugars of dietary fibre.     

The effects of silver nitrate and different carbohydrate sources on somatic embryogenesis in Coffea canephora

The response of five Coffea canephora Pierre genotypes with regard to somatic embryogenesis was tested on media containing silver nitrate (AgNO₃) and different carbohydrates (sucrose, fructose, maltose and glucose). The presence of AgNO₃ caused only small modifications to the ionic equilibrium of the media. At concentrations between 30–60 M, AgNO₃ improved embryo yield for the genotypes evaluated, while higher doses negatively affected the regenerative capacity. The substitution of maltose, glucose or fructose for sucrose produced different responses depending on the genotype. Fructose significantly increased somatic embryo production in genotypes N91 and N128, while maltose was highly effective for N75. In addition, more synchronous embryo development was observed in genotype N91 when glucose was used instead of sucrose.     

Effect of water stress on the reduction of nitrate and nitrite by soybean nodules

The effects of water stress on nitrate reductase and nitrite reductase activities in symbiotic nodules were examined in field-grown soybean plants (Glycine max L Merr. cv Clark). The in vitro assays of enzyme activity indicated that the nodule cytosol and bacteroids contained both nitrate reductase and nitrite reductase activities. The reduction of nitrate in bacteroids increased significantly as nodule water potential declined from −0.6 to −1.4 megapascals, and then decreased when −1.8 megapascals water potential was reached. On the contrary, the reduction of nitrate in nodule cytosol was inhibited as water stress progressed. Increases in water stress intensity also caused a significant inhibition in nitrite reductase activities of bacteroids and nodule cytosol within soybean nodules. The results show that nitrate reduction occurred both in the cytosol and bacteroids of water-stressed soybean nodules. The reduction of nitrate functioned at different physiological modes in these two fractions.     

Fine root production and carbohydrate concentrations of mature longleaf pine (Pinus palustris P. Mill.) as affected by season of prescribed fire and drought

The historical range of longleaf pine (Pinus palustris P. Mill) has been greatly reduced, in part, by lack of fire. Recently, the application of fire has become an accepted practice for the restoration of longleaf pine to former parts of its natural range. This study was designed to evaluate the effects of season of prescribed fire on the root growth and nonstructural carbohydrate concentrations of longleaf pine, and identify the time of year when fire has the least negative effect on longleaf pine root processes. We found that root growth was generally less on July-burned plots than on either March- or May-burned plots and we attribute these responses to the effect of fire on interaction between root processes and the soil environment. Specifically, soil water and temperature conditions may have been less favorable for root growth on the July-burned plots compared to the March- and May-burned plots. With two years of information on the seasonal dynamics of foliage production, root growth, and root carbohydrates, we determined that at our study site, optimal prescribed fire would impact tree growth less in November through March compared to other times of the year. We also observed that severe drought during the 1998 growing season was associated with a 3-month delay in peak root growth, and prolonged drought in 1999 through 2000 coincided with a reduction in root starch storage. We conclude that season of prescribed fire potentially affects root processes, but that severe or prolonged drought may either interact with or override these effects.     

Effect of nitrate on the organic Acid and amino Acid composition of legume nodules

Nitrate supplied to legume plants inhibits the activity of nitrogenase in Rhizobium bacteroids in root nodules. The accumulation of amino N which is known to occur in Glycine max (L.) Merr. nodules as nitrogenase activity declines was studied in more detail by analysis of changes in free amino acid composition in response to high nitrate supply. A 6-fold increase in asparagine concentration in Bradyrhizobium japonicum bacteroids was found about the time of maximum nitrogenase inhibition. However, the accumulation of amino acids in soybean nodules lagged behind the inhibition of nitrogenase. Furthermore, in studies of a second legume, Phaseolus vulgaris (L.) inoculated with two different strains of Rhizobium phaseoli, a high nitrate treatment inhibited nitrogenase but had no significant effect on amino acid composition of nodules. The possibility that nitrate may interfere with the supply of carbon substrates to bacteroids was examined by the analysis of organic acids in legume nodules supplied with nitrate. Nitrate had a small (10-20%) negative effect on the concentration of tricarboxylic acid cycle acids in P. vulgaris nodules. However, in G. max nodules, high nitrate treatment resulted in significant increases in the concentration of malate, succinate, fumarate, and citrate. Thus, carbon deprivation of bacteroids also seems unlikely as a cause of the inhibition of nitrogenase by nitrate. There was a transient increase in ammonium concentration in P. vulgaris nodules in response to high nitrate treatment. This effect was rapid relative to other effects of nitrate on nodule composition and was roughly coincident with the rapid decline in acetylene reduction activity.     

Effects of insecticides on acetylene reduction by Azotobacter vinelandii and soybean nodules

Summary Nine organophosphate and carbamate insecticides were tested for effects on ability of Azotobacter vinelandii to reduce acetylene. Only Gardona^R, at higher concentrations, was significantly inhibitory. The same pesticides were tested with soybeans (Glycine max L.). Some minor phytotoxic effects were noted, but there was no inhibition of the ability of the excised nodules of the plants to reduce acetylene.Published with the approval of the Director of the North Dakota Agricultural Experiment Station as Journal Article No. 726. Portion of a thesis presented by the senior author in partial fulfillment of the requirements for the M.S. degree in bacteriology at North Dakota State University.Published with the approval of the Director of the North Dakota Agricultural Experiment Station as Journal Article No. 726. Portion of a thesis presented by the senior author in partial fulfillment of the requirements for the M.S. degree in bacteriology at North Dakota State University.     

Effect of high nitrate concentrations on growth and nitrate uptake by free-living and immobilizedChlorella vulgaris cells

Growth and nitrate uptake were studied on free-living and immobilizedChlorella vulgaris cells cultivated in medium containing different nitrate concentrations. First, the effect of nitrate concentrations on growth indicated that cells can live in the presence of high concentrations as high as 97 mM. Although no lethal effect on cells was observed such concentration a slow down in growth and a decrease in biomass produced was observed. The rate of nitrate uptake increased with the nitrate concentration in the medium. The maximum uptake rate was reached in first days of culture in both free-living and immobilized cells. The rate dropped more rapidly for cells growing in 2 mM nitrate than for cells growing in higher nitrate concentration. The maximum rate was very much the same for free-living and immobilized and was within the order of 0.45 to 0.57 g NO₃ h^–1 10^–6 cells. Immobilization modified the changes of nitrate uptake rate for concentration higher than 2 mM.