The effect of nitrogen nutrition on cluster root formation and proton extrusion by Lupinus albus

Nitrogen nutrition can influence cluster root formation in many wild species, but the effect of N form on cluster root formation and root exudation by white lupin is not known. In a solution culture study, we examined the effect of N nutrition (ammonium, nitrate, both or N2 fixation) on cluster root formation and H+ extrusion by white lupin plants under deficient and adequate P supply. The number of cluster roots increased greatly when plants were supplied with I microM P compared with 50 microM P, the increase being 7.8-fold for plants treated with (NH4)2SO4, 3-fold for plants treated with KNO3 and NH4NO3, and 2-4-fold for N2-fixing plants. Under P deficiency. NH4+-N supply resulted in production of a greater number and biomass of cluster roots than other N sources. Dry weight of cluster roots was 30 % higher than that of non-cluster roots in P-deficient plants treated with (NH4)2SO4 and NH4NO3. In plants treated with sufficient P (50 microM), the weight of non-cluster roots was approx. 90 % greater than that of cluster roots. Both total (micromol per plant h(-1)) and specific (micromol g(-1) root d. wt h(-1)) H+ extrusions were greatest from roots of plants supplied with (NH4)2SO4, followed by those supplied with NH4NO3 and N2 fixation, whereas plants receiving KNO3 had negative net H+ extrusion between the third and fifth week of growth (indicating uptake of protons or release of OH- ions). The rate of proton extrusion by NH4+-N-fed plants was similar under P-deficient and P-sufficient conditions. In contrast, proton exudation by N2-fixing plants and KNO3-treated plants was ten-fold greater under P deficiency than under P sufficiency. In comparison with P deficiency, plants treated with 50 microM P had a significantly higher concentration of P in roots, shoots and youngest expanded leaves (YEL). Compared with the N2 fixation and KNO3 treatments, total N concentration was highest in roots, shoots and YEL of plants supplied with (NH4)2SO4 and NH4NO3, regardless of P supply. Under P deficiency, K concentrations in roots decreased at all N supplies, especially in plants treated with (NH4)2SO4 and NH4NO3, which coincided with the greatest H+ extrusion at these P and N supplies. In conclusion, NH4-N nutrition stimulated cluster root formation and H+ extrusion by roots of P-deficient white lupin.     

Root and nodule growth in Pisum sativum L. in relation to photosynthesis: analysis using 13C-labelling

The effect of the nitrogen source (gaseous nitrogen, N2, or nitrate ions, NO3-) on the use of carbon (C) for root and nodule growth of pea (Pisum sativum L.) was investigated using 13C-labelling of assimilated CO2 at various stages of growth. Nitrate supply and growing conditions (sowing dates, air CO2 concentration) were varied to alter photosynthetic rates. Nodules are the sink with the highest demand for C in both the vegetative and flowering stages, growing at the expense of shoot and root in the vegetative stage, but only at the expense of roots at flowering. Until flowering, the addition of C into root and nodule biomass was linearly related to pre-existing biomass, thus determining net sink strengths which decreased with root and nodule age. Nodule growth patterns did not depend on the N source, whereas root growth was increased by nitrate when nodule biomass was low. At seed filling, the increase in C of biomass of the root system was no longer related to pre-existing biomass and C was preferentially diverted to roots of plants assimilating nitrate, or to nodules for plants fixing N2.     

Low concentrations of ammonium inhibit specific nodulation (nodule number g-1 root DW) in soybean (Glycine max [L.] Merr.)

Experiments on peas (Gulden and Vessey, 1997) have indicated that NH ₄ ⁺ stimulates both whole plant (nodules plant^-1) and specific nodulation (nodules g^-1 root DW). The effect of low concentrations of NH ₄ ⁺ on the soybean/Bradyrhizobium symbiosis is unknown. The objectives of the current study were to determine the immediate and residual effects of NH ₄ ⁺ on nodulation and N₂ fixation in soybean (Glycine max [L.] Merr.) in sand culture. Soybean (cv. Maple Ridge) were exposed to 0.0, 0.5, 1.0 and 2.0 mM of ¹⁵N-labelled (NH₄)₂SO₄ for 28 days after inoculation (DAI). From 29 to 56 DAI the plants were grown on NH ₄ ⁺ -free nutrient solution. Plants were harvested at 7, 14, 21, 28 and 56 DAI for root, shoot and nodule dry weight (DW), total N content, nodule counts and ¹⁵N enrichment of plant composites. Nitrogenase activity was measured by gas exchange at 28 DAI. The plants in the control (0.0 mM NH ₄ ⁺ ) treatment had consistently lower relative growth rates than the plants in the NH ₄ ⁺ treatments during the first 28 DAI. Plant growth was also less at 2.0 mM NH ₄ ⁺ compared to growth at 0.5 and 1.0 mM NH ₄ ⁺ . At 28 DAI, plants exposed to 0.5 and 1.0 mM NH ₄ ⁺ had significantly more nodules per plant and larger individual nodules than either the NH ₄ ⁺ -free controls or the 2.0 mM NH ₄ ⁺ -supplied plants. However, specific nodulation (nodule number g^-1 root DW) and specific nitrogenase activity (nitrogenase activity g^-1 nodule DW) were on average approximately 286% and 60% higher in the control plants, respectively, than for plants in the NH ₄ ⁺ treatments at 28 DAI. Also at 28 DAI, specific nodule DW (nodule DW g^-1 root DW) were 17, 44 and 53% higher in control plants than plants that had been exposed to 0.5, 1.0 and 2.0 mM NH ₄ ⁺ . At 56 DAI, after an additional 4 weeks of NH ₄ ⁺ -free nutrition, the plants which had previously received 0.5 and 1.0 mM NH ₄ ⁺ still maintained the highest plant DW and N contents, however, specific nodule DW had become similar at 600 mg nodule DW g^-1 root DW among all treatments. It is concluded that NH ₄ ⁺ has a negative effect on the nodulation process in the soybean/Bradyrhizobium symbiosis (as best indicated by the negative effect of NH ₄ ⁺ on specific nodulation). Despite this negative effect on specific nodulation, 0.5 and 1.0 mM NH ₄ ⁺ resulted in higher whole plant nodulation and N₂ fixation due to a compensating, positive effect on overall plant growth (i.e. fewer nodules g^-1 root DW, but much larger roots). Once NH ₄ ⁺ was removed from all treatments, the soybean plants appeared to move toward a consistent level of nodule DW relative to root DW.     

The role of phosphorus in nitrogen fixation by young pea plants (Pisum sativum)

The influence of P on N₂ fixation and dry matter production of young pea ( Pisum sativum L. cv. Bodil) plants grown in a soil-sand mixture was investigated in growth cabinet experiments. Nodule dry weight, specific C₂H₂ reduction and P concentration in shoots responded to P addition before any growth response could be observed. The P concentration in nodules responded only slightly to P addition. A supply of P to P-deficient plants increased both the nodule dry weight, specific C₂H₂ reduction and P concentration in shoots relatively faster than it increased shoot dry weight and P concentration in nodules. Combined N applied to plants when N₂ fixation had commenced, increased shoot dry weight only at the highest P levels. This indicates that the smaller plant growth at the low P levels did not result from N deficiency. The reduced nodulation and N₂ fixation in P-deficient plants seem to be caused by impaired shoot metabolism and not by a direct effect of P deficiency of the nodules.     

The influence of phosphorus deficiency on growth and nitrogen fixation of white clover plants

The effects of P deficiency on growth, N(2)-fixation and photosynthesis in white clover (Trifolium repens L.) plants were investigated using three contrasting relative addition rates of P, or following abrupt withdrawal of the P supply. Responses to a constant below-optimum P supply rate consisted of a decline in N(2)-fixation per unit root weight and a small reduction in the efficiency with which electrons were allocated to the reduction of N(2) in nodules. Abrupt removal of P arrested nodule growth and caused a substantial decline in nitrogenase activity per unit root weight, but not per unit nodule mass. Similarly, the rate of photosynthesis per unit leaf area was unaffected by abrupt P removal, whereas CO(2) acquisition for the plant as a whole decreased due to a decline in total leaf area, leaf area per unit leaf weight and utilization of incoming radiation. These changes followed the decline in tissue P concentrations. The ratio between CO(2)-fixation and N(2)-fixation was maintained under short-term P deprivation but increased under long-term low P supply, indicating a regulatory inhibition of nodule activity following morphological and growth adjustments. It is concluded that N(2)-fixation did not limit the growth of clover plants experiencing P deficiency. A low P status induced changes in the relative growth of roots, nodules and shoots rather than changes in N and/or C uptake rates per unit mass or area of these organs.     

The stimulating effect of ammonium on nodulation in Pisum sativum L. is not long lived once ammonium supply is discontinued

Although mineral N generally has a negative effect on legume-rhizobia symbioses, experiments in hydroponic culture in our laboratory (Waterer et al., 1992) have shown that low concentrations of NH⁺ ₄ can stimulate nodulation in pea (Pisum sativum L.). The objectives of the current study were to determine the immediate and residual effects of NH⁺ ₄ on nodulation and N₂ fixation in pea in sand culture. Peas (cv. Express) were exposed to 0.0, 0.5, 1.0, and 2.0 mM of ¹⁵N-labelled (NH₄)₂SO₄ for 28 days after inoculation (DAI). From 28 to 56 DAI the plants were grown on a NH⁺ ₄-free nutrient solution. Plants were harvested at 7, 14, 21, 28 and 56 DAI and nitrogenase activity was measured by gas exchange at 28 and 56 DAI. Root, shoot, and nodule dry weight (DW) and total N content were obtained, in addition to nodule counts and ¹⁵N enrichment of plant composites. The 1.0 and 2.0 mM NH⁺ ₄ treatments consistently resulted in higher total plant DW accumulation than the control (0.0 mM NH⁺ ₄). At 28 DAI, plants exposed to 1.0 and 2.0 mM NH⁺ ₄ had 1.8 to 2.8 times more nodules plant^-1, respectively, and plants exposed to 2.0 mM NH⁺ ₄ had 1.7 fold higher specific nodulation (nodule number g^-1 root DW). However, individual nodule DW was greater in control plants, such that there were no differences in nodule DW per plant among treatments. Ammonium treatment resulted in more nitrogen derived from the atmosphere (NDFA) in peas early in the experiment, but by 28 DAI there were no treatment effects on NDFA. Whole plant and nodule specific nitrogenase activity (µmol H₂ g^-1 nodule DW h^-1) was higher in control plants at 28 DAI. However, by 56 DAI, after an additional 4 weeks of NH⁺ ₄-free nutrition, no differences in nitrogenase activity nor whole plant or specific nodulation were detectable. This study indicates that nodulation in pea is stimulated in sand culture while exposed to NH⁺ ₄. However, once NH⁺ ₄ is removed, relative growth rate, nodulation and nitrogenase activity becomes similar to plants that were never exposed to NH⁺ ₄.     

Concentration-dependent influence of quercetin on nodulation process and the main characteristics of soybean inoculated with Bradyrhizobium japonicum

Soybean plants cv. Corsoy were grown in greenhouse conditions on sterilized quartz sand. They were inoculated with Bradyrhizobium japonicum, strain 542. The plants were treated with different concentrations of quercetin (ranging from 10 nM to 1M) at regular intervals during the experiment. The experiment was terminated at flower development. The following parameters, important for symbiosis efficiency were determined: shoot, root and nodule weights, nodule number, total leghemoglobin in the nodules,total nitrogen and soluble protein concentrations in shoots and roots, as well as chlorophyll concentration in the leaves.The results obtained partly confirmed the earlier findings that quercetin inhibits nodulation since increasing quercetin concentration decreased the number of nodules. However, at very low concentrations, quercetin stimulated the number of nodules. Quercetin also exerted a stimulating influence on other characteristics of the plant and nodules which did not correlate with nodule number and quantity of N fixed. These are: nodule weight, leghemoglobin concentration, total soluble protein content in shoots and roots as well as shoot and root weight.     

P-deficiency increases the O2 uptake per N2 reduced in alfalfa

Nodulated alfalfa (Medicago sativa L. cv. Saranac) plants were grown in hydroponics at P-sufficient and P-deficient supply levels. After 5 weeks of growth, dry matter accumulation, nodulation, total N and P accumulation, as well as 15N2 uptake, were measured. Moreover, the response of nodule O2-uptake to raising external pO2 was determined in an open-flow measurement system and nodule permeability was calculated. Plants in the P-deficient supply treatment had a lower P concentration in all organs. In both treatments the highest P concentration was found in nodules. In the P-deficient supply treatment plants formed less dry matter, had a lower shoot/root ratio, less nodulation, decreased total N accumulation, and lower 15N2 uptake per dry matter nodule. Nodules in the P-deficient treatment were, on average, smaller and had a higher O2 uptake per N2 reduced, coinciding with increased nodule permeability and conductance. Thus increased oxygen uptake appears to be a mechanism to adjust nodule metabolism to P deficiency in indeterminate N2-fixing nodules such as in alfalfa, as has previously been shown for determinate nodule forms.     

The Physiology and Nitrogen-fixing Capability of Aquatically and Terrestrially Grown Neptunia plena: The Importance of Nodule Oxygen Supply

The aquatic legume Neptunia plena (L.) Benth. was grown in non-aeratedwater culture or vermiculite. Growth, nodulation, nitrogen fixationand nodule physiology were investigated. Over an 80-d period,plants grew and fixed nitrogen and carbon equally well in bothrooting media, although distribution of growth between plantparts varied. Total nodule dry weights and volumes were similarbut vermiculite-grown plants had three times as many (smaller)nodules than those grown in water. Oxygen diffusion resistanceof nodules exposed to 21% oxygen and 10% acetylene did not differsignificantly. Both treatments showed similar declines in rootrespiration and acetylene reduction activity (approx. 10%) whenroot systems were exposed to stepped decreases and increasesin rhizosphere oxygen concentration. However, nitrogenase activityof aquatically grown plants was irreversibly inhibited by rapidexposure of nodules to ambient air, whereas vermiculite-grownplants were unaffected. Aeration of water-cultured N. plenareduced stem length (but not mass) and number of nodules perplant. The concentration of nitrogen fixation by 163%. PossibleO₂ transport pathways from the shoot atmosphere to roots andnodules are discussed. Aquatic legume, diffusion resistance, Neptunia plena, nitrogen fixation, oxygen, root nodules     

Interactions Between High pH and Iron Supply on Nodulation and Iron Nutrition of Lupinus albus L. Genotypes Differing in Sensitivity to Iron Deficiency

Poor growth of white lupin (Lupinus albus L.) on alkaline soils may result from its sensitivity to iron deficiency and poor nodulation. This study examined interactive effects of iron supply and high pH on the growth and nodulation of three genotypes differing in their sensitivity to iron deficiency. Three genotypes (P27486, Ultra and WTD180) were grown for 17 days in buffered solutions with Fe supply of 0.2, 2 and 20 μM. Solution pH was adjusted to 5.2, 6.5 or 7.5. Plant growth, nodulation and nutrient concentrations in plants were measured. Decreasing Fe supply decreased chlorophyll concentration in young leaves by up to 92%. Increasing pH decreased chlorophyll concentration by an average of 40% at pH 6.5 and by 47% at pH 7.5. The decrease of chlorophyll was less obvious in P27485 than in Ultra or WTD180. Shoot biomass was reduced by up to 18% by Fe deficiency, with such decrease being less for P27486. Increasing pH exacerbated the effect of Fe deficiency on shoot biomass only of Ultra. Decreasing Fe supply decreased nodule number by an average of 54%, and increasing pH decreased nodule number by 80%. P27486 formed the greatest number of nodules while WTD180 the least. P27486 had high Fe uptake and low internal requirement. Irrespective of genotype, leaf chlorosis positively correlated with cluster root formation. The results suggest that a combination of Fe deficiency and high pH impaired nodulation in L. albus, and that selection of genotypes for both tolerance of iron deficiency and good nodulation at high pH is important for a successful lupin crop on alkaline soils.     

Nitrate uptake,nitrate reductase distribution and their relation to proton release in five nodulated grain legumes

Nitrate uptake, nitrate reductase activity (NRA) and net proton release were compared in five grain legumes grown at 0.2 and 2 mM nitrate in nutrient solution. Nitrate treatments, imposed on 22-d-old, fully nodulated plants, lasted for 21 d. Increasing nitrate supply did not significantly influence the growth of any of the species during the treatment, but yellow lupin (Lupinus luteus) had a higher growth rate than the other species examined. At 0.2 mM nitrate supply, nitrate uptake rates ranged from 0.6 to 1.5 mg N g(-1) d(-1) in the order: yellow lupin > field pea (Pisum sativum) > chickpea (Cicer arietinum) > narrow-leafed lupin (L angustifolius) > white lupin (L albus). At 2 mM nitrate supply, nitrate uptake ranged from 1.7 to 8.2 mg N g(-1) d(-1) in the order: field pea > chickpea > white lupin > yellow lupin > narrow-leafed lupin. Nitrate reductase activity increased with increased nitrate supply, with the majority of NRA being present in shoots. Field pea and chickpea had much higher shoot NRA than the three lupin species. When 0.2 mM nitrate was supplied, narrow-leafed lupinreleased the most H+ per unit root biomass per day, followed by yellow lupin, white lupin, field pea and chickpea. At 2 mM nitrate, narrow-leafed lupin and yellow lupin showed net proton release, whereas the other species, especially field pea, showed net OH- release. Irrespective of legume species and nitrate supply, proton release was negatively correlated with nitrate uptake and NRA in shoots, but not with NRA in roots.     

Evidence that P deficiency induces N feedback regulation of symbiotic N2 fixation in white clover (Trifolium repens L.)

Trifolium repens L. was grown to test the following hypotheses: when P is deficient (i) N2 fixation decreases as a result of the plant's adaptation to the low N demand, regulated by an N feedback mechanism, and (ii) the decrease in the photosynthetic capacity of the leaves does not limit N2 fixation. Severe P deficiency prevented nodulation or stopped nodule growth when the P deficiency occurred after the plants had formed nodules. At low P, the proportion of whole-plant-N derived from symbiotic N2 fixation decreased, whereas specific N2 fixation increased and compensated partially for poor nodulation. Leaf photosynthesis was reduced under P deficiency due to low Vc,max and Jmax. Poor growth or poor performance of the nodules was not due to C limitation, because (i) the improved photosynthetic performance at elevated pCO2 had no effect on the growth and functioning of the nodules, (ii) starch accumulated in the leaves, particularly under elevated pCO2, and (iii) the concentration of WSC in the nodules was highest under P deficiency. Under severe P deficiency, the concentrations of whole-plant-N and leaf-N were the highest, indicating that the assimilation of N exceeded the amount of N required by the plant for growth. This was clearly demonstrated by a strong increase in asparagine concentrations in the roots and nodules under low P supply. This indicates that nodulation and the proportion of N derived from symbiotic N2 fixation are down-regulated by an N feedback mechanism.     

The nodule conductance to O<Subscript>2</Subscript> diffusion increases with high phosphorus content in the <Emphasis Type="Italic">Phaseolus vulgaris-</Emphasis>rhizobia symbiosis

Although recent studies have addressed the effects of phosphorus (P) deficiency on nodule O₂ permeability, little attention has been given to the relationship between nodule P status and nodule permeability. To study these traits, four recombinant inbred lines, namely RILs 34, 83, 115, 147 and one local variety (Concesa) of common bean (Phaseolus vulgaris) were inoculated with RhM11 (a native rhizobial strain from Haouz area of Marrakesh), and grown in hydroaeroponic culture under P-sufficiency (250 μmol P plant⁻¹ week⁻¹) versus P-deficiency (75 μmol P plant⁻¹ week⁻¹) conditions. At the flowering stage, the biomass of plants and nodules and their P contents was determined after measuring O₂ uptake by nodulated roots (Conr) and nodule conductance to O₂ diffusion (g_n). The results showed that P-deficiency significantly decreased plant growth and nodulation, though there were differences between bean genotypes. P-deficiency also induced a decrease in nodule P content (31%) in both sensitive (83, 147) and tolerant lines (34, 115), a 42 and 27% reduction in shoots of sensitive and tolerant lines, respectively. These decreases were associated with significant variations in nodule surface and O₂ permeability among bean genotypes and P-nutrition. Under P-deficiency, g_n increased more for the sensitive (39%) than for the tolerant lines (27%). This increase was linked with a rise both in the P levels in nodules and shoots, as well as in the efficiency of symbiotic nitrogen fixation as determined by nodule-dependent biomass production for the sensitive lines. Furthermore, positive correlations were found between O₂ permeability, g_n and P content both in nodules and shoots (r ² = 0.94** and r ² = 0.96**). We conclude that nodule variations in Conr and g_n are related to nodule P content, and may contribute to the adaptation of energy metabolism in N₂-fixing bean nodules to P-deficiency.     

Role of phosphorus nutrition in development of cluster roots and release of carboxylates in soil-grown Lupinus albus

The present study examined the effect of phosphorus (P) limitation on cluster root formation and exudation of carboxylates by N₂-fixing white lupin (Lupinus albus L. cv. Kiev) grown in a P-deficient sandy soil. Plants received 10 (limited P) or 200 g P g^–1 soil as FePO₄ (adequate P) and were grown in a phytotron at 20/12 °C (12/12 h) for 76 days in soil columns. Cluster root formation was assessed and root exudates were collected at 9-day intervals. Shoot and root dry weights were higher in plants grown in the adequate-P compared to the limited-P treatment for 67 days. No clear difference in the total root length was observed between two P treatments before day 58. However, the specific root length increased rapidly from 17 m g^–1 DW at day 40 to 28 m g^–1 at day 49 in the P-limited plants, but decreased in the P-adequate plants. The effect of P limitation on enhancement of cluster root formation was observed from day 40 and reached the maximum at day 58. The number of cluster roots was negatively correlated with the P concentration in both roots and shoots. Phosphorus limitation increased exudation of citrate from day 40. The exudation of citrate displayed a cyclic pattern throughout the experiment, and appeared related to internal P concentration in plants, particularly P concentration in shoots. The sorption of exogenously added citrate in the soil was also examined. The amount of extractable citrate remained unchanged for 2 h, but decreased thereafter, suggesting that the soil had a low capacity to sorb citrate, and the rate of its decomposition by microorganisms was slow. Collecting solution leached through a soil column is a simple and reliable method to acquire root exudates from white lupin grown in soil. The results suggest that formation of cluster roots and exudation of citrate in white lupin are regulated by P concentration in shoots.     

Regulation of nodulation in the absence of N2 is different in actinorhizal plants with different infection pathways

Root nodulation in actinorhizal plants, like Discaria trinervis and Alnus incana, is subject to feedback regulatory mechanisms that control infection by Frankia and nodule development. Nodule pattern in the root system is controlled by an autoregulatory process that is induced soon after inoculation with Frankia. The final number of nodules, as well as nodule biomass in relation to plant biomass, are both modulated by a second mechanism which seems to be related to the N status of the plant. Mature nodules are, in part, involved in the latter process, since nodule excision from the root system releases the inhibition of infection and nodule development. To study the effect of N(2) fixation in this process, nodulated D. trinervis and A. incana plants were incubated under a N(2)-free atmosphere. Discaria trinervis is an intercellularly infected species while A. incana is infected intracellularly, via root hairs. Both symbioses responded with an increment in nodule biomass, but with different strategies. Discaria trinervis increased the biomass of existing nodules without significant development of new nodules, while in A. incana nodule biomass increased due to the development of nodules from new infections, but also from the release of arrested infections. It appears that in D. trinervis nodules there is an additional source for inhibition of new infections and nodule development that is independent of N(2) fixation and nitrogen assimilation. It is proposed here that the intercellular Frankia filaments commonly present in the D. trinervis nodule apex, is the origin for the autoregulatory signals that sustain the blockage of initiated nodule primordia and prevent new roots from infections. When turning to A. incana plants, it seems likely that this signal is related to the early autoregulation of nodulation in A. incana seedlings and is no longer present in mature nodules. Thus, actinorhizal symbioses belonging to relatively distant phylogenetic groups and displaying different infection pathways, show different feedback regulatory processes that control root nodulation by Frankia.     

Effect of inoculation and leaf litter amendment on establishment of nodule-forming Frankia populations in soil

High-N(2)-fixing activities of Frankia populations in root nodules on Alnus glutinosa improve growth performance of the host plant. Therefore, the establishment of active, nodule-forming populations of Frankia in soil is desirable. In this study, we inoculated Frankia strains of Alnus host infection groups I, IIIa, and IV into soil already harboring indigenous populations of infection groups (IIIa, IIIb, and IV). Then we amended parts of the inoculated soil with leaf litter of A. glutinosa and kept these parts of soil without host plants for several weeks until they were spiked with [(15)N]NO(3) and planted with seedlings of A. glutinosa. After 4 months of growth, we analyzed plants for growth performance, nodule formation, specific Frankia populations in root nodules, and N(2) fixation rates. The results revealed that introduced Frankia strains incubated in soil for several weeks in the absence of plants remained infective and competitive for nodulation with the indigenous Frankia populations of the soil. Inoculation into and incubation in soil without host plants generally supported subsequent plant growth performance and increased the percentage of nitrogen acquired by the host plants through N(2) fixation from 33% on noninoculated, nonamended soils to 78% on inoculated, amended soils. Introduced Frankia strains representing Alnus host infection groups IIIa and IV competed with indigenous Frankia populations, whereas frankiae of group I were not found in any nodules. When grown in noninoculated, nonamended soil, A. glutinosa plants harbored Frankia populations of only group IIIa in root nodules. This group was reduced to 32% +/- 23% (standard deviation) of the Frankia nodule populations when plants were grown in inoculated, nonamended soil. Under these conditions, the introduced Frankia strain of group IV was established in 51% +/- 20% of the nodules. Leaf litter amendment during the initial incubation in soil without plants promoted nodulation by frankiae of group IV in both inoculated and noninoculated treatments. Grown in inoculated, amended soils, plants had significantly lower numbers of nodules infected by group IIIa (8% +/- 6%) than by group IV (81% +/- 11%). On plants grown in noninoculated, amended soil, the original Frankia root nodule population represented by group IIIa of the noninoculated, nonamended soil was entirely exchanged by a Frankia population belonging to group IV. The quantification of N(2) fixation rates by (15)N dilution revealed that both the indigenous and the inoculated Frankia populations of group IV had a higher specific N(2)-fixing capacity than populations belonging to group IIIa under the conditions applied. These results show that through inoculation or leaf litter amendment, Frankia populations with high specific N(2)-fixing capacities can be established in soils. These populations remain infective on their host plants, successfully compete for nodule formation with other indigenous or inoculated Frankia populations, and thereby increase plant growth performance.     

Effects of drought stress on legume symbiotic nitrogen fixation: physiological mechanisms

Drought stress is one of the major factors affecting nitrogen fixation by legume-rhizobium symbiosis. Several mechanisms have been previously reported to be involved in the physiological response of symbiotic nitrogen fixation to drought stress, i.e. carbon shortage and nodule carbon metabolism, oxygen limitation, and feedback regulation by the accumulation of N fixation products. The carbon shortage hypothesis was previously investigated by studying the combined effects of CO2 enrichment and water deficits on nodulation and N2 fixation in soybean. Under drought, in a genotype with drought tolerant N2 fixation, approximately four times the amount of 14C was allocated to nodules compared to a drought sensitive genotype. It was found that an important effect of CO2 enrichment of soybean under drought was an enhancement of photo assimilation, an increased partitioning of carbon to nodules, whose main effect was to sustain nodule growth, which helped sustain N2 rates under soil water deficits. The interaction of nodule permeability to O2 and drought stress with N2 fixation was examined in soybean nodules and led to the overall conclusion that O2 limitation seems to be involved only in the initial stages of water deficit stresses in decreasing nodule activity. The involvement of ureides in the drought response of N2 fixation was initially suspected by an increased ureide concentration in shoots and nodules under drought leading to a negative feedback response between ureides and nodule activity. Direct evidence for inhibition of nitrogenase activity by its products, ureides and amides, supported this hypothesis. The overall conclusion was that all three physiological mechanisms are important in understanding the regulation of N2 fixation and its response of to soil drying.     

Symbiotic properties of Lotus pedunculitis root nodules induced by Rhizobium loti and Bradyrhizobium sp. (Lotus)

Vance, C. P., Reibach, P. H. and Pankhurst, C. E. 1987. Symbiotic properties of Lotus pedunculatus root nodules induced by Rhizobium loti and Bradyrhizobium sp. ( Lotus ).
Symbiotic properties of root nodules were evaluated in glasshouse-grown Lotus pedunculatus Cav. cv. Maku inoculated with either a fast-growing Rhizobium loti strain NZP2037 or a slow-growing Bradyrhizobium sp. ( Lotus ) strain CC814s. Although the nodule mass of plants inoculated with NZP2037 was twice that of plants inoculated with CC814s, the yield of NZP2037 shoots and roots was 50% that of CC814s shoots and roots. Nodules induced by Bradyrhizobium fixed substantially more N than nodules induced by R. loti. Glucose requirements [mol glucose (mol N₂ fixed)^-1] of nodules induced by CC814s and NZP2037 were 7.1 and 16.6, respectively. Nodule enzymes of carbon and nitrogen assimilation reflected the disparity of the two sym-bioses. Xylem sap of the symbiosis with the higher yield contained a higher concentration of asparagine [9.86 μmol (ml xylem sap)'] than did the lower yielding symbiosis [5.80 umol (ml xylem sap)"']. Nodule CO₂ fixation was directly linked to nodule N assimilation in both symbioses. The results indicate that the difference between the two symbioses extend to nodule N and C assimilation and whole plant N transport. The data support a role for host plant modulation of bacterial efficiency and assimilation of fixed N.     

Influence of Nematodes and Light Sources on Growth and Nodulation of Soybean

The influence of nematodes on nodulation of soybean varied according to their modes of parasitism. In the greenhouse, nodule formation was stimulated by the endoparasites, Meloidogyne hapla and Pratylenchus penetrans, but was inhibited slightly by the ectoparasite, Belonolaimus longicaudatus. In an experiment under controlled conditions in a phytotron, Heterodera glycines severely inhibited nodule formation, whereas plants inoculated with B. longicaudatus and P. penetrans had more nodules per g root than nematode-free plants. Nitrogen-fixing capacity, however, was inhibited by all three nematode species. Different light sources used in the phytotron experiment also influenced growth and nodulation of soybean. A fluorescent plus incandescent light regime resulted in plants with the greatest shoot weight, pod number, and nodules per g root. Plants grown under Lucalox lamps had excessive stem elongation.