Evaluation of symbiotic dinitrogen inputs of herbaceous legumes into tropical cover-crop systems

 Reliable estimates of symbiotically fixed N₂ in herbaceous legumes are important in order to determine their role in maintaining or improving N levels in tropical low-external-input farming systems. We have studied the effects of different management systems on the suitability of two non-N₂-fixing reference crops, imperata [Imperata cylindrica (L.) Rauescel] and maize (Zea mays L.), for estimating N₂ fixation in mucuna [Mucuna pruriens (L.) DC var. utilis (Wright) Bruck] and lablab [Lablab purpureus (L.) Sweet] in the field. The total-N-difference (TND) method of estimating N₂ fixation was compared to the ¹⁵N-isotope-dilution (ID) technique. The two methods did not differ with respect to estimates of N₂ fixation under in situ mulch (IM) systems. In contrast, under live-mulch (LM) systems the TND method underestimated N₂ fixed in mucuna by 29% and in lablab by 40% compared to estimates made with the ID method. Irrespective of the treatment, estimates of N derived from fixation in both herbaceous legumes were not influenced by either of the reference plants. Using the ID technique, the proportion of N₂ derived from fixation in mucuna and lablab at 12 weeks varied from 52% to 90% depending on whether the treatments were N fertilized, inoculated or uninoculated, cover-crop systems. In view of the nature of cover-crop systems in the derived savanna of West and Central Africa, where imperata is usually present as a weed or maize is grown in IM or LM systems, imperata or maize could be used to estimate N₂ fixation and N contributions of the legumes to soil fertility and subsequent crop improvements. Received: 16 October 1999     

Sesbania rostrata as a green manure for lowland rice: Growth,N2 fixation,Azorhizobium sp. inoculation,and effects on succeeding crop yields and nitrogen balance

Summary Root and stem nodulation, nitrogen fixation (acetylene-reducing activity), growth and N accumulation bySesbania rostrata as affected by season and inoculation were studied in a pot experiment. The effects ofS. rostrata as a green manure on succeeding wet-season and dry-season rice yields and total N balance were also studied.S. rostrata grown during the wet season showed better growth, nodulation, and greater acetylene-reducing activity than that grown during the dry season. Inoculation withAzorhizobium caulinodans ORS 571 StrSpc® (resistant to streptomycin and spectinomycin) on the stem alone or on both root and stem significantly increased N₂ fixation by the plants. Soil and seed inoculation yielded active root nodules under flooded conditions. Plants that were not inoculated on the stem did not develop stem nodules. The nitrogenase activity of the root nodules was greater than that of the stem nodules in about 50-day-oldS. rostrata. S. rostrata incorporation, irrespective of inoculation, significantly increased the grain yield and N uptake of the succeeding wet season and dry season rice crops. The inoculated treatments produced a significantly greater N gain (873 mg N pot^–1) than the noinoculation (712 mg N pot^–1) treatment. About 80% of the N gained was transferred to the succeeding rice crops and about 20% remained in the soil. The soil N in the flooded fallow-rice treatment significantly declined (–140 mg N pot^–1) but significantly increased in bothS. rostrata-rice treatments (159 and 151 mg N pot^–1 in uninoculated and inoculated treatments respectively). The N-balance data gave extrapolated values of N₂ fixed per hectare at about 303 kg N ha^–1 per two crops forS. rostrata (uninoculated)-rice and 383 forS. rostrata (inoculated)-rice.     

Field evaluation of lentil cultivars inoculated with Rhizobium leguminosarum bv. viciae strains for nitrogen fixation using nitrogen-15 isotope dilution

 A ¹⁵N isotope dilution technique was applied to quantify the extent of N₂ fixation in lentil (Lens culinaris Medik.) cultivars as influenced by Rhizobium leguminosarum bv. viciae strains in a field experiment in Pakistan. The experiment was conducted on a soil with a very small indigenous rhizobial population and where N was a limiting factor for crop production. Significant variations in number of nodules, dry weight of nodules, biomass yield, grain yield, total N yield, proportion of plant N derived from N₂ fixation (P_fix) and amount of N derived from the atmosphere (N_dfa) were observed among combined treatments of four rhizobial strains and six lentil varieties. In a field previously labelled with ¹⁵N, to which a basal dose of 75 kg P₂O₅ ha^–1 was applied as single super phosphate, N_dfa ranged from 15 to 24 kg N ha^–1 when calculated according to rhizobial strain and from 4 to 38 kg N ha^–1 when calculated according to lentil variety. Lc 26 was the most effective strain and fixed 243% more N than the indigenous population in the uninoculated control. In treatments with the lentil variety PL-406, N_dfa was 38 kg N ha^–1, which was 850% higher than with the lentil variety Precoz/F6-20-1×M-85. Generally, the varieties with greater P_fix produced a higher dry matter yield. Received: 26 May 1999     

A study on the improvement iron nutrition of peanut intercropping with maize on nitrogen fixation at early stages of growth of peanut on a calcareous soil

Abstract

A glasshouse study employing a split-root technique was conducted to investigate the influence of intercropping with maize (Zea mays L.) in a calcareous soil on N₂ fixation by peanut (Arachis hypogaea L.) at early stages of growth. In this intercropping system, competitive interactions between maize and peanut for N and improvement of Fe uptake were likely to be important factors affecting N₂ fixation of peanut. The experiment was comprised of three treatments which included treatment I: peanut monocropping; treatment II: maize/peanut intercropping (the major and the minor compartments with low N, 50 mg kg^?1); treatment III: maize/peanut intercropping (the major compartment with low N, 50 mg kg^?1 and the minor compartment with high, N 200 mg kg^?1). The minor compartment of treatment III was fertilized with 200 mg kg^?1 N for reducing or eliminating the competition of N coming from intercropping maize. Intercropping with maize corrected Fe chlorosis of peanut by significantly increasing plant Fe concentration and uptake. Compared with the monocropping treatment, iron uptake increased from intercropping treatment II and III by 22 and 24% per plant, 30 and 29% shoots, 38 and 60% nodules. Iron uptake by the root nodules was especially enhanced in the intercropping system. In contrast, intercropping with maize had little effect on NO₃ ^?1-N concentrations in the soil rhizosphere of peanut or on N concentrations and uptake by peanut compared with plants in monoculture. The results indicate that the improvement in Fe nutrition was an important factor promoting N₂ fixation by peanut in the intercropping system at the flowering stage of peanut growth, and that competition for N by intercropped maize had little effect on N₂ fixation by peanut under the experimental conditions.     

Evaluation of N2 fixation by applying 15N labeled plant material and ammonium sulfate

Effect of different ¹⁵N labeled sources on the estimation of N₂ fixation was investigated. The combination of ¹⁵N labeled ammonium sulfate, ¹⁵N labeled plant material, and ¹⁵N labeled ammonium sulfate with unlabeled plant material, was examined in pot experiments. Two cultivars of soybean (Glycine max) and one of mungbean (Vigna radiata) were used. No significant difference was observed among the treatments for the estimation of N₂ fixation. This was due to the homogeneity and stability of the ¹⁵N abundance in soil which resulted in a similar N uptake from the soil by the N² fixing and reference crops. The plant yield, total N uptake and amount of N₂ fixed were higher in the Yellow Soil than in the Andosol. The amount of N₂ fixed was strongly influenced by the plant growth and consequently it affected the plant yield. The slow decomposition of plant material in the Andosol resulted in a low yield in both the N₂ fixing and reference crops. Thus, the artificial decrease of the available N content in soil, by application of plant material, did not stimulate N, fixation but suppressed plant growth and N₂ fixation.     

Evaluation of nitrogen fixation by different strains of theAzolla-Anabaena symbiosis in the presence of a high level of ammonium

Summary The proportion of N derived from N₂ fixation for 99 strains ofAzolla spp. (comprising all known species) in the presence of ammonium (40 mg/1) was assessed using a¹⁵N-dilution technique. The percentage of N derived from air varied from 29.5% to 79.9%. Although the N concentration ofAzolla spp. was not correlated with fertilizer N, it correlated fairly well with N₂ fixation. Regression analysis suggests that the N yield ofAzolla spp. is more dependent on N₂ fixation than on ammonium assimilation. The high correlation between N yield and isotopically determined, fixed N₂ indicates that the N yield could be used as a parameter in the selection ofAzolla spp. strains that are capable of maintaining high N₂ fixation in the presence of a high level of ammonium.     

Drought and nitrogen source effects on nitrogen nutrition,seed growth,and yield in soybean

Drought in soybean [Glycine max (L.) Merr.] decreases yield‐related processes and N₂ fixation is more sensitive to drought than are many other of these processes. Therefore, application of nitrogen (N) fertilizer may increase drought tolerance over those plants primarily dependent on N₂ fixation. In a field experiment, NH₄NO₃ applications (+N) to drought‐stressed soybean resulted in biomass and N accumulation rates similar to those rates for an irrigated treatment without N fertilizer (‐N). In contrast, biomass and N accumulation rates were decreased for the ‐N treatment. N fertilization increased seed growth rate and decreased seed fill duration for irrigated and drought treatments. In the drought treatment, N application increased seed number per unit area, which resulted in higher yields. In a greenhouse experiment, fertilization with either KN0₃ or NH₄C1 increased biomass and N accumulation rates during drought over those of plants dependent solely on N₂ fixation. It was concluded that application of N fertilizer to soybean increases drought tolerance because of the extreme sensitivity of N₂ fixation to drought.     

Response of maize (Zea mays) to Azotobacter inoculation under fertilized and unfertilized conditions

Summary Inoculated seeds of maize (Zea mays) with 11 Azotobacter strains, sown in the fields receiving no fertilizer and fertilizers (N and P at the rate of 125 and 40 kg ha^–1 respectively) increased the grain yield by 19.63% and 15.89% respectively over the corresponding control. The effect was greater in unfertilized than in fertilized soil. The increase in yield due to fertilizers was 21.2% without inoculation and 37.09% with inoculation. The correlations between total yield, and N, P and K uptake were highly significant and comparable among themselves. This indicated that increase in yield due to inoculation was not due to N₂ fixation but that some other mechanisms like production of growth hormones by this bacterium may be responsible.     

Influence of mycorrhiza vs. soluble phosphate on growth,nodulation, and N2 fixation (15N) in alfalfa under different levels of water potential

Summary The legume Medicago sativa (+Rhizobium melilott) was grown under controlled conditions to study the interactions between soluble P in soil (four levels), or a mycorrhizal inoculum, and the degree of water potential (four levels) in relation to plant development and N₂ fixation. ¹⁵N-labelled ammonium sulphate was added to each pot for a qualitative estimate of N₂ fixation, in order to rank the effects of the different treatments.Dry-matter yield, nutrient content and nodulation increased with the amount of plant-available P in the soil, and decreased as the water stress increased, for each P-level. The mycorrhizal effect on dry matter, N yield, and on nodulation was little affected by the water potential. Since P uptake was affected by the water content in mycorrhizal plants, additional mechanisms, other than those mediated by P, must be involved in the mycorrhizal activity.There was a positive correlation between N yield and nodulation for the different P levels and the mycorrhizal treatment at all water levels. A high correlation between plant unlabelled N content and atom% ¹⁵N excess was also found for all levels of P. In mycorrhizal plants, however, the correlation between unlabelled N yield and ¹⁵N was lower. This suggests that mycorrhiza supply plants with other N sources in addition to those derived from the improvement on N₂ fixation.     

Nitrous oxide emissions from grain legumes as affected by wetting/drying cycles and crop residues

Grain legume production with rhizobial inoculation has drawn attention not only because of the economic value of nitrogen (N) fixation by grain legumes, but also because of the concern that N₂ fixation by grain legumes may enhance emissions of nitrous oxide (N₂O), a powerful greenhouse gas. However, the relationship between N₂O emissions and biological N₂ fixation by grain legumes is not well understood. The objective of this study was to quantify N₂O emissions associated with N₂ fixation by grain legumes as affected by wetting/drying cycles and crop residues. Two grain legumes, lentil (Lens esculenta Moench) and pea (Pisum sativum L.), either inoculated with two Rhizobium leguminosarum biovar viciae strains, 99A1 and RGP2, respectively, or fertilized with ¹⁵N-labeled fertilizer were grown in a controlled environment under three wetting/drying cycles. Profile N₂O concentrations and surface N₂O emissions were measured from the soil–plant systems, which were compared with those from a cereal, spring wheat (Triticum aestivum L. ac. Barrie). After harvest, crop residues were incorporated into soils that were seeded to spring wheat to evaluate the effect of crop residues on N₂O emissions. Results indicated that: (1) inoculating grain legumes with non-denitrifying rhizobia did not enhance N₂O emissions and the presence of grain legumes did not increase N₂O emissions compared with the cereal crop, and (2) profile N₂O accumulation and surface emissions were not related to the type of crop residues added to the soil, but related to the residual N applied previously as N fertilizer. This suggests that N₂O emissions are not directly related to biological N₂ fixation by grain legumes, and on a short time scale, N rich residues of N₂-fixing crops have a limited impact on N₂O emissions compared with N fertilization.     

Einfluß von NPK auf die Biomasseproduktion und Stickstoffbindung der stengelknöllchenbildenden Gründüngungsleguminosen Sesbania rostrata und Aeschynomene afraspera im Naßreisanbau

Influence of NPK on performance of the stem-nodulating legumes Sesbania rostrata and Aeschynomene afraspera in lowland rice The stem-nodulating tropical legumes S. rostrata and A. afraspera are promising green manure species for the low-input rice farming systems of lowland areas. Nutrient imbalances and soils low in available nutrients can considerably affect the use of biofertilizers. Use of mineral N, P, and K fertilizers in the growth of S. rostrata and A. afraspera as biofertilizers for lowland rice in the Philippines was evaluated. Applied P and K both stimulated growth, nodulation, and N₂ fixation. N accumulation in PK fertilized S. rostrata was about 40% higher than in nonfertilized green manure. Mineral N application (urea) depressed nodulation and N₂ fixation (ARA) in roots and resulted in an increased ARA in stem nodules. The legumes produced more N gains in the presence of small amounts of N. Use of S. rostrata and A. afraspera green manure increased in all cases significantly rice grain yield. Thus integration of mineral N, P, and K fertilizers in a green manurebased rice farming system can considerably improve biofertilizer production and increase rice grain yield.     

Inorganic nitrogen supply and symbiotic dinitro‐gen fixation in alfalfa

It may be desirable to minimize dinitrogen (N₂) fixation in alfalfa (Medicago saliva L.) when a source of inorganic nitrogen (N), such as manure, is readily available. Our objectives were to determine the N₂ fixation response of eight alfalfa germplasms to inorganic N and to characterize plant‐to‐plant variation for this trait. Seed was sown in vermiculite and irrigated with nutrient solution in growth chambers. Herbage was removed at 71 d and treatments of 1, 3, 5, or 10 mM N were applied as ¹⁵N‐depleted ammonium nitrate (NH₄NO₃). After 34 d of regrowth, herbage was removed and analyzed for dry mass, total N concentration, and N isotope ratio. Increased availability of inorganic N resulted in a linear increase in herbage weight, height, shoot number, and N concentration, and consistently decreased N₂ fixation for all germplasms. Estimated N₂ fixation was greater than zero at the highest rate of inorganic N, which we speculate was due, in part, to remobilized root and crown N, because nodules appeared to be nonfunctional. Across all treatments, N₂ fixation correlated best with herbage N concentration, but there was no relationship between these variables within a given N treatment concentration. Significant variation in reliance on N₂ fixation in the presence of inorganic N existed in all eight germplasms.     

Iron fertilizers applied to calcareous soil on the growth of peanut in a pot experiment

A greenhouse pot experiment was conducted with peanuts (Arachis hypogaea L., Fabceae) to evaluate iron compound fertilizers for improving within-plant iron content and correcting chlorosis caused by iron deficiency. Peanuts were planted in containers with calcareous soil fertilized with three different granular iron nitrogen, phosphorus and potassium (NPK) fertilizers (ferrous sulphate (FeSO₄)–NPK, Fe–ethylendiamine di (o-hydroxyphenylacetic) (EDDHA)–NPK and Fe–citrate–NPK). Iron nutrition, plant biomass, seed yield and quality of peanuts were significantly affected by the application of Fe–citrate–NPK and Fe–EDDHA–NPK to the soil. Iron concentrations in tissues were significantly greater for plants grown with Fe–citrate–NPK and Fe–EDDHA–NPK. The active iron concentration in the youngest leaves of peanuts was linearly related to the leaf chlorophyll (via soil and plant analyzer development measurements) recorded 50 and 80 days after planting. However, no significant differences between Fe–citrate–NPK and Fe–EDDHA–NPK were observed. Despite the large amount of total iron bound and dry matter, FeSO₄–NPK was less effective than Fe–citrate–NPK and Fe–EDDHA–NPK to improve iron uptake. The results showed that application of Fe–citrate–NPK was as effective as application of Fe–EDDHA–NPK in remediating leaf iron chlorosis in peanut pot-grown in calcareous soil. The study suggested that Fe–citrate–NPK should be considered as a potential tool for correcting peanut iron deficiency in calcareous soil.     

Yield,nodulation, and N2 fixation by cowpea cultivars at different phosphorus levels

A greenhouse experiment was conducted to investigate the effect of a P application (0 vs. 50 mg P kg^-1) on yield, nodulation, and N₂ fixation by three cowpea cultivars (Soronko, Amantin, and IT81D-1137) using the ¹⁵N isotope-dilution method. When P was not applied the inoculated cowpea genotypes showed significant differences (Soronko>Amantin> IT81D-1137) in N accumulation, in contrast to the uninoculated cowpea cultivars, which accumulated similar amounts of N. The differences in shoot N in inoculated plants were thus caused by differences in N₂ fixation. The average values of N fixed (for both P levels) were 74% in Soronko, 59% in Amantin, and 42% in IT81D-1137, corresponding to 80, 51, and 24 mg N plant^-1, respectively. Inoculation increased the total shoot-N accumulation in cv. Soronko by 270% without P and by 204% with P, cv. Amantin by 152 and 104%, and cv. IT81D-1137 by 74 and 58%, respectively. With P, the % N derived from atmosphere (%Ndfa) was 42% for IT81D-1137, 62% for Amantin, and 76% for Soronko. The high value for Soronko indicates that in a soil of medium fertility, certain cowpea cultivars are capable of satisfying their total N requirement through N₂ fixation. The P effect on N₂ fixation was mainly in the total amount of N fixed rather than on the percentage derived from the atmosphere.     

Subsurface drip irrigation and urea phosphate fertigation for vegetables on calcareous soils

Drip irrigation lines installed at 5 cm (shallow) or 15 cm (deep) below the soil surface and furrow irrigation were compared for vegetables grown on calcareous desert soils. Urea phosphate (UP) fertilizer (17–44–0) was injected twice in the drip irrigation lines during the growing season. Yields were compared to preplant fertilized and unfertilized furrows. Fall cabbage (Brassica oleracea var. capitata L.) gave comparable yields under the different irrigation treatments with the drip treatments using half the water used by the furrow treatment. Cabbage yield increased in all fertilized treatments as compared to the unfertilized furrow. Petiole P and NO₃‐N concentrations were higher from the drip than from the furrow treatments. Zucchini squash (Cucurbita pepo L.) had the highest yields under deep drip and fertilized furrow treatments, with the deep drip using half the water and P rate used by the furrow treatment. The deep drip increased squash yield by 34% over the shallow drip. The unfertilized furrow gave the lowest yield. Leaf tissue concentrations of P and NO₃‐N were comparable under deep drip and fertilized furrow treatments and were higher than the concentrations achieved from shallow drip and unfertilized furrow treatments.     

Effect of mineral nitrogen fertilizer forms on N2O emissions from arable soils in winter wheat production

Nitrogen fertilizers are supposed to be a major source of nitrous oxide (N₂O) emissions from arable soils. The objective of this study was to compare the effect of N forms on N₂O emissions from arable fields cropped with winter wheat (Triticum aestivum L.). In three field trials in North‐West Germany (two trials in 2011/2012, one trial in 2012/2013), direct N₂O emissions during a one‐year measurement period, starting after application of either urea, ammonium sulfate (AS) or calcium ammonium nitrate (CAN), were compared at an application rate of 220 kg N ha^?1. During the growth season (March to August) of winter wheat, N₂O emission rates were significantly higher in all three field experiments and in all treatments receiving N fertilizer than from the non‐fertilized treatments (control). At two of the three sites, cumulative N₂O emissions from N fertilizer decreased in the order of urea > AS > CAN, with emissions ranging from 522–617 g N ha^?1 (0.24–0.28% of applied fertilizer) for urea, 368–554 g N ha^?1 (0.17–0.25%) for AS, and 242–264 g N ha^?1 (0.11–0.12%) for CAN during March to August. These results suggest that mineral nitrogen forms can differ in N₂O emissions during the growth period of winter wheat. Strong variations in the seasonal dynamics of N₂O emissions between sites were observed which could partly be related to weather events (e.g., precipitation). Between harvest and the following spring (post‐harvest period) no significant differences in N₂O emissions between fertilized and non‐fertilized treatments were detected on two of three fields. Only on one site post‐harvest emissions from the AS treatment were significantly higher than all other fertilizer forms as well as compared to the control treatment. The cumulative one‐year emissions varied depending on fertilizer form across the three field sites from 0.05% to 0.51% with one exception at one field site (AS: 0.94%). The calculated overall fertilizer induced emission averaged for the three fields was 0.38% which was only about 1/3 of the IPCC default value of 1.0%.     

Influence of phosphorus or phosphorus-potassium fertilization on biomass and dinitrogen fixation of the stem-nodulating green-manure legume Sesbania rostrata in different marginally productive wetland rice soils

The performance of Sesbania rostrata varies widely from site to site. This makes it difficult to predict the N yield and biomass of this plant in marginally productive soils, and to arouse the interest of farmers in green manure technology. Three consecutive pot experiments were conducted in a greenhouse at the International Rice Research Institute (IRRI) to evaluate growth, nodulation, N₂ fixation (C₂H₂ reduction assay and ¹⁵N dilution method), and N yield of 6-week-old S. rostrata on 13 physicochemically different wetland rice soils of the Philippines and on three artificial substrates. The performance of S. rostrata on the unfertilized controls was compared with two fertilizer treatments containing either P (100 mg P kg^-1 dry soil) or P+K (100 mg P kg^-1 and 200 mg K kg^-1 dry soil). In the control soils and substrates, the N yield of S. rostrata varied between 20 and 470 mg N per pot, with the N rate from N₂ fixation ranging between 0 and 95%. In three of the nutritionally poor soils even Mn toxicity symptoms apparently occurred with S. rostrata. P application alleviated these symptoms and increased the overall N yield considerably, mainly through increased biological N₂ fixation. An additional increase in N yield was obtained by the PK treatment. Multiple regression analysis between soil characteristics and the N yield of S. rostrata showed that the original level of P (Olsen-extracted) and Mn in the soil accounted for 73% of the variance in biomass production by S. rostrata among the unfertilized soils and substrates.     

Effects of co-inoculation of Bradyrhizobium elkanii BLY3-8 and Streptomyces griseoflavus P4 on Rj4 soybean varieties

Co-inoculation of selected nitrogen-fixing bacteria with plant growth-promoting bacteria is the promising way for the improvement of soybean production through enhancing plant growth, nodulation, and N₂ fixation. Therefore, this experiment was conducted to study the effects of co-inoculation of Bradyrhizobium elkanii BLY3-8 with Streptomyces griseoflavus P4 on plant growth, nodulation, N₂ fixation, N uptake, and seed yield of Rj₄ soybean varieties. Two experiments with completely randomized design and three replicates were done in this study. N₂-fixation ability of soybean was evaluated by acetylene reduction activity (ARA) and relative ureide method. In the first experiment, synergetic effect in N₂ fixation and nodulation was occurred in co-inoculation treatment (BLY3-8 + P4) in Yezin-3 and Fukuyutaka. Based on these results, co-inoculation effect of BLY3-8 and P4 was assessed on Yezin-3 and Fukuyutaka varieties at three different growth stages, using Futsukaichi soil under natural environmental conditions. This study shows that co-inoculation of BLY3-8 and P4 significantly increased N₂ fixation at V6 stage; plant growth, nodulation, N₂ fixation, and N uptake at R3.5 stage; and shoot growth, N uptake, and seed yield at R8 stage, in Rj₄ soybean varieties compared with the control. Significant difference in plant growth, nodulation, N₂ fixation, N uptake, and yield between co-inoculation and control, not between single inoculation and control, suggests that there is a synergetic effect due to co-inoculation of BLY3-8 and P4.     

Inhibitory effects of acidic minesoil on the sericea lespedeza/Bradyrhizobium symbiotic relationship 1

Effects of acidic minesoil on sericea lespedeza [Lespedeza juncea (L.F.) var. sericea (Mig.)] and its nitrogen (N₂)‐fixing symbiotic relationship with Bradyrhizobium spp. were examined. Sericea lespedeza was grown in pots with N fertilization, without N fertilization, or with commercial Bradyrhizobium as a seed inoculant. Minesoil (pH 5.2) was fertilized with calcium (Ca), phosphorus (P), molybdenum (Mo), and potassium (K), and the pH level was adjusted to 4.8 or 4.5 with aluminum or iron sulfate [Al₂(SO₄)₃; Fe₂(SO₄)₃]. Minesoil was also limed to pH 6.1. Shoot dry weights, shoot N concentrations, nodule dry weights, and nodule numbers were significantly lower (P < 0.05) when inoculated plants were grown in soil at pH 4.5 and 4.8 compared to limed soil. Thus, the N₂ fixation process was adversely affected below pH 5.0. Nitrogen‐fertilized plants grew well in acidified soil, and there were no significant differences in shoot dry weights of such plants among the soil acidification treatments including limed soil. Thus, the N₂‐fixing symbiosis appeared to be more sensitive to acidified soil than the plant host. The effects of Al toxicity versus other factors could not be determined because Al₂(SO₄)₃‐ and Fe₂(SO₄)₃‐amended soils contained similar levels of toxic Al at the highest pH (4.8) that prevented N₂ fixation.

Time periods required for cells of Bradyrhizobium strains to multiply by a factor of 10⁴ were significantly longer (P ≤ 0.05) in extracts of Al₂(SO₄)₃‐amended soil (pH 4.8 and 4.5) than in extracts of calcium carbonate [CaCO₃]‐amended soil (pH 6.1). These increases suggested that reduced multiplication of Bradyrhizobium in acidified minesoils may have been at least partially responsible for the large decreases in nodulation and N₂ fixation observed in these soils. It was also reasoned that the inability of existing bacteria to infect and nodulate plant roots may also have been a factor, based on the high inoculation rates used and the abilities of Bradyrhizobium cells to survive and multiply (albeit at a reduced rate) in extracts of acidified soil. Sericea lespedeza is known to tolerate soils of pH 4.5. However, results of this study suggested sericea lespedeza may not fix appreciable N₂ in acidic soil below pH 5 when inoculated with commercial Bradyrhizobium, even after the establishment of lespedeza plants tolerant of such conditions.     

Growth and Nitrogen Fixation in Soybean as Affected by Phosphorus Fertilizer and Sheep Manure using 15N Isotopic Dilution

A field experiment was conducted to study the effect of adding different phosphorus (P) fertilizer levels [0, 40, and 80 kg phosphorus pentoxide (P₂O₅) ha^?1 (abbreviated as P0, P1, and P2, respectively)] and rates of sheep manure (M) [0, 20, and 40 ton ha^?1 (abbreviated as M0, M1, and M2, respectively)] on growth and nitrogen (N₂) fixation of soybean (Glycine max L.). Sorghum bicolor L. was employed as a reference crop to evaluate N₂ fixation using the ¹⁵N-isotpic dilution technique. Results showed that addition of P fertilizer or sheep manure had positive effects on dry-matter production, N accumulation, and seed yield. Such effects were more pronounced when adding sheep manure and P together than adding separately. Solely P fertilizer had a small impact on N₂ fixation. A tangible increase in the amounts of N₂ fixed due to manure addition occurred. The efficient use of N fertilizer (%NUE) increased significantly as the result of adding a high level of P fertilizer. However, a drastic decrease in %NUE was observed when sheep manure was added solely or in combination with P fertilizer. From productivity and ecological standpoints, P2M1 and P2M2 surpassed the other treatments in showing greater grain yield and greater N₂ fixation. However, considering the high cost of sheep manure, P2M1 was the optimal treatment for improving growth and N₂ fixation in soybean plants with minimal manure consumption. In conclusion, the integrated use of manure and P fertilizer could be considered a useful agricultural practice for improving the performance of soybean plants grown in an Aridisol. Their beneficial effects were mainly attributed to the enhancement of N₂ fixation through root growth and soil property improvements besides being a source of P and other nutrients that are essential for N₂-fixation process.