The effects of deep placement (supplied at 20 cm depth from soil surface below plants) of 100 kg N ha−1 of N fertilizers, urea, coated urea or calcium cyanamide (lime nitrogen) on the growth, nitrogen fixation activity, nitrogen absorption rate and seed yield of soybean (Glycine max L. Merr.) plants were examined by comparing them with control plots without deep placement of N fertilizer in sandy dune field. In addition, three different inoculation methods of bradyrhizobia were used for each N treatment: (1) transplantation of 10‐day‐old seedling in a paper pot with vermiculite inoculated with Bradyrhizobium japonicum USDA110, (2) direct transplantation of inoculated 10‐day‐old seedlings, and (3) transplantation of 10‐day‐old seedlings in a non‐inoculated paper pot. The deep placement of N fertilizers, especially calcium cyanamide and coated urea, markedly increased the growth and total N accumulation in shoot, roots and nodules, which resulted in an increase in seed yield. Daily N2 fixation activity and N absorption rate were estimated by relative abundance of ureide‐N analysed from the concentration of N constituents (ureide‐N, amide‐N and nitrate‐N) in root bleeding xylem sap and increase in total N accumulation in whole plants at R1, R3, R5 and R7 stages. The total amount of N2 fixation was about 50 % higher in the plants with calcium cyanamide and coated urea deep placements compared with control plants. Deep placement of slow release fertilizers kept nodule dry weight higher in the maturing stage of seed, possibly through abundant supply of photoassimilate to the nodules by supporting leaf area and activity until late reproductive stages. The results indicate that deep placement of calcium cyanamide or coated urea enhances N2 fixation activity, which ultimately increases the seed yield. The promotive effect was observed with the seedlings transplanted in paper pot with inoculum of bradyrhizobia within any treatments, although nodulation by indigenous rhizobia was observed in the plants transplanted with non‐inoculated paper pot.

Soybean is an important crop for human food and feed for livestock. World soybean production is increasing especially in North and South America. Soybean seeds contain a high percentage of protein about 35-40%, and they require a large amount of nitrogen compared with other crops. Soybean plants make root nodules with rhizobia, and rhizobia can fix atmospheric N 2 and give the fixed N to the host soybean plants. Also, soybean can absorb nitrogen usually nitrate from soil or fertilizers. The amount of total assimilated nitrogen in shoot is proportional to the soybean seed yield either from nitrogen fixation or from nitrogen absorption, and the nitrogen availability is very important for soybean cultivation. Maintenance of a high and long-term nitrogen fixation activity is very important for a high production of soybean. However, application of chemical nitrogen fertilizers usually depresses nodule formation and nitrogen fixation. Nitrate in direct contact with a nodulated part of roots causes severe inhibition of nodule growth and nitrogen fixation, although a distant part of nodules from nitrate application gives no or little effect. Deep placement of slow-release nitrogen fertilizers, coated urea, or lime nitrogen promoted the growth and seed yield and quality of soybean without depressing nitrogen fixation.

The long-term effect of the concentration and duration of application of nitrate from the lower part of soybean roots on the nodulation and nitrogen fixation in the upper part of roots was investigated using a two-layered pot system separating the upper roots growing in a vermiculite medium and the lower roots growing in a nutrient solution. Continuous absence of nitrate (hereafter referred to as "0-0 treatment"), and continuous 1 m~ (1-1 treatment) and 6 -(6-6 treatment) nitrate treatments were imposed in the lower pot from transplanting to the beginning of the maturity stage. In addition, 6 mM nitrate was supplied partially from the beginning of the pod stage till the beginning of the maturity stage (0-6 treatment) or from transplanting till the beginning of the pod stage (6-0 treatment). The values of the total plant dry weight and seed dry weight were highest in the 6-6 treatment, intermediate in the 1-1, 6-0, 0-6 treatments, and lowest in the 0-0 treatment. The values of the nodule dry weight and nitrogen fixation activity (acetylene reduction activity) were lowest in the 6-6 treatment. The value of the nodule dry weight in the upper roots was highest in the plants subjected to the 1-1 treatment and exceeded that in the 0-0 treatment. Total nitrogen fixation activity of the upper nodules per plant at the beginning of the pod stage was also highest in the 1-1 treatment. These results indicated that longterm supply of a low level of nitrate from the lower roots could promote nodulation and nitrogen fixation in the upper part of roots. Withdrawal of 6 m M nitrate after the beginning of the pod stage (6-0 treatment) markedly enhanced nodule growth and ARA per plant in the upper roots at the beginning of the maturity stage when the values of both parameters decreased in the other treatments. The nitrate concentration in the nodules attached to the upper roots was low, including the 6-6 treatment regardless of the stages of growth. This indicated that the inhibitory effect of 6 mM nitrate or promotive effect of 1 mM nitrate supplied from the lower roots was not directly controlled by nitrate itself, but was mediated by some systemic regulation, possibly by the C or/and N requirement of the whole plant.

In the present study, the effect and utilization of deep placement 15 N-labeled coated urea (CU) and lime nitrogen (LN) were investigated at various growth stages of soybean (Glycine max [L.]). Soybean (cv. Enrei) plants were planted in a rotated paddy field in Niigata, Japan. ) was supplied separately at a depth of 20 cm just below the seeding line. Deep placement of LN and CU significantly enhanced the dry weight and N content of the plants compared with the control treatment (Cont) without deep placement. As a result, the seed yield per plant in CU (67.2 g) and LN (70.6 g) was much higher than the Cont (37.1 g). 15N analysis of plants showed that the pattern of labeled N absorption tended to be lower with LN than CU at the R3 and R5 stages, but the recovery rate at R7 was higher in LN (70%) than CU (61%). Combining the 15 N analysis with a relative ureide analysis, the N derived from nitrogen fixation (Ndfa), from fertilizer (Ndff ) and from soil (Ndfs) were evaluated. At the R7 stage, the amount of Ndfa was higher in CU (32.1 g m -2 ) and LN (31.1 g m -2 ) than in Cont (21.4 g m -2 ). This positive response for N 2 fixation by the deep placement of CU and LN may result from the continuous supply of N from the lower parts of the roots, which promotes shoot growth and extends the photosynthetic activity of the green leaves, resulting in the promotion of nitrogen fixation and seed yield.

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