The application of combined nitrogen, especially nitrate, to soybean plants is known to strongly inhibit nodule formation, growth and nitrogen fixation. In the present study, we measured the effects of supplying 5 mM nitrate on the growth of nodules, primary root, and lateral roots under light at 28 °C or dark at 18 °C conditions. Photographs of the nodulated roots were periodically taken by a digital camera at 1-h intervals, and the size of the nodules was measured with newly developed computer software. Nodule growth was depressed approximately 7 h after the addition of nitrate under light conditions. The nodule growth rate under dark conditions was almost half that under light conditions, and nodule growth was further suppressed by the addition of 5 mM nitrate. Similar results were observed for the extending growth rate of the primary root as those for nodule growth supplied with 5 mM nitrate under light/dark conditions. In contrast, the growth of lateral roots was promoted by the addition of 5 mM nitrate. The 2D-PAGE profiles of nodule protein showed similar patterns between the 0 and 5 mM nitrate treatments, which suggested that metabolic integrity may be maintained with the 5 mM nitrate treatment. Further studies are required to confirm whether light or temperature condition may give the primary effect on the growth of nodules and roots.

Concern that depletion of fertilizer feedstocks, which are a finite mineral resource, threatens agricultural sustainability has driven the exploration of sustainable methods of soil fertilization. Given that microalgae, which are unicellular photosynthetic organisms, can take up nutrients efficiently from water systems, their application in a biological wastewater purification system followed by the use of their biomass as a fertilizer alternative has attracted attention. Such applications of microalgae would contribute to the accelerated recycling of nutrients from wastewater to farmland. Many previous reports have provided information on the physiological characteristics of microalgae that support their utility. In this review, we focus on recent achievements of studies on microalgal physiology and relevant applications and outline the prospects for the contribution of microalgae to the establishment of sustainable agricultural practices.

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 N2 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.

Non‐nodulated soybean (Glycine max (L.) Merr.) plants were cultivated hydroponically under N‐sufficient (5 mM NaNO₃) or N‐deficient (0.5 mM NaNO₃) conditions. ¹³N‐ or ¹⁵N‐ labelled nitrate was fed to the cut end of the stems, and the accumulation of nitrate‐derived N in the pods, nodes and stems was compared. Real‐time images of ¹³N distribution in stems, petioles and pods were obtained using a Positron Emitting Tracer Imaging System for a period of 40 min. The results indicated that the radioactivity in the pods of N‐deficient plants was about 10 times higher than that of N‐sufficient plants, although radioactivity in the stems and nodes of N‐deficient versus N‐sufficient plants was not different. A similar result was obtained by supplying ¹⁵ to cut soybean shoots for 1 h. The fact that the N translocation into the pods from fed to the stem base was much faster in N‐deficient plants may be due to the strong sink activity of the pods in N‐deficient plants. Alternatively, the redistribution of N from the leaves to the pods via the phloem may be accelerated in N‐deficient plants. The temporal accumulation of ¹³ in nodes was suggested in both N‐sufficient and N‐deficient plants. In one ¹³ pulse‐chase experiment, radioactivity in the stem declined rapidly after transferring the shoot from the ¹³ solution to non‐labelled NO₃; in contrast, the radioactivity in the node declined minimally during the same time period.

Leguminous plants form root nodules with rhizobia that fix atmospheric dinitrogen (N₂) for the nitrogen (N) nutrient. Combined nitrogen sources, particular nitrate, severely repress nodule growth and nitrogen fixation activity in soybeans (Glycine max [L.] Merr.). A microarray-based transcriptome analysis and the metabolome analysis were carried out for the roots and nodules of hydroponically grown soybean plants treated with 5 mM of nitrate for 24 h and compared with control without nitrate. Gene expression ratios of nitrate vs. the control were highly enhanced for those probesets related to nitrate transport and assimilation and carbon metabolism in the roots, but much less so in the nodules, except for the nitrate transport and asparagine synthetase. From the metabolome analysis, the concentration ratios of metabolites for the nitrate treatment vs. the control indicated that most of the amino acids, phosphorous-compounds and organic acids in roots were increased about twofold in the roots, whereas in the nodules most of the concentrations of the amino acids, P-compounds and organic acids were decreased while asparagine increased exceptionally. These results may support the hypothesis that nitrate primarily promotes nitrogen and carbon metabolism in the roots, but mainly represses this metabolism in the nodules.