Allantoin production and its utilization in relation to nodule formation in soybeans

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Abstract
A remarkable accumulation and utilization of allantoin are observed in soybean plant bearing modules (Glycine max variety A62-1). To study ita physiological role, changes in activities and distribution of the enzymes concerning the purine catabolism (i.e., xanthine oxidase, uricase, allantoinase and allantoicase) were measured during development. Uricase activity was found in the radicles in the early stage of seedling development, and no difference of the enzyme activity was detected between the radicles of the nodulating variety (A62-) and non-nodulating variety (A62-2). On maturation the activity disappeared rapidly in radicles and appeared in leaves, pods and nodules. The activity in nodules was much higher than that in the leaves and the pods throughout the culture period. Xanthine oxidase activity was found in the radicles and nodules. Allantoinase activity was present in all regions of both varieties, and the specific activity in the nodules Was 2-10 times that in other regions. Allantoicase activity was detected in the leaves, but not in the nodules. The distribution of these enzymes suggested that allantoin is formed in the nodules and utilized mainly in the leaves after translocation. Changes in the allantoin concentration and in the activities of uricase and allantoicase were studied in non-nodulating soybean (A62-2) grown in a culture solution containing various nitrogenous compounds. The allantoin concentration in the leaves increased as the plants were cultured in the solution containing 100 ppm of nitrate-N, urea-N or allantoin-N, whereas no uricase activity in the root was detected. The specific activity of uricase decreased in the process of plant development in all cultures. Allantoicase activity increased in the roots and leaves when the plants were cultured in this solution containing 100 ppm of urea-N or allantoin-N, indicating that allantoin is utilized easily in the soybean plants. Intracellular localization of uricase in the nodules was studied by the method of linear sucrose density gradient centrifugation. The uricase activity was associated with the bacteroids, density 1.25. The uricase activity peak was coincident with the peak of fonnic dehydrogenase activity (bacteria type), and not with peaks of catalase activity, cytochrome oxidase activity and absorbance at 280 nm. This evidence together with the observation by electron microscopy suggested that at uricase is associated with bacteroids.

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