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JXB Advance Access originally published online on March 12, 2004
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Journal of Experimental Botany, Vol. 55, No. 398, pp. 867-877, April 1, 2004
© 2004 Oxford University Press

Cell and Molecular Biology, Biochemistry and Molecular Physiology

Soybean cultivars ‘Williams 82’ and ‘Maple Arrow’ produce both urea and ammonia during ureide degradation

Received 12 November 2003; Accepted 20 January 2004

Christopher D. Todd and Joe C. Polacco*

Department of Biochemistry and Interdisciplinary Plant Group, 117 Schweitzer Hall, University ofMissouri–Columbia, Columbia, MO 65211, USA

* To whom correspondence should be addressed. Fax: +1 573 882 5635. E-mail: polaccoj{at}missouri.edu

The ability of two soybean (Glycine max L. [Merrill]) cultivars, ‘Williams 82’ and ‘Maple Arrow’, which were reported to use different ureide degradation pathways, to degrade the ureides allantoin and allantoate was investigated. Protein fractions and total leaf homogenates from the fourth trifoliate leaves of both cultivars were examined for the ability to evolve either 14CO2 or [14C]urea from 14C-labelled ureides in the presence of various inhibitors. 14CO2 evolution from [2,7-14C]allantoate was catalysed by 25–50% saturated ammonium sulphate fractions of both cultivars. This activity was inhibited by acetohydroxamate (AHA), which has been used to inhibit plant ureases, but not by phenylphosphorodiamidate (PPD), a more specific urease inhibitor. Thus, in both cultivars, allantoate may be metabolized by allantoate amidohydrolase. This activity was sensitive to EDTA, consistent with previous reports demonstrating that allantoate amidohydrolase requires manganese for full activity. Total leaf homogenates of both cultivars evolved both 14CO2 and [14C]urea from [2,7-14C] (ureido carbon labelled) allantoin, not previously reported in either ‘Williams 82’ or in ‘Maple Arrow’. In situ leaf degradation of 14C-labelled allantoin confirmed that both urea and CO2/NH3 are direct products of ureide degradation. Growth of plants in the presence of PPD under fixing and non-fixing conditions caused urea accumulation in both cultivars, but did not have a significant impact on total seed nitrogen. Urea levels were higher in N-fixing plants of both cultivars. Contrary to previous reports, no significant biochemical difference was found in the ability of these two cultivars to degrade ureides under the conditions used.

Key words: Allantoate, allantoin, amidinohydrolase, amidohydrolase, nitrogen metabolism, N-fixation, urea, urease, ureide.


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