Todd Christopher D, Polacco Joe C
Department of Biochemistry and Interdisciplinary Plant Group, 117 Schweitzer Hall, University ofMissouri-Columbia, Columbia, MO 65211, USA.
J Exp Bot. 2004 Apr;55(398):867-77. doi: 10.1093/jxb/erh100. Epub 2004 Mar 12.
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 (14)CO(2) or [(14)C]urea from (14)C-labelled ureides in the presence of various inhibitors. (14)CO(2) evolution from [2,7-(14)C]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 (14)CO(2) and [(14)C]urea from [2,7-(14)C] (ureido carbon labelled) allantoin, not previously reported in either 'Williams 82' or in 'Maple Arrow'. In situ leaf degradation of (14)C-labelled allantoin confirmed that both urea and CO(2)/NH(3) 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.
研究了两个大豆(Glycine max L. [Merrill])品种‘Williams 82’和‘Maple Arrow’降解尿囊素和尿囊酸这两种脲类化合物的能力,据报道这两个品种采用不同的脲类降解途径。检测了两个品种第四片三出复叶的蛋白质组分和总叶匀浆在各种抑制剂存在下从14C标记的脲类化合物中释放14CO2或[14C]尿素的能力。两个品种25 - 50%饱和度硫酸铵组分催化了[2,7-(14)C]尿囊酸释放14CO2。该活性受到用于抑制植物脲酶的乙酰氧肟酸(AHA)的抑制,但不受更特异的脲酶抑制剂苯基磷二酰胺(PPD)的抑制。因此,在两个品种中,尿囊酸可能由尿囊酸酰胺水解酶代谢。该活性对EDTA敏感,这与先前报道一致,即尿囊酸酰胺水解酶需要锰才能完全发挥活性。两个品种的总叶匀浆都能从[2,7-(14)C](脲基碳标记)尿囊素中释放14CO2和[14C]尿素,这在‘Williams 82’或‘Maple Arrow’中均未见先前报道。14C标记尿囊素的原位叶片降解证实尿素和CO2/NH3都是脲类降解的直接产物。在固定和非固定条件下,PPD存在时植物的生长导致两个品种都积累尿素,但对种子总氮没有显著影响。两个品种的固氮植物中尿素水平更高。与先前报道相反,在所用条件下,这两个品种降解脲类化合物的能力未发现显著的生化差异。