Silvente Sonia, Camas Alberto, Lara Miguel
Centro de Investigación sobre Fijación de Nitrógeno, Universidad Nacional Autónoma de México, Apartado Postal 565-A, Cuernavaca, Morelos, México.
J Exp Bot. 2003 Feb;54(383):749-55. doi: 10.1093/jxb/erg086.
Sucrose synthase (SS), the key sucrose hydrolytic enzyme (EC 2.4.1.13), plays an important role in N(2)-fixing nodule metabolism. It has also been proposed that N(2) fixation in soybean nodules could be mediated by the potential to metabolize sucrose. The isolation and characterization of a nodule-enhanced SS full-length cDNA clone from the bean Phaseolus vulgaris is reported here. Southern blot analysis indicated that there are at least two SS genes in beans. Using a 3' specific probe from this SS cDNA clone, it was possible to identify a nodule-enhanced SS gene (PvSSn), which is expressed almost exclusively in nodules. A second gene (PvSS), which is expressed in all tissues tested, was detected using a coding region probe. Nodule-enhanced PvSSn transcript levels, but not the enzyme activity or protein amount, is reduced during nodule development. These data indicated that this reduction could be due to a limitation on the carbon availability in the nodule. PvSSn expression is reduced in the asparagine-treated nodules. By contrast, PvSSn transcript levels in nodules increased in the presence of glutamine, allantoin and allopurinol. This result suggests a relationship between ureide transport and SS regulation and could help in understanding why the ureide transport mechanism is activated during nitrogen fixation in bean.
蔗糖合酶(SS)是关键的蔗糖水解酶(EC 2.4.1.13),在固氮根瘤代谢中起重要作用。也有人提出,大豆根瘤中的固氮作用可能由蔗糖代谢潜力介导。本文报道了从菜豆中分离和鉴定一个根瘤增强型SS全长cDNA克隆。Southern杂交分析表明,菜豆中至少有两个SS基因。使用该SS cDNA克隆的3'特异性探针,能够鉴定出一个根瘤增强型SS基因(PvSSn),它几乎只在根瘤中表达。使用编码区探针检测到另一个在所有测试组织中都表达的基因(PvSS)。在根瘤发育过程中,根瘤增强型PvSSn转录水平降低,但酶活性和蛋白量未降低。这些数据表明,这种降低可能是由于根瘤中碳供应的限制。在天冬酰胺处理的根瘤中,PvSSn表达降低。相比之下,在谷氨酰胺、尿囊素和别嘌呤醇存在的情况下,根瘤中PvSSn转录水平升高。这一结果表明了酰脲转运与SS调节之间的关系,有助于理解为什么在菜豆固氮过程中酰脲转运机制会被激活。