Department of Biochemistry and Interdisciplinary Plant Group, University of Missouri-Columbia, Columbia, Missouri 65211.
Plant Physiol. 1990 Oct;94(2):681-9. doi: 10.1104/pp.94.2.681.
Roots of young soybean (Glycine max [L.] Merr.) plants (up to 25 days old) contain two distinct urease isozymes, which are separable by hydroxyapatite chromatography. These two urease species (URE1 and URE2) differ in: (a) electrophoretic mobility in native gels, (b) pH dependence, and (c) recognition by a monoclonal antibody specific for the seed ("embryo-specific") urease. By these parameters root URE1 urease is similar to the abundant embryo-specific urease isozyme, while root URE2 resembles the "ubiquitous" urease which has previously been found in all soybean tissues examined (leaf, embryo, seed coat, and cultured cells). The embryo-specific and ubiquitous urease isozymes are products of the Eu1 and Eu4 structural genes, respectively. Roots of the eu1-sun/eu1-sun genotype, which lacks the embryo-specific urease (i.e. ;seed urease-null'), contain no URE1 urease activity. Roots of eu4/eu4, which lacks ubiquitous urease, lack the URE2 (leaflike) urease activity. From these genetic and biochemical criteria, then, we conclude that URE1 and URE2 are the embryo-specific and ubiquitous ureases, respectively. Adventitious roots generated from cuttings of any urease genotype lack URE1 activity. In seedling roots the seedlike (URE1) activity declines during development. Roots of 3-week-old plants contain 5% of the total URE1 activity of the radicle of 4-day-old seedlings, which, in turn, has approximately the same urease activity level as the dormant embryonic axis. The embryo-specific urease incorporates label from [(35)S]methionine during embryo development but not during germination, indicating that there is no de novo synthesis of the embryo-specific (URE1) urease in the germinating root. We conclude that the seedlike urease (URE1) found in roots of young soybean plants is a remnant of the Eu1-encoded, abundant, embryo-specific urease which accumulates in the embryonic root axis during seed development.
大豆幼根(25 天龄以下)中含有两种不同的脲酶同工酶,可通过羟磷灰石层析分离。这两种脲酶(URE1 和 URE2)在以下几个方面有所不同:(a)天然凝胶中的电泳迁移率,(b)pH 依赖性,以及(c)对一种针对种子(“胚性”)脲酶的单克隆抗体的识别。通过这些参数,根 URE1 脲酶与丰富的胚性脲酶同工酶相似,而根 URE2 类似于先前在所有检查过的大豆组织(叶、胚、种皮和培养细胞)中发现的“普遍”脲酶。胚性和普遍存在的脲酶同工酶分别是 Eu1 和 Eu4 结构基因的产物。eu1-sun/eu1-sun 基因型的根,缺乏胚性脲酶(即种子脲酶缺失),不含有 URE1 脲酶活性。eu4/eu4 基因型的根,缺乏普遍存在的脲酶,缺乏 URE2(叶状)脲酶活性。因此,根据这些遗传和生化标准,我们得出结论,URE1 和 URE2 分别是胚性和普遍存在的脲酶。任何脲酶基因型的不定根都缺乏 URE1 活性。在幼苗根中,种性(URE1)活性在发育过程中下降。3 周龄植物的根含有 4 天大幼苗胚根总 URE1 活性的 5%,而后者的脲酶活性水平与休眠胚轴大致相同。胚性脲酶在胚胎发育过程中会从 [(35)S]蛋氨酸中掺入标记物,但在萌发过程中不会,这表明在萌发的根中没有新合成胚性(URE1)脲酶。我们得出的结论是,在大豆幼根中发现的种性脲酶(URE1)是 Eu1 编码的、丰富的胚性脲酶的残留物,该脲酶在种子发育过程中在胚胎根轴中积累。