Goldraij A, Polacco J C
Department of Biochemistry and Interdisciplinary Plant Group, 117 Schweitzer Hall, University of Missouri, Columbia, Missouri 65211, USA.
Plant Physiol. 1999 Jan;119(1):297-304. doi: 10.1104/pp.119.1.297.
Arginase (EC 3.5.3.1) transcript level and activity were measured in soybean (Glycine max L.) embryos from the reserve deposition stage to postgermination. Using a cDNA probe for a small soybean arginase gene family, no transcript was detected in developing embryos. However, arginase transcripts increased sharply on germination, reaching a maximum at 3 to 5 d after germination. There was low but measurable in vitro arginase specific activity in developing embryos (less than 6% of seedling maximum). During germination arginase specific activity increased in parallel with the sharply increasing arginase transcript level. Seedling arginase activity was largely localized in cotyledons. Arginase activity was assayed in vivo by measuring urea accumulation in a urease-deficient mutant. No urea was detected in developing embryos, whereas accumulated urea paralleled arginase specific activity and transcript level in germinating seedlings. As in planta embryos, cultured cotyledons did not accumulate urea when arginine (Arg) was provided with other amino acids in a "mock" seed-coat exudate. Arg as the sole nitrogen source was converted to urea but did not support cotyledon growth. There appeared to be a lack of recruitment of the low-level arginase activity to hydrolyze free Arg in developing embryos, thus avoiding a futile urea cycle.
在大豆(Glycine max L.)胚胎从储备积累阶段到萌发后的过程中,对精氨酸酶(EC 3.5.3.1)的转录水平和活性进行了测定。使用针对一个小的大豆精氨酸酶基因家族的cDNA探针,在发育中的胚胎中未检测到转录本。然而,精氨酸酶转录本在萌发时急剧增加,在萌发后3至5天达到最大值。发育中的胚胎中存在低水平但可测量的体外精氨酸酶比活性(不到幼苗最大值的6%)。在萌发过程中,精氨酸酶比活性与急剧增加的精氨酸酶转录水平平行增加。幼苗精氨酸酶活性主要定位于子叶中。通过测量脲酶缺陷型突变体中尿素的积累来体内测定精氨酸酶活性。在发育中的胚胎中未检测到尿素,而在萌发的幼苗中积累的尿素与精氨酸酶比活性和转录水平平行。与植物胚胎一样,当在“模拟”种皮渗出物中向培养的子叶提供精氨酸(Arg)和其他氨基酸时,子叶不会积累尿素。以Arg作为唯一氮源时会转化为尿素,但不支持子叶生长。在发育中的胚胎中,似乎缺乏对低水平精氨酸酶活性的招募以水解游离Arg,从而避免了无效的尿素循环。