Yalpani N, Balke N E, Schulz M
Department of Agronomy, University of Wisconsin, Madison, Wisconsin 53706.
Plant Physiol. 1992 Nov;100(3):1114-9. doi: 10.1104/pp.100.3.1114.
A UDP-glucose:salicylic acid 3-O-glucosyltransferase (EC 2.4.1.35) (GTase) from oat (Avena sativa L. cv Dal) root extracts was assayed in vitro using [(14)C]salicylic acid (SA) and an ion exchange column to separate SA from beta-glucosylsalicylic acid. The GTase, present at a very low constitutive level, was inducible to 23 times the constitutive level. When excised roots were exposed to SA at pH 6.5, the specific activity of the enzyme increased within 1.5 h, peaked after 8 to 10 h, and then declined. The increase in specific activity depended on the concentration of SA in the induction medium. Among 16 phenolics and phenolic derivatives tested, GTase induction showed high specificity toward SA and acetylsalicylic acid. Specific activity of the enzyme was induced to higher levels in roots from 7-d-old seedlings than roots from younger plants. GTase activity was less inducible in basal compared with median or apical root sections. Induction of GTase activity was a result of de novo RNA and protein synthesis. Candidate peptides for the GTase were identified by comparison of two-dimensional electrophoresis gels of proteins labeled with [(35)S]methionine during incubation of roots in the presence or the absence of SA and a gel of a partially purified GTase preparation.
利用[¹⁴C]水杨酸(SA)和离子交换柱从β-葡萄糖基水杨酸中分离出SA,对燕麦(Avena sativa L. cv Dal)根提取物中的UDP-葡萄糖:水杨酸3-O-葡萄糖基转移酶(EC 2.4.1.35)(GTase)进行了体外测定。该GTase以非常低的组成水平存在,可被诱导至组成水平的23倍。当将离体根置于pH 6.5的SA中时,该酶的比活性在1.5小时内增加,在8至10小时后达到峰值,然后下降。比活性的增加取决于诱导培养基中SA的浓度。在所测试的16种酚类和酚类衍生物中,GTase诱导对SA和乙酰水杨酸表现出高度特异性。与较幼龄植株的根相比,7日龄幼苗根中的该酶比活性被诱导到更高水平。与根中段或根尖相比,根基部的GTase活性诱导性较低。GTase活性的诱导是从头RNA和蛋白质合成的结果。通过比较在有或没有SA存在的情况下根培养期间用[³⁵S]甲硫氨酸标记的蛋白质的二维电泳凝胶和部分纯化的GTase制剂的凝胶,鉴定了GTase的候选肽段。