Inouhe M, Nevins D J
Department of Vegetable Crops, University of California, Davis, California 95616.
Plant Physiol. 1991 Jun;96(2):426-31. doi: 10.1104/pp.96.2.426.
Polyclonal antibodies were raised in rabbits in response to the administration of purified exo- and endoglucanases extracted from cell walls of maize (Zea mays L. B37 x Mo17) coleoptiles. Since the antibodies formed specific conjugates when challenged with the glucanase antigens in immunoblot assays they were employed to evaluate the participation of glucanases in tissue growth. Indole-3-acetic acid induced cell elongation of abraded coleoptile segments was inhibited when the antibodies were supplied as a short term pretreatment (25-200 microgram/milliliter of serum protein). The extent of inhibition of IAA induced cell elongation was additive when endo- and exoglucanase antibodies were applied together. The results suggest that both enzymes have a role in mediating IAA-induced cell elongation. Pretreatment with exo- and endoglucanases antibodies also inhibited IAA induced degradation of noncellulosic beta-d-glucans and the increased level of cellulosic polymers in maize coleoptiles. Antibodies also inhibited the expression of the autohydrolytic degradation of glucans in isolated cell walls. The extent of inhibition was dependent on the antibody concentration applied. The results support the contention that enzymatic processes mediated by exo- and endoglucanases are responsible for cell wall autolytic reactions and that these reactions are linked to the mechanism for expressing auxin induced cell elongation in maize coleoptiles.
通过给兔子注射从玉米(Zea mays L. B37 x Mo17)胚芽鞘细胞壁中提取的纯化外切葡聚糖酶和内切葡聚糖酶,产生了多克隆抗体。由于在免疫印迹分析中,这些抗体与葡聚糖酶抗原反应时形成了特异性结合物,因此被用于评估葡聚糖酶在组织生长中的作用。当作为短期预处理(血清蛋白浓度为25 - 200微克/毫升)提供抗体时,吲哚 - 3 - 乙酸诱导的磨损胚芽鞘切段细胞伸长受到抑制。当同时应用内切葡聚糖酶抗体和外切葡聚糖酶抗体时,吲哚乙酸诱导的细胞伸长抑制程度具有加和性。结果表明这两种酶在介导吲哚乙酸诱导的细胞伸长中都起作用。用外切葡聚糖酶抗体和内切葡聚糖酶抗体进行预处理也抑制了吲哚乙酸诱导的玉米胚芽鞘中非纤维素β - d - 葡聚糖的降解以及纤维素聚合物水平的升高。抗体还抑制了分离细胞壁中葡聚糖自水解降解的表达。抑制程度取决于所用抗体的浓度。这些结果支持了这样的观点,即由外切葡聚糖酶和内切葡聚糖酶介导的酶促过程负责细胞壁的自溶反应,并且这些反应与玉米胚芽鞘中生长素诱导细胞伸长的表达机制相关。