Gijzen M., Van Huystee R., Buzzell R. I.
Agriculture Canada, London Research Centre, 1391 Sandford Street, London, Ontario, Canada N5V 4T3 (M.G.).
Plant Physiol. 1993 Dec;103(4):1061-1066. doi: 10.1104/pp.103.4.1061.
Peroxidase activity in the seed coats of soybean (Glycine max [L.] Merr.) is controlled by the Ep locus. We compared peroxidase activity in cell-free extracts from seed coat, root, and leaf tissues of three EpEp cultivars (Harosoy 63, Harovinton, and Coles) to three epep cultivars (Steele, Marathon, and Raiden). Extracts from the seed coats of EpEp cultivars were 100-fold higher in specific activity than those from epep cultivars, but there was no difference in specific activity in crude root or leaf extracts. Isoelectric focusing of root tissue extracts and staining for peroxidase activity showed that EpEp cultivars had a root peroxidase of identical isoelectric point to the seed coat peroxidase, whereas roots of the epep types were lacking that peroxidase, indicating that the Ep locus may also affect expression in the root. In seed coat extracts, peroxidase was the most abundant soluble protein in EpEp cultivars, whereas this enzyme was present only in trace amounts in epep genotypes, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Histochemical localization of peroxidase activity in seed coats of EpEp cultivars shows that the enzyme occurs predominately in the cytoplasm of hourglass cells of the subepidermis. No obvious difference in the gross or microscopic structure of the seed coat was observed to be associated with the Ep locus. These results suggest that soybean seed coat peroxidase may be involved in processes other than seed coat biosynthesis.
大豆(Glycine max [L.] Merr.)种皮中的过氧化物酶活性受Ep位点控制。我们比较了三个EpEp品种(哈罗索伊63、哈罗温顿、科尔斯)和三个epep品种(斯蒂尔、马拉松、雷登)种皮、根和叶组织的无细胞提取物中的过氧化物酶活性。EpEp品种种皮提取物的比活性比epep品种高100倍,但粗根或叶提取物的比活性没有差异。对根组织提取物进行等电聚焦并对过氧化物酶活性进行染色,结果表明EpEp品种的根过氧化物酶与种皮过氧化物酶具有相同的等电点,而epep类型的根缺乏该过氧化物酶,这表明Ep位点可能也影响根中的表达。在种皮提取物中,过氧化物酶是EpEp品种中最丰富的可溶性蛋白质,而通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定,该酶在epep基因型中仅以痕量存在。EpEp品种种皮中过氧化物酶活性的组织化学定位表明,该酶主要存在于亚表皮沙漏细胞的细胞质中。未观察到种皮的大体或微观结构与Ep位点有明显差异。这些结果表明,大豆种皮过氧化物酶可能参与种皮生物合成以外的过程。