稻瘟菌侵染诱导的水稻过氧羟基脂肪酸羟化环氧化酶的抗血清制备

 通过选择性抽提、盐析、阴阳离子交换柱层析等步骤从非亲和性稻瘟菌侵染的120g稻叶中获得了诱导性过氧羟基脂肪酸羟化环氧化酶(POG)的电泳纯蛋白250μg。利用这些纯化蛋白,制备了POG的抗血清。点免疫杂交结果表明,该抗血清效价大于1:5000,其抗原检测灵敏度可达0.05ng,并具有较好的特异性。蛋白印迹结果显示,稻瘟菌侵染可诱导POG在水稻细胞膜上的累积。

PREPARATION OF ANTISERUM AGAINST PEROXYGENASE FROM RICE LEAVES INFECTED BY BLAST FUNGUS Magnaporthe grisea

A novel rice peroxygenase induced by blast fungus was purified through selective extraction, salting out, anion and cation chromatographies. From 120 grams of the infected rice leaves, about 250 μg of peroxygenase was obtained with electrophoretic homogeneity. By immunizing rabbit with the purified protein, antiserum was raised. The antiserum had a titre over 1∶5000 and its antigen sensitivity reached up to 0.05 ng in an immunological dot hybridization. Western blotting showed that infection of blast fungus can induce accumulation of pero xygenase protein in rice cell membrane.