杂交竹梢枯病菌毒素蛋白的质膜结合位点  

Binding site of toxic protein from Arthrinium phaeospermum on plasmalemma of hybrid bamboo

作　　者：李姝江[1] 朱天辉[1,2] 

机构地区：[1]四川农业大学林学院,四川雅安625014 [2]四川农业大学 长江上游林业生态工程省级重点实验室,四川雅安625014

出　　处：《浙江大学学报（农业与生命科学版）》2012年第4期355-361,共7页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家自然科学基金资助项目(31070578)

摘　　要：为探明杂交竹梢枯病菌毒素蛋白的质膜结合位点及在不同品种的结合活性,在低压层析分离纯化毒素蛋白的基础上,以该蛋白免疫新西兰大白兔制备毒素特异性抗血清,并将该蛋白用不同浓度的抗体吸附后处理杂交竹幼嫩枝条。结果表明:毒素所引起的症状有不同程度的减轻,说明所制备的抗体在与毒素发生特异性免疫学反应的同时,可部分封闭毒素分子上与毒素受体结合的位点;利用竞争酶联免疫吸附试验(ELISA)测定杂交竹嫩枝的质膜制剂与毒素蛋白的结合活性显示,质膜制剂与毒素结合后能部分阻断毒素与其抗体的免疫学反应,即质膜制剂中含有毒素的结合位点,且不同品种的结合活性有差异;用胰蛋白酶或加热处理质膜制剂后,质膜制剂对毒素与其抗体反应的抑制作用消失,证实质膜制剂中与毒素结合的是蛋白类物质。To explore the binding site of toxic protein from Arthriniurn phaeospermum on plasmalemma of hybrid bamboo and the binding activities of different hybrid bamboo varieties, based on the separation and purification of toxic protein by a low-pressure chromatography, the specific antiserum against the toxin was prepared by injecting the New Zealand white rabbit with the purified toxin. The results showed that the symptom was alleviated to different degree when the toxin absorbed by different concentration antibodies was inoculated the hybrid bamboo, illuminating that the prepared antibody not only immunologically reacted with the toxin, but also partly blocked some sites of toxin molecular which could recognize the binding sites on the hybrid bamboo cells. The binding activity of plasmalemma preparation of tender branches with toxic protein was determined by a competitive enzyme-linked immunosorbent assay （ELISA）. The results revealed that the immunological reaction of the toxin and the antibody could be inhibited by the plasmalemma preparation, which hinted that the plasmalemma preparation contained the binding sites of the toxin. But the binding activities of different varieties had different. After treated by trypsin or heating, the plasmalemma preparation could not inhibit the immunological reaction of the toxin and the antibody, verifying that the protein in the plasmalemma preparation was responsible for binding with the toxin.

关 键 词：杂交竹 暗孢节菱孢菌 毒素 质膜 竞争酶联免疫吸附试验 

分 类 号：Q936[生物学—微生物学] S462.1[农业科学—植物保护]