杂交竹梢枯病菌毒素蛋白的质膜结合位点.pdfVIP

浙江大学学报(农业与生命科学版) 38(4)：355～361，2012 Journal of Zhejiang University(Agric． Life Sci． 文章编号 ：1008-9209(2012)04—0355—07 DOI：10．3785／j．issn．1008—9209．2012．04．001 杂交竹梢枯病菌毒素蛋白的质膜结合位点 李妹江 ，朱天辉 (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 625O14) 摘要 为探明杂交竹梢枯病茵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住点，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． 关键 词 杂交竹 ；暗孢 节菱孢 茵 中图分类号 Q 936；S 462．1 毒素；质膜 ；竞争酶联免疫吸附试验 文献标志码 A LI Shu-jiang ，ZHU Tian—hui ’。(1．College of Forestry，Sichuan Agricultural University，Yaan，Sichuan 625014，China；2．Forestry Ecological Engineering in the Upper Reaches of Yangtze River Key Laboratory of Sichuan Province，Sichuan Agricultural University，Yaan，Sichuan 62501 4，Ch ina ) Binding site of toxic protein from Arthrinium phaeospermum on plasmalemma of hybrid bamboo．Journal of Zhejiang University(Agric． Life Sci．)，2012，38(4)：355—361 Abstract To explore the binding site of toxic protein from Arthrinium 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 aleviated to diferent degree when the toxin absorbed by different concentration antibodies was inoculated the hybrid bamboo，iluminating that the prepared antibody not only immunologicaly reacted wi th the toxin，but also partly blocked some sites of toxin molecular which could recognize the binding sites on the hybrid bamboo cels．The binding activity of plasma lemma preparation of tender branches with toxic protein was determined by a competitive enzym