甘薯青枯菌粗毒素的制备及热、紫外线、酶对粗毒素生物活性的影响
Preparation of Crude Toxins of Ralstonia solanacearum in Sweet Potatoes and Influence of Heat、Ultraviolet Ray、Enzyme to the Biological Activity of Crude Toxins
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摘要: 将有致病力的甘薯青枯菌在液体肉汁胨培养基中28~30℃培养48~72h,在无菌条件下经2000r/min离心;取上清液再经过0.45μm的细菌过滤膜过滤,获得甘薯青枯菌粗毒素(代谢产物).粗毒素对热效应较为敏感,在50℃下水浴1h,生物活性不变;70℃下水浴1h,生物活性降低28.7%;100℃下水浴1h,生物活性几乎完全丧失。粗毒素对紫外线不敏感,在25W紫外光下照射30min、60min、120min均不影响生物活性。粗毒素对蛋白酶和溶菌酶敏感,加入木瓜蛋白酶4~16u/ml或加入胰蛋白酶10~40u/ml时生物活性均受破坏,丧失毒力;加入溶菌酶15000~60000u/ml时生物活性降低了91.6%~92.0%,几乎丧失了毒力。Abstract: The strains with pathogenicity of Ralstonia solanacearum isolated from sweet potatoes were grown in PYG medium at 28-30℃ for 48-72 hours After sterile centrifugation (2000 r/min ),the culture fluids were filtered through 0 45 μm Millipore filters The crude toxins (metabolite) were obtained The crude toxins of Ralstonia solanacearum in sweet potatoes is sensitive to thermal treatment and it’s biological activity decreased separately 0%、28 7%、100% at 50℃、 70℃ and 100℃ waterbath for one hour The crude toxins is not sensitive to ultraviolet ray and the biological activity is unchanged under ultraviolet light for 30min、60min and 120min The crude toxins is sensitive to Pronase and lysozyme,the biological activity was destroyed and the toxicity was lost while adding papayain (4-16 u/ml) or trypsin (4-16 u/ml) to the crude toxins It’s biological activity decreased 91 6%-92 0% while adding lysozyme(15000-60000 u/ml) to the crude toxins.