用扫描电镜观察DFRIKN-1分离物菌体的方法研究

[目的]选择合适的样品处理方式,在扫描电镜下清晰地观察DFRKN-1分离物菌体的表面结构特征。[方法]以不经过固定液处理的菌体为对照,将使用戊二醛、卢戈氏碘液、福尔马林固定的DFRKN-1分离物菌体细胞采用离心法、直接干燥法收集,进行酒精梯度逐级脱水,干燥并真空喷金后在扫描电镜下观察。[结果]不经过固定液处理的菌体形态脱水变形,经过固定液固定后的样品仍然保持原细胞形态,且戊二醛的固定效果优于其他2种固定剂;离心法不能收集到游动孢子,而直接干燥法收集则弥补了这一缺陷。使用戊二醛固定,直接干燥法收集菌体是处理DFRKN-1分离物样品的最优选择。在电镜下观察到DFRKN-1分离物表面的假根、游动孢子以及孢子囊的释放孔。[结论]通过扫描电镜观察DFRKN-1分离物的结构特征,为研究其与根结线虫的作用机制提供参考。

Research on the Observation Method of the Bacterial Isolate of DFRKN-1 with Scanning Electron Microscope

[Objective] The surface structure of the bacterial isolate of DFRKN-1 would be clearly observed under the scanning electron microscope through the selection of the appropriate method of sample treating.[Method] The isolate of DFRKN-1 without the treatment of fixation solution being taken as CK,the sample,which was treated with fixation solution:glutaraldehyde,Luge Shi iodine,formalin,then collected after centrifugation and direct drying and finally gradually dehydrated with ethanol,dried and sprayed in vacuum condition,respectively,was checked under scanning electron microscopy.[Results] The result indicated the material of CK was deformed and the treated material could keep its original cell morphology.The efficiency of glutaraldehyde fixation was better than the other 2 agents.The isolated spore could not be collected with centrifugation method but the directly drying method might make up for the defect in the procedure of sample treatment.The optimal treatment choice was the sample was fixed with glutaraldehyde and collected with directly drying method.The rhizoid in the surface of isolates,the release of zoospores and the sporangia hole of DFRKN-1 could be observed under electron microscopy.[Conclusion] in order to the and the role of The reference for the study on the reaction mechanism between the isolate and root-knot nematode was provided through the understanding of structural feature of DFRKN-1 isolate.