间接免疫荧光显微术检测泡桐丛枝病原MLO的研究  

Detection of Mycoplasma-like Organism in the Infected Paulownia by Indirect Immunofluorescence Microscopy

作　　者：田国忠[1] 张锡津[1] 朱水方 张成良[1] 罗飞[1] 黄文胜[1] 

机构地区：[1]中国林业科学研究院森林保护研究所,农业部植物检疫实验所

出　　处：《林业科学研究》1996年第1期1-6,共6页Forest Research

基　　金：国家"八五"攻关项目

摘　　要：以感染类菌原体ＭＬＯ的泡桐组培苗为病原繁殖材料，采用差速离心和Ｐｅｒｃｏｌｌ不连续密度梯度离心来提纯ＭＬＯ作为免疫抗原制备兔抗血清。抗血清经健康汁液吸收后，作为第一抗体，用异硫代氰酸荧光素ＦＩＦＣ标记的羊抗兔免疫球蛋白作为第二抗体，进行间接免疫荧光显微镜检查染病组织中的ＭＬＯ特异性荧光。经与ＤＡＰＩ染色技术比较，找出了适宜的徒手切片和减少非特异性反应的途径。此技术具有灵敏度高、特异性强和测定简便等特点。适用于ＭＬＯ组织定位、组培苗脱毒效果评估、病原株系鉴定和病害检疫等。Using paulownia witches 'broom MLO extracted and concerntrated from in-fected paulownia tissue culture plantlets by differential and discontinuous Percoll density cen-trifugation as antigen,the antiserum was produced.After the absorption with healthypaulownia extract,the antiserum could effectively detect paulownia witches'broom MLO indifferent tissues of infected plants with FITC Iabelled sheep-anti-rabbit IgG by indirect im-munofluoresence microscopy. Compared with DAPI staining test,a more specific,sensitiveand handy method for detecting MLO was developed,which would facilitate the detection ofMLO in situ,identification of MLO strains and the diagnosis of plant diseases associatedwith MLOs.

关 键 词：泡桐 丛枝病原MLO 间接免疫荧光 显微术 

分 类 号：S763.724.3[农业科学—森林保护学]