柑橘溃疡病菌免疫荧光和生物学检测技术研究  

Study on immunofluorescent and biological identification of Xanthomonas axonopodis pv. citri.

作　　者：罗志萍 洪霓[2] 

机构地区：[1]湖北出入境检验检疫局,430022 [2]华中农业大学柑橘脱毒中心

出　　处：《植物检疫》2006年第5期272-274,共3页Plant Quarantine

摘　　要：本研究通过对7个不同柑橘品种的离体叶片接种及致病性观察筛选出了柑橘溃疡病菌敏感寄主材料,并建立了柑橘溃疡病菌的生物学检测方法;还初次尝试建立了一种简便易行的柑橘溃疡病菌免疫荧光快速鉴定方法。柑橘离体叶片接种实验结果表明:福本、红肉脐橙、纽荷尔和枳壳对柑橘溃疡病菌表现出一定的感病性,而粗柠檬和邓肯葡萄柚未表现出明显感病迹象,反而出现了坏死性应激反应。免疫荧光试验结果显示:浓度为103cfu/mL的菌液在30℃下用BSA封闭2h,30℃下抗体结合1h,荧光抗体浓度为4μg/mL,室温放置40min后镜检,就可以在荧光显微镜下清晰地看到绿色的柑橘溃疡病菌体,而用该方法检测不到水稻白叶枯病菌。用免疫荧光抗体检测柑橘溃疡病菌操作简便,仅需一台荧光显微镜,整个检测过程仅需要4h。The study tried to screened out the sensitive host material through inoculating on the leaves of seven citrus varieties in vitro , and set up the methods of quickly immunofluorescent identification. Considering that the in vitro_test of orange leaves supports that Fuben,sweet orange[ Citrus sinensis （L.） Osbesk cv. Cara Cara] ,Newhall and Bittet Orange_appear a little susceptible to CBCD while rough lemon （Citrus jambhiri Lush） and Duncan grapefruit do not but show the hypersensitive reactions which are generally considered as signals of host resistance, it is more feasible to assess the pathogenesis of CBCD using cultivars like trifoliate orange and sweet orange in practical test. Immunofluorescence experiments demonstrate that green bacteria bodies can be seen explicitly under fluorescent microscope after enclosed by BSA under 30℃ for two hours, combined with antibody for one hours, with the fluorescent antibody concentration of 4 μg/mL in room temperature for forty minutes while X. not. Testing CBCD bacteria with immunofluorescence antibodies by this mean is quite simple that only a fluorescent microscope is needed and the whole process only takes four hours with an extremely strong specificity in antigen-antibody reaction .

关 键 词：柑橘溃疡病菌 致病性试验 免疫荧光 柑橘 

分 类 号：S41[农业科学—植物保护] S436.661