小西葫芦黄化花叶病毒罗汉果分离株抗血清的制备和应用  

Preparation and Application of Antiserum Against Siraitia grosvenorii Isolate of Zucchini yellow mosaic virus

作　　者：朱英芝[1,2] 韩英 蒙姣荣[2,4] 廖咏梅[1,2] 邹承武[2,4] 陈保善[2,4] 

机构地区：[1]广西大学农学院,广西南宁530004 [2]亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004 [3]河南出入境检验检疫局,河南郑州450008 [4]广西大学生命科学与技术学院,广西南宁530004

出　　处：《热带作物学报》2011年第9期1683-1686,共4页Chinese Journal of Tropical Crops

基　　金：广西自然科学基金项目(桂科自No.0832045);广西创新能力建设项目(桂科能No.05112001-1B;桂科能No.05112001-1)

摘　　要：将分离自罗汉果(Siraitia grosvenorii)的小西葫芦黄化花叶病毒(Zucchini yellow mosaic virus,ZYMV-LHG)在西葫芦上繁殖,利用差速离心法与PEG沉淀法相结合提纯病毒粒子,用于免疫新西兰大白兔,成功制备出效价为1∶10 240的抗血清。分别用健康西葫芦叶片总蛋白及健康西葫芦叶片汁液吸附处理抗血清,比较2种处理后抗血清背景反应,以健康西葫芦汁液吸附法较佳。用间接ELISA对罗汉果果园及附近共46个科114种植物进行检测,其中9个科25种植物有阳性反应,以葫芦科、苋科、豆科、锦葵科等阳性率较高。田间病毒病发病率调查表明,宿根苗发病率比组培苗高,发病时间更早,是田间罗汉果病毒病的主要初侵染来源之一。The Luohanguo （Siraitia grosvenorii）-infected by Zucchini yellow mosaic virus （ZYMV-LHG）, was propagated on Cucurbita pepo and purified by differential centrifugation in combination with Polyethylene Glycol （PEG）precipitation. The purified ZYMV-LHG preparation was injected into New Zealand rabbits. The titer of the antiserum was 1：10 240 assayed by the indirect ELISA. The antiserum was treated either by the leaf extract or total protein of zucchini squash to remove non specific reaction. The former was found better than the latter treatment. The treated antiserum was used to detect plants collected from the Luohanguo field and nearby lands with indirect enzyme-linked immunosorbent assay（ELISA）. Of 114 plant species belonging to 46 families, 25 species from nine families were with positive reaction, among which plants of families Cucurbitaceae, Amaranthaceae, Amaranthaceae, Leguminosae and Malvaceae showed higher positive rates. A survey on the virus disease in the field revealed that the disease incidence of ratoon Luohanguo was higher than that of the tissue culture seeding planted annually, and the symptoms develops earlier in the ratoon. It suggested that the infected ratoon Luohanguo serve as the major primary source of the viral disease.

关 键 词：罗汉果 小西葫芦黄化花叶病毒 抗血清 寄主检测 

分 类 号：R282.71[医药卫生—中药学]