利用免疫磁性分离-PCR检测安祖花细菌性枯萎病病原菌  

Detection of Xanthomonas axonopodis pv. dieffenbachiae in Authurium andreanum by Immunomagnetic Separation-PCR

作　　者：王钊[1] 储丽红[2] 赵凯[1] 彭佳佳[3] 明军[1] 袁素霞[1] 刘春[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]南京林业大学风景园林学院,南京210037 [3]宁夏大学农学院,银川750021

出　　处：《园艺学报》2013年第8期1600-1608,共9页Acta Horticulturae Sinica

基　　金：北京市科技计划研发攻关项目(YLHH201200109);中国农业科学院与北京市大兴区科技合作项目(YLHH200900106);农业部园艺作物生物学与种质创制重点实验室项目

摘　　要：根据地毯草黄单胞菌花叶万年青致病变种（Xanthomonas axonopodis pv.dieffenbachiae）的RAPD序列设计特异性引物,并对羧基化磁珠的结合性能进行检验,从而建立和优化了免疫磁性分离–PCR体系,对安祖花细菌性枯萎病进行早期检测。结果表明：1mg磁珠对多克隆抗体最大吸附值为0.268mg,进行免疫磁性捕获时免疫磁珠的最佳浓度是0.566~0.741mg·mL-1。免疫磁性分离–PCR可以减少PCR反应中的抑制物质,对X.axonopodispv.dieffenbachiae的检测灵敏度可以达到10~100cfu·mL-1,比常规PCR检测灵敏至少100倍。According to the RAPD of Xanthomonas axonopodis pv. dieffenbachiae,specific primers were designed,and binding ability of carboxyl magnetic beads were tested. The system of optimization of immunomagnetic separation-PCR（IMS-PCR）protocol for detecting the Bacterial Blight of Anthurium were establisted. The results showed that the saturate absorption of 1 mg carboxyl magnetic beads to the polyclonal antibodys is 0.268 mg. The optimal concentration of immunomagnetic beads（IMB）is 0.566– 0.741 mg · mL -1 when X. axonopodis pv. dieffenbachiae is captured. The detection sensitivity for X. axonopodis pv. dieffenbachiae,by IMS-PCR which can reduce the inhibitory substances in reaction,is 10–100 cfu · mL -1 . It is 100 times more sensitive than ordinary PCR at least.

关 键 词：安祖花 地毯草黄单胞菌花叶万年青致病变种 免疫磁性分离-PCR 最大吸附值 捕获率 

分 类 号：S682.14[农业科学—观赏园艺]