沙田柚黄龙病病原β-操纵子DNA片段的克隆与序列分析  

Cloning and sequencing of huanglongbing pathogen in shatianyou pomelo

作　　者：单振菊[1] 郭恒[1] 冯震[1] 邓晓玲[1] 

机构地区：[1]华南农业大学柑桔黄龙病研究室,广东广州510642

出　　处：《仲恺农业技术学院学报》2005年第4期45-48,共4页Journal of Zhongkai Agrotechnical College

基　　金：广东省自然科学基金(032262);广东省重点科技计划(2004B20901010)资助项目

摘　　要：采集表现斑驳症状的沙田柚叶片,根据柑桔黄龙病特异保守区域β-操纵子序列设计引物,通过对其病叶脉总DNA进行PCR扩增、扩增产物纯化、与pMD18-T Vector连接、转化大肠杆菌(Escherichia coli)DH 5a菌株、筛选阳性克隆重组子、重组子测序,并将测序结果与柑桔黄龙病病原β-操纵子部分DNA相应序列进行比较.结果表明,引起沙田柚斑驳症状的病原与亚洲柑桔黄龙病原(Liberobacter asiaticum)相应序列的同源率为99%,而与非洲柑桔黄龙病病原的同源率仅为79.9%.因此,从分子水平上证明沙田柚斑驳症状是由黄龙病病原侵染所致,该病原属于亚洲种中的一个成员.The amplification of leaf samples on shatianyou pomelo with yellow symptom was carried out using HLB - specific primers. The products were ligated into pMD18 - T vector after purified, and then were transferred into DH5a strain of Escherichia coli respectively. The positive recombinant plasmids were sequenced and compared with β- operon of the citrus Huanglongbing pathogen. The results showed that the cloned DNA fragment shared 99% and 79.9% homogeneity of sequence with those of Liberobacter asiaticum and Liberobacter africanum respectively. This indicated on level of molecule that the shatianyou pomelo disease was the same as citrus Huanglongbing and that the pathogen belonged to Liberobacter asiaticum.

关 键 词：沙田柚 黄龙病 多聚酶链式反应 克隆 序列分析 

分 类 号：S436.66[农业科学—农业昆虫与害虫防治]