摘要
【目的】鉴定侵染鬼针草引起花叶或斑驳的病原。【方法】利用电镜技术观测病原粒子;间接ELISA方法鉴定病原与马铃薯Y病毒属病毒的血清学关系;分子生物学技术克隆了病原3′-端序列;BLAST,Clustal_X,BioEdit和MEGA4.0等生物信息学软件用于所得序列的序列分析。【结果】在电镜下可观察到长约650-700nm×13nm左右的线状病毒粒子和风轮状内含体;利用Potyvirus PathoScren试剂盒检测呈阳性;用马铃薯Y病毒科病毒简并引物可扩增获得一条约1800bp的片段;序列分析表明该序列与鬼针草斑驳病毒相应序列高度相似,外壳蛋白的氨基酸同源性及3′-UTR核苷酸同源性均达96%以上。【结论】侵染云南鬼针草引起花叶或斑驳症状的病原为鬼针草斑驳病毒,这是该病毒侵染中国鬼针草属植物的首次报道。
[Objective] This study was carried out to determine the pathogen causing the mosaic and mottle symptom on Bidenspilosa L.. [Method] Viral particles in the infected leaves were detected by electron microscopy. Serological relation between the pathogenic virus and genus Potyvirus was examined by indirect ELISA. Moreover, the sequence of the 3＇-part genome of the virus was cloned by RT-PCR and sequenced by common molecular biological methods. For bioinformational analysis, programs of BLAST at NCBI, Clustal_X, BioEdit and MEGA 4.0 were used for alignment and phylogenetic tree construction. [Result] Electron microscopy of leaf dips from infected plants revealed flexuous particles 650-700 nm in length and 13 nm in width. Cytoplasmic laminar aggregates and pinwheel inclusions were observed in ultrathin sections. Serological reaction between the virus and the antibody ofpotyvirus was positive. A DNA fragment of 1.8 kb with high identity to the sequence of Bidens mottle virus （BiMoV） could be amptified through the universal primers of family Potyviridae virus. Analysis of the sequence revealed that the amino acid identity of CP and nucleotide identity of 3＇UTR to corresponding sequences of BiMoV were both more than 96%. [ Conclusion ] The evidences suggests that the pathogen causing mosaic and mottle symptom on Bidenspilosa L. is BiMoV, which is the first report of BiMoV infecting a member of the Bidens L. in China.
出处
《中国农业科学》
CAS
CSCD
北大核心
2009年第5期1849-1853,共5页
Scientia Agricultura Sinica
基金
云南省科技攻关项目(2006NG02)
