甘薯羽状斑驳病毒和甘薯褪绿矮化病毒复合侵染甘薯引起的细胞病理学研究

           甘薯羽状斑驳病毒和甘薯褪绿矮化病毒复合侵染甘薯引起的细胞病理学研究

谢 礼¹, 吕明芳¹, 王 芳², 季志仙³, 吴列洪³, 严成其^1*, 陈剑平^1*

（1. 浙江省植物有害生物防控重点实验室—省部共建国家重点实验室培育基地，农业部植保生物技术重点实验室, 浙江省农业科学院病生所，浙江 杭州310021；2. 宁波市农业科学院,浙江 宁波315101；3. 浙江省农业科学院作物与核技术研究所,浙江 杭州310021）

摘 要：2011~2012年在浙江杭州连续发现感病甘薯植株具有甘薯病毒病SPVD的典型症状，透射电镜观察发现其叶片包含有长度为600~900nm，直径为12nm的线状病毒粒子。使用特异性引物进行RT-PCR扩增及测序，其病原为甘薯羽状斑驳病毒（Sweet potato feathery mottle virus, SPFMV）和甘薯褪绿矮化病毒（Sweet potato chlorotic stunt virus, SPCSV）复合侵染。运用透射电镜研究SPVD引起的细胞病理学变化，观察到叶肉细胞的细胞质中含有马铃薯Y病毒属特征性柱状内含体，其结构分类为Ⅳ型；线状病毒样粒子以束状平行排列成聚集体的方式存在于细胞质中；叶绿体伸出伪足状结构包裹线粒体。在维管束细胞的筛管和伴胞中观察到弯曲线状病毒样粒子形成的聚集体，其粒子排列交错，与叶肉细胞中平行排列的病毒样粒子聚集体不同，符合毛形病毒属特征。另外还在维管束和叶肉细胞结合的部位发现复合侵染的细胞病理学特征。本研究表明危害甘薯生产的SPVD已扩散蔓延到浙江省。

关键词：甘薯病毒病（SPVD）；甘薯羽状斑驳病毒；甘薯褪绿矮化病毒；复合侵染；细胞病理学

中图分类号：S432.1；S432.4；Q336   文献标识码：A  

doi:10.3969/j.1000-6281.2013.06.006

Cyto-pathology study of sweet potato induced by synergetic infection of Sweet potato feathery mottle virus and Sweet potato chlorotic stunt virus

Xie Li¹, LÜ Ming-fang¹,Wang Fang², Ji Zhi-xian³, Wu Lie-hong³, Yan Cheng-qi^1*, Chen Jian-ping^1*

（1. State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Ministry of Agriculture Key Laboratory of Biotechnology in Plant Protection. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou Zhejiang 310021；2. Ningbo Academy of Agricultural sciences, Ningbo Zhejiang 315101；3. Institute of Crop andNuclear Technology Utilization, Zhejiang Academy of Agricultural Sciences, Hangzhou Zhejiang 310021, China）

Abstract: In 2011 and 2012, cultivars of sweet potato with the likely syptom of sweet potato virus disease (SPVD) were found in Zhejiang, Hangzhou. The infected leaf was taken under transmission electron microscopy (TEM) for pathogen observation. Linear virus particles with the length of 600-900 nm, diamater of 12 nm was observed. For further molecular pathogen identification, specific primers were used to carry out RT-PCR and sequencing. At last, Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) were identified as the pathogen of the infected sweet potato, namely the synergetic infection by SPVD. Cytopathology induced by SPVD in sweet potato was also studied by TEM after ultra-thin section. Distinctive cylindrical inclusion bodies of subdivisionⅣwere observed in mesophyll cytopalsm. Aggregates of linear virus particles arranged paralleled as bundles were also observed in mesophyll cytoplasm. All of the cytopathology above represents the infection of Potyvirus. Abnormal chloroplasts containing membrane-bound vesicles packing or encircling mitochodria were also observed. In phloem cells including sieve elements and companion cells, aggregates of linear virus particles arranged not paralleled were observed. The patterns of particle arrangement in phloem (not paralleled) and in mesophyll (paralleled bundles) are different. Linear virus particles distributing in phloem cell is the characteristic of Crinivirus infection. In addition, cytopathology of synergetic infection was observed in the boundary of phloem and mesophyll. This paper was the first report of SPVD outbreak in Zhejiang province.

Keywords:sweet potato virus disease (SPVD)；Sweet potato chlorotic stunt virus；Sweet potato feathery mottle virus；synergetic infection；cytopathology

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