蝴蝶兰中建兰花叶病毒和齿兰环斑病毒复合侵染的鉴定及其vsiRNAs特征分析  

Characterization of vsiRNAs derived from Cymbidium Mosaic/Odontoglossum Ringspot Viruses in co-infected Phalaenopsis equestris

作　　者：兰汉红 LAN Hanhong(School of Biological Sciences and Biotechnology,Minnan Normal University/Key Laboratory of Landscape Plants with Fujian and Taiwan Characteristics of Fujian,Zhangzhou,Fujian 363000,China)

机构地区：[1]闽南师范大学生物科学与技术学院/闽台特色园林植物福建省高校重点实验室,福建漳州363000

出　　处：《福建农业学报》2024年第9期1086-1093,共8页Fujian Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31601613);福建省自然科学基金项目(2018J01465);漳州市自然科学基金项目(ZZ2017J03)。

摘　　要：【目的】探明蝴蝶兰和病毒(建兰花叶病毒、齿兰环斑病毒)之间的相互作用,进而为制定蝴蝶兰病毒性病害的有效防控措施提供理论基础。【方法】首先通过RT-PCR特异扩增建兰花叶病毒(cymbidium mosaic virus,CymMV)和齿兰环斑病毒(odontoglossum ringspot virus,ORSV)外壳蛋白基因片段,通过电镜负染和切片技术明确CymMV和ORSV在蝴蝶兰细胞中的存在;然后通过小RNA深度测序技术鉴定分析病毒来源的小干扰RNA(vsiRNAs)的丰度、长度、碱基偏好性和正负义来源等特征。【结果】RT-PCR能够特异性扩增到病毒外壳蛋白基因片段,电镜负染和超薄切片结果中都能够观察到长约300 nm线状的CymMV病毒粒子和长约500 nm杆状的ORSV病毒粒子的存在;小RNA深度测序分别获得7563892和6133689个读数的CymMV和ORSV来源的vsiRNAs,vsiRNAs在丰度、长度、碱基偏好性和正负义来源等方面具有一定的普遍性和特异性。【结论】CymMV和ORSV两种病毒在蝴蝶兰植株中存在复合侵染;CymMV和ORSV vsiRNAs的丰度、长度、悬挂、碱基偏好性、正负义链分布、Hotspot和Coldspot等特征具有普遍性和特异性。【Objective】Interactions between Phalaenopsis equestris and cymbidium mosaic virus(CymMV)and odontoglossum ringspot virus(ORSV)were studied to aid the effort in developing effective preventive and control means against the diseases caused by the pathogens.【Method】The coat protein(CP)genes of CymMV and ORSV were amplified using RT-PCR.Under an electron microscope,viral morphology and size of CymMV and ORSV particles in P.equestris cells were examined.Abundance,length,base preference and origin of virus-derived vsiRNAs were analyzed applying the small RNA deep sequencing technology.【Result】The amplifications of CP genes of CymMV and ORSV were specifically obtain by RT-PCR.The electron microscopy revealed the lengths of the rod-like CymMV to be approximate 300 nm,while the linear ORSV,500 nm.The small RNA deep sequencing yielded 7563892 CymMV-derived and 6133689 ORSV-derived vsiRNAs exhibiting the universality and specificity in abundance,length,base preference and sense strand distribution.【Conclusion】Coinfections of CymMV and ORSV on P.equestris were clearly demonstrated in this study.The vsiRNAs of CymMV and ORSV displayed characteristic patterns in abundance,length,base preferences and sense strand distribution.

关 键 词：蝴蝶兰 病毒 vsiRNAs 互作 

分 类 号：S682[农业科学—观赏园艺]