侵染肥城桃的病毒和类病毒的分子检测与鉴定

为明确山东肥城桃种植区桃树上主要存在的病毒和类病毒及其发生情况,采集具有花叶、斑驳和皱缩典型症状的肥城桃样品,提取叶片总RNA后,分别选用桃树上已报道的啤酒花矮化类病毒Hopstuntviroid(HSVd)、桃潜隐花叶类病毒Peach latent mosaic viroid(PLMVd)、苹果褪绿叶斑病毒Apple chlorotic leaf spot virus(ACLSV)、樱桃锉叶病毒Cherry rasp leaf virus(CRLV)、桃花叶病毒Peach mosaic virus(PMV)、李属坏死环斑病毒Prunus necrotic ringspot virus(PNRSV)、李痘病毒Plum pox virus(PPV)、李矮缩病毒Prunus dwarf virus(PDV)、樱桃绿环斑驳病毒Cherry green ring mottle virus(CGRMV)、杏假褪绿叶斑病毒Apricot pseudo-chlorotic leaf spot virus(APCLSV)、李树皮坏死茎纹孔伴随病毒Plum bark necrosis stem pitting-associated virus(PBNSPaV)和小樱桃病毒1号Little cherry virus 1(LchV1)的特异性引物进行RT-PCR检测。PCR结果显示仅HSVd、PLMVd、ACLSV、PNRSV和PBNSPaV的扩增产物中得到了预期大小的目的片段,将目的片段克隆测序后,经NCBI BLAST比对发现,山东肥城桃分离物HSVd、PLMVd、ACLSV、PNRSV和PBNSPaV与GenBank已报道分离物序列一致性均达90%以上。表明山东肥城桃已感染HSVd、PLMVd 2种类病毒和ACLSV、PNRSV、PBNSPaV 3种病毒。

Molecular detection and identification of viruses and viroids infecting Feicheng peach

To investigate field occurrence and causing agents of virus and viroid diseases on Feicheng peach in the main planting areas in Shandong Province, peach leaf samples with typical symptoms of mosaic, mottled and shrinking reminiscent of virus and viroid infection were collected from Feicheng peach orchard. After extracting total RNA, specific primers of Hop stunt viroid (HSVd), Peach latent mosaic viroid (PLMVd), Apple chlorotic leaf spot virus (ACLSV), Cherry rasp leaf virus (CRLV), Peach mosaic virus (PMV), Prunus necrotic ringspot virus (PNRSV), Plum pox virus (PPV), Prunus dwarf virus (PDV), Cherry green ring mottle virus (CGRMV), Apricot pseudo-chlorotic leaf spot virus (APCLSV), Plum bark necrosis stem pitting-associated virus (PBNSPaV) and Little cherry virus 1 (LchV1 ) were applied for RT-PCR detection. Only five expected segments specific to HSVd, PLMVd, ACLSV, PNRSV and PBNSPaV were obtained. PCR products were cloned and sequenced. BLAST analysis showed that sequences of Feicheng peach isolates of HSVd, PLMVd, ACLSV, PNRSV and PBNSPaV shared similarities higher than 90% with those of the corresponding domestic isolates deposited in GenBank. Our results demonstrated that Feicheng peach trees were infected by HSVd, PLMVd, ACLSV, PBNSPaV and PNRSV.