中国樱桃花变叶病相关植原体的分子检测及鉴定  

Molecular Detection and Identification of Subgroup 16SrV-B Phytoplasma Associated with Chinese Cherry Phyllody Disease in China

作　　者：高瑞 杨淑珂[2] 王洁[1] 路兴波[2] 孙玉刚[1] 田延平[3] 汪卫星[4] GAO Rui;YANG Shuke;WANG Jie;LU Xingbo;SUN Yugang;TIAN Yanping;WANG Weixing(Shandong Institute of Pomology,Tai’an,Shandong 271000,China;Institute of Plant Protection,Shandong Academy of Agricultural Sciences,Jinan 250100,China;College of Plant Protection,Shandong Agricultural University,Tai’an,Shandong 271018,China;College of Horticulture and Landscape,Southwest University,Chongqing 400716,China)

机构地区：[1]山东省果树研究所,山东泰安271000 [2]山东省农业科学院植物保护研究所,济南250100 [3]山东农业大学植物保护学院,山东泰安271018 [4]西南大学园艺园林学院,重庆400716

出　　处：《园艺学报》2019年第7期1249-1256,共8页Acta Horticulturae Sinica

基　　金：国家自然科学基金青年基金项目（31401718,31701899）;山东省农业科学院青年英才培养计划项目

摘　　要：从重庆市采集到表现花变叶症状的中国樱桃枝条样品,利用透射电镜在感病樱桃枝条韧皮部筛管细胞内观察到直径为200~500nm的圆形或者近圆形的植原体粒子。提取感病和健康花瓣的总DNA,利用植原体16S rRNA基因及延伸因子rp基因通用引物进行PCR扩增,从感病植株中扩增得到了长度分别为1.4kb的16SrRNA基因和1.2kb的rp基因。序列一致率分析表明,樱桃花变叶植原体16SrRNA基因和rp基因均与四川甜樱桃花变绿植原体相应基因的核苷酸一致率最高,分别为100%和99.8%;16SrRNA基因相似系数分析表明,樱桃花变叶植原体与四川甜樱桃花变绿植原体、枣疯植原体的相似系数均为1.00;基于16SrRNA基因和rp基因构建系统进化树时发现,樱桃花变叶植原体均与16SrV-B亚组成员聚为一簇,该植原体属于16SrV组的B亚组。To confirm phytoplasma infection,samples of Chinese cherry(Prunus pseudocerasus L.)plants showing phyllody symptom were collected from an orchard in Chongqing,China. By using transmission electron microscope technology,structures resembling phytoplasmal cells,about 200–500 nm,were observed in the phloem sieve elements of symptomatic but not in healthy plant tissues. Total DNAs were extracted from symptomatic and asymptomatic plants for PCR amplification. Specific fragments of 1.4 kb and 1.2 kb in length were amplified from diseased cherry tissue,but not from healthy plant,using universal primers for phytoplasmal 16 S rRNA gene and rp gene,respectively. The phytoplasma associated with the disease was designated as Chinese cherry phyllody phytoplasma Chongqing isolate(ChP-CQ). Sequence analysis indicated that the 16 S rRNA gene and rp gene of ChP-CQ shared the highest similarities of 100% and 99.8%,respectively,to those of cherry virescence phytoplasma Sichuan isolate(MF848961). i PhyClassifier analysis for 16 S rRNA gene showed that ChP-CQ has the same restriction enzyme pattern with reference strains of 16 SrV-B. Phylogenetic analysis based on 16 S rRNA and rp genes further revealed that ChP-CQ belonged to 16 SrV-B subgroup. In conclusion,the phytoplasma associated with Chinese cherry phyllody disease in Chongqing,China was classified as a member of the subgroup B of the elm yellows group(16 SrV).

关 键 词：中国樱桃 樱桃花变叶病 植原体 枣疯病植原体 

分 类 号：S662.5[农业科学—果树学]