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以田间采集的来源于我国湖北省枣树产业主产区随州市随县种植的表现为“枣疯病”症状的枣树分离株为试材,对其16S rDNA和核糖体蛋白(ribosomal protein,rp)基因采用Nested-PCR进行扩增以及序列分析。结果表明,湖北JWB-Hubei植原体分离物16S rDNA基因的核苷酸序列与我国山东、河南等地的分离株一致率均为99%以上,在进化树中位于同一亚组的不同进化分支;虚拟RFLP图谱分析表明,JWB-Hubei属于16SrV-B亚组一个成员,与其进化树分组结果一致。JWBHubei分离株rp基因的核苷酸序列也与我国山东、陕西等地区的分离株一致率均为99%以上,在进化树中聚为同一亚组,与报道的基于RFLP分类属于rpV-C亚组的中国枣疯病分离物(JWB)聚集于同一亚组不同分支。该研究结果明确了湖北省枣疯病植原体的分类地位以及与来源于我国不同地区枣疯病分离株之间的遗传进化关系,为进一步研究植原体的株系划分、基因遗传变异研究提供了理论基础。
The jujube isolated from the jujube tree in Suixian, Suizhou County, the main producing area of jujube tree industry in Hubei Province, was collected from the field. The 16S rDNA and ribosomal protein protein, rp) gene was amplified by Nested-PCR and sequenced. The results showed that the nucleotide sequences of the 16S rDNA gene of the JWB-Hubei phytoplasma isolate in Hubei Province and those in Shandong and Henan of China were all more than 99% and were located in different evolutionary branches of the same subgroup in the phylogenetic tree. The virtual RFLP pattern analysis showed that JWB-Hubei belonged to a member of 16SrV-B subgroup, which was consistent with its phylogenetic tree grouping. The nucleotide sequence of the rp gene of JWBHubei isolates was also consistent with that of isolates from Shandong, Shaanxi and other regions in China. All of them were clustered in the same subgroup in the phylogenetic tree, and the reported rpV-C subtypes based on RFLP Groups of Chinese jujube broom isolates (JWB) clustered in different branches of the same subgroup. The results of this study clearly identified the taxonomic status of the jujube palmatum in Hubei Province and the genetic evolution between jujube baker’s disease isolates from different regions in China. In order to further study the phylogenetic relationships of the phytoplasmas, Theoretical basis.