来源于湖北省枣疯病植原体分离物16S rDNA和rp基因的序列分析  

Sequences analysis of 16S rDNA and rp genes of the phytoplasma associated with jujube witches'-broom(JWB ) from Hubei Province

作　　者：王利平[1,2] 洪霓[1,2] 王国平[1,2] 

机构地区：[1]华中农业大学湖北省作物病害监测与安全控制重点实验室,武汉430070 [2]华中农业大学农业微生物学国家重点实验室,武汉430070

出　　处：《植物病理学报》2014年第1期1-7,共7页Acta Phytopathologica Sinica

基　　金：湖北省自然科学基金(2011CDB150);教育部新教师基金(20100146120037);中央高校基本科研业务费专项资金资助(2010QC033)

摘　　要：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。16S rDNA and ribosomal protein gene （rp） of JWB-Hubei phytoplasma isolates from jujube trees collected in Sui county, Suizhou city, Hubei Province were amplified by Nested-PCR. The PCR products were purified, cloned and sequenced. The size of cloned fragments from 16S rDNA and rp gene were 1 247 bp and 1 196 bp, respectively. The phylogenetic analysis on nucleotide （nt） sequences of 16S rDNA and rp were constructed. The results of 16S rDNA gene nucleotide sequences analysis showed that the JWB-Hubei phytoplasma isolates had the higher 99% nucleotide sequence identity with the isolates from Shandong, Henan Province and so on, and were grouped into a separate sub-branch of the same sub-group. Virtual RFLP analysis results showed that JWB- Hubei belonged to a member of 16SrV-B subgroup, which was in accordance with the phylogenetic tree. The nucleotide sequences based on rp122-rps3 gene from JWB-Hubei phytoplasma isolates showed that the higher 99% nucleotide sequence identity with the isolates from Shandong, and Shannxi Province. The phylogenetic tree analysis indicated that JWB-Hubei phytoplasma isolate was clustered into a separate sub-branch of the same sub-group with the JWB phytoplasma, belonging to rpV-C subgroup. The above results identify the classification status and genetic evolution relationship of the JWB-Hubei isolate, which lay theoretical foundation to further study on classification and genetic evolution of the phytoplasma.

关 键 词：枣疯病 16S RDNA rp基因 进化树分析 

分 类 号：S436.6[农业科学—农业昆虫与害虫防治]