| 摘要: |
| 【目的】利用分子生物学方法分析采自海南儋州的苦楝丛枝病植原体核糖体蛋白(rp)基因,从亚组水平上确定苦楝丛枝病植原体的分类地位。【方法】利用植原体核糖体蛋白基因通用引物对rpF1/rpR1,应用PCR技术对苦楝丛枝病植原体的核糖体蛋白基因进行扩增,对其核苷酸序列进行测定并分析。【结果】通过PCR扩增得到约1.2 kb的特异片段,测序结果表明该片段长1 240 bp,包括rps19基因的3′区域,rpl22和rps3基因的全部区域。进一步分析发现,该片段与苦楝丛枝病贵州株系(Chinaberry witches'-broom,CWB-GZh)的亲缘关系最近,核苷酸同源性达99.9%。基于核糖体蛋白基因核苷酸序列,构建了海南苦楝丛枝病植原体与其他18个已知分类地位植原体的系统分类树状图,结果表明,引起海南苦楝丛枝病的植原体与16SrⅠ-B亚组植原体越南苦楝黄化、苦楝丛枝贵州株系和苦楝丛枝云南株系在同一条进化枝上。【结论】报道了海南苦楝丛枝病植原体的核糖体蛋白基因序列,海南苦楝丛枝病植原体属于16SrⅠ-B。 |
| 关键词: 苦楝丛枝病 植原体 核糖体蛋白基因 序列分析 |
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| 基金项目:中国热带农业科学院科技基金项目（Rky0709） |
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| Sequence analysis of ribosomal protein gene of phytoplasma associated with Chinaberry witches'-broom in Hainan Province |
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| Abstract: |
| 【Objective】 The study analyzed the phytoplasma rp gene of Chinaberry witches′-broom (CWB) samples which were collected in Danzhou of Hainan Province.This study can differentiate and classify phytoplasma of CWB from subgroup. 【Method】 Universal primers(rpF1/rpR1) for phytoplasma ribosomal protein gene were used to amplify DNA samples extracted from infected Chinaberry tree,then the amplified fragment was sequenced and analysed. 【Result】 A 1.2 kb or so fragment was amplified in DNA samples extracted from diseased chinaberry tree samples,but not in those extracted from healthy plants.Sequence analysis of the amplified fragment indicated that the amplified ribosomal protein gene fragment consisted of 1 240 bp,containing part of rps19,rpl22 and rps3 genes.It had the highest sequence identity (99.9%) with chinaberry witches-broom phytoplasma strain Guizhou.Homologic tree constructed by analysis of ribosomal protein gene nucleotide sequences from 19 strains of phytoplasmas showed that Chinaberry witches′-broom strain CWB-Hn,Vietnam chinaberry yellows,Chinaberry witches′-broom strain CWB-GZh,Chinaberry witches′-broom strain CWB-WSh clustered in the same clade. 【Conclusion】 This article reported ribosomal protein gene nucleotide sequences of Chinaberry witches'-broom phytoplasmas strain CWB-Hn,Chinaberry witches′-broom phytoplasmas strain CWB-Hn belonged to subgroup 16SrⅠ-B. |
| Key words: Chinaberry tree witches′-broom phytoplasma ribosomal protein gene sequence analysis |
