枣疯病植原体tuf和rp基因的克隆与序列分析

Cloning and analysis of tuf and rp gene of the phytoplasma associated with Jujube Witches’ -Broom

摘要:

摘要：【目的】枣疯病是一种重要的植原体病害，本研究旨在明确北京及河北地区枣疯病植原体的分类地位，为枣疯病在亚组水平上分类提供一定的参考依据。【方法】利用植原体通用引物fTufu/rTufu和rp(v)F1A/rp(v)R1A对北京和河北地区枣疯病植原体延伸因子tuf基因和核糖体蛋白基因(rp)进行PCR扩增并进行核苷酸序列测定及相似性分析。【结果】获得北京地区JWB-XFSZ株系、JWB-XFDO株系以及河北地区JWB-TXSZ株系的tuf基因片段均为824 bp；北京地区JWB-XFSZ株系的rp基因片段为1196 bp。经序列相似性比较表明：tuf基因与16SrV组的葡萄黄叶病（Flavescence dorée）相似性最高，为92.84%，而与已经公布的其它地区（陕西杨凌）的枣疯病植原体tuf基因相似性较低，为57.29%；关于rp基因，北京地区枣疯病JWB-XFSZ株系与16SrV组的枣疯病泰山株系（JWB-Taishan）以及大麻丛枝病植原体（HFWB）相似性最高，均为99.83%，与16SrV组的成员相似性均在96%以上。【结论】北京与河北地区枣疯病植原体具有较高的相似性，而在tuf基因水平上，与陕西地区枣疯病植原体具有较大的差异；本研究中北京与河北两地区枣疯病植原体归属于16SrV组。

Abstract:

Abstract: [Objective] Jujube witches’-broom (JWB) is an important plant disease caused by phytoplasma. The major objective of our research was to classify JWB in Beijing and Hebei districts and to provide reference for classification in subgroup level. [Methods] By use of PCR, the elongation factor Tu (tuf gene) and ribosomal protein (rp) gene of phytoplasma associated with JWB in Beijing and Hebei districts were amplified separately with universal primer pairs fTufu/rTufu and rp(v)F1A/rp(v)R1A. Partial tuf gene and rp gene were sequenced and similarity analysed with other phytoplasmas. [Results] We obtained partial tuf gene sequence (824bp) and complete rp gene (1196bp) from the diseased sample. In tuf gene, JWB in Beijing shared most similarity (92.84%) with Flavescence dorée (FD) phytoplasma (Candidatus Phytoplasma vitis) , however, shared a low similarity (57.29%) with JWB in Shaanxi district which had been already reported .The similarity analysis for sequences of rp gene showed a high identity(﹥96 %) with members of the 16SrV group phytoplasmas. It shared most identity (99.83%) with JWB strain Taishan and Hemp fiber witches’-broom phytoplasma (HFWB) of the 16SrV group. [Conclusion] The JWB strains in Beijing and Hebei are members of 16Sr V; JWB in Beijing and Hebei share high similarity, and show a diversity with JWB in Shaanxi.