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secYnusA基因在泡桐丛枝植原体亚组分类中的应用
张 磊1, 韩 翔2, 李正男3
1.西北农林科技大学 林学院;2.西北农林科技大学 理学院;3.西北农林科技大学 植物保护学院
摘要:
【目的】利用分子生物学方法分析泡桐丛枝植原体转运蛋白secY基因及转录延伸因子nusA基因,从亚组水平上确定陕西泡桐丛枝植原体的分类地位。【方法】采集陕西杨凌、彬县、旬邑、永寿、周至和宝鸡等6个泡桐栽培区的泡桐丛枝样品,提取病样总核酸;设计引物对secY-F/R和nusA-F/R,对泡桐丛枝植原体secYnusA基因进行PCR扩增及核苷酸序列测定与系统发育分析。【结果】获得了泡桐丛枝病植原体secY基因序列的全长,大小为1 358 bp,编码414个氨基酸;获得了部分nusA基因序列,大小为946 bp,编码315个氨基酸。序列同源性分析结果表明,陕西各县区泡桐丛枝植原体的secYnusA基因序列均一致。一致性分析和系统发育分析表明,陕西泡桐丛枝植原体与台湾泡桐丛枝植原体的亲缘关系最近,secYnusA基因序列的同源性分别为99.9%和99.6%。【结论】首次报道了泡桐丛枝植原体转运蛋白secY基因及转录延伸因子nusA基因的序列,并以其作为分类标准,将陕西泡桐丛枝植原体归属到16SrⅠ-D亚组。
关键词:  泡桐丛枝植原体  secY基因  nusA基因  序列分析
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基金项目:林业公益性行业科研专项(201004003-4);教育部长江学者和创新团队发展计划项目
Application of secY gene and nusA gene to classification of paulownia witches’ broom associated phytoplasma
ZHANG Lei1, HAN Xiang2, LI Zhengnan3
1.College of Forestry ,Northwest A&F University;2.College of Sciences, Northwest A&F University?;3.College of Plant Protection, Northwest A&F University?
Abstract:
【Objective】The research was conducted to divide the paulownia witches’broom phytoplasma-Shaanxi (PaWB-Shaanxi) into subgroups based on the phytoplasma secY gene and nusA gene.【Method】The paulownia withces’broom samples were collected in Yangling,Binxian,Xunyi,Yongshou,Zhouzhi and Baoji,Shaanxi Province.Total nucleic acid of each sample was extracted and used as template to amplify the secY gene and nusA gene by PCR with designed primer pairs secY-F/R and nusA-F/R.The PCR products were sequenced and analyzed.【Result】Complete secY gene of PaWB-Shaanxi was 1 358 bp,and contained protein of 414 amino acids while the partial nusA was 946 bp with 315 amino acids.The identity analyses and phylogenetic analyses revealed that PaWB-Shaanxi was most closely related to PaWB-Taiwan,and the similarity of secY gene and nusAgene between them was 99.9% and 99.6%,respectively.【Conclusion】This was the first study of secY gene and nusA gene of paulownia witches’broom phytoplasma in China.Based on the two genes PaWB-Shaanxi was further classified into subgroup 16SrⅠ-D.
Key words:  paulownia witches’broom phytoplasma  secY gene  nusAgene  sequence analysis