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利用分子生物学技术对天津滨海新区泡桐丛枝病病原进行分类鉴定。采用植原体16S rDNA通用引物R16mF2/R16mR1对患病植株总DNA进行PCR扩增,得到约1.4 kb特异性片段。克隆测序、Blast比对和iPhyClassifier分析结果表明,天津滨海新区泡桐丛枝植原体16S rDNA基因片段长1 432 bp,与国内泡桐丛枝植原体PY株系相似性最高,达99.86%,归属于16SrI组(aster yellows group,翠菊黄化组)D亚组。系统树构建与分析显示,泡桐丛枝病天津滨海株PaWB-TJBH与16SrI其他亚组亲缘关系较近,同在16SrI组进化枝上,与16Sr I-D组亲缘关系最近;16S rDNA序列RFLP电子酶切图谱表明,PaWB-TJBH属于16SrI-D组一个成员,与同源性比较和系统进化分析结果一致。
Using Molecular Biology Techniques to Identify Pathogens of Paulownia Witches’ Broom in Binhai New Area, Tianjin. The total DNA of the diseased plants was amplified by PCR using the 16S rDNA universal primer R16mF2 / R16mR1, and the specific fragment of about 1.4 kb was obtained. The results of cloning and sequencing, Blast analysis and iPhyClassifier showed that the 16S rDNA gene fragment of Phyllosticta flora in Tianjin Binhai New Area was 1 432 bp in length, which had the highest similarity (99.86%) with the PY strain of P. paulownia, which belonged to 16SrI Group (Aster yellows group, Aster yellow group) D subgroup. The construction and analysis of phylogenetic tree showed that PawB-TJBH was closely related to other 16SrI subgroups of paulownia witches’ broom disease strain, and had the closest genetic relationship with 16Sr ID group on the clades of 16SrI group. The 16S rDNA sequence of RFLP Atlas showed that PaWB-TJBH belongs to a member of 16SrI-D group, which is consistent with homology comparison and phylogenetic analysis.