基于Dickeya dadantii特有标志基因检测甘薯茎腐病菌

Detection of stem and root rot pathogen of sweet potato based on conserved signature genes specific to Dickeya dadantii

甘薯细菌性茎腐病是由达旦提狄克氏菌(Dickeya dadantii)引起的一种检疫性病害,近年来在我国多地发生,严重威胁我国甘薯产业的发展。建立特异灵敏的检测D. dadantii的方法对于鉴定检疫病原菌、田间监测病原菌和防控病害有重要意义。本研究对Dickeya属菌株的全基因组序列进行比较基因组学分析,筛选到D. dadantii特有的标志基因,针对标志基因设计引物,其中针对编码登录号为WP077245517未知蛋白基因的1对引物Dad1-F(5’-CATATCAACCAGACCAGCCGTT-3’)和Dad1-R(5’-CGGCCTGCTTTTAAACAACGTATTA-3’)能只从D. dadantii扩增到167 bp目的片段。由此建立了特异灵敏的常规PCR和实时荧光定量PCR检测D. dadantii的方法,为鉴定检疫甘薯茎腐病病原菌和田间监测病害提供了有效方法。

Bacterial stem and root rot of sweet potato caused by Dickeya dadantii is a new quarantine plant disease in China.Recent outbreaks of this disease threaten sweet potato production in China.Specific and sensitive detection of D.dadantii is critical for the pathogen identification,distribution monitoring and the disease control.Here,conserved signature genes specific to D.dadantii were found by the whole genome sequences analysis of the strains belonging to the genus Dickeya.Primers targeting to the conserved signature genes were designed and tested by PCR.One set of primers Dad1-F(5’-CATATCAACCAGACCAGCCGTT-3’)and Dad1-R(5’-CGGCCTGCTTTTAAACAACGTATTA-3’),which targets to a conserved signature gene encoding an unknown protein with the accession number WP077245517,specifically amplified a 167 bp fragment from D.dadantii by conventional PCR and real-time fluorescent quantitative PCR,providing a specific and sensitive method for identification and monitoring of D.dadantii in the field.