Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology, Beijing 100097, People's Republic of China.
Plant Dis. 2020 Apr;104(4):1158-1166. doi: 10.1094/PDIS-05-19-1033-RE. Epub 2020 Feb 13.
, a causal agent of vegetable soft rot, contains three valid subspecies: subsp. (), subsp. (), and subsp. (). Using 16S rDNA sequencing and genus-specific PCR, we identified 72 strains from Chinese cabbage, bok choy, and celery and assessed their pathogenicity on Chinese cabbage petioles and potato tubers. Based on phylogenetic analysis of sequences and confirmation by subspecies-specific PCR, the strains were divided into 18 , 29 , and 25 Several characteristic features were also assessed and supported the distinctiveness of the strains. All strains caused soft rot symptoms on Chinese cabbage and potato, but the strains exhibited the greatest severity. We developed a conventional PCR and a quantitative PCR (qPCR) assay for the identification of based on its specific gene encoding sorbitol-specific phosphotransferase. These two methods could specifically amplify the expected products of 674 and 108 bp, respectively, from all of the strains. The assays demonstrated high sensitivity and could detect as little as 1 and 100 pg/µl of bacterial genomic DNA, respectively. Both assays could also detect the pathogens directly from plant tissues infected with as little as 2.5 × 10 CFU/mg of , even before external symptoms appeared. These assays constitute effective tools for disease diagnosis and the rapid identification of soft rot pathogens.
软腐欧文氏菌是一种引起蔬菜软腐病的病原菌,包含三个有效亚种:subsp. ()、subsp. ()和 subsp. ()。我们使用 16S rDNA 测序和属特异性 PCR 从白菜、油菜和芹菜中鉴定出 72 株菌株,并评估了它们对白菜叶柄和土豆块茎的致病性。根据序列的系统发育分析和亚种特异性 PCR 的确认,这些菌株分为 18 个、29 个和 25 个 。还评估了一些特征,这些特征支持了 菌株的独特性。所有菌株都能引起白菜和土豆的软腐病症状,但 菌株的严重程度最高。我们基于其特异性编码山梨醇特异性磷酸转移酶的 基因,开发了一种常规 PCR 和一种定量 PCR(qPCR)检测方法,用于鉴定 。这两种方法都可以分别从所有 菌株中特异性扩增预期的 674 和 108 bp 产物。这些检测方法具有很高的灵敏度,分别可以检测到低至 1 和 100 pg/µl 的细菌基因组 DNA。这两种检测方法也可以直接从感染量低至 2.5×10 CFU/mg 的植物组织中检测到病原体,甚至在出现外部症状之前。这些检测方法构成了疾病诊断和快速鉴定软腐病病原菌的有效工具。