{"title":"Development of a species-specific PCR assay for identification and detection of Dickeya solani","authors":"Alireza Salimi Darani, Masoud Bahar","doi":"10.1007/s13313-023-00935-7","DOIUrl":null,"url":null,"abstract":"<div><p><i>Dickeya solani</i> associated with other members of <i>Pectobacterium</i> spp. and <i>Dickeya</i> spp. (Pectobacteria) causes the destructive potato blackleg in the Iranian province of Isfahan. The seed tuber dispersal ability and better adaptation of <i>D. solani</i> to warm climates raise major concerns about the spread and establishment of the pathogen across the country. Therefore, monitoring of the target pathogen on potato seeds and diseased plants is necessary to make effective decisions to limit the spread of the pathogen. However, the similarity in symptom development and phenotypic characteristics of Pectobacteria makes it difficult to distinguish <i>D. solani</i> in the collected samples. The aim of the present study was to develop a reliable PCR-based method for the specific detection of <i>D. solani</i> in naturally infected samples. In ERIC-PCR genotyping of different species and subspecies of Pectobacteria, a distinct PCR product around 950 bp was amplified only in <i>D. solani</i> strains. The amplicon was cloned, sequenced, analyzed and found to be highly homologous to the <i>ugpC-1</i> gene sequence. Based on the sequenced fragment, a primer pair (DSF1/DSR2) was designed that allowed the specific amplification of a 895 bp band from the isolates of <i>D. solani</i>, but not from non-target Pectobacteria and other bacterial species tested in the present study. The approximate limit of detection of the PCR assay has been estimated to be approximately 2.7 × 10<sup>4</sup> CFU/ml. The developed <i>D. solani</i> specific PCR assay can likely serve as a valuable method for various purposes.</p></div>","PeriodicalId":8598,"journal":{"name":"Australasian Plant Pathology","volume":null,"pages":null},"PeriodicalIF":0.9000,"publicationDate":"2023-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Australasian Plant Pathology","FirstCategoryId":"97","ListUrlMain":"https://link.springer.com/article/10.1007/s13313-023-00935-7","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
引用次数: 0
Abstract
Dickeya solani associated with other members of Pectobacterium spp. and Dickeya spp. (Pectobacteria) causes the destructive potato blackleg in the Iranian province of Isfahan. The seed tuber dispersal ability and better adaptation of D. solani to warm climates raise major concerns about the spread and establishment of the pathogen across the country. Therefore, monitoring of the target pathogen on potato seeds and diseased plants is necessary to make effective decisions to limit the spread of the pathogen. However, the similarity in symptom development and phenotypic characteristics of Pectobacteria makes it difficult to distinguish D. solani in the collected samples. The aim of the present study was to develop a reliable PCR-based method for the specific detection of D. solani in naturally infected samples. In ERIC-PCR genotyping of different species and subspecies of Pectobacteria, a distinct PCR product around 950 bp was amplified only in D. solani strains. The amplicon was cloned, sequenced, analyzed and found to be highly homologous to the ugpC-1 gene sequence. Based on the sequenced fragment, a primer pair (DSF1/DSR2) was designed that allowed the specific amplification of a 895 bp band from the isolates of D. solani, but not from non-target Pectobacteria and other bacterial species tested in the present study. The approximate limit of detection of the PCR assay has been estimated to be approximately 2.7 × 104 CFU/ml. The developed D. solani specific PCR assay can likely serve as a valuable method for various purposes.
期刊介绍:
Australasian Plant Pathology presents new and significant research in all facets of the field of plant pathology. Dedicated to a worldwide readership, the journal focuses on research in the Australasian region, including Australia, New Zealand and Papua New Guinea, as well as the Indian, Pacific regions.
Australasian Plant Pathology is the official journal of the Australasian Plant Pathology Society.