Detection of Pectobacteria Causal Agents of Potato Soft Rot in North Western Provinces of Iran

Abstract

Various bacteria belonging to Enterobacteriaceae are known to be agents causing blackleg and soft rot in potato. In this study, a rapid and specific detection method was used to investigate bacteria in some northwestern provinces of Iran (Zanjan, Kordestan and eastern Azarbaijan). 26 strains were selected for the study that had been identified in previous studies as representative pathogens, they were as follows: 14 Pectobactrium carotovorum subsp carotovorum, 7 Dickeya chrysanthemi and 5 Pectobactrium atrosepticum. Primers Y1/Y2, Expccf/r, ADE1/2 and Eca1f/r, published as specific primers for the pathogens. They were tested to determine if they could be used for specific amplification of DNA in the strains. The expected specific fragment was not amplified in many strains or several non-specific bands, some close to those expected fragments were amplified. Specific primers for amplification of the DNA of pathogens were designed by sequencing the ITS region from representative strains and a PCR test was developed. In the PCR test a 300 bp fragment was amplified from all strains collected from the provinces. In specificity tests, no PCR products were obtained from other bacteria belonging to other genera.

Detection and identification of bacterial soft rot of potato Pectobacterium carotovorum subsp. carotovorum using specific PCR primers in Jordan

“…Our results are also in agreement with previous finding of Azadmanesh et al (2013) where none of the 12 Iranian Pcc tested isolates produced the 550 bp products in PCR in contrast to two standard Pcc isolates that produced the desired bands and they found that these 500 bp M 1 2 3 4 5 6 7 8 9 10 C+ C-B 380 bp 500 bp bp 380 bp bpbpb p M 1 2 3 4 5 6 7 8 9 10 11 C+ B two isolates could not be identified by PCR using Pectobacterium subsp. specific primers.…”

“…Also, Kang et al (2003) found that only genomic DNA of 29 strains of Pcc out of 54 bacterial strains which is equal to about 54% yielded the expected 550 bp amplified product following PCR with EXPCC specific primers. These results could be due to sequence variation among strains of Pectobacterium isolated from different regions of the world (Azadmanesh et al, 2013). Whereas the speciesspecific EXPCC was generated from the nucleotide sequence of a Pcc universal rice primer (URPs) PCR product, although URPs were developed from repetitive sequences in the rice genome that have been used to fingerprint genomes of diverse organisms (Kang et al, 2003).…”

PCR-identification of bacteria belonging to the genus Pectobacterium – agents of cucumber soft rot and wilting in Ukraine

“…Тому, останнім часом для швидкої і коректної діагностики збудників використовується підходи, що базуються на полімеразно-ланцюговій реакції (ПЛР). Порівняно з іншими ці методи значно спрощують і підвищують чутливість виявлення збудника та базуються на ампліфікації конкретної цільової нуклеотидної послідовності, яка є унікальною для даного виду бактерій [2,5]. Так Darrasse A et al, сконстру-йовано специфічні праймери (Y1, Y2) для детектування pel-гену відповідального за синтез пектатліази у наступних видів пектолітичних бактерій P. carotovorum (subsp.…”

“…Поєднання розроблених Darrasse A et та S.H. de Boer з колегами методик застосовується для ідентифікації представників видів P. carotovorum і P. atrosepticum [5].…”

Identification and genetic diversity of Jordanian potato soft rot isolates, Pectobacterium carotovorum subspecies carotovorum (DYE 1969)

“…Within the past 30 years, there has been a shift from microbiological and serological methods to molecular approaches (PalacioBielsa et al, 2009;Czajkowiski et al, 2014). DNA markers have been used for the rapid detection of different strains of P. carotovorum subspecies carotovorum (Pcc) (Kang et al, 2003;Zhu et al, 2010;Rahmanifer et al, 2012;Azadmanesh et al, 2013). DNA sequence analysis of gene pmrA, which is linked to pathogenicity (response regulator), was used to evaluate the relationship among Pcc isolates collected from different regions of Morocco and results were equivalent to enterobacterial repetitive intergenic consensuspolymerase chain reaction (ERIC-PCR) and 16S rDNA sequencing (Kettani-Halabi et al, 2013).…”