Identificação de Pectobacterium carotovorum subsp. brasiliensis através de PCR-RFLP do gene recA

Tassa, Samira O. M. El; Duarte, Valmir

doi:10.1590/s0100-41582006000100004

Cited by 9 publications

(2 citation statements)

References 12 publications

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“…Knowledge of diversity is an important prerequisite for the identification of phytobacteria, as well as for taxonomic classifications, to support epidemiological studies and the development of strategies for plant diseases control, especially regarding selection of resistant varieties in breeding programs. This is particularly important when different closely related species and subspecies cause diseases in the same host, as is the case of pectobacteria in potatoes (EL TASSA; DUARTE, 2006).…”

Section: Resultsmentioning

confidence: 99%

Diversity of Pectobacterium strains by biochemical, physiological, and molecular characterization

et al. 2020

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Pectobacterium is a complex taxon of strains with diverse characteristics. It comprises several genera, including Erwinia, Brenneria, Pectobacterium, Dickeya, and Pantoea. Pectobacterium and Dickeya cause diseases in a wide range of plants, including potatoes, where they are causative agents of soft rot in tubers and blackleg in field-grown plants. Characterizing Pectobacterium species allows for the analysis of the diversity of pectinolytic bacteria, which may support control strategies for plant bacterial diseases. The aim of this study was to perform biochemical, physiological, and molecular characterizations of Pectobacterium spp. from different sites and host plants. The isolated strains were characterized by the glucose fermentation test, Gram staining, catalase activity, oxidase activity, growth at 37 ºC, reducing substances from sucrose, phosphatase activity, indole production, acid production from different sources (sorbitol, melibiose, citrate, and lactose), pathogenicity in potato, and hypersensitivity reactions. Molecular characterization was performed with species-specific primers ECA1f/ECA2r and EXPCCF/EXPCCR, which identify P. atrosepticum and P. carotovorum subsp. carotovorum (Pcc), respectively, and with primers 1491f/L1RA/L1RG and Br1f/L1RA/L1RG that differentiate Pcc from Dickeya chrysanthemi and from P. carotovorum subsp. brasiliensis. The strains were identified as belonging to the genus Pectobacterium, though they did not fit the biochemical nor the molecular classification standards for subspecies differentiation, indicating significant diversity among the strains.

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Section: Resultsmentioning

confidence: 99%

Diversity of Pectobacterium strains by biochemical, physiological, and molecular characterization

et al. 2020

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“…Although 16S rDNA is relatively short and contains fewer phylogenetically informative characters than housekeeping genes often used in taxonomical studies (Ma et al , ), this gene sequences have several advantages, including being a highly conserved multicopy gene and easy to amplify. Genes encoding for rRNAs, heat‐shock proteins, ATPases, RNA polymerase and housekeeping genes, acnA , gapA , icdA , mdh , mtlD , pgi , proA and recA , have been successfully used to delineate phylogenetic relationships among bacteria (Ludwig & Schleifer, ; Santos & Ochman, ; El Tassa & Duarte, ; Ma et al , ). Waleron et al () demonstrated that the restriction fragment length polymorphism (RFLP) analysis of recA gene can act as a useful marker for identification of Pectobacterium and Dickeya , at both species and subspecies levels.…”

Section: Introductionmentioning

confidence: 99%

Multiplex detection and identification of bacterial pathogens causing potato blackleg and soft rot in Europe, using padlock probes

Slawiak

Doorn²,

Szemes

et al. 2013

Annals of Applied Biology

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The objective of this study was to develop a multiplex detection and identification protocol for bacterial soft rot coliforms, namely Pectobacterium wasabiae (Pw), Pectobacterium atrosepticum (Pba) and Dickeya spp., responsible for potato blackleg and tuber soft rot. The procedures were derived from the phylogenetic relationships of these and other Enterobacteriaceae based on recA sequences. The group of Pw strains was highly homogeneous and could be distinguished from the other species. A ligation-based method for detection of Pw was developed. Five padlock probes (PLPs) were designed, targeting recA sequences to identify the Pw, Pba or Dickeya spp., whereas a sixth probe recognised recA sequences of all soft rot coliforms including Pectobacterium carotovorum subsp. carotovorum (Pcc). Two PLP-based applications were developed: one using real-time PCR and one using universal microarrays. Assay sensitivity and specificity were demonstrated using 71 strains of Pw, Pcc, Pba and Dickeya spp. Both multiplex methods can be potentially used for seed testing and in ecological studies, but further validation is required.

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PCR-based methods for detection of Erwinia psidii on guava

Silva

Torres

Oliveira

et al. 2015

Trop. plant pathol.

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Erwinia psidii causes bacterial blight of guava (Psidium guajava), one of the most important diseases of this crop in Brazil. Control measures are not effective, and dissemination often occurs through contaminated but asymptomatic propagating plant material. Considering the need for a reliable and sensitive method for detecting the pathogen in asymptomatic plant material, E. psidii-specific PCR primers were designed from a 355-bp fragment of the recombinase A gene (recA) amplified from E. psidii type strain. Primer pair Ep2L/2R only amplified DNA from E. psidii and its detection limit was 10 −5 ng/μL of purified DNA and 10 CFU (colony forming units) of bacterial cell suspension/mL. Three methods, conventional PCR, IC-PCR, and BIO-PCR were evaluated with the selected primers for their potential to detect E. psidii on guava leaves. BIO-PCR and conventional PCR were more sensitive and less time-consuming than IC-PCR. The detection limits on extracts of macerated guava leaves spiked with bacterial suspensions at different concentrations were 10 and 10 3 CFU/mL for BIO-PCR and conventional PCR, respectively. The PCR method here described could be useful for developing a protocol for early detection of this pathogen in asymptomatic guava plants.

show abstract

Identificação de Pectobacterium carotovorum subsp. brasiliensis através de PCR-RFLP do gene recA

Cited by 9 publications

References 12 publications

Diversity of Pectobacterium strains by biochemical, physiological, and molecular characterization

Diversity of Pectobacterium strains by biochemical, physiological, and molecular characterization

Multiplex detection and identification of bacterial pathogens causing potato blackleg and soft rot in Europe, using padlock probes

PCR-based methods for detection of Erwinia psidii on guava

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