Characterization of Dickeya and Pectobacterium species by capillary electrophoretic techniques and MALDI-TOF MS
- 388 Downloads
Dickeya and Pectobacterium species represent an important group of broad-host-range phytopathogens responsible for blackleg and soft rot diseases on numerous plants including many economically important plants. Although these species are commonly detected using cultural, serological, and molecular methods, these methods are sometimes insufficient to classify the bacteria correctly. On that account, this study was undertaken to investigate the feasibility of three individual analytical techniques, capillary zone electrophoresis (CZE), capillary isoelectric focusing (CIEF), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), for reliable classification of Dickeya and Pectobacterium species. Forty-three strains, representing different Dickeya and Pectobacterium species, namely Dickeya dianthicola, Dickeya dadantii, Dickeya dieffenbachiae, Dickeya chrysanthemi, Dickeya zeae, Dickeya paradisiaca, Dickeya solani, Pectobacterium carotovorum, and Pectobacterium atrosepticum, were selected for this purpose. Furthermore, the selected bacteria included one strain which could not be classified using traditional microbiological methods. Characterization of the bacteria was based on different pI values (CIEF), migration velocities (CZE), or specific mass fingerprints (MALDI-TOF MS) of intact cells. All the examined strains, including the undetermined bacterium, were characterized and classified correctly into respective species. MALDI-TOF MS provided the most reliable results in this respect.
KeywordsDickeya Pectobacterium CZE CIEF MALDI
This work was supported by a grant from the Ministry of the Interior of the Czech Republic (No. VG20112015021) and with the institutional support (RVO: 68081715).
- 1.Samson R, Legendre JB, Christen R, Fishcer-Le Saux M, Achouak W, Gardan L (2005) Transfer of Pectobacterium chrysanthemi (Burkholder et al. 1953) Brenner et al. 1973 and Brenneria paradisiaca to the genus Dickeya gen. nov. as Dickeya chrysanthemi comb. nov. and Dickeya paradisiaca comb. nov. and delineation of four novel species, Dickeya dadantii sp. nov., Dickeya dianthicola sp. nov., Dickeya dieffenbachiae sp. nov. and Dickeya zeae sp. nov. Int J Syst Evol Microbiol 55:1415–1427CrossRefGoogle Scholar
- 12.Van der Wolf JM, Nijhuis EH, Kowalewska MJ, Saddler GS, Parkinson N, Elphinstone JG, Pritchard L, Toth IK, Lojkowska E, Potrykus M, Waleron M, de Vos P, Cleenwerck I, Pirhonen M, Garlant L, Hélias V, Pothier JF, Pflüger V, Duffy B, Tsror L, Manulis S (2014) Dickeya solani sp. nov., a pectinolytic plant-pathogenic bacterium isolated from potato (Solanum tuberosum). Int J Syst Evol Microbiol 64:768–774CrossRefGoogle Scholar
- 13.Janse JD (2012) Bacterial diseases that may or do emerge, with (possible) economic damage for Europe and the Mediterranean basis: notes on epidemiology, risk, prevention and management on first occurrence. J Plant Pathol 94:S4.5–S4.29Google Scholar
- 16.Sasser M (1990) Technical note 102: tracking a strain using the Microbial Identification System. MIS, North NewarkGoogle Scholar
- 20.Slawiak M, van Doorn R, Szemes M, Speksnijderm AGCL, Waleron M, van der Wolf JM, Lojkowska E, Schoen CD (2013) Multiplex detection and identification of bacterial pathogens causing potato blackleg and soft rot in Europe, using padlock probes. Ann Appl Biol 163:378–393Google Scholar
- 23.Šalplachta J, Kubesová A, Moravcová D, Vykydalová M, Süle S, Matoušková H, Horký J, Horká M (2013) Use of electrophoretic techniques and MALDI-TOF MS for rapid and reliable characterization of bacteria: analysis of intact cells, cell lysates, and “washed pellets”. Anal Bioanal Chem 405:3165–3175CrossRefGoogle Scholar
- 24.Horká M, Šalplachta J, Karásek P, Kubesová A, Horký J, Matoušková H, Šlais K, Roth M (2013) Combination of capillary isoelectric focusing in a tapered capillary with MALDI-TOF MS for rapid and reliable identification of Dickeya species from plant samples. Anal Chem 85:6806–6812CrossRefGoogle Scholar
- 25.Subirats X, Blaas D, Kenndler E (2011) Recent developments in capillary and chip electrophoresis of bioparticles: viruses, organelles, and cells. Electrophoresis 32:1579–1590Google Scholar
- 40.Nassar A, Darrasse A, Lemattre M, Kotoujansky A, Dervin C, Vedel R, Bertheau Y (1996) Characterization of Erwinia chrysanthemi by pectinolytic isoenzyme polymorphism and restriction fragment length polymorphism analysis of PCR-amplified fragments of pel genes. Appl Environ Microbiol 62:2228–2235Google Scholar