Genotyping of Bacillus spp. isolate collection from natural samples
2017
Draganić, Veselin (Obstetrics and Gynaecology Clinic Narodni front, Belgrade (Serbia)) | Lozo, Jelena (Faculty of Biology, Belgrade (Serbia)) | Biočanin, Marjan (Faculty of Biology, Belgrade (Serbia)) | Dimkić, Ivica (Faculty of Biology, Belgrade (Serbia)) | Garalejić, Eliana (Obstetrics and Gynaecology Clinic Narodni front, Belgrade (Serbia)) | Fira, Đorđe (Faculty of Biology, Belgrade (Serbia)) | Stanković, Slaviša (Faculty of Biology, Belgrade (Serbia)) | Berić, Tanja (Faculty of Biology, Belgrade (Serbia))
The aim of this study was genotyping and identification of collection of 164 Bacillus spp. isolates, from samples of soil, manure, and straw gathered from across Serbia, using Pulse field gel electrophoresis (PFGE) combined with sequencing of tuf gene, one of the housekeeping genes. The PFGE analysis with NotI enzyme was used to determine phylogenetic relationships of isolates and referent strains. Four large groups of Bacillus spp. were distinguishable: cereus, subtilis, pumilus and megaterium and within enormous genetic diversity. Bacillus subtilis Marburg referent strain did not group with rest of the strains from the subtilis group (Bacillus subtilis ATCC6633 and Bacillus atrophaeus ATCC9372). Strains from the cereus group were distinguished and closely grouped together. One representative isolate from each of 21 distinct PFGE groups was identified by sequencing of tuf gene. Eight different species were identified among chosen isolates: B. amyloliquefaciens, B. subtilis, B. pumilus, B. safensis, B. megaterium, B. cereus, B. anthracis and B. thuringiensis. Our results showed that PFGE analysis combined with sequencing of one of the housekeeping genes could be used for characterization of large collections of Bacillus isolates. The determination of tuf gene recommended itself to be an adequate and sufficient analysis for obtaining very clear and unambiguous results, with high resolution of separation of Bacillus species.
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