Open AccessGenotyping of Bacillus spp. isolate collection from natural samples
Author(s) -
Veselin Draganic,
Jelena Lozo,
Maijan Biocanin,
Ivica Dimkić,
Eliana Garalejić,
Djordje Fira,
Slavka Stanković,
Tanja Berić
Publication year - 2017
Publication title -
genetika
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.24
H-Index - 15
eISSN - 1820-6069
pISSN - 0534-0012
DOI - 10.2298/gensr1702445d
Subject(s) - biology , pulsed field gel electrophoresis , bacillus pumilus , genotyping , housekeeping gene , microbiology and biotechnology , bacillus anthracis , bacillus amyloliquefaciens , cereus , bacillus subtilis , genetics , bacillus cereus , genotype , gene , bacteria , gene expression
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.