The genetic diversity of genus Bacillus and the related genera revealed by 16S rRNA gene sequences and ardra analyses isolated from geothermal regions of turkey

Abstract

Previously isolated 115 endospore-forming bacilli were basically grouped according to their temperature requirements for growth: the thermophiles (74%), the facultative thermophiles (14%) and the mesophiles (12%). These isolates were taken into 16S rRNA gene sequence analyses, and they were clustered among the 7 genera: Anoxybacillus, Aeribacillus, Bacillus, Brevibacillus, Geobacillus, Paenibacillus, and Thermoactinomycetes. Of these bacilli, only the thirty two isolates belonging to genera Bacillus (16), Brevibacillus (13), Paenibacillus (1) and Thermoactinomycetes (2) were selected and presented in this paper. The comparative sequence analyses revealed that the similarity values were ranged as 91.4-100 %, 91.8- 99.2 %, 92.6- 99.8 % and 90.7 - 99.8 % between the isolates and the related type strains from these four genera, respectively. Twenty nine of them were found to be related with the validly published type strains. The most abundant species was B. thermoruber with 9 isolates followed by B. pumilus (6), B. lichenformis (3), B. subtilis (3), B. agri (3), B. smithii (2), T. vulgaris (2) and finally P. barengoltzii (1). In addition, isolates of A391a, B51a and D295 were proposed as novel species as their 16S rRNA gene sequences displayed similarities ≤ 97% to their closely related type strains. The AluI-, HaeIII- and TaqI-ARDRA results were in congruence with the 16S rRNA gene sequence analyses. The ARDRA results allowed us to differentiate these isolates, and their discriminative restriction fragments were able to be determined. Some of their phenotypic characters and their amylase, chitinase and protease production were also studied and biotechnologically valuable enzyme producing isolates were introduced in order to use in further studies

Morphological, enzymatic screening, and phylogenetic analysis of thermophilic bacilli isolated from five hot springs of Myagdi, Nepal

“…Thus, all strains could be classified as thermophiles [38,39]. The temperature of hot springs was below 60°C except at Bhurung and Paudwar hot springs which show that any microbes do not necessarily have the same optimum temperature as their natural habitats [40].…”

Thermolongibacillus altinsuensis gen. nov., sp. nov. and Thermolongibacillus kozakliensis sp. nov., aerobic, thermophilic, long bacilli isolated from hot springs

“…Of those endospore-forming bacteria, 115 of the bacilli were subjected to 16S rRNA gene sequencing and their phylogenetic analyses revealed the presence of two novel thermophilic, endospore-forming, aerobic, long, rod-shaped bacteria: strains E265 T and E173a T , isolated from soil and sediment samples collected from two different hot springs of Altinsu and Kozakli provinces in Nevsehir. Although these new isolates showed distant phylogenetic relationships with some of the members of the genera Geobacillus and Anoxybacillus , they did not fit into any of the thermophilic genera previously described as belonging to the family Bacillaceae as they displayed very low (<95.8 %) 16S rRNA gene sequence similarities (Cihan et al , 2011, 2012). Therefore, they were taken into consideration for further analyses and this paper describes the taxonomic positions of strains E265 T and E173a T by presenting evidence on the level of phenotypic, chemotaxonomic, phylogenetic and genotypic characteristics with the proposal of the novel genus, Thermolongibacillus gen. nov., with two species, Thermolongibacillus altinsuensis sp.…”

Molecular characterization and PCR-based screening of cry genes from Bacillus thuringiensis strains

“…The PCR was performed with Taq polymerase for 30 cycles as follows: 94 °C for 1 min, 60 °C for 45 s and 72 °C for 1 min., the final extension was performed for 10 min at 72 °C. The 16S rDNA amplicons were digested using four different restriction endonucleases viz Hinf I, Hae III, Alu I and Taq I (Cihan et al 2012). The restriction-digested products were analyzed on 2% agarose gel prepared in 1× TAE buffer containing 0.5 µg/ml of ethidium bromide.…”