Identification of IS elements in Acidithiobacillus ferrooxidans strains grown in a medium with ferrous iron or adapted to elemental sulfur.

IS elements were identified in the genomes of five Acidithiobacillus ferrooxidans strains isolated from various media. IST2 elements were revealed in all the strains grown in a medium with ferrous iron, ISAfe1 elements were detected in four strains (TFBk, TFL-2, TFV-1 and TFO). Three strains (TFV-1, TFN-d and TFO) were found to contain IS elements, approximately 600 bp long. These were named preliminary as ISAfe600. Partial sequencing of the 5'- and 3'-terminal nucleotide stretches of an ISAfe1 element in TFBk and TFL-2 strains and complete sequencing of the ISAfe1 element in the TFBk strain has revealed nucleotide substitutions as compared to the prototype, i.e., the ISAfe1 element of an ATCC 19859 strain. Partial sequencing of the 5'- and 3'-terminal nucleotide stretches of the IST2 elements in TFO, TFBk and TFL-2 strains has shown numerous nucleotide substitutions when compared to the IST2 element of an ATCC 19859 strain. Complete sequencing of the IST2 element in the TFBk strain has revealed: the divergence between the IST2 elements in the TFBk strain and the prototype was 21.2%. Southern hybridization of EcoRI fragments of the chromosomal DNA from five A. ferrooxidans strains grown in a medium with ferrous iron using an internal region of ISAfe1, a full-length ISAfe1 or a full-length IST2 as probes has shown them to differ in the number of copies of IS elements and their localization on the chromosomes. Adaptation to elemental sulfur in A. ferrooxidans strains caused changes in the number, intensity and localization of hybridization bands. The authors discuss the role of IS elements in the adaptation of A. ferrooxidans to the new energy substrate.

Phylogenetic heterogeneity of the species Acidithiobacillus ferrooxidans.

Polyphasic genotypic analysis of 25 Acidithiobacillus ferrooxidans strains isolated from ores and ore concentrates collected in different regions of the world showed considerable strain heterogeneity. Restriction patterns of the chromosomal DNA of these strains obtained by PFGE were specific for each strain. According to the degree of DNA relatedness, 17 of the 23 strains studied were divided into four genomovars. Six independent, considerably divergent strains could not be assigned to any of the genomovars. A comparison of nearly complete nucleotide sequences of the 16S rDNA of five representatives of the genomovars (including the type strain of A. ferrooxidans, ATCC 23270T) with those of species of the genus Acidithiobacillus available from GenBank showed that most of the A. ferrooxidans strains, together with the type strain and some other strains of the species Acidithiobacillus thiooxidans, comprised a monophyletic cluster. Within this major cluster, A. ferrooxidans strains fell into four phylogenetic groups that were equidistant from the phylogenetic group of A. thiooxidans strains. In general, the distribution of strains among the phylogenetic groups correlated with their distribution among the genomovars, except that the representatives of two different genomovars fell into one phylogenetic group. Thus, at least two levels of phylogenetic heterogeneity for A. ferrooxidans have been found. The phylogenetic heterogeneity of A. ferrooxidans strains, which are phenotypically indistinguishable, suggests the occurrence of microevolutionary processes in different econiches. This should be taken into account in the biohydrometallurgical applications of A. ferrooxidans strains.