Genetic Fingerprint To Study Relatedness Of Acidithiobacillus Ferrooxidans Strains

Acidithiobacillus ferrooxidans has worldwidely used in the field of bioleaching as an important microorganisms metallurgy bacteria. Althought Acidithiobacillus ferrooxidans has been deeply studied, strains are difficult to be seperated and purified by its nature, and to make rapid identification within species in traditional methods.This paper studies the the maximum tolerated ability of the20strains of A.ferrooxidans to copper ions, and make phenotypic classification. The results showed that the copper ions maximum tolerated concentration(MTC) of the20strains is from60mmol/L to200mmol/L variedly. The20Acidithiobacillus ferrooxidans strain tolerance in the order of DY26, DY30, DY32, GF, BY-01, BY-0502, GD-A, GD-B, GD-9, DXS, DY15, DY18, JYC-2, JYC-3, DX5, YTW, JYC-1, BY-03, DC, GD-0. High copper ions tolerated strains descending in the order of DY26, DY30, DY32, GF, BY-01, BY-0502, GD-A, GD-B, GD-9, DXS, DY15, DY18, JYC-2, JYC-3, while the MTC range from160mmol/L to200mmol/L. Low copper ions tolerated strains descending in the order of DX5, YTW, JYC-1, BY-03, DC, GD-0, while the MTC range from60mmol/L to120mmol/LTo study the genome diversity of23Acidithiobacillus ferrooxidans strains in China, and the relationship between the genome classification and physiological-biochemical characteristic of Acidithiobacillus ferrooxidans, DNA was amplified by PCR and40random primers, amplification products were separated by gel electrophoresis and photographed under UV light. The RAPD (Randomly Amplified Polymorphic DNA) profiles were compared on the basis of the presence or absence of each DNA band, and strain diversity was calculated using the Jaccard’s coefficient and UPGMA (Unweighted Pair-Group Arithmetic Average Clustering) cluster analysis. A total of518polymorphic bands were generated. Similarities between the strains ranged from42.8%to92.1%. OPC primers produce more specific genetic fingerprinting, and are better in classification and identification of strains of Acidithiobacillus ferrooxidans. In the strains with high MTCs, similarity range from61.2%to91.4%, while in the strains with low MTCs, similarity range from53.3%to88.8%.To study the possibility for rep-PCR to classify and identificate of strains of Acidithiobacillus ferrooxidans, two primers,(GTG)s and BOXA1R, are used in this study. Primer (GTG)5produces a total of12reproducable electrophoretic bands in different lengh, and there are no non-specific bands among the23strains. Bands are range from0.2kb to1.6kb. Primer BOXA1R produces a total of13reproducable electrophoretic bands in different lengh, and there are no non-specific bands among the23strains. Bands are range from0.3kb to1.4kb.Results above show that23strains have a high degree of genomic diversity and the RAPD markers related to characteristics such as metal resistance and geographical locations. Rep-PCR can be used in genotype classification of Acidithiobacillus ferrooxidans strains within a certain range, but is weak in the identification of strains. These studies can provide a theoretical basis for further standardized genetic fingerprint of technical conditions, and developing into a fast identification strains of Acidithiobacillus ferrooxidans.