"Biotechnology" journal V. 5, No. 3, 2012
Р. 53-61, Bibliography 32, Ukrainian.
Universal Decimal classification: 576.871.155. 557
Institute of Plant Physiology and Genetics of National Academy of Sciences of Ukraine, Kyiv
Institute of Molecular Biology and Genetics of National Academy of Sciences of Ukraine, Kyiv
Plazmid vector pSUP2021::Tn5 for transposon mutagenesis in Rhizobium leguminosarum bv. trifolii strain 348a was used. The frequency of KmR-mutants formation after transfer this vector from Escherichia coli to Rhizobium was 1.9 • 10–7. The presence of Tn5 in mutants DNA was verified using polymerase chain reaction. By the symbiotic traits and Eff++ -phenotype of clover plants the Tn5-mutants strains were screened in vegetative and field conditions and rhizobia with increased synthesis of nitrogenase and economical valuable properties were selected. Tn5-mutants T2k, T28k, T72k were significantly higher than other rhizobia by the nodulation activity. In the phase of clover flowering on roots of plants through their involvement it was formed 1.4–1.7 times as much nodules which dominated by mass. Aboveground plant weight increased by 29,6–41,8% at strengthening ryzohenezis. Intensification of physiological processes in clover ensured by increase of nitrogenase activity of symbiotic system which in the early flowering phase exceeded at an average of 2.5 times the results of plants inoculated by the original (parent) strain 348a. Obtained data indicate the possibility of reating highly efficient symbiotic systems clover — Tn5-mutants Rhizobium leguminosarum bv. trifolii.
Key words: transposone mutagenesis, Tn5-mutants, polymerase chain reaction, clover, symbiosis, nitrogen-fixing activity, efficiency.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008