Genetic Grouping of Certain Rhizobial Strains Native to Iranian Soils by 16S-23S IGS PCR-RFLP and Plasmid Profile Techniques

Document Type : Research Paper

Authors

1 Assistant Professor,Department of Soil Science, Faculty of Soil and Water Science Engineering, University of Tehran, Karadje, Iran

2 Assistant Professor, National Institute for Genetic Engineeing and Biotechnology, Tehran, Iran

3 Professor, Agriculture Faculty Instructor of Laval university, Canada

Abstract

Among prokaryotes, there are some soil bacteria called Rhizobia that are very important owing to their N2 fixation ability in legumes. The sequence between two 16S and 23S fragments of rDNA genes complex is called 16S-23S rDNA IGS. It differs by size and nucleotide order, therefore, it is very useful for grouping of rhizobial strains. In this research 16S-23S IGS PCR-RFLP and Plasmid Profile (modified Eckhardt method) grouped 52 superior strains which were found as plant growth promoting Rhizobacteria (PGPR). For cluster analysis, phylip version 3.6a3 software was used. The results show that the16S-23S IGS fragments of different rhizobial strains are very diversified in terms of size (600-1486 bp), and the number of IGS fragment differs from 1 to 3 copies. Based on 16S-23S IGS PCR-RFLP method, all 52 rhizobial strains were located in 46 groups, of which 11 groups had 70% similarity. By this method all strains (77%) except 12 strains (Bj53, Bj54; Rlv27, Rlv28; Rlv23, Rlv24; Rlp16, Rlp17; Sm10, Sm11, Sm12, and Sm13) were separated into different groups. Therefore, the mentioned method is very reliable and practical, besides being simple and having few laboratory requirements (in comparison with other molecular methods). In this research, The grouping based on Plasmid Profile of different rhizobial strains agrees with the results of IGS-RFLP to some extent, for example: Bj53, Bj54; Rlv23, Rlv24 also Rlp16, Rlp17 and Sm11, Sm12, Sm13 in both methods were completely similar and were located in separate cluster groups. 

Keywords

References

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Iranian Journal of Soil Research
Volume 21, Issue 1
May 2007
Pages 89-100

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