Transfer of the epsps gene to chickpea (Cicer arietinum) for resistance to glyphosate herbicide

Chickpea (Cicer arietinum) is very sensitive in competition with weeds and suffers from a decrease in yield and product quality. Therefore, it is useful to produce plants resistant to general herbicides such as glyphosate. In this research, Agrobacterium tumefaciens strain LBA4404 was used. This bacterium contains plasmid pBI121 and mutated epsps gene of Escherichia coli to create resistance to glyphosate herbicide. In this plasmid nptII gene is placed under 35S promoter as a selective gene. In order to transfer the gene, embryonic axes of chickpea varieties (Samin and Bivanij) were used. Chickpea embryonic axes were placed on B5 co-culture medium after co-cultivation with bacterial suspension for 3 days and then transferred to MS culture medium with appropriate concentration of BAP, Kin and NAA hormones for regeneration. Finally, after successive selections using kanamycin antibiotic, rooting and establishment of T0 plants were done in hydroponic environment. Then, PCR amplified epsps and nptII genes in 15 plants and their transgenecity was confirmed. Non-contamination of DNA in putative transgenic plants was confirmed by using the primers of the bacterial ITS regions. Also the expression of the target gene verified using RT-PCR test and the transgenic percentage was 0.75% in this study.