Fusion of a chitin-binding domain to an antibacterial peptide to enhance resistance to Fusarium solani in tobacco (Nicotiana tabacum )

3 Biotech
Azam BadrhadadAhmad Ismaili

Abstract

An antibacterial peptide-encoding gene from alfalfa seeds, alfAFP, was fused to the C-terminal part of chitin-binding domain (CBD) of the rice chitinase-encoding gene (CBD-alfAFP) and introduced to tobacco by Agrobacterium-mediated transformation. Polymerase chain reaction (PCR) technique was used to confirm the integration of the recombinant CBD-alfAFP encoding gene in transgenic tobacco plants. A number of transgenic lines and a non-transgenic control plant were selected for further molecular analyses. The result of analyzing the transgenic plants by semi-quantitative RT-PCR showed that the recombinant gene is expressed in transgenic plants and there is a difference between the transgenic plants in terms of the level of CBD-alfAFP expression. The total protein was extracted from a few selected transgenic plants and used to evaluate the antibacterial/antifungal of recombinant protein activity against some important plant and human pathogens. The results of this experiment showed that the total protein extract obtained from transgenic lines significantly (P < 0.05) inhibited the growth of various bacteria and fungi compared to the non-transgenic plants. Transgenic lines showed a significant (P < 0.01) difference considering the...Continue Reading

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