In this study, the DNA fragment (AFPFP) including antifungal gene AFPF and specific promoter AFP was successfully isolated from genomic DNA of chickpea (Cicer arietinum L.) by PCR techniq4e. This fragment was cloned in plasmid pGEM-T Easy and sequenced. The sequencing of AFPFP fragment was totally matched to sequences submitted on genbank Acc. Number DQ342338. The fragment was 0.9 kb in length, in which specific promoter was 596 bp and antifungal gene was 304 bp. AFPF gene was included 2 exons (58 bp and 167 bp in length, respectively) and 1 intron (79 bp), encoding for a peptide of 74 amino acid. Plant expression vectors pCAMBIA1300polyA AFPFP and pCAMBIA1300polyA AFPFP were constructed by two steps: firstly, the fragment of 355 polyA derived from pFF19 was cloned in pCAMBIA1300 and pCAMBIA2300
secondly, the AFPFP fragment was cloned in these vectors at SalI. site. Number of copy and orientation of insertion were checked by EcolR. restriction enzyme digestion. Plant expression vectors were transferred in Agrobacterium. A tumefaciens LBA4404 harboring pCAMBIA1300polyA AFPFP was used for transformation in tobacco. Molecular analysis indicated that antifungal gene under promoter AFP control was integrated in genomic DNA and overexpressed in To transgenic tobacco clones. Analysis of bioassays in vitro demonstrated a significantly enhanced resistance of the transgenic tobacco lines to fungal pathogen Altemaria altemata. Thus, constructed plant expression vectors harboring antifungal gene and specific promoter could be used for transformation in other crops.