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Background   Plants have various defense mechanisms such as production of antimicrobial peptides, particularly pathogenesis related proteins (PR proteins). PR10 family is an essential member of this group, with antifungal, antibacterial and antiviral activities.Objective   The goal of this study is to assess the antifungal activity of maize PR10 against some of fungal phytopathogens.Materials and Methods    Zea mays PR10  gene (TN-05-147) was cloned from genomic DNA and cDNA and overexpressed in  Escherichia coli . The existence of a 77- bp intron and two exons in  PR10  was confi rmed by comparing the genomic and cDNA sequences. The  PR10  cDNA was cloned in pET26b (+) expression vector and transformed into  E. coli  strain Rosetta DE3 in order to express PR10 recombinant protein. Expression of the recombinant protein was checked by western analysis. Recombinant PR10 appeared as insoluble inclusion bodies and thus solubilized and refolded. PR10 was isolated using Ni- NTA column. The activity of the refolded protein was confi rmed by DNA degradation test. The antifungal activity of PR10 was assessed using radial diff usion, disc diff usion and spore germination. The hemolytic assay was performed to investigate the biosafety of recombinant PR10.Results   Recombinant maize PR10 exerted broad spectrum antifungal activity against   Botrytis cinerea, Sclerotinia sclerotiorum, Fusarium oxysporum, Verticillium dahlia   and  Alternaria solani . Hemolysis biosafety test indicated that the protein is not poisonous to mammalian cells.Conclusions   Maize PR10 has the potential to be used as the antifungal agent against diff erent fungal phytopathogens. Therefore, this protein can be used in order to produce antifungal agents and fungi resistance transgenic plants.

Cloning, Overexpression and in vitro Antifungal Activity of Zea Mays PR10 Protein