INSERTION OF CHITINASE GENE TO ATTENUATE EARLY BLIGHT DISEASE IN SOME POTATO VIRUS RESISTANT LINES

Abstract

Potato (Solanum tuberosum L.) an agro-economically important food crop in the world, is sensitive to many fungal pathogens including Alternaria solani, the causal agent of early blight disease. Chitinase is cell wall degrading enzyme which has been shown to have high antifungal activity against a wide range of phytopathogenic fungi. In the present study, plasmid pRI 201-AN binary vector, containing the kanamycine selectable marker in plant and the chitinase gene was used in potato transformation. Leaves of Desriee cultivar, PVY5 and PVY15 lines were transformed with the pRI 201-AN construct via the Agrobacterium tumefaciens delivery system. Transformed leaves were incubated for 5 days in dark on callus induction media which contained MS with 5 mg/l 2-4, D, 1 mg/l cefatoxine (200mg/ml) and 1 mg/l kanamycine (25mg/ml). After that callus was transferred to regeneration media which contained MS with 1 mg/l IAA, 1 mg/l BA, 10 mg/l GA3, 1 mg/l cefatoxine (200mg/ml) and 250 µg/l kanamycine (25mg/ml) and their expression at the transcriptional level was confirmed by polymerase chain reaction (PCR) by using chitinase and vector primers. After the transformed plants were evaluated, the positive transgenic plants were detected by using forward and reverse primer of kanamycine.