Transformation of Canola (R line Hyola 308) by pgip1 Gene from Bean cv. Daneshjoo to Improve Resistance to Sclerotina sclerotiorum

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During plant infection, fungal pathogens secret polygalacturonase (PG) that are capable of degrading cell wall of susceptible plant tissues. Polygalacturonase-inhibiting proteins (PGIPs) are able to specifically inhibit fungal polygalacturonases (PGs) activity. The inhibition of fungal PGs by PGIPs suggests that PGIPs have a role in plant tolerance to fungal infections. In this study bean (Daneshjoo cultivar) pgip1 gene was overexpressed under the control of the CaMV 35S constitutive promoter in Brassica napus, R line Hyola 308. Transformation of cotyledonary petioles was achieved via Agrobacterium tumefaciens LBA4404. After infection with Agrobacterium, the explants were transferred to selected regeneration medium. The transformed explants were screened in kanamycin containing media. The explants were excised and rooted using appropriate growth regulator. Putative transgenic lines obtained from independent transformation events transferred to the greenhouse for hardening. The presence of pgip1 in putative transgenic lines confirmed by the polymerase chain reaction (PCR) using specific primers. Also, the results of bioassay demonstrated that the lesion sizes occurred on leaves of transgenic canola by Sclerotina sclerotiorum (causal agent of canola stem rot) were significantly retarded in compared to non transgenic canola plant using leaf detached assay.

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