The Optimized Method of Agrobacterium Mediated Transformation in Nitraria schoberi (Ghar-e-Dagh in Persian)

Abstract

Ghar-e-Dagh (Nitraria schoberi) is a native perennial plant profoundly resistant to salinity and drought. This plant, having a well-developed deep rooting system can act as sand dunes stabilization agent and also, due to its special alkaloids may perform a very significant role in pharmaceutical industries. Considering the importance of genetic engineering as a novel method to generate genetic variation in practicing precise and purposeful breeding of plants in terms of increasing their resistance against biotic and abiotic stresses and also, to increase the production of specific secondary metabolites, the present investigation was performed as a prerequisite of any transformation. In this research, the issues of pre-culture time, co-culture time, acetosyringone and kanamycin concentrations were investigated. The results obtained suggest that the interaction of pre-culture and co-culture times and also, acetosyringone concentration is significant at 1% probability level. The mean comparisons also showed that the maximum transformation rate was gained in 48 hours pre-culture time, at 100 µM acetosyringone density and 72 hours co-culture. The ultimate confirmation of β- glucuronidase (GUS) gene presence in transgenic plants was performed by performing polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) and histochemical assay. The method presented here can be used for further genetic engineering purposes of N. schoberi.

Keywords

• nitraria schoberi
• agrobacterium
• acetosyringone
• pre-culture
• co-culture