In Vitro Screening and Identification of Putative Sunflower (Helianthus annus L.) Transformants Expressing ECNAC1 Gene by Salt Stress Method

A simple and effective screening methodology to identify transformants at plant level under NaCl stress has been established. Leaf disc of putative transformants (T₁ individual, 6–8 weeks old) and wild type plants were placed on filter paper inthepresence of NaCl (200mM) for 48hr at room temperature. The leaf discs of EcNAC1 gene transformed plants showed salt tolerance by retaining green colour after 48 hours, while leaf discs fromnon-transformed plants turned dark brown or black in colour. Further, molecular analysis of these salt tolerant plants showed PCR positive to EcNAC1 gene specific and HPT II specific primers, and digestion of amplified EcNAC1 gene product with Sac I restriction enzyme showed expected bands size on agarose gel. Our data suggest that in vitro screening strategy at plant level based on the target gene incorporated would result in the initial identification of promising transformants for further analysis.