Agrobacterium mediated transformation of anti-stress genes into cauliflower (Brassica oleracea var. botrytis L.). 2. Transformation and confirmation of stress tolerance.

Abstract

Integration of APX and SOD anti-stress genes into cauliflower (Brassica oleracea var. botrytis L.) plants was achieved by using Agrobacterium tumefaciens - mediated transformation method. Cauliflower explants (hypocotyls and cotyledons) and Agrobacterium tumefaciens strains (APX, SA, TA) were used in this experiment. The procedure utilizes polymerase chain reaction (PCR) amplification of insert DNA directly after isolation of individual colonies without the necessity of separate procedures for DNA isolation and purification. Integration of the introduced stress gene (APX and SOD) in the plants was confirmed by using β-glucuronidase gene (GUS) and leaf disc assays as a gene fusion and diagnostic marker, respectively. The stable integration of the APX and SOD gene at 478 bp was detected by using polymerase chain reaction (PCR) of the putative transgenic plants. Analysis of APX and SOD gene expression under salt treatment showed that putative transgenic cauliflower survived the salinity stress comparing with the control plants.