A high yield method for protoplast isolation and ease detection of rol B and C genes in the hairy roots of cauliflower (Brassica oleracea L.) inoculated with Agrobacterium rhizogenes

Protoplasts represent a unique experimental system for the circulation and formation of genetically modified plants. Here, protoplasts were isolated from hairy root tissues Brassica oleracea L. induced by Agrobacterium rhizogenes strain (ATCC13332). The concentration enzyme solutions utilized protoplast isolation was 1.5 % Cellulase YC 0.1 Pectolyase Y23 in 13% mannitol solution, which resulted high efficiency within 8 hours, yield 2 x 104 cells ml-1 percentage viability 72%. Each has one nucleus with nucleation 48%. A polymerase chain reaction (PCR) assay verified presence rol B C genes detaching single bundle DNA replication these roots using specific pair primers. current study demonstrated that A. (ATCC13332) is vector incorporation T-DNA into cauliflower plants, as well success retention transferred to it.