انتقال ژن هاي هايگرومايسين فسفوترانسفراز و بتاگلوكورونيداز به برنج به روش زيست پرتابي

Biolistic Transformation of Rice (Oryza saliva L.) Using hpt and Gus Genes

Different callus - induction, and regeneration media were used for varieties Nemat, Neda, and Khazar Highest callus -induction and regeneration frequency was obtained for variety Nemat. Nemat as a commercial rice variety in Iran was transformed through biolistic approach. Embryogenic portion of calli derived from mature seeds were bombarded with gold particles coated with Beta- glucuronidase gene under the control of Actl promoter. Analysis of transformed calli for the presence of 13- glucoronidase activity was conducted in calli, using staining solution for Gus assay. After optimization of this method for rice by use Beta- glucoronidase, embryogenic calli derived from mature seeds of variety Nemat were bombarded with gold particles coated with barley chitinase, and hpt genes under the control of Actl, and CaMV35S promoters, respectively. The bombarded calli were selected on callus - induction media supplemented with 50 mg/l hygromycin B. Resistant calli were regenerated on regeneration media containing hygromycin B. PCR analysis of Putative transgenic plants derived from variety Nemat showed the integration of at least one copy of hpt gene into the rice genome.