Transformasi Genetika Padi Indica dengan Gen OsMADS18 Menggunakan Agrobacterium Tumefaciens untuk Merakit Varietas Padi Umur Pendek

Abstract

Rice is the staple food of more than half the world's human population. The need of rice each year continues to increase with increasing population. This study aimed to assemble a short-lived rice varieties by introducing OsMADS18 gene into the genome of plants using Agrobacterium tumefaciens. OsMADS18 gene is a short age gene which has been successfully constructed to pCAMBIA1301 vector under CaMV35S constitutive promoter. This is evidenced by the emergence of 12561 bp DNA band size, this measure is a combination among vector pCAMBIA 1301 (11819 bp) and OsMADS18 genes (742 bp). Then the vector pCAMBIA 1301 that contains OsMADS18, was used to transform Agrobacterium tumefaciens strain EHA105 through Heat-shock method. The success of transformation marked by the growth of the white colonies Agrobacterium tumefaciens on solid media LB, this media contained antibiotic, X-gal and IPTG. Then the bacteria are used to transforms 50 calli Indica rice by soaking the calli into colonies for 20 minutes. The calli moved to selection media that contains hygromycin. After 14 days, calli that resistant to hygromycin moved to regeneration media R3B grown for transgenic plant. 18 transformants plantlets were obtained. Gus-A successful test on the show with the appearance of blue color in leaf tissues. While PCR analysis of the 18 transgenic plantlets produced 742 bp sized bands.