Optimization of Medium Putative Transgenic Shoots Containing Metallothionin Type II In Controlled Round Bottom During Acclimatization of Kappaphycus alvarezii
Emma Suryati , Rosmiati, U. Widyastuti , A. Tenriulo and Rohama Daud 2014

Abstract
Gene transformation of metallothionin (Mamt) typ Gene transformation of metallothionin (Mamt) type II into seaweed Kappaphycus alvarezii mediated with Agrobacterium tumefacien was successfully carried out. K.alvarezii explants were inoculated with Agrobacterium tumefacien containing PlG6- MaMt2 through PES selection medium containing of 10 and 20 mg/L hygromisin with the transformant efficiency of. alvarezii at the selection medium I was 82.07%, and medium II was 84.21%. Meanwhile, the transformant efficiency of putative transgenic K. alvarezii was 98.43%. Acclimization of putative transgenic shoots containing Mamt 2 gene was carried out by using round bottom placed in a culture chamber. Culture medium as liquid PES medium with the salinity of 20-30 ppt, pH of 5-8, ratio of dark and light of 16:8; 12:12, and 8: 16, light intensity of 1500 lux. The addition of ZPT stimulated the growth of transgenic shoots. The evaluation of MaMt2 gene transformation was done by PCR under promotor ubiquitin control and nos terminator. The purpose of this study wasThis study aims to determine the optimal condition at the acclimization of transgenic shoots using controlled round bottom placed in cultur cumber. The result showed that the best survival rate was obtained at the salinity and pH of 30 ppt and 6, respectively whilst, the best daily growth rate was found in transgenic shoots cultured intoliquid PES medium enriched with IAA and BAP with the ratio of 1:2. The PCR analysis exhibited the presence of band at 500 kb indicating the existence of MaMt2 gene in K.alvarezii putative transgenic shoots.