ACCLIMATIZATION AND PRESERVATION OF SEAWEED Kappaphycus alvarezii AS PRODUCT OF PaCS (Cytrat cintase) TRANSFORMATION  FOR ENHANCED  RESISTANCE TO ENVIRONMENTAL STRESS   IN FLOATING NET CAGE IN PANGKEP WATER

PaCS (cytrat synthase) gene transformation in seaweed K.alvarezii has been successfully performed in vitro with the aim to improve resistance on environmental stress especially extreme environmental changes causing death and damage to the tissue cultivated seaweed. Nevertheless, the death often occurs during the regeneration of the flask cultures and acclimatization in the bath recirculation. Therefore, it is requred an effort to improve the survival rate of gene PaCS (Cytrat synthase) inserted seaweed through the regeneration using the flask cultures continued with acclimatization and rearing of gene PaCS inserted seaweed on floating net cage (KJA), which is done in stages, namely: (1) the transformation of genes PaCS (cytrat synthase) into seaweed K.alvarezii followed by regeneration on the flask cultures in the laboratory; (2) acclimatization in cage on a net green (mess size of 100 um) with the size of 50x50x50 cm with the density of 200 explants and reared for 2 weeks, the explants were then after transferred to the net blue with the size of 50x50x50 cm (mess size of 200 um) with the density of 200 explants. After 4 weeks of the rearing period, regeneration of seaweed resulted of transformation  was carried out by using long line method at the floating net cage, being tied up with a double ris rope with the distance of 15 cm. The measurement of weight, shoot length, and water quality was conducted at intervals of 2 weeks of rearing perid

Analysis of gene PaCS integration was done by PCR using primers PaCS-F and PaCS-R. Findings demonstrated that the daily growth rate (DGR) on regeneration in the culture flask was ranged from 0.11 to 0.18%, while the acclimatization using a net green exhibited  the daily growth rate of 0, 62 to 1.3%, and the raring using net blue areas showed DGR increasing to 2.18% per day. Further enlargement of transgenic seaweed using a long line method showed DGR 1.42-4,3%. The integration of gene PaCS (cytrat cynthase) into seaweed showed the tape at the position of 1300 bp as the promoter fragment of PaCS-F and PaCS-R at CaMV 35S vector. Based on

these results, it proved that the PaCS gene has been successfully transformed to the seaweed K.alvarezii