Induksi Kalus Jeruk Siam Triploid dan Transformasi Genetik pada Jeruk Siam (Citrus nobilis L.).

Abstract

This study aims to obtain the best medium composition for callus induction of triploid tangerine from different kind of explant and regeneration of transformant citrus callus. This study is divided into 3 activities. The first aimed to induce embryogenic callus of triploid tangerine with explant from internodes, nodes, and leaves isolated from in vitro triploid tangerine buds. The explants were cultured in medium MS + MW vitamins + EM 500 mgL-1suplemented with BA 1 mgL-1, 2 mgL-1 and 3 mgL-1. The second, to introduce the gene constructs containing a selection marker (gus gene and kanamycin antibiotic) vector A. tumefaciens to triploid tangerine embryogenic callus. The third, to regenerate embryogenic callus transformant candidates in several formulations of regeneration medium. Transformant callus grown on media containing MS vitamins and MW with the addition of organic matter EM 500 mgL-1 and 10; 20; and 30% of coconut milk. Transformant callus that have been through the proliferation is regenerated on MS medium with a combination of Plant Growth Regulator:0.01 mgL-1IAA, 0.01 mgL-1 BA, 0.01 mgL-1 TDZ and 0.01 mgL-1 Kinetin. The results showed that the optimal concentration of BA for callus induction was 3 mgL-1, while different types of explants not give different results. Frequention of antibiotics application to sterilized the A. tumefaciens bacteria from callus showed the best results when done everyday. The best callus growth results of transformant callus occurs in medium MS containing vitamin MW with the addition of organic material EM 500 mgL-1. The combination of medium used for regeneration of transformant tangerine callus has not succesful yet in regenerating callus into planlet.