TRANSFORMASI GEN ILP (Increasing Level of Polyploidy) PADA TOMATâMICROTOMâ
Sp Anung Wahyudi, Aziz Purwantoro
Abstract
Sp Anung Wahyudi, Aziz Purwantoro
Abstract
Transgenic plants can be created with the technique of gene transformation using Agrobacterium tumefaciens mediators. The aim of this research was to determine efficiency of gene transformation method using the ILP1, ILP2 and ILP5 genes using the 35S promoter of CaMV (Cauliflower Mosaic Virus) and 2A11. Agrobacterium tumefaciens strain GV3101 and RK 2013 were used in this study. The promoter used in this study is 35S promoter (over-express) and 2A11 (specific promoter for fruit). Due to their ability to increse the ploidy of plants, genes ILP1, ILP2 and ILP5 (Increasing levels of Poliploidy) were chosen. Transformation protocol using in vitro culture includes seed sterilization and germination, callus induction, shoot induction, rooting induction, induction of the final rooting and planting in pots (acclimatization). Detection of transformants was conducted using PCR analysis (Polymerase Chain Reaction) and electrophoresis gel. The results showed that 'MicroTom' callus can be inserted with the gene. Percentage of successful transformation using the 35S promoter was 18.42% (35S::ILP1), 21.43% (35S::ILP2) and 10.26% (35S::ILP5). Percentage of successful transformation using 2A11 promoter (35S::ILP1) were 19.07%, 15.56% (35S::ILP2), and 12.50% (35S::ILP5). The efficiency of gene transformation ILP1, ILP2, and ILP5 using 35S promoter and promoter 2A11 was 10.26% as the lowest to 21.43% as the highest.
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Transgenic plants can be created with the technique of gene transformation using Agrobacterium tumefaciens mediators. The aim of this research was to determine efficiency of gene transformation method using the ILP1, ILP2 and ILP5 genes using the 35S promoter of CaMV (Cauliflower Mosaic Virus) and 2A11. Agrobacterium tumefaciens strain GV3101 and RK 2013 were used in this study. The promoter used in this study is 35S promoter (over-express) and 2A11 (specific promoter for fruit). Due to their ability to increse the ploidy of plants, genes ILP1, ILP2 and ILP5 (Increasing levels of Poliploidy) were chosen. Transformation protocol using in vitro culture includes seed sterilization and germination, callus induction, shoot induction, rooting induction, induction of the final rooting and planting in pots (acclimatization). Detection of transformants was conducted using PCR analysis (Polymerase Chain Reaction) and electrophoresis gel. The results showed that 'MicroTom' callus can be inserted with the gene. Percentage of successful transformation using the 35S promoter was 18.42% (35S::ILP1), 21.43% (35S::ILP2) and 10.26% (35S::ILP5). Percentage of successful transformation using 2A11 promoter (35S::ILP1) were 19.07%, 15.56% (35S::ILP2), and 12.50% (35S::ILP5). The efficiency of gene transformation ILP1, ILP2, and ILP5 using 35S promoter and promoter 2A11 was 10.26% as the lowest to 21.43% as the highest.
Key concepts: Cauliflower mosaic virus, Agrobacterium tumefaciens, Transformation (genetics), Biology, Callus, Transformation efficiency, Agrobacterium, Gene