TRANSFORMASI GEN ILP (Increasing Level of Polyploidy) PADA TOMAT ‘MICROTOM’

ANUNG WAHYUDI, SP, Dr. Ir. Aziz Purwantoro, M.Sc.

2011 | Tesis | S2 Ilmu Pemuliaan Tanaman

Perakitan tanaman transgenik dapat dilakukan dengan teknik transformasi gen menggunakan mediator Agrobacterium tumefaciens. Penelitian ini bertujuan untuk mengetahui tingkat keberhasilan atau efisiensi metode transformasi gen ILP1, ILP2 dan ILP5 dengan menggunakan promoter 35S dari CaMV (Cauliflower Mozaic Virus) dan 2A11 dalam upaya memperbesar ukuran buah tomat. Agrobacterium tumefaciens yang digunakan dalam penelitian ini adalah strain GV 3101 dan RK 2013. Protokol transformasi menggunakan kultur invitro yang meliputi tahapan sterilisasi dan perkecambahan benih, induksi kalus, induksi tunas, induksi perakaran, induksi perakaran akhir dan penanaman dalam pot (aklimatisasi). Deteksi transforman menggunakan analisis PCR (Polimerase Chain Reaction) dan elegtroforesis gel. Hasil penelitian menunjukkan bahwa Kalus ‘MicroTom’ dapat ditransformasi dengan gen ILP1, ILP2 dan ILP 5. Persentase keberhasilan transformasi menggunakan promoter 35S sebesar 18,42% (35S::ILP1), 21,43% (35S::ILP2 ) dan 10,26% (35S::ILP5). Persentase keberhasilan transformasi menggunakan promoter 2A11 sebesar 19,07% (35S::ILP1), 15,56% (35S::ILP2), dan 12,50% (35S::ILP5). Dari data persentase keberhasilan transformasi tersebut maka dapat ditarik kesimpulan bahwa efisiensi transformasi gen ILP1, ILP2, dan ILP5 menggunakan promoter 35S dan promoter 2A11 dengan menggunakan mediator Agrobacterium tumefaciens terendah adalah 10,26% dan tertinggi 21,43%.

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.

Kata Kunci : Transformasi gen ILP, Agrobacterium tumefaciens, ‘MicroTom’