Studi Ekspresi Gen Penyandi AGAMOUS dan LEAFY Tanaman Kelapa Sawit (Elaeis guineensis Jacq.) Normal dan Abnormal Hasil Kultur Jaringan

Abstract

Tissue culture of oil palm has been established, but its commercialization has been hampered by reproductive organ abnormality. The development of flowers and fruits is influenced by expression of genes controlling organ identity. The function AGAMOUS (AG) gene is to induce organ identity, whereas the function LEAFY (LFY) gene is to induce AG expression. A research was conducted to study AG and LFY transcript accumulation (expression) in several developmental reproductive organs of normal and abnormal oil palm derived from tissue culture. Total RNA was isolated from flowers and fruits of normal and abnormal oil palm trees derived from tissue culture. Synthesis of first strand cDNA was carried out using total RNA. Amplification of AG and LFY gene fragment were conducted by means of RT-PCR technique using specific primers. The RT-PCR product was isolated, cloned, sequenced and analysed to confirm its homology to the target gene. Accumulation of AG and LFY transcript was analysed by quantitative program of UN-SCAN-IT GelTM connected to gel doc. The results revealed that RT-PCR product of AG is highly homologous with AG gene sequence of oil palm from the GenBank. Reproductive organ of normal and abnormal oil palm accumulated AG, but LFY could not be detected. There was difference of quantity on AG transcript accumulation between normal and abnormal oil palm. The biggest difference was detected between normal and abnormal fruits. AG fragments of normal oil palm has maximum identity of 100% with that of oil palm AG sequence from the GenBank, whereas AG fragments of abnormal oil palm has maximum identity of 99%. Difference in one nucleotide was detected between AG DNA sequence of normal and abnormal oil palm.