Isolasi dan Karakterisasi Fragmen Gen LEAFY dari Bantalan Bunga Kakao

Abstract

Cocoa plant productivity in Indonesia is still low. Information and knowledge at the molecular level about the process of development of regenerative organs of cocoa can be an effective solution to problems associated with the regenerative low productivity of cocoa plants.LEAFY gen is key gene in the shoot meristem transition to flower one. The purposes of the research were to isolate LEAFY gene from cacao’s flower cushion and analyse its expresion in several cacao’s tissues. Isolation of LEAFY gene using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) technique showed that three spesific primer pairs designed based conserved area can amplify the target gene. Futhermore this fragment were inserted into pGEM-T Easy vector then transformed into competent cell E. Coli XL-1 Blue. The result of bioinformatic analysis showed that the fragment of target gene has a high homology to LEAFY gene of other plants in gene bank’s database. Characterization of target gene was initiated with RNA isolation phase from several cacao’s tissues then those were amplified with RT-PCR technique. Amplification at 62 ºC of annealing temperature for 33 cycles was successful to show the existence of LEAFY gene in several cacao’s tissues isolated.