TRANSFORMASI GEN XILOGLUKANASE PADA Eucalyptus pellita F.Muell MELALUI Agrobacterium tumefaciens

Oleh :
Andriyani Puspitaningrum - M0404023 -

ABSTRACT Eucalyptus pellita is one of the forestry species which support the raw materials of pulp and paper industry. The improvement of wood quality especially high cellulose content wood could support the ever-increasing pulp and paper demand. Genetic engineering approach trough Agrobacterium-mediated transformation could be applied to improve wood quality and higher transformation rate could be reached by SAAT (Sonication-assisted Agrobacterium-mediated Transformation) method. The aim of this research was to study the xyloglucanase gene transfer rate of E. pellita obtained trough Agrobacterium method. The pAa XEG 300 plasmid encoding xyloglucanase gene was introduced to the germinated seed of E. pellita 006 and 06A variety. The 8 and 15 days old in vitro germinated seed grown on MS hormone free medium were used as material for transformation. Two seedling explant groups which has different age were dipped in Agrobacterium solution (OD 600=1) with 1 minute sonication and without sonication. The co-cultivated explant were selected in M1 medium (MS+ 0,465 mg/L NAA + 0,5 mg/L TDZ) containing 25 mg/l kanamysine and 50 mg/l cefotaxime, and then transfer to MM medium (MS+ 0,2 mg/l BAP + 10µg/l NAA) containing 25 mg/l kanamysine and 25 mg/l cefotaxime. Gene integration was confirmed trough PCR method using specific primer. The result showed that 15 days old E. pellita 006 variety seedlings used as transformation material has highest transformation of efficiency value (72%). The sonication method could not increase the transformation rate. Two of eight tested plant samples showed the positive PCR result indicating that xyloglucanase gene has integrated successfully. Key word : transformation, E. pellita, Xyloglucanase