Optimizing agrobacterium-mediated transformation parameters of melastoma decemfidum ROXB EX JACK

Abstract

Melastoma decemfidum is a tropical ornamental plant belongs to family of Melastomataceae and widely used in traditional medicine. The plant was reported to contain bioactive flavonoids that exhibited antioxidant and anticancer effects. To date, there is no report on transformation system for M. decemfidum. In this study, the effects of transformation parameters on the transformation efficiency of Agrobacterium-mediated transformation of M. decemfidum were investigated. Parameters such as bacterial concentration, infection time and acetosyringone concentration were optimized using histochemical GUS assay. Results showed that the highest transformation efficiency (96.7%) for M. decemfidum with an average of 479.6 ± 10.2 blue spots per explant was obtained when leaf explants were treated with bacterial concentration of OD600nm = 0.6. Forty-five minutes of infection time gave the highest percentage (96.7%) of positive transformants and an average of 316.9 ± 7.6 blue spots per explant. Addition of 100 µM acetosyringone was optimum (100%) for transforming M. decemfidum with an average of 509.8 ± 26.7 blue spots per explant. In conclusion, an efficient Agrobacterium-mediated transformation protocol for M. decemfidum was successfully established. Thus, the developed protocol will facilitate the delivery of desirable genes into M. decemfidum via Agrobacterium-mediated transformation.