In Vitro Culture and Agrobacterium Tumefaciens - Mediated Transformation of Precious Patumma (Curcuma sp.)

Abstract

Precious Patumma (Curcuma sp.) was a promising ornamental plant that could benefit from further development using modern biotechnology. In this study, In vitro retarded shoots culture of Precious Patumma was conducted on MS medium supplemented with cytokinin of TDZ at 0.0, 0.1 and 0.5 mg/l and IMA at 0, 2 and 4 mg/l . The results indicated that the highest number of retarded shoots (12 retarded shoots per explant) could obtain when cultured on MS medium supplemented with 0.5 mg/l TDZ combination with 4 mg/l IMA and developed the Agrobacterium-mediated transformation protocol into Precious Patumma. The A. tumefaciens strain AGLO harboring the binary vector pBI121 containing the neomycin phosphotransferase gene (nptII) as a selectable marker and the α-glucuronidase (gus) gene as a reporter was cultured in MS medium supplemented with 0.5 mg/l TDZ combination 4 mg/l IMA . The bacteria were diluted incubated with in vitro propagated Precious Patumma retarded shoots in MS medium supplemented with 0.5 mg/l TDZ combination 4 mg/l IMA for 30 minutes. Treated retarded shoots were selected in MS medium containing 100 mg/l kanamycin for 6 weeks. GUS bioassay was used to observe gus gene expression in leaves of transgenic plants. The results revealed that the highest percentage of transformed Precious Patumma (73.33%) was obtained when in vitro Precious Patumma retarded shoots were inoculated with A. tumefaciens at 30 minutes.