Abstract

BONILLA BETANCOURT, Martha Liliana; MUNOZ FLOREZ, Jaime Eduardo  and  ANGEL SANCHEZ, Fernando. Transient gene expression in sugarcane using Agrobacterium tumefaciens. Acta Agron. [online]. 2008, vol.57, n.3, pp.161-166. ISSN 0120-2812.

The aim of the present study was to develop a transformation method mediated by Agrobacterium in Colombian cultivars of sugarcane. Transformation was evaluated in each step through transient GUS expression. Embryogenic calli and meristematic explants of CC85-92, CC84-75 y CC87-505 cultivars, were transformed using Agrobacterium tumefaciens AGL-1, LBA 4404 and EHA 105 strains, harboring pCambia 1305.2 plasmid. Furthermore, strains LBA 4404 and EHA 105 harboring pCambia 2301 were also tested. Bacterian activator medium, named infiltration media (IM) with acetosyringone was used. Co-cultivation time and bacteria optical density before induction were tested. Sugarcane cultivars evaluated showed differential response to different strain-plasmid combinations, obtaining higher GUS expression when CC85-92 was transformed using AGL-1 strain harboring the vector pCambia 1305.2. CC84-75 and CC87-505 cultivars showed higher GUS expression using EHA 105 strain harboring pCambia 1305.2 vector. The most efficient expression was evident after 7 days of co-cultivation and when bacteria optical density was 0.2_600nm before induction in IM medium. This study demonstrated that meristematic explant is more suitable than embryogenic callus as a target to be transformed by Agrobacterium tumefaciens.

Keywords : Agrobacterium tumefaciens; Saccharum spp; sugarcane; genetic transformation.

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