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Figure 3 | BMC Plant Biology

Figure 3

From: Regeneration and transformation of Crambe abyssinica

Figure 3

Histochemical and molecular evidence for the transgenic nature of putative genetically modified crambe shoots. A, GUS staining of leaf samples from a candidate transformant obtained from cotyledonary node explants inoculated by AGL0 (pJS-M14) with a shifting selection scheme; B, GUS staining of a leaf sample from the WT control; C, PCR using either primers for NPTII (upper panel) or vir-G (lower panel). DNA samples from four randomly chosen candidate transformants obtained from cotyledonary node explants inoculated by AGL0 (pJS-M14) with a shifting selection scheme. DNA from wild type (WT) crambe was used as negative control and Agrobacterium as positive control. The fluorescent shoot in panel D was obtained from the transformation of axillary bud explants with AGL1 carrying pBinGlyRed (AscI), while panel E shows the absence of fluorescence in a regenerated shoot of WT. Panel F showed the Southern blotting analysis of transformants obtained from cotyledonary node explant based transformation protocol with a shifting selection scheme. DNA samples from four transgenic candidates (coded as F2, F6-1, F5 and F1 from left to right) of pBinGlyRed (AscI) transformation are presented. The restriction enzyme was DraI and the NPTII probe was generated using the same primers as used for PCR. Next to the molecular weight marker lane control DNA from WT crambe is loaded. The lower size limit of fragments to be visualized is 2.3 kb.

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