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Fig. 5. Analysis of CO function in the phloem by in situ hybridisation and confocal
microscopy of GFP:CO fusion proteins. (A) RT-PCR analysis of CO and
FT mRNA abundance in emerging leaves of Ler, co-2, 35S::CO
Ler and SUC2::CO co-2 plants. (B) In situ hybridisation of
CO and FT expression in the leaf vasculature of plants grown
in LDs (10-hours light/6-hour day extension/8-hours dark). For SUC2::CO
co-2 transverse sections are also shown. Scale bar: 25 µm. (C)
Confocal images of GFP fluorescence in whole leaf (a,b; using a 5x lens)
and leaf epidermis (c; 40x lens) of SUC2::GFP plants; in
epidermal cells (d, 40x lens) and vascular tissues (g,h; 63x oil
immersion lens) of SUC2::GFP:CO plants; and in vascular tissues of
CO::GFP:CO plants (e,f; 63x oil immersion lens). The GFP
fluorescence channel is overlaid with red and the transmissible light channels
in a,d,e and g. GFP emission fingerprinting is shown in b,f and h. Plants were
grown on MS plate in LDs. (D) Confocal image of the apex of a SUC2::GFP:CO
plant (using a 10x lens). GFP fluorescence is detected in the vascular
tissue (a), but not in the meristem (b).
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