181 AN ULTRACELLULAR LOOK AT LIGHT-TREATED IPT-TRANSGENIC TOBACCO

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  • 1 Horticulture Department, Clemson University, Clemson, SC 29634

Similarities exist between the effects of phytochrome and cytokinins on plant growth and development (e.g., chloroplast development. amaranthin synthesis, seed germination, photomorphogenesis). It is unclear. however, if and how these two systems interact.

To determine the effects of phytochrome activity on cytokinin synthesis and ultracellular plant development, we utilized tobacco transformed with the Agrobacterium tumefaciens isopentenyl transferase (ipt) gene. This gene encodes for isopentenyl transferase (iptase) which is an enzyme active in cytokinin biosysthesis.

Ipt-transgenic tobacco cultures were treated with end-of-day red or far-red light for 15 minutes. After 15-30 days of treatment, the plant tissue was harvested and ipt expression was verified by SDS-PAGE and western blot analysis. Polyclonal antibodies specific to iptase were used as a primary antibody. Colloidal gold conjugated to goat. anti-rabbit antiserum served as an electron dense, secondary antibody and a probe to light-influenced iptase synthesis and distribution within the cell.

A Hitachi 600AB transmission electron microscope was used to determine the influence of phytochrome/light treatments on the ultrastructure of ipr-transgenic cells.

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