The Carnation Ethylene-responsive element-Binding Protein (CEBP), a nuclear-encoded transcription factor found in plants, is believed to be involved in the senescence of flowers. Previous studies have elucidated the putative role of this protein in the nucleus, however the role of CEBP in the chloroplast remains unknown. An RNAi construct to silence CEBP mRNA expression was developed under control of the Clarkia breweri linalool synthase (LIS) promoter, a flower specific promoter, in addition to Kanamycin resistance. The construct was transformed into E. coli, and then the plasmid containing the construct was then transferred into Agrobacterium. Next, Agrobacterium mediated transformation was utilized to insert the RNAi into the petunias using leaf disk infiltration. Four independent transgenic lineages were transferred from tissue culture to the greenhouse. QRTPCR confirmed expression of the RNAi construct as well as knock down effect on CEBP expression. Senescence timeframes, physiological changes, and downstream effects will be reported.
