Abstract
Chrysanthemum is a short-day herbaceous perennial plant. Their flowering time is strongly affected by the photoperiod and temperature. To understand the molecular mechanisms of flowering in Chrysanthemum, we applied a diploid wild chrysanthemum, Chrysanthemum seticuspe f. boreale as a model system for molecular-genetic study. We constructed a full-length cDNA library derived from the leaves and shoot apex of Chrysanthemum seticuspe (accession: NIFS-3) grown under various photoperiod and temperature conditions. After normalization of the cDNA library, 3'EST-, 5'EST- and fragment library were constructed and sequenced by Genome Sequencer FLX (Roche), to obtain total sequence tags of about 2,724,000. These fragments were grouped into 42,450 clusters and 60,306 contigs. Based on these 60,306 contig sequence data, we designed 60K and 180K custom array (Agilent). To analyze the functions of flowering related genes, we developed Agrobacterium-mediated transformation protocol for Chrysanthemum seticuspe (NIFS-3). At this moment, we have succeeded in generating transgenic plants with transformation efficiency of 2~5%.
