Development of Agrobacterium-mediated Transformation Method for Domestically Bred Chrysanthemmn Cultivar 'Moulinrouge' and Genetic Change of Leaf Morphology Using AtSICKLE Gene

摘要

'Moulinrouge' was selected as the best regenerating cultivar among 18 different spray-type chrysanthemum cultivars bred in the Gyeongnam Flowers Breeding Research Institute. When the leaf explants from standard- and spray-type chrysanthemum 'Jinba' and 'Moulinrouge' were incubated on MS basal medium supplemented with 0.5 mg.L~(-1) BA and 1.0 mg-L~(-1) NAA, both 'Jinba' and 'Moulinrouge' induced adventitious shoots that can be regenerated into plantlets. Based on these regeneration conditions, we developed an efficient Agrobacterium-madiated chrysanthemum 'Moulinrouge' fransformation method by using sequential selection of shoots from low (10 mg-L~(-1)) to high (30 mg-L~(-1)) concenfrations of kanamycin after co-cultivation of leaf explants with Agrobacterium for 10 days and induction of shoots. All kanamycin resistant plants investigated with genomic PGR analysis carried the report gene, AtSICKLE, in their genome. Although expression levels of the report gene in the transgenic plants investigated with RT-PCR were relatively low because of inefficiency of CaMY 35S promoter in chrysanthemum, transgenic lines expressing AtSICKLE efficiently showed leaf epinasty phenotype. We expect that our results will provide a useful method that can perform a high-throughput investigation of genes isolated and studied well in model plants for molecular breeding of chrysanthemum.