아그로박테리움를 이용한 국내개발 국화품종 ‘무랑루즈’의 형질전환 기술 및 AtSICKLE 유전자를 이용한 엽형 변화 국화 형질전환체 개발

‘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 ㎎ㆍL?¹ BA and 1.0 ㎎ㆍL?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-mediated chrysanthemum ‘Moulinrouge’ transformation method by using sequential selection of shoots from low (10 ㎎ㆍL?¹) to high (30 ㎎ㆍL?¹) concentrations of kanamycin after co-cultivation of leaf explants with Agrobacterium for 10 days and induction of shoots. All kanamycin resistant plants investigated with genomic PCR 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 CaMV 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.