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Development of anAgrobacterium-mediated transformation system for regenerating garland chrysanthemum (Chrysanthemum coronarium L.)

한국식물학회지 2005년 48권 1호 p.136 ~ 141
정귀미, 박영두,
소속 상세정보
정귀미 ( Chung Kwi-Mi ) - Kyung Hee University Graduate School of Biotechnology
박영두 ( Park Young-Doo ) - Kyung Hee University Graduate School of Biotechnology

Abstract


Although efficient shoot regeneration and selection are essential for genetic transformation mediated byAgrobacterium, success has been limited with the garland chrysanthemum (Chrysanthemum coronarium L.). In this study, we developed a useful protocol for shoot regeneration with leaf disk explants. The optimal concentrations of NAA and BA were 0.2 mg L?1 and 0.5 mg L?1, respectively. To optimize the selection system for regenerating plants from genetically transformed tissues, we tested the effects of four antibiotics (kanamycin, hygromycin, carbenicillin, and cefotaxime). Among them, 5 mg L-1 hygromycin proved adequate as a selectable marker, whereas 500 mg L-1 carbenicillin was effective in eliminating excessiveAgrobacterium after co-cultivation. Transgenic plants were obtained by first co-culturing garland chrysanthemum leaf disks withA. tumefaciens strain EHA105, which harbors plasmid pRCVII containing the hygromycin resistance (hpt) and β-glucuronidase (GUS) genes. After the transgenic plants were confirmed via Southern analysis, they were rooted in soil and appeared phenotypically normal. Our report is the first to describe the optimum conditions for producing transgenic plants of this species.

키워드

Agrobacterium tumefaciens; antibiotics; β-glucuronidase gene; garland chrysanthemum; hygromycin resistance gene

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