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KMID : 0903619980390030360
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Journal of the Korean Society for Horticultural Science
1998 Volume.39 No. 3 p.360 ~ p.366
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Transformation of Chrysanthemum by Agrobacterium tumefaciens with Three Different Types of Vectors
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엄보영/Um, Bo Young
박천호/신정섭/김주영/박선정/정영수/Pak, Chun Ho/Shin, Jeong Sheop/Kim, Joo Young/Park, Sun Jung/Chung, Young Soo
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Abstract
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Two cultivars of Chrysanthemum (Dendranthema grandiflora ´Fashion Yellow´ and ´Golden Glory´) were tested on a wide range of shoot regeneration media to select the most efficient regeneration medium for Agrobacterium-mediated transformation. These cultivars were transformed using Agrobacterium LBA4404 with three different vectors, pBI121, pCMAsCP121-123, and pTOK233. In shoot regeneration experiment, the most efficient medium for ´Fashion Yellow´ and ´Golden Glory´ was the MS basal media supplemented with 2.0 ㎎/L NAA and 0.5 ㎎/L BA. Rate of chlorosis in Chrysanthemum explants cultured in the MS medium added with 20-100 ㎎/L kanamycin were 100.0%. In control, frequency of tissue formation was 0% at all the level of kanamycin concentrations. In the media containing 20 ㎎/L kanamycin, Chrysanthemum ´Fashion Yellow´ showed 30.0% and 38.1% of tissue formation rates when transformed with pBI121 and pCMAsCP121-123, respectively. In Chrysanthemum ´Golden Glory´, however, tissue formation rates were 77.8%, 65.1 %n, and 98.6% in transformation experiments with pBI121, pCMAsCP121-123 and pTOK233, respectively. PCR showed the presence of NPTII genes in the genome of all transformants except the control. PCR-Southern analysis confirmed that the band observed in the transgenic plants were originated from T DNA transfer with strong hybridization. The genomic Southern analysis showed the same result and indicated multiple copy of T DNA integration.
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