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http://dx.doi.org/10.5010/JPB.2008.35.2.147

Agrobacterium-mediated transformation of Bacillus thuringiensis cry1Ac gene in chrysanthemum (Dendranthema grandiflorum Kitamura) 'Linneker Salmon'  

Han, Bong-Hee (Horticultural Biotechnology Division, National Horticultural Research Institute)
Lee, Su-Young (Horticultural Biotechnology Division, National Horticultural Research Institute)
Lim, Jin-Hee (Floriculture Division, National Horticultural Research Institute)
Publication Information
Journal of Plant Biotechnology / v.35, no.2, 2008 , pp. 147-153 More about this Journal
Abstract
Cry1Ac gene was introduced into chrysanthemum (Dendranthema grandiflorum Kitamura) 'Linneker Salmon' through Agrobacterium-mediated gene transformation to develop new lines showing resistance to tobacco cutworm (Spodoptera litura). Cry1Ac gene was transferred into chrysanthemum by Agrobacterium C58C1 containing pCAMBIA2301. After infection of Agrobacterium C58C1 with leaf segments, the segments were cultured on regeneration medium (MS + 1.0 mg/L BA + 0.5 mg/L IAA) containing 10 mg/L kanamycin for the first selection, on the same medium containing 20 mg/L kanamycin for the second selection, and on rooting medium (MS basal medium) containing 20 mg/L kanamycin for the third selection. Until the third selection, sixty nine plantlets (1.6%) were survived and rooted. Thirty six ones (0.8%) among them were confirmed as putative transformants with nptll gene by nptll primer PCR, and 35 (0.8%) of 36 ones as transformants with nptll gene and cry1Ac gene by Southern analysis. The gene transformation efficiency of cry1Ac gene was favorable with 0.8%. The resistance of tobacco cutworm (Spodoptera litura) in chrysan-themum transformant introduced cry1Ac gene was tested in green house. Three transformants were confirmed to have resistance to tobacco cutworm.
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