국화 (Dendranthema grandiflorum Kitamura) ‘Linneker Salmon’에 Agrobacterium을 이용한 Bacillus thuringiensis cry1Ac 유전자의 형질전환
Agrobacterium-mediated transformation of Bacillus thuringiensis cry1Ac gene in chrysanthemum (Dendranthema grandiflorum Kitamura) ‘Linneker Salmon’
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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 nptII gene by nptII primer PCR, and 35 (0.8%) of 36 ones as transformants with nptII 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 chrysanthemum transformant introduced cry1Ac gene was tested in green house. Three transformants were confirmed to have resistance to tobacco cutworm.
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 nptII gene by nptII primer PCR, and 35 (0.8%) of 36 ones as transformants with nptII 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 chrysanthemum transformant introduced cry1Ac gene was tested in green house. Three transformants were confirmed to have resistance to tobacco cutworm.
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