农杆菌介导Cry1Ac基因转化菊花

Transformation of Chrysanthemum with a Synthetic Cry1Ac Gene Mediated by Agrobacterium tumefaciens

用农杆菌介导法,以茎段(节间)为外植体,将含有苏云金芽孢杆菌(Bt)毒蛋白Cry1Ac基 因的植物遗传载体导入菊花(切花菊)品种‘日本黄’中,得到126株抗性植株,PCR检测及基因组DNA 的Southern杂交分析表明Cry1Ac基因已整合到菊花总DNA中,共获得6株转基因植株。利用棉铃虫做转基 因植株的盆栽和叶片平皿试验,发现其中2株对棉铃虫具有很高抗性,校正死亡率达90%以上;1株对棉 铃虫具有高抗性,校正死亡率达80%以上;3株对棉铃虫抗性较明显,校正死亡率44.7%～60.9%。

A synthetic Bt(Bacillus thuringiensis)Cry1Ac gene was introduced into the genome of cut flower chrysanthemum(Chrysanthemum morifolium)cultivar‘Japanese Yellow’ in vitro with explants of internodal segments mediated by Agrobacterium tumefaciens. Results of PCR and Southern blot analysis indicated that Cry1Ac gene were integrated as one copy into the genome of chrysanthemum. Six transgenic lines were obtained in total. Upon insect bioassay using Heliothis armigera(Hubner) in the pot-culture and in petri dishes, two of the screened plants were demonstrated to be very highly resistant to the insect larvae, the corrected mortality of insect larvae was higher than 90% in five days. One individual line showed strong resistance, the corrected mortality of insect larvae was higher than 80%. Three transformed lines showed relative strong resistance, the corrected mortality of insect larvae was ranged from 44.7% to 60.9%.