农杆菌介导的ACO基因RNAi载体转化百合抗衰老研究

摘要

With the scale of asiatic lily as explants, the RNAi vector with ACO anti-aging gene was transferred into lily with Agrobacterium-mediated transformation to study the factors which affected the transformation and to provide theoretical and technical basis for anti-aging of plants. Hie results showed that the highest transformation efficiency was obtained through the following transformation procedure: after preculture for 3 d, 25 mg/L acetosyringone (AS) was added to Agrobacterium and co-cultivation medium respectively, the explants were inoculated for 8min with diluted Odjoo = 0.8 recombinant Agrobacterium tumefacien culture liquid, and then co-cultivated for 3 d. PCR detection showed that some resistant plants were positive, which indicated preliminarily that the ACO gene was integrated into the genome of trans-genic plants.%本研究旨在通过根癌农杆菌介导法将ACO基因转入亚洲百合植株中,为植物抗衰老提供理论依据和技术基础.以亚洲百合‘精粹’无菌苗离体小鳞片为外植体,利用农杆菌介导法将携带有抗衰老基因ACO的RNAi载体转化到百合中,观测分析影响百合遗传转化的因素.结果表明:在遗传转化过程中,预培养3d后,在菌液和共培养基中均添加25 mg/L乙酰丁香酮,将OD600=0.8的根癌农杆菌侵染小鳞片8min,再共培养3d,有利于获得最高遗传转化效率,抗性植株经PCR检测部分呈现阳性,初步证明ACO基因已经导入到百合基因组中.