根瘤农杆菌介导ACO基因转化延长石竹花期的研究
Extending the flowering period of Dianthus chinensis with Agrobacterium-mediated ACO gene transformation

以石竹（Dianthus chinensis）叶片为外植体，通过根癌农杆菌介导的遗传转化体系，转ACO基因获得花期延长的石竹材料。结果表明：以MS为基本培养基，BA和NAA质量浓度分别为1.0、0.3 mg/L时，供试材料H58II、X82I和H68Ⅲ分化率较高，分别为56.9%、31.7%、93.2%；遗传转化筛选过程中，Kan、Hyg、Cef的最适筛选质量浓度分别为40、4和400 mg/L；染色检测表明，转化成功的叶片或植株染色后呈现蓝色；PCR检测转ACO基因植株，阳性率达到89%。田间观察转ACO基因植株的瓶插花期和单朵花期可达到12 d，比未转ACO基因植株的花期（6 d）增加1倍；转ACO基因植株的盆栽群体花期可达到45 d，比未转ACO基因植株的盆栽群体花期（30 d左右）延长15 d。
The leaves of Dianthus chinensis were used to transform ACO gene with Agrobaceriumtumefaciens-mediated system to obtain the new Dianthus with extending flowering period.Results showed that H58Ⅱ，X82I and H68Ⅲ had high differentiation rates of 56.9%,31.7% and 93.2% when the concentrations of BA and NAA in the MS medium were 1.0 mg/L and 0.3 mg/L.The optimal concentration of kanamycin，hygromycin and cephalosporin was 40.0 mg/L，4.0 mg/L and 400.0 mg/L in the selective medium during selective culture of transformation.Transformed leaves and plants presented blue color after being stained.Molecular test showed that the positive rate of transgenic plants was 89%.The vase life for a single flower of plants transformed with ACO gene was 12 days compared with the wild plants flowers of 6 days.The population flowering period of transformed plants in pots was 45 days while that of wild plants was 30 days.