苹果转化细胞的悬浮培养及植株再生

以试管苗叶片为外植体，将绿色荧光蛋白基因（GFP）转入苹果品种昌红中，研究发现抗生素筛选压对苹果转化效率的影响主要表现为对转化细胞再生植株的抑制作用。虽然降低筛选压（10或20 mg/L卡那霉素）可获得较高频率（分别为62.0%和57.8%）的转化愈伤组织，但其对转化细胞再生植株的抑制仍然明显。通过建立转化细胞悬浮培养系，在无筛选压下获得大量的由悬浮转化细胞增殖形成的愈伤组织，该愈伤组织再转到无筛选压的再生培养基上，6周以后15.8%的愈伤组织可再生转化植株。研究结果显示，可通过建立悬浮培养系的方式消除抗生素筛选压的抑制作用，快速大量获得苹果转化植株，为进一步开展苹果转化基因的整合机制与表达特性研究，推动苹果转基因育种的应用奠定基础。

Regeneration of Transgenic Plants from Cell Suspensions of Transformed Apple

Experiments were conducted with different types of leaves of apple (Malus pumila Borkh.cv.Fuji) to study the effects of antibiotic concentration on the efficiency of apple transformation and on the plant regeneration of apple. Although shoot regeneration was highly sensitive to antibiotic selection, a high percentage of callus formation (62.0% or 57.8%) was observed under selection pressure (10 or 20 mg/L Kanamycin). Over 80.0% or 95.6% of these calli were transgenic, although the efficiency of transgenic shoot regeneration was low. The vitality of transgenic cells from shoot leaves was strong and small cell aggregates could originate from these single cells when the transgenic calli were cultured in liquid medium. After transferred onto regeneration medium without kanamycin selection pressure, 15.8% of these transgenic cell aggregates could regenerate into plantlets after 6 weeks.