葡萄糖氧化酶( GO ) 基因在番茄中的遗传转化

 将来自黑曲霉的葡萄糖氧化酶( glucose oxidase, GO ) 基因与病原诱导启动子Prp1-1融合,构建了植物表达载体pCPGO, 该载体具有抗除草剂PPT选择标记。经农杆菌介导转入番茄自交系, 获得了阳性转基因植株58株, Southern杂交结果表明, GO基因已整合到番茄基因组中。通过淀粉- KI显色反应和定量检测, 表明GO基因已经表达。转基因植株花器及开花正常, 种子一旦萌发, 其生长、发育各时期均正常。但结实率有所减少, 种子萌发率有所降低。用番茄叶霉病( Fulvia fulva) 病原菌1.2.3.4生理小种侵染T₁ 代转基因植株, 结果显示转基因番茄植株的抗病性有不同程度的提高, 多数发病时间推迟, 病情明显减轻。经测定与抗病性相关的几个指标, 结果转基因番茄叶片中SOD、POD、CAT的活性和蛋白质含量均增高。

Transformation of Glucose Oxidase Gene in Tomato

The plant expression vector pCPGO, comprising a GO gene from Aspergillus niger driven by prp1-1 promoter and containing a selectable marker bar gene for resistance to the herbicide phosphinothricin, was constructed. Five tomato inbred lines were transformed via Agrobacterium tumefaciens mediated method and 58 transgenic tomato plants were obtained. PPT resistance, PCR detection and Southern blot analysis showed that the GO gene was integrated into tomato genome. Expression of the GO gene was verified by KI-Starch staining and measurement of H_2O_2 content. The transgenic tomato flowered normally, whereas compared with wild-type plants, the seed set in transgenic tomato was fewer and the germination rate of seeds was less. After germination, the transgenic plants showed normal development. T_1 tomato plants infected with Fulvia fulva showed enhanced disease resistance, delayed disease symptoms and reduced leaf lesions to different extent. The activity of SOD, POD and CAT and the content of soluble protein were increased. These results indicated that overexpression of GO gene in transgenic tomato plants improved disease resistance.