农杆菌介导的ACC氧化酶反义基因转化甜瓜品种河套蜜瓜的研究

Genetic Transformation of the ACC Oxidase Antisense Gene to Cucumis melo L.cv.Hetao Via Agrobacterium Tumefaciens

将从甜瓜品种河套蜜瓜中克隆的ACC氧化酶基因cDNA片段反向构建到植物表达载体pROKⅡ中，获得重组质粒pRACO1，用三亲融合法将其导入根癌农杆菌LBA4404中，转化河套蜜瓜子叶外植体，在芽诱导培养基MS＋IAA0．8mg／L＋BA1．0mg／L＋ABA0．26mg／L＋Cef500mg／L＋Kan75mg／L中诱导生芽，待芽长到2cm左右转入MS＋IBA0．5mg／L＋Cef300mg／L＋Kan50mg／L的培养基中诱导生根，1～2周后可诱导产生大量的根，形成完整的转化小植株．经PCR检测证明，目的基因已整合到河套蜜瓜的基因组中．

The cDNA of the ACC oxidase gene cloned from melon Hetao was constructed reversedly into plant expression vector pROKⅡ. The recombinant plasmid pRACO1 was introduced into melon Hetao cotyledon explants by Agrobacterium-mediated transformation. After shoot inducing culture and root inducing culture,the regenerated plantlets were obtained from the cotyledons on MS medium with 0.5 mg/L IBA, 50 mg/L kanamycin and 300 mg/L cefotaximine. The PCR detection showed that the antisense gene of ACC oxidase has been integrated into melon cultivar Hetao genome.