摘要
建立有效的遗传转化体系是基因工程手段改良植物性状的重要前提。本研究对西兰花遗传转化体系进行了初步研究，成功地建立了农杆菌介导的西兰花遗传转化体系。以7日龄无菌苗的下胚轴为外植体，预培养2—3d后进行农杆菌的侵染，共培养后将外植体转移至无筛选压力的脱菌培养基中，暗培养7d后置于含有10mg·L-1卡那霉素的筛选培养基中继续培养，待不定芽生长至2—3cm时转入0．1mg·L-1IBA的生根培养基中诱导不定根，最后进行驯化和移栽，PCR分子鉴定及外源基因表达水平检测结果证明，我们已经成功地将芥子油苷合成关键酶基因CYP79F1基因转入西兰花商业品种“青秀”中，利用HPLC检测转基因植株中短链脂肪族芥子油苷的含量显著增加。
Abstract Establishment of effective transformation system is an important precondition to improve plant traits by genetic engineering. In this study, an agrobacterium-mediated transformation system of Brassica oleracea var. italica was established. Hypocotyls of 7-day-old seedlings were used as explants and preincubated for 2 - 3 d and infected by agrobacterium-tumefaciens. After co-incubation, the explauts were transferred to degerming medium without selective agent to induce adventitious buds. Following the incubation in darkness for 7 d, the explants were transferred to selective medium containing 10 mg· L-1 kanamycin. When the adventitious buds grew 2 -3 cm long, they were moved to rooting medium with 0. 1 mg· L-1 IBA to induce adventitious roots. Finally, the domestication and transplantation were performed. PCR detection demonstrated that glucosinolate biosynthetic gene
出处
《植物研究》
CAS
CSCD
北大核心
2014年第4期516-523,共8页
Bulletin of Botanical Research
基金
黑龙江省留学归国科学基金项目(LC2009C31)
关键词
西兰花
农杆菌
遗传转化体系
芥子油苷
CYP79F1
Brassica oleracea var. italica
Agrobacterium-tumefaciens
genetic transformation system
glucosinolate
CYP79F1
