CYP79F1基因对西兰花的遗传转化的研究
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2~3 d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7 d后置于含有10 mg·L-1卡那霉素的筛选培养基中继续培养，待不定芽生长至2~3 cm时转入0.1 mg·L-1 IBA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。
关键词: 西兰花, 农杆菌, 遗传转化体系, 芥子油苷, CYP79F1
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 explants 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 CYP79F1 was successfully transferred into the genome of B.oleracea var. italica commercial cultivar “Qingxiu”. HPLC determination proved that short chain aliphatic glucosinolates significantly increased in transgenic plants.
Key words: Brassica oleracea var. italica, Agrobacterium-tumefaciens, genetic transformation system, glucosinolate, CYP79F1
中图分类号:
Q789
相关知识
植物开花基因FT的遗传转化及其参与开花调控的研究
根癌农杆菌介导的F3’5’H基因对菊花遗传转化的初步研究
花色相关基因的克隆鉴定及兰花遗传转化研究
大豆抗病相关基因SR1正反义植物表达载体的构建及遗传转化研究
菊花遗传转化
根癌农杆菌介导的花魔芋遗传转化体系的研究
兰花遗传转化技术研究进展
植物抗病防卫相关基因的克隆及其转化烟草的研究
农杆菌介导的TFL1基因转化菊花的研究
类黄酮3',5'羟化酶基因的克隆及转化矮牵牛的研究
网址: CYP79F1基因对西兰花的遗传转化的研究 https://m.huajiangbk.com/newsview432097.html
| 上一篇: 神奇的植物基因工程 |
下一篇: 基因工程辅助萝卜硫苷在十字花科作 |
