转几丁质酶基因和β-1,3-葡聚糖酶基因烟草的研究

Transgenic Tobacco Containing Chitinase and β-1,3-Glucanase

为选育出高抗真菌病害的烟草新品种 ,利用叶盘法 ,通过农杆菌介导 ,将来自芥菜和橡胶的抗真菌蛋白基因几丁质酶基因和 β 1,3 葡聚糖酶基因转入烟草品种K32 6和红花大金元 ,在含卡那霉素的MS培养基 (MS +NAA0 .1mg/L +6 BA 1.0mg/L为芽诱导培养基 ,根诱导培养基为不含激素的MS)上进行不定芽和根诱导 ,共获耐卡那霉素再生苗 10 1株 .经PCR扩增检测 ,TB 1等 5 7株呈阳性 ,即已成功导入外源目的基因 .初步接种炭疽病菌和黑胫病菌结果表明 ,转基因植株后代具有良好的抗病能力 .

Fungal diseases have been seriously affecting tobacco quality and yield for a long time. Since conventional breeding and chemical control became less effective, plant generic engineering has offered an alternative approach for the control of these fungal diseases.The anti-fungi protein chitinase and β-1,3-glucanase genes were transferred into tobacco varieties K326 and Honghuadajinyuan, by agrobacterium mediated-leaf disc method. After regeneration of leaf explants on basal MS with NAA 0.1 mg/L, 6-BA 1.0 mg/L and Kanamycin 75 mg/L, the shoots were excised on hormone-free MS with Kanamycin 75 mg/L to induce roots. 101 small plants with developed roots were obtained, and 57 plants of them showed PCR positive. Brimary results indicate that the offsprings of transgenic plants have good resistance to Collectrichum and phytophthora.