农杆菌介导法获得草地早熟禾转Bt基因植株

Acquirement of Bt Transgenic Plants by Agrobacterium tumefaciens in Kentucky Bluegrass (Poa pratensis L.)

以草地早熟禾品种超级伊克利成熟种子为外植体材料,当愈伤组织诱导培养基中琼脂的用量增加至16g/L时,诱导产生的颗粒状胚性愈伤组织占愈伤组织总数的40%以上。愈伤组织经携带pGBI4AB农杆菌LBA4404的感染和共培养后,在继代培养基上添加50mg/L筛选剂G418,可较好地筛选出具有卡那霉素抗性的愈伤组织,进而获得具有卡那霉素抗性的再生植株。PCR分子检测结果表明,Bt基因已导入转化植株。生物学抗虫性鉴定结果显示,转Bt基因当代植株对1～2龄棉铃虫具有不同程度的抗性。

The rate of pellet embryogenic calli induced from mature seeds of Poa pratensis L. cv. Total eclipse was up to 40% by increasing agar quantity to 16 g/L over the control quantity in the callus induction medium. Through Agrobacterium-mediated transformation, the recombinant plasmid carrying Bt gene and NPT Ⅱ gene was introduced into Poa pratensis L. After infected with Agrobacterium tumefaciens strain LBA4404/pGBI4AB, 50 mg/L Geneticin (G-418) was added to the callus proliferation medium. Regenerated plants with kanamycin resistance were obtained. PCR analysis confirmed that Bt gene had been intergrated into the transgenic plant. The insect resistance bioassay showed that Bt transgenic plants had resistance to the first or second instar larvae of Helicoerpa armigera.