Sujatha M, Lakshminarayana M, Tarakeswari M, Singh P K, Tuli Rakesh
Directorate of Oilseeds Research, Rajendranagar, Hyderabad, 500 030, India.
Plant Cell Rep. 2009 Jun;28(6):935-46. doi: 10.1007/s00299-009-0699-x. Epub 2009 Apr 1.
Castor (cv. DCS-9) has been transformed through Agrobacterium-mediated and particle gun bombardment methods using appropriate vectors containing the Bt chimeric gene cry1EC driven by enhanced 35S promoter. About 81 and 12 putative transformants were regenerated following selection on hygromycin and kanamycin, respectively. Southern analysis of DNA extracted from T(0) plants confirmed integration of the introduced gene in castor genome. The integration and inheritance of the introduced genes was demonstrated up to T(4) generation by PCR and Southern analysis. Southern analysis of two events having single and two copies showed the same pattern of integration in the subsequent generations. Insect feeding experiments conducted in the laboratory by releasing neonate larvae of castor semilooper and S. litura on leaf tissues excised from transgenic and control plants showed varying degrees of larval mortality and slow growth in larvae fed on transgenic leaf tissue. Field bioassays against Spodoptera litura and castor semilooper conducted for eight events in T(1)-T(4) generations under net confinement were more informative and events conferring resistance to the two major defoliators were identified.
已使用含有由增强型35S启动子驱动的Bt嵌合基因cry1EC的合适载体，通过农杆菌介导法和粒子枪轰击法对蓖麻（品种DCS - 9）进行了转化。在分别用潮霉素和卡那霉素进行筛选后，分别再生出约81个和12个推定的转化体。对从T(0)代植株提取的DNA进行的Southern分析证实了导入基因已整合到蓖麻基因组中。通过PCR和Southern分析，证明了导入基因在T(4)代之前的整合和遗传情况。对具有单拷贝和双拷贝的两个事件进行的Southern分析表明，在后代中整合模式相同。在实验室中，通过在从转基因植物和对照植物上切下的叶片组织上释放蓖麻半环饰夜蛾和斜纹夜蛾的新生幼虫进行昆虫取食实验，结果显示取食转基因叶片组织的幼虫有不同程度的死亡率且生长缓慢。在网室条件下，对T(1)-T(4)代的8个事件针对斜纹夜蛾和蓖麻半环饰夜蛾进行了田间生物测定，结果更具参考价值，并且鉴定出了对这两种主要食叶害虫具有抗性的事件。
