抗菌核病转基因油菜植株的获得

Obtaining Transgenic Rape Plants Resistant to Sclerotinia Sclerotinorium

将携带菜豆几丁质酶基因的植物表达载体pBch 通过农杆菌介导法导入甘兰型油菜H165 中。经卡那霉素对T0 、T1 和T2 代植株进行连续的筛选和菌核病(sclerotiniasclerotinorium) 接种试验, 共获得33 株抗菌核病的T2 代植株。对其中的22 株进行PCR 检测, 从6 株扩增得到与菜豆几丁质酶基因大小对应的DNA 带型,Southern 杂交证实其中2 株呈现阳性结果, 表明外源基因已整合到油菜基因组中。对转化植株抗菌核病的接种试验及在菌核病高发区的种植试验表明, 含几丁质酶基因的转化体对菌核病具有抗性, 并已得到T3

The plant expression vector pBch containing chitinase gene from bean was introduced into Brassica napus (variety H165) by Agrobacterium infection. After continuous screening of T 0, T 1 and T 2 plants by Km and fungal inoculation, 33 transgenic T 2 plants were obtained. While PCR analysis was carried out on 22 transgenic plants, 6 plants gave the DNA band corresponding to the size of chitinase gene. Southern hybridization further confirms that, of the 6 plants, 2 plants showed positive signals, indicating the foreign genes have been integrated into the rape genome. Fungal challenge and field testing of the transgenic plants in the area which suffers from the disease seriously displayed that the transgenic plants containing chitinase gene expressed the resistance to Sclerotinia sclerotinorium, and the seeds of T 3 were obtained.