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利用农杆菌介导法将大白菜启动子BcA9与Barnase的融合基因对烟草叶片进行遗传转化,经含Basta的筛选培养基连续筛选,获得了转基因再生植株.转基因植株经PCR扩增,表明BcA9-Barnase基因已整合到烟草基因组中.对转基因植株和野生型植株的花药发育进行细胞学比较观察,显示野生型植株的花药发育正常,花粉粒形态正常,活力高;而转基因植株则发生花药绒毡层细胞提前降解,花粉母细胞退化,成熟花药中无正常花粉粒产生等系列现象.转基因烟草植株最终表现为花药瘦小、自交不能正常结籽等.同时,在高温条件下转基因植株不育性稳定,无育性恢复现象.
Agrobacterium-mediated transformation of Chinese cabbage promoter BcA9 Barnase fusion gene on tobacco leaves genetic transformation by Basta-containing screening medium continuous screening of transgenic regenerated plants obtained by PCR amplification of transgenic plants showed that BcA9- The Barnase gene has been integrated into the genome of tobacco.The cytological comparison of the anther development of transgenic plants and wild type plants showed that the anther development of wild type plants was normal and the morphology of pollen grains was normal and the vitality was high.And anther tapetum Layer cells prematurely degraded, pollen mother cells degenerated, and no normal pollen grains were produced in mature anthers.Transgenic tobacco plants eventually showed small anther size and selfed self-pollination, etc. At the same time, the transgenic plants were sterile at high temperature Stable, infertility recovery phenomenon.