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以冬小麦品种 8901、5-98、99-92和 104等品种的幼穗和幼胚为材料,用基因枪转化含逆境诱导转录因子 DREB 和bar 基因的质粒 pBAC128F(7 024 bp)。经筛选与植株再生,共获得 70 多个转基因小麦植株及其后代株系。转基因株系经PCR 分析和 RNA 点杂交检测,结果表明外源转录因子 DREB 基因已稳定整合到转基因植株及其后代株系中,并且在部分后代株系中获得了表达。叶片脯氨酸含量测定表明,有 16 个转基因株系的脯氨酸含量与非转基因对照相比,增加相当显著,其中10个株系的脯氨酸含量在1 100 μg/g以上,比对照提高了2倍多。室内抗旱模拟实验表明,转基因株系停止浇水15 d 后,叶片仍然表现绿色,而对照叶片则失绿、枯干;复水 10 d 后,转基因株系恢复活力,对照则死亡。研究表明,利用逆境诱导型启动子(rd29B)来增强外源DREB基因的表达,能显著改良小麦的抗旱性。
The pBAC128F (7 024 bp) plasmid containing the stress-responsive transcription factors DREB and bar genes was transformed with young shoots and young embryos of winter wheat cultivars 8901, 5-98, 99-92 and 104, respectively. After screening and plant regeneration, a total of more than 70 transgenic wheat plants and their offspring strains were obtained. The transgenic lines were detected by PCR and RNA dot blot. The results showed that the foreign transcription factor DREB gene was stably integrated into the transgenic plants and their offspring lines, and was expressed in some progeny lines. Proline content of leaves showed that the proline content of 16 transgenic lines increased significantly compared with non-transgenic control, of which the proline content of 10 lines was above 1 100 μg / g, More than doubled. Drought simulation experiments showed that the transgenic leaves were green after 15 days of watering, while the control leaves were green and withered. After 10 days of rewatering, the transgenic lines were revitalized and the control died. Studies have shown that the use of stress-inducible promoter (rd29B) to enhance the expression of exogenous DREB gene can significantly improve the drought resistance of wheat.