外源脱水应答转录因子CBF4基因转化玉米的获得

Obtainment of Transformed Maize with Dehydration-Responsive Transcription Factor CBF4 Gene

用PCR方法克隆了拟南芥脱水应答转录因子CBF4基因,以逆境诱导表达基因rd29A的启动子为驱动,构建了逆境诱导表达载体pBAC146。用基因枪转化法转化玉米优良自交系的幼胚和胚性愈伤组织,轰击后的愈伤组织经过筛选、分化和植株再生过程,共获得36棵转基因植株。经PCR、PCR-Southern和Southern检测表明,外源目的基因已成功整合到部分转基因玉米株系的基因组中。人工干旱处理下,抗旱生理指标测定显示,一个转基因株系的脯氨酸含量和叶绿素含量比野生型对照提高一倍,间接表明转基因株系的抗旱能力在某种程度上有所提高。

The expression vector of CBF4 gene cloned from Arabidopsis thaliana by PCR and driven with promoter of rd29A gene, a preceding stress responsive gene, was constructed. Immature embryo and embryogenic callus of several excellent maize inbred lines were transformed with biolistic bombardment. After in vitro culture steps of callus induction, resistance selection, differentiation and plant regeneration, thirty six transgenic plants were obtained. PCR, PCR-Southern, and Southern blot analysis indicated that successful integration of alien target gene into genome of transgenic maize lines was happened. Measurement of drought physiological indexes under artificial drought treatment showed that either proline content or chlorophyll content of a transgenic line increased over one time than that of its wild type, suggesting that the drought resistance ability of transgenic maize might be improved to a certain extent.