延缓叶片衰老ZmIPT2基因的玉米遗传转化及功能验证

发布时间：2018-09-11 21:06

【摘要】：从玉米自交系郑58中克隆Zm IPT2基因并构建单子叶植物表达载体,通过农杆菌侵染萌动胚方法将其转入玉米自交系K10中,对转基因后代进行分子检测和功能验证。结果表明,Zm IPT2基因c DNA全长969 bp,成功构建其单子叶植物表达载体p CAMBIA5300-ubi-Zm IPT2。农杆菌侵染萌动胚法共转化K10种子5 163粒,获得T0代PCR阳性幼苗48株,其中13株结实收获种子;获得的3个T2代株系PCR阳性率符合3∶1的分离比,且RT-PCR检测呈阳性;2个T2代转基因株系的成熟期叶绿素含量和细胞分裂素含量极显著高于对照,相对绿叶面积和叶面积持绿期极显著或显著高于对照,百粒重和小区产量显著高于对照。结果初步证明,Zm IPT2基因在玉米中的过表达可延缓叶片衰老,提高玉米产量。
[Abstract]:The Zm IPT2 gene was cloned from maize inbred line Zheng58 and the expression vector of monocotyledonous plant was constructed. The gene was transferred into maize inbred line K10 by Agrobacterium tumefaciens infection and germinating embryo method. The results showed that the full length of c DNA of Zm IPT2 gene was 969 bp, and its monocotyledonous expression vector p CAMBIA5300-ubi-Zm IPT2. was successfully constructed. A total of 5,163 K10 seeds were transformed by Agrobacterium tumefaciens infecting germinating embryo, 48 T0 generation PCR positive seedlings were obtained, 13 of them harvested seeds, and the PCR positive rate of 3 T2 lines was in accordance with the isolation ratio of 3:1. The chlorophyll content and cytokinin content of 2 T2 transgenic lines were significantly higher than that of the control, and the relative green leaf area and leaf area were significantly or significantly higher than those of the control. 100 seed weight and plot yield were significantly higher than the control. The results showed that the overexpression of Zm IPT2 gene in maize could delay leaf senescence and increase maize yield.
【作者单位】：东北农业大学农学院;巴彦淖尔市农牧业科学研究院;重庆市农业科学院;中国农业科学院作物研究所;
【基金】：国家转基因生物新品种选育重大专项(2016ZX08003003-005) 黑龙江省留学归国人员科学基金(LC201411)
【分类号】：S513

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