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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。
The Zm IPT2 gene was cloned from maize inbred line Zheng 58 and the monocot plant expression vector was constructed. Agrobacterium tumefaciens was used to infect maize inbred line K10 through inoculation of embryo germinant, and the molecular test and functional verification of transgenic progenies were carried out. The results showed that the full length cDNA of Zm IPT2 gene was 969 bp, and its pMCBIA5300-ubi-Zm IPT2 vector was successfully constructed. Agrobacterium tumefaciens was used to inoculate 5 163 K10 seeds to obtain 48 PCR-positive seedlings in T0 generation, of which 13 were harvested for seed harvesting. The PCR positive rate of the 3 T2 generation lines was in accordance with the segregation ratio of 3: 1, And the RT-PCR test was positive. The chlorophyll content and cytokinin content of the two T2 generation transgenic lines were significantly higher than those of the control at the mature stage, and the relative green leaf area and the leaf area were significantly or significantly higher than the control Heavy and plot yield was significantly higher than the control. Preliminary results showed that overexpression of Zm IPT2 gene in maize delayed leaf senescence and increased maize yield.