Improvement of the Hydroponic Growth and Waterlogging Tolerance of Petunias by the Introduction of vhb Gene  

粪透明颤菌血红蛋白基因促进转基因矮牵牛在水培条件下生长并增强其抗涝能力(英文)

作　　者：毛自朝[1] 胡鸢雷[1] 钟瑾[1] 王立霞[1] 郭俊毅[1] 林忠平[1] 

机构地区：[1]北京大学蛋白质工程和植物基因工程国家重点实验室,北京100871

出　　处：《Acta Botanica Sinica》2003年第2期205-210,共6页Acta Botanica Sinica（植物学报：英文版）

摘　　要：The coding sequence of Vitreoscilla hemoglobin (vhb) was cloned with PCR technique from Vitreoscilla stercoraria Pringsheim. The plant expression vector with vhb gene under the control of CaMV 35S promoter was constructed and used in the transformation of Petunia hybrida Vilm by the Agrobacterium mediated procedure. The results of PCR amplification and Southern hybridization indicated that the vhb gene had been integrated into the petunia genome and the vhb gene expression had been detected by RT-PCR amplification. In hydroponic culture the transgenic petunias grew much better than non-transgenic controls. For further analysis of hypoxia tolerance of transgenic petunia, the petunia plants with vhb gene were submerged into liquid MS medium. The transgenic plants survived in hypoxic condition and grew out of the liquid surface in a few weeks, while non-transgenic plants were still submerged and suffocated in culture solution without ability to grow out of liquid medium in submersed culture for four to five weeks. The vhb gene transformed petunia plants had been planted and tested in a simulated flooding condition, and showed obvious tolerance to water-logging. It seen is that hemoglobin gene from Vitreoscilla might have the potential use in molecular breeding for the improvement of plant resistance to hypoxia and flooding.通过PCR程序克隆粪透明颤菌 (VitreoscillastercorariaPringsheim)血红蛋白基因 (vhb)的编码区 ,将其置于CaMV 35S启动子的驱动下导入矮牵牛 (PetuniahybridaVilm)。PCR和Southern杂交分析证明vhb基因已整合到受体基因组中 ,而RT_PCR检测证实了转基因矮牵牛中vhb基因转录水平的表达。观察了转基因株系在液体培养基中对淹水和缺氧的反应 ,发现vhb的表达明显促进了水培植物的生长。为进一步研究转基因植物在低氧条件的耐受能力 ,将上述的转基因株系在静置的液体培养基中进行完全的淹没培养 ,结果显示转基因植株具有较强的低氧耐受能力 ,培养两周后能从淹没状态长出液面 ,并由此而在液体中较正常地生长 ,而未转基因的对照不能长出淹没的培养基表面 ,在 4～ 5周后因缺氧而窒息死亡。另将上述转基因株系在温室中盆栽并进行抗涝分析 ,在模拟的持续淹水胁迫中 ,转基因株系比对照表现出较强的忍受能力。这些结果预示vhb基因在抗涝作物培育和提高水培植物缺氧耐受能力的分子育种方面具有较良好的应用前景。

关 键 词：Vitreoscilla hemoglobin transgenic petunia hypoxic tolerance WATERLOGGING 

分 类 号：Q943[生物学—植物学]