摘要
RT-PCR扩增得水稻质膜H+-ATPase全长cDNA(PM-H+-ATPase),构建了植物表达载体获得质粒pBI121-PM-H+-ATPase,农杆菌介导、真空渗入法转化PM-H+-ATPase基因入拟南芥,得35株T1代卡那抗性植株。抗性植株PCR分析表明,抗性植株整合了目的基因。Northern杂交分析进一步表明T3代转基因拟南芥表达了目的基因。抗盐(NaHCO3和NaCl)性分析表明:转基因植株的耐盐能力明显高于野生型;盐碱胁迫下转基因植株开花优先于野生型植株。
The complete coding region of rice plasma membrane H^＋-ATPase （PM-H^＋-ATPase） was amplified by RT-PCR, and was fused into pBI121 plant expression vector to construct the pBI121-PM-H^＋-ATPase plasmid. The resultant plasmids were introduced into Arabidopsis by Agrobacterium tumefaciens （strain EHA105）-mediated transformation using the vacuum infiltration method. Thirty-five individual kanamycin resistant plants were obtained. PCR analysis showed that kanamycin resistant plants had integrated the rice PM-H^＋-ATPase. Furthermore, Northern blot analysis showed that T3 generation transgenic Arabidopsis seedlings have overexpressed rice PM-H^＋-ATPase. Salt tolerance oftransgenic A rabidopsis seedlings was analyzed, and the results showed that transgenic A rabidopsis plants have greater salt tolerance at the seedling stage than wild-type plants under salt stress. Aslo, transgenic A rabidopsis plants flowering have precedence over wild-type plants under NaHCO3 stress.
出处
《分子植物育种》
CAS
CSCD
2007年第4期451-454,共4页
Molecular Plant Breeding
基金
国家高技术研究发展863计划(2002AA241111)
黑龙江省科技计划项目(GB04B606-02-02)资助。
