南林895杨离体培养及耐盐基因GmNHX1的转化

摘要：【目的】建立南林895杨杂交无性系最适遗传转化体系，为创新耐盐南林895杨种质资源奠定基础。【方法】以南林895杨叶片和茎段为外植体，对其再生体系和耐盐基因GmNHX1遗传转化的部分影响因子进行分析。【结果】使用NaClO消毒处理10～15min的外植体褐化率和污染率均较低；以叶片为外植体产生的不定芽状态良好，而茎段分化的不定芽容易白化死亡。PCR检测结果表明，转GmNHX1基因植株可扩增获得符合大小的特异性条带（1641bp）；通过荧光显微镜能够观察到转基因植株中的SGFP激发出的绿色荧光，证明GmNHX1基因已经高效整合到南林895杨的基因组中。【结论】叶片是南林895杨离体培养再生体系的最佳外植体来源，其离体培养所得的植株完全可以满足根癌农杆菌介导基因转化对受体的要求，可用于耐盐基因GmNHX1的遗传转化。

Culture in vitro of Nanlin 895 poplar and transformation of salt-tolerance related gene GmNHX1

The present study was conducted to establish optimizing genetic transformation system of Nanlin 895 poplar in order to lay the foundation for innovating its new germplasm resources. Method]The leaf and stem of Nanlin 895 polar were used as explants to study the affecting factors on regeneration system and genetic transformation of salt-tolerance related gene GmNHX1. Result] Browning rate and pollution rate were at low level when explants was sterilized with NaClO for 10-15 min. The adventitious buds induced from leaf were in good condition. However, those adventitious buds induced from stem was easy to albefaction and dead. PCR detection results showed that specific bands （1641 bp） could be amplified from GmNHX1 transgenic plantlets. The green fluorescence of sGFP inspired could observed by fluorescence microscopy, proving that the GmNHX1 gene had been efficiently integrated into Nanlin 895 poplar genome. Conclusion ] Leaf was the best explants resource for regeneration system of Nanlin 895 polar The induced plantlets in this way could meet the requirements for receptor of Agrobacterium-mediated gene transformation and be used in transformation of tolerance related GmNHX1 gene.