野罂粟花粉管通道法转双价抗真菌病基因方法研究

Pollen Tube Pathway Anti-Fungus Gene Transformation of Papaver nudicaule

以野罂粟开放花蕾为受体,以载有几丁质酶和β-1,3-葡聚糖酶基因双价植物表达载体pBLGC的质粒为供体,在11%浓度的蔗糖溶液中,加入花粉和含有目的基因的质粒DNA.得到药粉处理液。然后辅以人工授粉的方法将花粉处理液滴注野罂粟柱头上获得种子,将种子经卡那霉素筛选和PCR检测,有目的基因特异带出现。获得T1植物。实践证明:在野罂粟上,利用花粉管通道法进行几丁质酶和β-1,3-葡聚糖酶基因转化方法可行,为特殊中药材转基因技术提供一个新方法。

Plasmid DNA of pBILGC containing Chitinase and β-1,3-glucanase gene and fresh pollen of Pa- paver nudicaule were mixed in 11% sucrose solution, then the pollen suspension were pollinated on stigmas of the Papaver nudicaule by artificial pollination. Transformed plants were primarily confirmed by PCR amplification and kanamycin selection, trangenetic T1 plant were obtained.The experimental results indicated that exog- enous gene could be transformed by the Pollen tube pathway Transformation, and the new breeding method of Papaver nudicaule was provided.