用gfp基因标记法研究大豆根瘤菌在大豆根部定殖结瘤情况

Adoption of gfp Marker Gene Method to Study Soybean Rhizobium and the State of Nodulation Residence on Soybean Root

采用三亲本杂交的方法,将绿色荧光蛋白基因(gfp)转入高效、抗逆、广适应性的快生大豆根瘤菌Sinorhizobium frediiCCBAU 01287中,获得含gfp基因的转基因菌株CBAU 01287(G);平板传代和共生检测表明:外源质粒在CCBAU 01287中能够自我复制,稳定遗传。进一步研究表明,gfp标记菌CCBAU 01287(G)可用于实时监测根瘤菌在大豆根部的早期定殖情况和定殖密度的测定;标记菌株对大豆的生长及生物量的积累与出发菌株的效果无显著差异。

Adopting triple-parent hybridizing method, green-fluorescent protein gene （gfp） was transferred into Sinorhizobium fredii CCBAU 01287, a fast growing soybean rhizobium strain with efficient nitrogen fixation, wide adaptability, and resistance, and obtained a transgenic strain named CCBAU 01287 （G） containing gfp. The generation transferring on flat plate and symbiotic determination indicated that the exogenous plasmid could auto-replicate and stably transfer to next generations. Further study also indicated that CCBAU 01287 （G） marked with gfp could inspect early residence of the rhizobium on soybean root, and determine the residence density. The marked strain has no effect on the growth of soybean and there are not notable differences between it and the starting strain on biomass accumulation.