转基因杨树中农杆菌残存状况分析

Analysis on the status of agrobacterium translated into poplar mediated by Agrobacterium rhizogenes

以转发根农杆菌741杨为材料,在继代和移栽过程中对其残存农杆菌进行追踪检测。结果表明:培养基中附加头孢噻钨纳300 mg/L可将农杆菌基本抑制;随着转化植株在含抗生素培养基中继代时间的延长,残存农杆菌的数量和活性逐步下降,在继代18个月后,无法培养出目的菌落,但转化植株一旦脱离抗生素环境,农杆菌数量和活性又将得到恢复;残存农杆菌主要分布于植株体表和茎中,且影响组培苗PCR检测结果;将转化株系组培苗移栽到花盆半月和田间6个月后,植株体表、体内均未检测到目的农杆菌,但在室内培养半个月后2个株系根际土壤中检测到目的菌落。

Taking poplar 741 mediated by Agrobacterium rhizogenes as material, an examination was made concerning the survival of agrobacterium in transgenic plants during the process of transplanting and subculturing on the nutrient medium. Result shows that the agrobacterium on transgenic plant can be controlled basically when the transgenic plants cultured in medium with 300 mg/ L ceftiofur; the quantity and activity of the survival agrobacterium on transgenic plant will decline as time progresses, and when the transgenic plant has been cultured 18 months, the agrobacterium on transgenic plant cannot be cultured on YEB medium; while the agrobacterium on transgenic plant will resume when the transgenic plant cultured in medium without ceftiofur; the transgenic plants all have the survival agrobacterium, which distribute transgenic plant surface and stem inside and effect the result of PCR to the transgenic plants; when the transgenic plants with agrobacterium are placed into flowerpot half a month and into farm half of year, the agrobacterium on transgenic plant cannot be detected on the transgenic plant, whereas the agrobacterium can be detected in half a month in 2 rhizospher.