甘蔗转基因甘露糖筛选系统的建立

Establishment of Mannose Selection System in Sugarcane Transformation

以甘露糖作为筛选底物,对甘蔗品种新台糖22号进行临界筛选浓度测定,获得愈伤组织的继代、分化、生根的临界筛选浓度。而后应用含有甘露糖筛选标记基因pmi及绿色荧光蛋白报告基因GFP的植物表达载体,通过农杆菌介导法对新台糖22号的愈伤组织进行遗传转化。用所测定的临界筛选浓度先后进行继代、分化、生根筛选培养,获得抗性植株。对获得的抗性植株分别进行pmi基因和GFP基因的PCR检测,以及GFP显微镜荧光检测,结果证实已成功建立了高效的甘蔗转基因甘露糖筛选系统。

In this research the killing curve of mannose was tested at the subculture,differentiation and rooting stages of sugarcane cultivar ROC22. Then the expression cassette containing a pmiselection marker gene and a GFP report gene was transformed into sugarcane embryonic callus byAgrobacterium-mediated method. Resistant plants were obtained after selection by Mannose. The PCR detection result and GFP microscope observation result both show that our lab had established a high efficiency PMI/Mannose selection system in sugarcane transformation.