甘露糖筛选系统的建立及在甘蔗转基因中的应用

Establishment of a Novel Selection System by Mannose and Its Application in Sugarcane Transformation

用甘露糖作为筛选底物,对甘蔗品种福农95-1702外植体的再生进行研究,建立福农95-1702的甘露糖筛选体系;在甘蔗愈伤组织的继代、分化、生根三个阶段进行抗性筛选时,甘露糖占总糖的比例分别是60%、60%和30%。在以上筛选体系的基础上,将福农95-1702作为受体材料,通过基因枪轰击法,将含有cryIA(c)抗虫基因的基因表达盒与筛选标记基因(pmi)进行共转化,通过甘露糖筛选,最终获得26株抗性再生植株,经PCR检测,其中4株呈阳性。初步说明cryIA(c)基因已经整合到福农95-1702的基因组中,获得了4株转基因阳性甘蔗植株。

The novel selection system for the regeneration of sugarcane of FN95-1702 by mannose was established.With mannose screening, the proportion of the mannose at the subculture, differentiation and rooting stages of sugarcane callus were 60%, 60%, and 30%, respectively.Based on the screening system established, the cassette containing cryIA （c） gene and screening mark gene of pmi was transformed into FN95-1702 by co-transformation of bombardment, and 26 regenerated plants were obtained by selection with mannose.Among them, four transformed plants were detected by PCR to be positive and the cryIA（c） gene was assumed to be integrated into sugarcane genome.From above, it indicated that the novel selection system is feasible in the transformation of sugarcane and four transgenic plants of anti-insect sugarcane were obtained.