高粱BTx623幼胚遗传转化及再生体系的建立

Establishment of Sorghum BTx623 Immature Embryos Genetic Transformation and Regeneration System

高粱是世界上仅次于小麦、水稻、玉米和大麦的重要作物之一,虽然高粱基因组已经完成了测序,但是针对高粱测序品种BTx623,高效、稳定的遗传转化和再生体系的缺乏,阻碍了高粱遗传育种和功能基因组研究发展。本研究以高粱基因组测序品种BTx623幼胚为外植体材料,利用农杆菌介导的方法以抗草铵膦的Bar基因为筛选标记进行高粱遗传转化。通过筛选愈伤组织对不同浓度草铵膦的适应性,确定了高粱品种BTx623遗传转化中合适的草铵膦浓度为2.5 mg/L。以BTx623幼胚为外植体转化材料,通过农杆菌遗传转化方法,获得抗性愈伤组织。通过在分化培养中添加浓度为0.0067 mg/L的ZNC,建立了高粱品种BTx623的植株再生体系,获得了再生植株。因此,本研究成功获得了抗性愈伤组织和再生植株,建立了高粱品种BTx623遗传转化及再生体系,这对高粱功能基因组研究和遗传育种技术发展具有重要意义。

Sorghum is one of the most important crops in the world after wheat,rice,maize,and barley.Although the sorghum genome has been sequenced,the lack of efficient and stable genetic transformation and regeneration system for the sorghum sequenced variety BTx623 has hindered the development of sorghum genetic breeding and functional genome research.In this study,the immature embryos of sorghum genome-sequencing variety BTx623 was used as the explants material,and the Bar gene resistant to glufosinate ammonium was used as the screening marker for sorghum genetic transformation by Agrobacterium-mediated method.By screening the adaptability of callus to different concentrations of glufosinate ammonium,the appropriate concentration of glufosinate ammonium in the genetic transformation of sorghum variety BTx623 was determined to be 2.5 mg/L,and BTx623 immature embryo was used as explants to obtain resistant callus.Callus regeneration was obtained by adding ZNC with concentration of 0.0067 mg/L in differentiation culture.Therefore,this study successfully obtained resistant callus and regenerating plants,and established a genetic transformation and regeneration system of sorghum variety BTx623,which is of great significance to the research of functional sorghum genome and the development of genetic breeding technology.