作物学报 ›› 2018, Vol. 44 ›› Issue (10): 1423-1432.doi: 10.3724/SP.J.1006.2018.01423
• 作物遗传育种·种质资源·分子遗传学 • 下一篇
陈倩楠1,2,王轲1,汤沙1,杜丽璞1,智慧1,贾冠清1,赵宝华2,叶兴国1,刁现民1,*()
Qian-Nan CHEN1,2,Ke WANG1,Sha TANG1,Li-Pu DU1,Hui ZHI1,Guan-Qing JIA1,Bao-Hua ZHAO2,Xing-Guo YE1,Xian-Min DIAO1,*()
摘要:
农杆菌介导的谷子遗传转化效率一直是制约谷子功能基因研究及转基因育种效率的主要因素。本研究以冀谷11谷子幼穗为外植体, 选取0.5~1.0 cm的幼穗, 切成0.5 cm左右的小段, 置改良后的MS培养基上诱导15~20 d形成胚性愈伤组织3120块。愈伤组织侵染转化前用侵染液悬浮浸泡, 并经45°C热激预处理3 min, 可以有效提高26.1%的瞬时遗传转化效率。将转化后的愈伤组织经草丁膦(PPT)筛选后获得513块抗性愈伤组织, 抗性愈伤组织获得率为16.4%。抗性愈伤组织经分化、壮苗培养后获得7株抗性植株, 经PCR和Southern杂交检测得到6株T0代转基因阳性株, 对T3代转化植株叶片进行PPT抗性分析, 并结合Bar蛋白抗体试纸条鉴定, 表明Bar基因已稳定整合到谷子基因组中。本研究建立了农杆菌介导转化谷子优良品种的遗传转化体系, 对提高谷子转基因育种效率和谷子模式研究系统的建立有重要意义。
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