【作者】 邹晶晶； 曾祥玲； 蔡璇； 陈洪国； 王彩云；

【Author】 ZOU Jing-jing;ZENG Xiang-ling;CAI Xuan;CHEN Hong-guo;WANG Cai-yun;School of Nuclear Technology and Chemistry & Biology,Hubei University of Science and Technology;Key Laboratory for Biology of Horticultural Plants,Ministry of Education,College of Horticulture & Forestry Sciences,Huazhong Agricultural University;

【机构】 湖北科技学院核技术与化学生物学院； 华中农业大学园艺植物生物学教育部重点实验室；

【摘要】 本研究以结实桂花（Osmanthus fragrans Lour.）早期子叶期的未成熟合子胚为外植体材料,在MS+0.5mg·L^-12,4-D+1.0mg·L^-16-BA初始培养基上暗培养30天后诱导得到的胚性愈伤组织作为遗传转化的受体,利用根癌农杆菌介导的方法,通过对不同的侵染及培养条件的摸索,初步探索了桂花的遗传转化体系的条件。结果表明,桂花胚性愈伤组织的卡那霉素的选择敏感性浓度为200mg·L^-1,头孢霉素浓度为300mg·L^-1。GUS瞬时表达表明最优的转化条件为:农杆菌菌株为EHA105时,侵染菌液浓度为0.6,侵染时间为20min,共培养天数为2d,共培养时培养基中添加100μM AS时效率最高。选择培养基为MS+0.5mg·L^-1NAA+1.0mg·L^-1BA+200mg·L^-1Km+300mg·L^-1Cef。下一步将继续优化选择培养基抗生素的种类和浓度,解决反菌问题,以获得完整转化植株。更多还原

【Abstract】 The embryogenic callus induced by the cotyledon segments from the ZEs explants collected at the early cotyledonary stage,with MS medium suppled with 1.0mg·L^-1BA and 0.5mg·L^-12,4-D as the initial medium,were used for the Agrobacterium-mediated transformation in O.fragrans.The selective concentration of kanamycin（Km）was 200mg·L^-1,and cefotaxime（Cef）was 300mg·L^-1.According to the GUS transient assay,the opmizated transformation conditions were preliminary abtained:the strain of Agrobacterium tumefaciens was EHA105;the infection time was 20 min;the co-culture day was 2d;the co-culture medium was MS+100μM acetosyringone（AS）.The selective medium was MS+0.5 mg·L^-1NAA+1.0mg·L^-1BA+200mg·L^-1Km+300mg·L^-1Cef.However,embryonic callus was infected by the agrobacterium during the selective culture process 20 days later.Type and concentration of antibiotics will be optimized in the later study to obtain the complete transformated plant更多还原