尾赤桉DH201-2遗传转化体系建立的研究  

Establishment of Genetic Transformation of Eucalyptus urophylla × E. camaldulensis Clone DH201-2

作　　者：范春节[1] 曾炳山[1] 裘珍飞[1] 刘英[1] 李湘阳[1] 陈李花[1,2] 

机构地区：[1]中国林科院热带林业研究所,广东广州510520 [2]广西大学林学院,广西南宁530005

出　　处：《浙江林业科技》2009年第4期15-20,共6页Journal of Zhejiang Forestry Science and Technology

基　　金：国家林业局"948"引进项目"桉树转基因技术及抗病育种基因质粒引进"(2006-4-70);热林所基础科研项目(2007-23)

摘　　要：以建立的尾赤桉(Eucalyptus urophylla×E.camaldulensis)DH201-2无性系高频再生体系为基础,利用GUS染色组织分析法研究了根癌农杆菌菌株、农杆菌预培养时间、浸染时间和共培养时间等因素对尾赤桉茎段和叶片外植体遗传转化的影响,探讨了适宜尾赤桉转化的卡那霉素和抑菌抗生素头孢塞污钠的使用浓度。结果表明,较优的转化条件是:采用农杆菌菌株GV3101,以茎段为受体材料,经过预培养6 d,在光密度OD600≈0.5的菌液中浸泡30 min,然后转移到共培养培养基中共培养4 d,再转移到含90 mg/L卡那霉素和300 mg/L头孢噻肟钠的筛选培养基上,进行转化植株的再生,同时在共培养培养基和菌液中添加100μM的乙酰丁香酮能够提高茎段外植体的转化效率。Based on the established regeneration system, an efficient kanamycin and cefotamicine system of Eucalyptus urophylla× E. camaldulensis clone DH201-2 mediated by Agrobacterium was developed. Stems and leaves were served as explants for transformation, and the factors affecting the transformation frequency, such as pre-culture time, infection time, co-cultured time and pH value of co-culture medium were studied by GUS coloration analysis. Experiments were conducted on concentration of kanamycin and cefotamicine for better transformation effect Of E. urophylla × E. carnaldulensis clone DH201-2. The results indicated that better transformation system was as following： using Agrobacterium strains GV3101 and stem segment pre-cultured for 6 days, infected 30 minutes in solution of bacterial concentration OD600≈0.5, adding 100 BM of AS in co-culture medium and bacterial solution, co-cultured for 4 days, and then transferred to culture medium with 90mg/L of kanamycin and 300 mg/L of cefotamicine for the regeneration oftransgenic plantlets.

关 键 词：尾赤桉 茎段 叶片 农杆菌介导法 遗传转化 GUS基因 

分 类 号：S792.39[农业科学—林木遗传育种]