农杆菌介导的黄瓜未受精子房培养遗传转化体系的建立  

Transformation System Through Agrobacterium tumefaciens Mediated Unfertilized Ovary Culture in Cucumber

作　　者：魏爱民[1] 杜胜利[1] 韩毅科[1] 刘楠[1] 

机构地区：[1]天津科润黄瓜研究所,天津300192

出　　处：《中国瓜菜》2014年第5期5-9,共5页China Cucurbits And Vegetables

基　　金：国家科技计划项目(2012AA100202-6);天津市科技计划项目(12ZCDZNC03700)

摘　　要：为了建立一种直接获得转基因纯系的转化方法，首次开展了农杆菌介导的黄瓜未受精子房培养遗传转化体系研究。试验以除草剂草胺膦为选择标记，胚胎发生阶段，最适除草剂质量浓度为0.5~1.0 mg＆#183;L-1，幼苗生根阶段除草剂质量浓度为2.0 mg＆#183;L-1；以羧苄青霉素为抑菌剂，最适抑菌质量为450 mg＆#183;L-1；最佳浸染方法为OD600=0.3农杆菌工程菌，浸染10 min；共培养时间为3 d；在浸染菌液和共培养基中加入0.01%的silwet-77，有利于获得较多抗性胚胎及抗性植株。该遗传转化体系的建立，为黄瓜转基因方法研究开辟了新途径。A new transformation technology through Agrobacterium tumefaciens mediated cucumber unfertilized ovary cul-ture was studied. Herbicide glufosinate-ammonium was used as the selection marker,the optimum herbicide selection pres-sure during embryogenesis was 0.5-1.0 mg＆#183;L-1,and optimum selection pressure during plantlet rooting was 2.0 mg＆#183;L-1;the optimum carbenicillin concentration for bacteria control was 450 mg ＆#183; L-1;the optimum infection method was soaking ovary slices into A. tumefaciens solution with OD600 at 0.3 for 10 min,and then co-cultured for 3 days;0.01%surfactant silwet-77 in bacteria solution and co-culture medium was beneficial for transformation. The establishment of this new technology pro-vided a new way for cucumber transformation research.

关 键 词：黄瓜 未受精子房 农杆菌 转化体系 Cucumber(Cucumis SATIVUS L.) 

分 类 号：S642.2[农业科学—蔬菜学]