Three main parameters were selected to study their importance in transformation by budmicroinjection in non-head Chinese cabbage [Brassica campestris ssp. chinensis (L.) Makinovar. communis Tsen et Lee]. The results s...Three main parameters were selected to study their importance in transformation by budmicroinjection in non-head Chinese cabbage [Brassica campestris ssp. chinensis (L.) Makinovar. communis Tsen et Lee]. The results showed that the developmental stage of floral bud, theconcentrations of sucrose and surfactant Silwet L-77 were critical for the successfultransformation by this new method. The suitable bud size is 2-3 mm in length, the favorableconcentration of sucrose and surfactant Silwet L-77 are 8 and 0.02% respectively. When thesucrose concentration was greater than 10% or that of Silwet L-77 was above 0.1%, the treatedbuds became yellow and finally blighted. 4/6 T1 seedlings resistant to kanamycin were positiveby PCR analysis, and T2 progeny of all these positive T1 plants have one or more hybridizingbands by Southern blot. Under 5% sucrose, 0.02% Silwet L-77 and grade 2 bud (2-3 mm in itslength) parameters, the most favorable transformation efficiency is about 0.56%, and meanefficiency reaches 0.16% in all experiments indicating that bud microinjection is potentialtransformation way in non-head Chinese cabbage.展开更多
1材料与方法
1．1材料
1．1．1供试材料。白菜型油菜苏州青，市售。
1．1．2菌株及质粒。采用根癌农杆菌LBA4404，该菌株含有植物双元表达载体pCAMBIA1390，油菜油体基因与bFGF的融合基因已整合到该载体上，该融合基因由油菜油体基...1材料与方法
1．1材料
1．1．1供试材料。白菜型油菜苏州青，市售。
1．1．2菌株及质粒。采用根癌农杆菌LBA4404，该菌株含有植物双元表达载体pCAMBIA1390，油菜油体基因与bFGF的融合基因已整合到该载体上，该融合基因由油菜油体基因启动子启动。植物基因组提取试剂盒购自北京百泰克生物技术有限公司，Southern检测试剂盒为DIG High Prime DNA Labeling and Detection Starter Kit I，购于Roche公司。展开更多
文摘Three main parameters were selected to study their importance in transformation by budmicroinjection in non-head Chinese cabbage [Brassica campestris ssp. chinensis (L.) Makinovar. communis Tsen et Lee]. The results showed that the developmental stage of floral bud, theconcentrations of sucrose and surfactant Silwet L-77 were critical for the successfultransformation by this new method. The suitable bud size is 2-3 mm in length, the favorableconcentration of sucrose and surfactant Silwet L-77 are 8 and 0.02% respectively. When thesucrose concentration was greater than 10% or that of Silwet L-77 was above 0.1%, the treatedbuds became yellow and finally blighted. 4/6 T1 seedlings resistant to kanamycin were positiveby PCR analysis, and T2 progeny of all these positive T1 plants have one or more hybridizingbands by Southern blot. Under 5% sucrose, 0.02% Silwet L-77 and grade 2 bud (2-3 mm in itslength) parameters, the most favorable transformation efficiency is about 0.56%, and meanefficiency reaches 0.16% in all experiments indicating that bud microinjection is potentialtransformation way in non-head Chinese cabbage.
基金Supported by Bioreactor Important Special Item of 863-Program inthe "Eleventh Five-Year" Plan (No. 2007AA100503)Science and Technology Development Key Plan of Jilin Province( No.20070922)+1 种基金Cultivation Fund of Scientific and Technical Innovation Project Major Program of Higher Education Institutions ( No.70S018)Science and Technology Plan of Changchun City (No.06GG150)~~
文摘1材料与方法
1．1材料
1．1．1供试材料。白菜型油菜苏州青，市售。
1．1．2菌株及质粒。采用根癌农杆菌LBA4404，该菌株含有植物双元表达载体pCAMBIA1390，油菜油体基因与bFGF的融合基因已整合到该载体上，该融合基因由油菜油体基因启动子启动。植物基因组提取试剂盒购自北京百泰克生物技术有限公司，Southern检测试剂盒为DIG High Prime DNA Labeling and Detection Starter Kit I，购于Roche公司。
