甘蓝型黄籽油菜自交系转化体系的建立  

Establishment of Transformation System Using Inbred Line of Yellow-Seeded Brassica napus

作　　者：林呐[1] 刘列钊 殷家明[1] 王瑞[1] 柴友荣[1] 李加纳[1] 

机构地区：[1]西南大学农学与生物科技学院/重庆市油菜工程技术研究中心/南方山地农业教育部工程研究中心,重庆400716

出　　处：《作物学报》2012年第12期2185-2191,共7页Acta Agronomica Sinica

基　　金：高等学校学科创新工程计划"111"项目(B12006);国家自然科学基金项目(31071450)资助

摘　　要：利用黄籽甘蓝型油菜自交系建立和优化了遗传转化系统。首先构建了由质粒pCNR与Δ6-脂肪酸脱氢酶基因插入到植物的高效表达载体pCAMBIA2301G。利用在Murashige和Skoog培养基（含有200μmolL–1乙酰丁香酮）培养5~7d的下胚轴外植体与农杆菌株LBA4404共培养63~69h（pCNR）,再于芽诱导培养基上培养3个月诱导芽再生。在最佳条件下,平均转化效率约为1.3%。转化植株的GUS分析和PCR分析结果表明,外源基因成功导入甘蓝型油菜。Southern杂交表明,这些转化子含有目标基因1~2个拷贝。用气相色谱分析转基因植物种子的脂肪酸,γ-亚麻酸含量达8.2%。In this study, we established a transformation system using an inbred line of yellow-seeded Brassica napus. Hypocotyl explants precultured for 5-7 d on Murashige and Skoog medium containing 200 μmol L-1 acetosyringone were cocultured with Agrobacterium tumefaciens strain LBA4404 （pCNR） for 63-69 hours. The plasmid pCNR was constructed by inserting Δ6-fatty acid desaturase gene from Rhizopus stolonifer into plant high-efficient expression vector pCAMBIA2301G. Kanamycin-tolerant shoots were regenerated on shoot induction medium for three months after Agrobacterium inoculation. The average transformation efficiency was about 1.3% under optimal conditions. Results from GUS assay and PCR analysis of transformed plants indicated that the introduced gene was integrated into B. napus genomes. The result of Southern blot revealed that those transformants carried one or two copies of the goal gene. The fatty acids of the transgenic plant seeds were analyzed by GC, and the γ-linolenic content was 8.2%.

关 键 词：甘蓝型油菜 Δ6-脂肪酸脱饱和酶 转化 

分 类 号：S565.4[农业科学—作物学]