南方农业学报2012，Vol.43Issue(8)：1086-1089,4.DOI:10.3969/j:issn.2095-1191.2012.08.1086
融合杀虫基因对甘蔗的遗传转化
Fusion insecticidal gene for genetic transformation of sugarcane
摘要
Abstract
[ Objective ]The aim of this experiment is to produce transgenic sugarcane plants with high insect-resistance by using MAR sequence mediated transgenic expression and fusion insecticidal gene to overcome the exogenous gene inactivation,strengthen gene expression and delay insect resistance. [ Method ] Using the embryogenic calli sugarcane variety ROC25 as the acceptors,its sensitivity of hygromyein to callus proliferation,bud differentiation and rooting were tested. Fusion insecticidal gene (AVAc-CpTI) flanked on both sides of the MARs were introduced into sugarcane by A. tumefaciens infection. [ Result ]The optimum screening concentration of hygromyein for callus proliferation and budding was 50 mg/L; as for rooting,the concentration was 30 mg/L. Thirteen transgenic lines were attained when the vector ligated with MARs in the same direction on both sides of the fused gene,and resulted in 48 transgenic positive sugarcane plants by methods of southern blotting; which was 62.5% higher than that of the control. On the other hand,8 transgenic lines without MARs,produced only 7 confirmed transgenic sugarcane plants in the control group. [Conclusion]The screening system of hygromyein resistance for sugarcane cultivar ROC25 was established. Fusion insecticidal gene (AVAc-CpTI) that flanked on both sides of the MARs were successfully integrated into sugarcane genome by A .tumefaciens infection and transgenic plants were obtained. In the genetic transformation of sugarcane,fusion insecticidal gene that flanked on both sides of the MARs could increase the number of transgenic lines and improve the transformation efficiency of exogenous gene.关键词
甘蔗/融合杀虫基因/野苋菜凝集素基因/豇豆胰蛋白酶抑制剂基因/MAR序列Key words
sugarcane/fusion insecticidal gene/Amaranthus viridis agglutinin/CpTI/MAR sequence分类
农业科技引用本文复制引用
邓智年,魏源文,黄诚梅,潘有强,吕维莉,李杨瑞..融合杀虫基因对甘蔗的遗传转化[J].南方农业学报,2012,43(8):1086-1089,4.基金项目
国家科技支撑计划项目(2007BAD30B02) （2007BAD30B02）
广西科技基础条件平台建设项目(09-007-08) （09-007-08）
广西科学研究与技术开发计划项目(桂科能0815011-6-1,桂科攻10100005-9) （桂科能0815011-6-1,桂科攻10100005-9）
广西科学基金项目(2010GXNSFA013102,桂科基0991021) （2010GXNSFA013102,桂科基0991021）
广西农业科学院基本科研业务专项项目[200803Z(基),200902Z(基)] （基）
广西农业科学院科技发展基金项目(2007023) （2007023）
