棕色棉茎尖基因枪遗传转化.pdfVIP

棉 花学 报 Cotton Science 20 16, 28(2): 129―137 色棉茎尖基因枪遗传转化 云鹏 冯鸿杰 程文瀚 张新宇 孙杰 刘永昌* ( 河子大学农学院/ 新疆兵团绿洲生态农业重点实验室 新疆石河子832003) 摘要院为了建立高效的棉花茎尖遗传转化体系 以棕色棉品种新彩棉5 号为材料 采用茎尖培养和基因枪遗传 转化方法 通过对凝固剂种类尧活性炭含量尧卡那霉素浓度尧渗透处理时间尧轰击压力尧轰击距离等因素的优化 建立了棕色棉新彩棉5 号茎尖基因枪遗传转化技术体系遥 结果表明 不 凝固剂种类尧活性炭含量尧卡那霉素 浓度尧渗透处理时间尧轰击压力和轰击距离对棕色棉茎尖转化率有明显影响遥 最佳的遗传转化体系为院前渗处 理4 h 轰击压力7.586 MPa 渊1100 psi冤 轰击距离9 cm 轰击后恢复培养4 d 然后在MSB+8 g 窑L 1 琼脂+ 125 mg 窑L 1 卡那霉素筛选培养基中筛选25 d遥 获得抗性芽后 在添加1 g窑L 1 活性炭的生根培养基中诱导抗性芽 生根 并获得抗性再生植株遥 通过PCR 和qPCR 检测 5200 个茎尖共获得16 株阳性转基因植株 转化率为 0.31%遥 该技术体系的建立为棕色棉的遗传转化奠定了基础 为彩色棉育种提供了新的材料遥 关键词院棕色棉曰茎尖曰基因枪遗传转化曰转化率 中图分类号院S562.03 文献标志码院A 文章编号院1002-7807(2016)02-0129-09 10.11963/issn.1002-7807.20 1602005 Hou Yunpeng, Feng Hongji e, Cheng Wenhan, Zhang Xinyu, Sun Jie, Liu Yongchang* ( 832000, To genetically transform cotton shoot tips, we established a biolistic genetic transformation system though the plant shoot tip culture technique and the gene gun-mediated transformation method, using the shoot tips of brown cotton Xincaimian No.5, a L. variety cultured in Xinji ang as materials. Several critical factors of the system were assessed, including the type of coagulant, the activated charcoal concentration, the kanamycin concentration, the penetration processing time, and the bombardment pressure and distance. Results showed that each factor clearly influenced the transformation rate of brown cotton shoot tips. An optimal biolistic genetic transformation technology system was determined: (1) shoot tips were cultured for 4 h before being bombarded in the osmotic medium; (2) the shoot tips were bombarded with helium pressure of 7.586 MPa ( 1100 psi), and a bombardment distance of 9 cm; (3) after