园艺学报 ›› 2005, Vol. 32 ›› Issue (02): 307-309.

• 研究报告 • 上一篇    下一篇


张应华1, 2;王小佳1   

  1. (1 西南农业大学园林园艺学院, 重庆市蔬菜学重点实验室, 重庆400716; 2 云南农业大学园林园艺学院, 昆明650201)
  • 收稿日期:2004-06-17 修回日期:2004-08-24 出版日期:2005-04-25 发布日期:2005-04-25

Transformation of Cabbage with In sulin-like Growth Factor-ⅠRegulated byMatrix Attachment Regions

Zhang Yinghua1, 2;Wang Xiaojia1   

  1. (1College of Horticulture and Landscape, Southwest Agricultural University, Key Lab in Vegetable Research of Chongqing,Chongqing 400716, China; 2College of Horticulture and Landscape, Yunnan Agricultural University, Kunming 650201, China)
  • Received:2004-06-17 Revised:2004-08-24 Online:2005-04-25 Published:2005-04-25

摘要: 采用根癌农杆菌介导法, 将表达框两翼构建了核基质结合区(matrix attachment region, MAR)的胰岛素样生长因子-Ⅰ ( Insulin-like Growth Factor-Ⅰ, IGF-Ⅰ) 基因导入甘蓝中, 在含卡那霉素的培养基上连续4次筛选, 通过10个批次的转化试验, 获得25株抗性植株, PCR检测均呈阳性, 随机选取5株进行基因组Southern杂交, 结果有3株出现了杂交带, 表明IGF-Ⅰ基因已成功整合到甘蓝基因组中。

关键词: 胰岛素样生长因子-Ⅰ, 核基质结合区, 甘蓝, 转化

Abstract: Hypocotyl explants of 7 day-old cabbage (Brassica oleracea L. var. capitata L. ) seedlings were pre-cultured on bud induction medium (MS +BA 3 mg/L +NAA 0.1 mg/L ) for two days. The explants were then infected with Agrobacteriumtum efaciens strain EHA105 harboring a plasmid vector containing IGF-Ⅰ gene that flanked bymatrix attachment regions (MARs) isolated from the genome of tobacco (Nicotiana tabacum L. ). After two days of co-culture, the explants were transferred onto agrobacterium suppressive medium (MS +BA 3 mg/L +NAA 0.1 mg/L +Cb 400 mg/L) and cultivated for six days. Through four times of kanamycin-resistant selected culture (MS +BA 3 mg/L +NAA 0.1 mg/L + Km 5 mg/L + Cb 400 mg/L) , 25 resistant shootswere obtained from 10 transformation experiments with total 15 200 explants. Some of the
resistant regenerated plants were tested by PCR analysis and Southern blot, which showed IGF-Ⅰ gene was integrated into cabbage genome.

Key words: Insulin-like growth factor-Ⅰ, Matrix attachment region, Cabbage, Transformation