中山大学学报自然科学版 ›› 2011, Vol. 50 ›› Issue (4): 84-90.

• 研究论文 • 上一篇    下一篇

拟南芥多效性基因CPR5转化水稻中花11的研究

王亚琴1,2,陈兴瑶1,阳成伟2

  

  1. (1.华南理工大学生物科学与工程学院,广东 广州 510006;2. 华南师范大学生命科学学院//广东省植物发育生物工程重点实验室,广东 广州 510631)
  • 收稿日期:2010-10-30 修回日期:1900-01-01 出版日期:2011-07-25 发布日期:2011-07-25

Transformation of Pleiotropic Gene CPR5 from Arabidopsis into Zhonghua 11 (Oryza sativa L. subsp. japonica)

WANG Yaqin1,2,CHEN Xingyao1,YANG Chengwei 2   

  1. (1. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China; 2. Guangdong Key Lab of Biotechnology for Plant Development, College of Life Science, South China Normal University, Guangzhou 510631,China)
  • Received:2010-10-30 Revised:1900-01-01 Online:2011-07-25 Published:2011-07-25

摘要: 利用根癌农杆菌介导法将拟南芥多效性基因CPR5转化水稻中花11,同时优化了其再生体系和遗传转化体系。结果表明:相比27 ℃、暗培养,32 ℃、持续光照培养可使诱导率提高到100 %,且缩短了诱导周期;〖JP2〗预分化培养后,在附加5 mg/L 6BA和1 mg/L NAA的分化培养基上分化率和再生率可分别达到100 %和90.50 %。同时探索了较高转化频率的条件为 A600≈ 0.05~0.1的农杆菌菌液浸染5 min,共培养时间为3 d,同时添加1张用AAI培养液浸湿的无菌滤纸。对部分转化植株进行PCR、Southern blot和半定量RTPCR检测,结果表明AtCPR5基因已经整合到水稻基因组中,在所试验的转化植株中均为单拷贝,但表达程度存在差异。

关键词: 中花11(Oryza sativa L. subsp. japonica), AtCPR5, 多效性基因, 遗传转化

Abstract: This study investigated the transformation of pleiotropic gene AtCPR5 into Zhonghua 11(Oryza sativa L. subsp. japonica) mediated by Agrobacterium tumefaciens, and optimized the regeneration system and genetic transformation system. It was showed the induction rate of callus could be increased to 100 % and the period of induction could be shortened under the condition of continuous light and 32 ℃ compared with the condition of dark and 27 ℃. After predifferentiation, the differentiation rate and regeneration rate could be raised to 100 % and 90.50 %, respectively, by adding 5 mg/l 6BA and 1 mg/l NAA in the differential medium. Additionally, the higher transformation efficiency was obtained with the infected concentration of A.tumefaciens to an A600 of 0.05~0.1, 5 minutes of infection and 3 days of cocultivation by placing a filter moistened with AAI medium. Furthermore, the results of PCR, Southern blot and semi-quantitative RTPCR indicated AtCPR5 gene has been integrated into rice genome with single copy, however, the expression level 5 are different in transgenic plantlets.

Key words: Zhonghua 11(Oryza sativa L. subsp. japonica), AtCPR5, pleiotropic gene, genetic transformation

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