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园艺学报 ›› 2004, Vol. 31 ›› Issue (5): 691-693.

• 研究报告 • 上一篇    下一篇

基因枪介导的高羊茅基因转化体系的建立

马生健1,3;曾富华1*;徐碧玉2;卢向阳3*;吴志华1,3   

  1. (1 湛江师范学院生物系, 湛江524048 ; 2 中国热带农业科学院热带作物国家重点实验室, 海口571101 ; 3 湖南农业大学生物技术系, 长沙410128)
  • 收稿日期:2004-02-02 修回日期:2004-04-22 出版日期:2004-10-25 发布日期:2004-10-25

Protocol Establ ishment of Tall Fescue Transgene by Biolistics

Ma Shengjian1,3;Zeng Fuhua1*; Xu Biyu2; Lu Xiangyang3*;Wu Zhihua1,3   

  1. (1 Department of Biology , Zhanjiang Normal College,Zhanjiang 524048, China; 2 The State Key L aboratory of Tropical Crops Biotechnology,Chinese Academy of Tropical Agricultural Sciences , Haikou 571101,China;3 Department of Biotechnology , Hunan Agricultural University , Changsha 410128 , China)
  • Received:2004-02-02 Revised:2004-04-22 Online:2004-10-25 Published:2004-10-25

摘要: 用高羊茅( Festuca arundinacea Schreb. ) 种胚离体诱导的胚性愈伤组织作受体进行了基因枪的轰击试验, 确定了以轰击2 枪, 子弹距可裂圆片6 cm , 子弹距轰击受体6 cm 为最佳轰击参数, 建立了基因枪优化转化方案, 获得了遗传稳定的抗除草剂PPT 的转化植株。随机抽取的转化植株的Bar 基因PCR检测、总DNA 点杂交, 皆成阳性, 除草剂PPT 抗性试验表明, 转化植株比对照有了更强的抗性。

关键词: 草坪草, 愈伤组织, 基因枪, 转化, 抗除草剂

Abstract: In this paper , excised mature embryos of tall fescue were used as explants to induce embryonic calli , which were used as materials to optimize bombardment parameters of biolistics , and the most suitable delivery parameters for PDS-1000/ He were 6 cm shoot distance , 6 cm distance between particles and rupture disks , 1100 psi helium pressure and ombarding 2 shots. An optimized t ransformation protocol was established and transgenic turfgrass varieties resistant to herbicide with stable heredity were acquired.All tested tall fescues selected randomly were positive by PCR analysis and by dot blotting analysis for bar gene. The results of daubing tests with herbicide showed t ransgenic plants had more resistance than plants
cont rol did.

Key words: Turfgrass, Callus, Biolistics, Transformation, Herbicide resistance

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