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中国农学通报 ›› 2010, Vol. 26 ›› Issue (7): 177-180.

• 林学 园艺 园林 • 上一篇    下一篇

沙田柚遗传转化再生体系的建立

胡新喜1,2,敖小平2,邓子牛1,3,熊兴耀1,2   

  • 收稿日期:2009-11-26 修回日期:2009-12-13 出版日期:2010-04-05 发布日期:2010-04-05
  • 基金资助:

    国家转基因植物研究与产业化开发专项

Establishment of Regeneration System for Genetic Transformation of Citrus grandis Osbeck cv. Shatianyou

  • Received:2009-11-26 Revised:2009-12-13 Online:2010-04-05 Published:2010-04-05

摘要:

以沙田柚的实生苗不同外植体为对象,进行了沙田柚遗传转化再生体系建立研究。研究结果表明:以实生苗上胚轴作为外植体,以MS无机盐+100 mg/L肌醇+0.2 mg/L维生素B1+1 mg/L维生素B6+1 mg/L烟酸+3%蔗糖+0.8%琼脂(pH5.8)+5 mg/L 6-BA为不定芽诱导培养基,暗培养7 d后,转移至光/暗周期16/8 h的条件下培养,不定芽的分化率高达91.1%;不定芽在1/2MS基本培养基+3%蔗糖+0.7%琼脂+1.5 mg/L IBA+0.2%活性炭的生根培养基,生根率高达80%;以卡那霉素作选择剂时,选择浓度为50mg/L。

关键词: 辽宁地区菜豆炭疽病生理小种初鉴报告, 辽宁地区菜豆炭疽病生理小种初鉴报告

Abstract:

Transgenic plants appear to be a new way to citrus improvement and a powerful tool for studying gene functions. The establishment of an efficient in vitro plant regeneration system suitable for genetic transformation is the first step in this approach. Using epicotyl and hypocotyl pieces and cotyledons of Citrus grandis Osbeck cv. Shatianyou seedlings grown in vitro as explants,a efficient regeneration system was established. The results were obtained as follow. When the epicotyl pieces were explants were cultured on the adventitious shoot regeneration medium (MS salts +100 mg/L inositol +0.2 mg/LV-B1+1 mg/LV-B6+1 mg/L niacin + 3% sucrose + 5 mg/L6- BA + 0.8% agar,pH5.8) and incubated in darkness for 7 days,then in the condition of 16/8 h light/darkness cycle,adventitious shoot regenerated from 91.1% of the epicotyl explants. When the adventitious shoots were cultured on the medium (1/2MS+1.5 mg/L IBA +3%sucrose +0.7%agar,pH5.8) , adventitious root regenerated from 80% of the adventitious shoot after grown in vitro for 40d. A s a selectable agent, concentration of Kanamycin is 50 mg/L for epicotyl pieces.