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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (1): 89-92.

• 林学 园艺 园林 • 上一篇    下一篇

拟青山海关杨高效遗传转化系统的建立

陈玉玉,苏乔,祖勇,纪纯阳   

  • 收稿日期:2008-10-30 修回日期:2008-11-23 出版日期:2009-01-05 发布日期:2009-01-05

Establishment of High Efficient System for Genetic Transformation of Populus pseudo-cathayana ×P. deltodides Barry CV. ‘Shan Hai Guan’)

  • Received:2008-10-30 Revised:2008-11-23 Online:2009-01-05 Published:2009-01-05

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摘要： 本实验以叶片为外植体，建立了拟青山海关杨的高频再生系统；在此基础上，利用GUS瞬时表达法研究了菌种、预培养时间、侵染菌液浓度、侵染时间和乙酰丁香酮（AS）浓度5个因素对拟青山海关杨叶片遗传转化的影响。结果表明，拟青山海关杨叶片最适不定芽诱导培养基为MS添加0.5mg/L的6-BA 和0.1mg/L的NAA，叶片再生频率为95%，平均不定芽数为10.07个；该杨树的高效遗传转化体系为：叶片省去预培养，菌种使用GV3101，侵染液OD600为 0.4～0.6，侵染10min，共培养培养基中添加乙酰丁香酮 200μmol/L，在此条件下，GUS荧光定量分析结果显示叶片的GUS活性达到最大值。

关键词: 高产粮田, 高产粮田, 土壤, 重金属, 土壤环境质量, 评价

Abstract: The high frequency regeneration system of Populus pseudo-cathayana ×P. deltodides Barry CV.‘Shan Hai Guan’ was established with leaf explants in our study. Furthermore, the factors related to transformation including Agrobacterium strains, pre-culture time, Agrobacterium concentration, inoculation time and acetosyringone (AS) concentration were evaluated via GUS transient expression. The results indicated that, the optimal adventitious induction culture was MS medium supplemented with 0.5mg.L-1 6-BA and 0.1mg.L-1 NAA, on which the regeneration rate reached 95% and 10.07 average adventitious buds were obtained. The application of Agrobacterium GV3101, no pre-culture, bacterium concentration at OD600=0.4~0.6, 10 min inoculation and supplement of 200μmol/L AS in co-culture medium produced the highest transient GUS expression detected by fluorescence quantification method, which were demonstrated optimal for high frequency transformation of ‘Shan Hai Guan’.