Department of Biological Sciences, University of Lethbridge, 4401 University Drive, Lethbridge, AB T1K 3M4, Canada.
Plant Cell Rep. 2011 Apr;30(4):505-18. doi: 10.1007/s00299-010-0960-3. Epub 2010 Dec 5.
Plant transformation efficiency depends on the ability of the transgene to successfully interact with plant host factors. Our previous work and the work of others showed that manipulation of the activity of host factors allows for increased frequency of transformation. Recently we reported that exposure of tobacco plants to increased concentrations of ammonium nitrate increases the frequency of both homologous recombination and plant transgenesis. Here we tested the influence of KCl and salts of rare earth elements, Ce and La on the efficiency of Agrobacterium-mediated plant transformation. We found that exposure to KCl, CeCl(3) and LaCl(3) leads to an increase in recombination frequency in Arabidopsis and tobacco. Plants grown in the presence of CeCl(3) and LaCl(3) had higher biomass, longer roots and greater root number. Analysis of transformation efficiency showed that exposure of tobacco plants to 50 mM KCl resulted in ~6.0-fold increase in the number of regenerated calli and transgenic plants as compared to control plants. Exposure to various concentrations of CeCl(3) showed a maximum increase of ~3.0-fold in both the number of calli and transgenic plants. Segregation analysis showed that exposure to KCl and cerium (III) chloride leads to more frequent integrations of the transgene at a single locus. Analysis of transgene intactness showed better preservation of right T-DNA border during transgene integration. Our data suggest that KCl and CeCl(3) can be effectively used to improve quantity and quality of transgene integrations.
植物转化效率取决于转基因与植物宿主因子成功相互作用的能力。我们之前的工作和其他人的工作表明,宿主因子活性的操纵允许提高转化的频率。最近我们报道,暴露于增加浓度的硝酸铵的烟草植物增加同源重组和植物转基因的频率。在这里,我们测试了 KCl 和稀土元素盐,Ce 和 La 对农杆菌介导的植物转化效率的影响。我们发现,暴露于 KCl、CeCl(3)和 LaCl(3)导致拟南芥和烟草的重组频率增加。在 CeCl(3)和 LaCl(3)存在下生长的植物具有更高的生物量、更长的根和更多的根。转化效率分析表明,与对照植物相比,暴露于 50mM KCl 的烟草植物再生愈伤组织和转基因植物的数量增加了约 6.0 倍。暴露于不同浓度的 CeCl(3)导致愈伤组织和转基因植物的数量最大增加约 3.0 倍。分离分析表明,暴露于 KCl 和氯化铈导致转基因在单个基因座上更频繁地整合。转基因完整度分析表明,在转基因整合过程中右 T-DNA 边界更好地保存。我们的数据表明,KCl 和 CeCl(3)可有效用于提高转基因整合的数量和质量。