水曲柳DNA浓度及滴加时间对杨树花粉管通道法结实率的影响

Effect on Concentration of Fraxinus mandshurica DNA and Time of dripping on Seed Setting Rate via Pollentube Pathway in Poplar

[目的]对适宜的水曲柳DNA浓度及滴加时间进行初步探索,获得具有水曲柳优势性状的杨树植株。[方法]通过花粉管通道法,将携带水曲柳基因组的DNA导入杨树。人工授粉后一定时间在花柱基部进行横切,把不同浓度DNA溶液滴加到子房切口端,10 d后统计结实率。[结果]在同一时间点,随着水曲柳DNA浓度的增加,结实率呈现逐渐降低趋势;随着授粉与滴加DNA时间的延长,同一浓度下呈现出结实率先降低后升高趋势。当水曲柳DNA溶液滴加时间控制在人工授粉后6 h左右,DNA浓度为50μg/ml时是在杨树上运用花粉管通道法转化水曲柳基因最适宜的条件,结实率可达17%。[结论]滴加时间与外源DNA不同梯度水平对转化结实率有很大影响。

[Objective] The aim was to study appropriate Fraxinus mandshurica DNA concentration and time of dripping in order to obtain poplar plant with advantage of Fraxinus mandshurica.[Method] Using pollen tube pathway transformation,DNA containing Fraxinus mandshurica gene was introduced into P.deltoides × P.cathayana.First step was taking artificial pollination among P.deltoides × P.cathayana,then crosscutting the base of style at four different levels and dripping five different concentrations of DNA solution into ovary incision end at every time level.Seed setting rate statistics were gotten 10 days later.[Result] The results showed that,at the same level,with the increasing of DNA concentration of Fraxinus mandshurica,seed setting rate presented a gradual decreasing trend ; with extending of the time between pollination and dripping DNA solution,seed setting rate appeared first low ebb and then rising.When the time of dripping was controlling in 6 hours so after artificial pollination,with the concentration of DNA of 50 μg/ml,the seed setting rate got 17％ that was the best consequence in the P.deltoides × P.cathayana of using pollen tube pathway transformation.[Conclusion] Time of dripping and exogenous DNA level had a great influence on the conversion rate of different gradient.