基于液-固交替培养的水曲柳快速微繁系统研究

Rapid Micropropagation of Fraxinus mandshurica Rupr.Based on Liquid and Solid Alternate Culture

以茎部木质化、叶片老化、顶芽休眠的水曲柳组培苗为材料,开发了一种液体—固体交替培养的水曲柳(Fraxinus mandshurica Rupr.)快速高效的再生系统,该技术通过液体悬浮培养在短时间内使腋芽萌发,并在固体培养中腋芽离体再生获得新的组培苗。在补充不同植物生长调节剂的WPM液体和固体培养基上,可以诱导水曲柳腋芽萌发并伸长成苗。发现在添加了0.6 mg·L^(-1) TDZ的WPM液体培养基中暗培养,7~15 d之内可促使水曲柳腋芽100%萌发,将萌发的嫩芽切下后接种到0.05 mg·L^(-1) TDZ和0.6 mg·L^(-1) BA的WPM固体培养基中光照培养,腋芽在1~2个继代内可以伸长成苗,苗平均高为2.64 cm,增殖系数达到4.04。将生根的苗移栽,50 d后存活率为90%。该技术的建立有助于水曲柳的大规模繁殖,并且液体—固体交替循环培养,简单、可控、易操作,适用于不同的生产条件,减少成本。

A rapid and efficient regeneration system of liquid-solid alternate culture was developed for the tissue culture seedlings of Fraxinus mandshurica with lignificated stalk,aged leaves and dormant apical buds,and the axillary buds were germinated in a short time by liquid suspension culture,and the axillary buds were regenerated in vitro in solid culture to obtain the new tissue culture seedlings.On liquid and solid media supplemented of WPM with different plant growth regulators,axillary buds of F.mandshurica were induced to germinate and grow into seedlings.It was found that the axillary bud germination rate of F.mandshurica could reach 100%within 7-15 d after dark culture in WPM liquid medium supplemented with 0.6 mg·L^(-1) TDZ.After cutting the sprouted buds and inoculating them into WPM solid medium of 0.05 mg·L^(-1) TDZ and 0.6 mg·L^(-1) BA,axillary buds could grow into shoots after 1-2 subcultures with an average height of 2.64 cm and a multiplication coefficient of 4.04.After transplanting,the survival rate of rooting seedlings was 90%after 50 d.The establishment of the method could be conducive to the large-scale propagation of F.mandshurica,and the liquid-solid alternate circulation culture,simple,controllable,easy to operate,suitable for different production conditions and reduces the cost.