柳兰组织培养和快速繁殖体系优化研究

Research on Optimization System on Tissue Culture and Rapid Propagation of Geranium pratense L.

以抑制柳兰褐化和建立柳兰离体快繁体系为目的,试验以柳兰种子为起始材料,获得柳兰无菌苗,在此基础上研究不同添加物对柳兰褐化的影响和不同培养基对柳兰生长、增殖的影响,并筛选获得适宜柳兰生长、增殖培养的培养基。试验结果表明,用20%NaClO对柳兰种子灭菌1.5 min抑菌效果最好,种子萌发率100%;活性炭和长时间继代培养可有效缓解柳兰褐化现象并诱导获得不定根,植株生长状况较好,不同浓度Vc和Na2S2O3没有有效抑制柳兰褐化,植株生长量小且没有不定根生成;不同培养基对柳兰生长影响不同,其中1/2 MS和WPM培养基中高生长显著高于其他培养基中植株高生长,但植株生长较细弱,1/2 MS+AC 2 g/L培养基中植株生长矮小但健壮;1/2 MS+AC 2 g/L培养基中主根长度、须根数量显著高于其他培养基,不同培养基中主根数量、须根长度没有显著差异;适宜柳兰生根、壮苗的培养基是1/2MS+AC 2 g/L;适宜柳兰增殖培养的培养基是1/2 MS+6-BA 0.5 mg/L+NAA 0.05 mg/L,增殖系数为5.4。试验结果为柳兰快速繁殖和规模化生产提供了依据。

In order to establish Geranium pretense＇ s culture in vitro, the seeds of Geranium pretense were used as donor materials. The effect of different accretion on browning of Geranium pretense was studied and the opti- mal medium composition for rooting and proliferation of Geranium pretense were selected by comparison the function of different medium culture on growing of Geranium pretense. These results showed that, the seeds of Geranium pratense were sterilizated by 20% NaC10 for 1.5 min was efficiency and the rate of germination was 100%; Browning was alleviated by active carbon and longer timing culture in vitro and adventitious roots creat- ed. Vc and Na2S203 didn＇t alleviate browning effectively and no adventitious roots were induced in it, also the seedlings didn＇t grow nearly; Different medium culture had different effecting on Geranium pretense. The height growth of Geranium pretense in 1/2 MS and WPM was higher than other treatments markedly, but these seed- lings were slim and fragile. Conversely, they were stocky in 1/2 MS＋AC 2 g/L medium; The length of taproot and number of fibril in 1/2 MS＋AC 2 g/L were higher than other treatments markedly, and the number of tap- root and the length of fibril had no markedly different in different medium culture; The optimal medium compo- sition for rooting was 1/2MS＋AC 2 g/L and 1/2MS＋ 6-BA 0.5 mg/L ＋NAA 0.05 mg/L was suit for proliferation of Geranium pretense, the multiplication was 5.4. These results provided theoretical basis for rapid propagation and production in enlarged scale of Geranium pretense.