金线莲和铁皮石斛无糖组培与传统组培的对照试验

A Comparative Experiment on Sugar-free Tissue Culture and Traditional Tissue Culture of Anemone roxburghii and Dendrobium candidum

为了对比无糖组培和传统组培之间的差异,以金线莲和铁皮石斛为例,探索无糖微环境中CO2的最适宜浓度和通气时间。试验在光照强度4600 lx的可控范围内,按照浓度梯度通入CO2,通过CO2响应曲线综合分析后得出光合有效辐射的最佳CO2浓度,在该微环境下进行无糖组培培养,同时进行叶片细胞的形态、气孔开闭程度的测量及酶活性的检测比较。结果表明经过无糖组培培育出来的植株比传统组培苗更健壮、气孔功能更完善、抵御外界胁迫环境能力更强、成活率更有保障。在生产上铁皮石斛最适宜的CO2浓度为1200~1400μmol/mol,金线莲最适宜的CO2浓度为800~1000μmol/mol,通气时间均应选在黑暗环境下,三种不同培育环境下SOD、POD、CAT的活性:无糖组培>传统组培>驯化苗。该试验为无糖组培在林下中草药工厂化生产中降低污染,控制成本的同时提高成活率提供技术支持。

In order to compare the differences between sugar-free tissue culture and traditional tissue culture,the optimal concentration and aeration time of CO2 in sugar-free microenvironment were explored by taking Anemone roxburghii and Dendrobium candidum as examples.The experiment was conducted in the controllable range of light intensity of 4600 lx,CO2 was introduced based on concentration gradient,and the optimal CO2 concentration for the photosynthetic effective radiation was obtained through the comprehensive analysis of the CO2 response curve.The sugar-free tissue culture was carried out under this microenvironment.Meanwhile,the morphology of leaf cells,the degree of stomatal opening and closing and the enzyme activity were measured and compared.The results showed that the plants cultured in sugar-free tissue culture were more robust than those cultured in conventional tissue culture,with more perfect stomatal function,stronger ability to resist external stress and more secure survival rate.The optimum CO2 concentration for Dendrobium candidum and Anemone roxburghii is 1200~1400μmol/mol and 800~1000μmol/mol respectively.The aeration time should be chosen in dark environment.Activities of SOD(Superoxide Dismutase),POD(Peroxidase)and CAT(Catalase)under three different breeding environments:Sugarfree tissue culture>traditional tissue culture>acclimated seedlings.This research could provide technical support for reducing pollution,controlling cost and improving survival rate of sugar-free tissue culture in the factory production of undergrowth Chinese herbal medicine.