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目的:诱导玄参愈伤、不定根产生,分离内生菌,并比较其中有效成分哈巴俄苷的含量。方法:玄参嫩叶片消毒切成小块接种于含不同激素水平的MS,N6培养基诱导愈伤;不定根诱导采用液体培养:将愈伤小块先转入不含激素MS液体培养基,100 r.min-1室温震荡培养,待开始出现不定根后转入含0.05 mg.L-1NAA+2 mg.L-16-BA的MS液体培养基继续震荡培养,内生菌分离采用常规方法。哈巴俄苷含量测定采用紫外分光光度法,测定波长255 nm。结果:MS培养基+NAA0.05,0.2,2 mg.L-1+6-BA 2 mg.L-1均能诱导出质地较疏松、生长较快的愈伤组织;接种1.5 g愈伤30 d左右可得到100 mL满瓶不定根;从玄参鲜块根分离出4株可产哈巴俄苷的内生菌。玄参愈伤组织、不定根及四株内生菌发酵液的哈巴俄苷含量依次分别为:0.411,0.099 5,0.451,0.444,0.489,0.440 g.L-1.结论:玄参愈伤组织中哈巴俄苷含量是不定根的4倍,内生菌发酵液哈巴俄苷含量可达到与愈伤相当水平,具有开发应用的潜力。同时为研究玄参与内生菌相互作用打下基础。
Objective: Induced Scrophulariaceae callus injury, adventitious roots produce endophytic bacteria isolated, and compare the content of the active ingredient Harpagoside. Methods: The young leaves of Scrophularia ningpoensis were disinfected and cut into small pieces and inoculated on MS and N6 medium containing different hormone levels to induce callus. Adventitious root induction was induced by liquid culture: the callus was first transferred to hormone-free MS liquid medium r.min-1 at room temperature shaking culture, to be started after adventitious root into MS liquid medium containing 0.05 mg.L-1NAA + 2 mg.L-16-BA continued shaking culture, endophyte separation using conventional methods. Determination of Harpagoside using UV spectrophotometry, determination of wavelength 255 nm. Results: MS medium + NAA0.05, 0.2, 2 mg.L-1 + 6-BA 2 mg.L-1 could induce more loose and fast-growing callus; 1.5 g callus 30 d about 100 mL of adventitious roots can be obtained full root; Scrophulariaceae callus, adventitious roots and four endophytic fermentation broth of carbidoside levels were: 0.411,0.099 5,0.451,0.444,0.489,0.440 gL-1 .Conclusion: Scrophulariaceae callus Huabu Russia Glucosinolate content is adventitious root 4 times, endophytic fermentation broth hydrabasidin content can reach a considerable level with the callus, with the potential of development and application. At the same time to lay the foundation for the study of mysterious participation and endophyte interaction.