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目的研究凹叶厚朴愈伤组织的诱导,并对其产生的有效成分厚朴酚与和厚朴酚总量进行检测,以期获得生产药用成分的新途径,补充药材资源的不足。方法以不同母株种子幼苗的不同营养器官为外植体,采用不同种类及不同质量浓度的植物调节剂诱导愈伤组织,并在继代培养基中加入前体化合物L-苯丙氨酸及D,L,β-苯丙氨酸对愈伤组织进行诱导培养;HPLC法检测不同培养途径获得的愈伤组织中厚朴酚与和厚朴酚量的总和。结果筛选出最佳愈伤组织诱导培养基为:B5+6-BA 2.0 mg/L+2,4-D 1.5 mg/L;不同母株种子幼苗诱导的愈伤组织总酚量存在显著差异,质量分数变化范围为0.004%~0.228%;不同营养器官作为外植体诱导的愈伤组织总酚量存在差异,以幼茎为外植体诱导的愈伤组织总酚量最高可达0.25%;继代培养基中添加前体化合物D,L,β-苯丙氨酸,能有效提高愈伤组织总酚量8~10倍。结论凹叶厚朴愈伤组织中含有少量的厚朴酚及和厚朴酚,不同的母株种子幼苗及不同幼苗器官诱导的愈伤组织总酚量都有差异,D,L,β-苯丙氨酸能有效促进愈伤组织中总酚的合成。
Objective To study the induction of Callus of Magnolia officinalis and to detect the total amount of honokiol and honokiol which are the active ingredients in order to obtain new ways of producing medicinal ingredients and to supplement the shortage of medicinal materials. Methods Different vegetative organs of seedlings of different parental plants were used as explants. Callus tissues were induced by plant modifiers with different kinds and concentrations of different concentrations. L-phenylalanine and its precursor compound D, L, β-phenylalanine were used to induce the callus. HPLC method was used to determine the total amount of honokiol and honokiol in callus obtained from different culture ways. Results The optimum medium for callus induction was B5 + 6-BA 2.0 mg / L + 2,4-D 1.5 mg / L. The total phenols content of callus induced by seedlings of different mother plants was significantly different, The mass fraction varied from 0.004% to 0.228%. The total phenols of callus induced by different vegetative organs were different. The total phenols of callus induced by young stems were up to 0.25%. Subculture medium added precursor compounds D, L, β-phenylalanine can effectively improve the total phenolic callus 8 to 10 times. Conclusions Calluses of Magnolia officinalis contains a small amount of magnolol and honokiol, and the total phenols content of callus induced by different seedlings of different parental plants and organs of different seedlings are different. D, L, β-amphetamine Acid can effectively promote the synthesis of total phenols in callus.