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目的构建国槐Sophora japonica种胚愈伤组织培养体系,并对愈伤组织中的异黄酮量进行初步分析。方法通过添加不同浓度不同种类的植物生长调节剂,得到国槐种胚愈伤组织诱导、悬浮细胞培养、异黄酮量最高的最佳培养基;通过对愈伤组织和悬浮细胞在生长中的生长量和异黄酮量进行测定,得到悬浮细胞最佳收获期。结果国槐种胚愈伤组织诱导的最佳培养基为B5+2,4-D(1.0 mg/L)+6-BA(0.2 mg/L)+蔗糖(20 mg/L)。固体培养基上,愈伤组织培养7 d之后组织进入迅速生长期,第35天,愈伤组织的生长量达到最高(鲜质量7.413 7 g),但其异黄酮量均无显著变化。悬浮培养中,愈伤组织培养24 d达到最高生长量(鲜质量11.563 8g),24 d后进入衰亡期,继续培养细胞其鲜质量、干质量均呈现下降趋势,且细胞逐渐变褐,最终死亡,其中异黄酮量的变化趋势与生长曲线相反,在24 d达到最低(4.826 mg/g)。结论国槐种胚愈伤组织诱导、培养和异黄酮量与植物生长调节剂种类及浓度均具较大关联。
Objective To construct the embryo callus culture system of Sophora japonica, and to analyze the amount of isoflavones in callus. Methods By adding different concentrations of different plant growth regulators, the optimal medium for inducing callus induction, suspension cell culture and highest isoflavone concentration of Sophora japonica was obtained. Through the growth of callus and suspension cells in growth The amount of isoflavones and determination of the amount of suspension cells obtained the best harvest. Results The optimal medium for the induction of embryo callus was B5 + 2,4-D (1.0 mg / L) + 6-BA (0.2 mg / L) and sucrose (20 mg / L). On the solid medium, the tissue entered the rapid growth stage after 7 days of callus culture. On the 35th day, the callus grew to the highest level (fresh weight 7.413 7 g), but its isoflavone content did not change significantly. In suspension culture, the callus reached the highest growth (fresh weight 11.563 8g) after 24 d of culture, and entered the decay stage after 24 d. The fresh weight and the dry weight of the cells continued to decline, and the cells gradually brown and eventually died , And the trend of isoflavone content was contrary to the growth curve, reaching the lowest (4.826 mg / g) at 24 d. Conclusions The induction, culture and isoflavone content of the seed embryo of Sophora japonica are closely related to the species and concentration of plant growth regulators.