国槐种胚愈伤组织培养与异黄酮量的分析

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7d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.

Callus culture of embryos in Sophora japonica and analysis on isoflavones content

Objective To establish the callus culturing system of embryos in Sophora japonica, and to analyze the contents of isoflavones in callus. Methods Through adding the plant growth regulators of different kinds and at different concentration, the optimal media for callus induction, suspension cell culture, and the highest isoflavone content were obtained. Results The optimal medium for callus induction of embryos in S. japonica was B5 + 2, 4-D (1.0 mg/L) + 6-BA (0.2 mg/L) + sucrose (20 mg/L). In solid medium, the callus was growing rapidly after 7 d, and the yield reached to the highest on the day 35 (fresh weight 7.413 7 g), but the isoflavone content had no significant changes. In suspension culture, the yield of callus reached to the highest on the day 24 (fresh weight 11.563 8 g), and the callus turned into decline phase after 24 d, in which the fresh and dry callus weights were descending. With continuous culture, the cells were browning gradually, aging, and dying at last, but the isoflavone content was opposite to growth curve, and reached its lowest point (4.826 mg/g) on the day 24. Conclusion The induction and cultivation of embryos callus in S. japonica and the isoflavone content have a larger correlation with the plant growth regulators of the kinds and the concentration.