北柴胡移栽试管植株与种子植株中皂苷的动态分析

Dynamic analysis of saikoside in test-tube plants of Bupleurum chinense compared with seeding plants

目的为应用离体快速繁殖技术进行北柴胡优良种质选育提供实验依据。方法应用分光光度法测定北柴胡移栽试管植株不同生长期的根、茎叶中柴胡总皂苷的量,应用柱前衍生化HPLC-UV法测定柴胡皂苷a和d的量,并以同型、同时的种子植株作为对照。结果北柴胡试管植株根、茎叶中柴胡总皂苷及皂苷a和d的量在试管苗阶段最低,在一年生植株中最高,在两年生植株中的营养生长期次高,在花期、果期、枯萎期依次降低;在根中的量远高于茎、叶;引种自山西左权的移栽试管植株的量均高于引种自山西陵川和万荣的类型;移栽试管植株根、茎叶中的量均高于相同类型的种子植株中的量。结论离体快速繁殖是纯化北柴胡种质和提升药材品质、提高药材产量的有效方法;快速繁殖植株具有优良的遗传稳定性,选择综合性状好的种质作为繁殖的起始材料,能够保证后代性状的优良和整齐一致;果期二级花序的种子成熟期为移栽试管植株根的适宜采收期。

Objective To provide a scientific basis for breeding on Bupleurum chinense by rapid propagation method in vitro.Methods Pre-column derivatization HPLC-UV and visible-spectrophotometry methods were used to determine the contents of saikosaponin a,saikosaponin d,and saikosaponins in the root and aerial part of test-tube plants of B.chinense in different growing periods,and compared with seeding plants of same breeding and growing periods.Results The contents of saikosaponins,saikosaponin a,and saikosaponin d were minimum in roots,stems,and leaves of plantlets.They were the uppermost in annual test-tube plants,and were the second at vegetative growth phase of biennial test-tube plants.The contents were reduced ordinally at bud forming phase,seed ripening phase,and wilting phase of biennial test-tube plants.The content in the root was much higher than that in aerial part of test-tube plants.The contents of test-tube plants which were introduced from Shanxi Zuoquan were higher than which from Shanxi Wanrong and Lingchuan.The content in method of B.chinense breeding was an effective way to maintain and improve the yield in roots,stems,and leaves of test-tube plants were higher than those in seeding plants.Conclusion The rapid propagation in vitro is an effective method for purifying germplasm of B.chinense and ascending honewort medicinal material quality and yield.The genetic stability of rapid propagation plants are excellent,so the starting material should have excellent comprehensive traits to ensure offspring with good characters and uniformity.The seed maturing stage in the secondary inflorescence is suitable harvest time for the root.