天山雪莲组织培养体系的优化

Optimization of Tissue Culture System of Saussurea involucrata

通过研究天山雪莲种子消毒萌发条件、愈伤组织培养和继代培养条件,优化出了一套能获得总黄酮成分含量较高的天山雪莲愈伤组织的组织培养体系。结果表明:将种子用70%乙醇处理30s、0.1%升汞溶液处理10min,并种植于MS培养基,可获得生长状态良好的雪莲苗。比较了7种培养基对天山雪莲无菌苗叶片愈伤组织的诱导情况,愈伤诱导率均能达到100%,其中4种培养基获得愈伤组织适合继代培养。在继代培养中,①号培养基的愈伤组织易分化出根,②号培养基、③号培养基的愈伤组织易分化出芽;④号培养基可得到较多黄绿色致密愈伤组织。对①号、②号、③号、④号培养基获得愈伤组织的总黄酮含量检测,发现其总黄酮含量为1.14%~7.37%,远高于《中华人民共和国药典(2015版)》不少于0.15%的规定,以②号培养基的愈伤组织总黄酮含量最高。

By studying the germination conditions, callus culture conditions and secondary culture conditions of Saussurea involucrata, we optimized the tissue culture system to obtain the callus with high content of total flavonoids. The results showed that the seeds were treated with 70% ethanol for 30 seconds and 0.1% mercury solution for 10 minutes, and planted on MS medium, the seedlings of Saussurea involucrata in good growth condition could be obtained. The induction of callus from leaves of Saussurea involucrata Sterile seedlings on 7 media was compared. The induction rate of callus could reach 100%. Among them, 4 the obtained callus were suitable for subculture. In subculture, callus of No. 1 medium was easy to differentiate into roots, callus of No. 2 and No. 3 were easy to differentiate into buds, and callus of No. 4 medium could obtain more yellow-green dense callus. The content of total flavonoids in callus obtained from medium No. 1, No. 2, No. 3 and No. 4 was determined. It was found that the content of total flavonoids was 1.14%~7.37%, which was much higher than that stipulated in Chinese Pharmacopoeia (2015 edition).The content of total flavonoids in callus obtained from callus of No. 1 medium was the highest.