老瓜头悬浮细胞培养及基质消耗规律的初探

目的]为老瓜头资源利用提供前期的生物技术试验参考。方法]以老瓜头愈伤组织为外植体,运用正交设计试验,进行细胞悬浮培养,对接种量及悬浮培养基进行筛选试验研究,并测定了老瓜头细胞悬浮培养过程中硝酸盐、氨盐、磷酸盐、蔗糖及可溶性总糖含量的消耗。结果]老瓜头愈伤细胞最佳液体培养基为不添加任何激素的MS培养基;其最适接种量为40 g/L;黑暗培养条件适合其细胞生长,其生长周期较短,生物量大;在老瓜头细胞培养初期(0~9 d),培养中的硝酸盐、氨盐、磷酸盐、蔗糖含量均呈急剧下降趋势,分别为初始含量的7.19%、21.53%、0.28%、1.53%,之后各自变化趋于平缓。结论]初步建立了老瓜头悬浮细胞培养技术体系。

Study on the Suspension Cell Culture of Cynanchum komarrovii AI Iljiniski and Its Matrix Consumption Laws

Objective] The research aimed to provide the biotechnology test reference for the utilization of Cynanchum komarrovii resources.Method] With the calli of C.komarrovii as explants,the orthogonal design test was used to make the cell suspension culture.The screening test was made on the inoculation amount and suspension culture media.And the consumption of nitrate,ammonia salt,phosphate,sucrose and total soluble sugar during the cell suspension culture process of C.komarrovii were determined.Result] The optimum liquid medium for the calli of C.komarrovii was MS medium without adding any hormone and its suitable inoculation amount was 40 g/L.The dark culture conditions were suitable for the cell growth and it had shorter growth period and greater biomass.The content of nitrate,ammonia salt,phosphate,sucrose all showed the sharply decreasing trend during the initial cell suspension culture of C.komarrovii(0-9 d),being 7.19%,21.53%,0.28% and 1.53% of the initial content respectively.And then the respective changes slowed down.Conclusion] The suspension cell culture technology system of C.komarrovii was preliminarily established.