牛大力毛状根生长及培养液中碳源、氮源与磷源利用研究

Research on the Growth of Callerya speciosa Hairy Root and Utilization of Carbon Source, N and P Source in Medium

目的探讨牛大力毛状根的生长曲线研究,研究不同培养基对其生物量积累的影响,以及在其生长过程中对培养液中碳源(蔗糖)、氮源(NH_4^+与NO_3^-)与磷源(PO_4^(3-))的吸收利用情况。方法在1/2 MS培养基中,每5 d取样称量鲜重与干重考察毛状根的生长曲线。使用5种培养基(MS、1/2MS、1/4 MS、B_5、WPM)培养毛状根,30 d后统计生物量。利用离子色谱分析方法检测培养液中蔗糖、NH_4^+、NO_3^-与PO_4^(3-)含量。结果毛状根的生长表现为"慢-快-慢"的趋势,呈"S"型曲线,生长30d进入稳定期。不同培养基对牛大力毛状根生物量的积累具有显著差异,1/2 MS培养基的营养成分及比例最有利于毛状根生物量的积累。在毛状根生长过程中,1/2 MS培养基中的碳源、氮源与磷源均被毛状根充分吸收利用,呈"快-慢"或"慢-快-慢"规律。当毛状根进入稳定期时,蔗糖、NO_3^-与PO_4^(3-)均被全部消耗殆尽,仅检测到少量的葡萄糖、果糖与NH_4^+。结论牛大力毛状根的生长周期为30 d,在1/2 MS培养基中最有利于生物量积累,能充分吸收代谢培养液中的蔗糖、氮源与磷源等营养成分。

Objective To study the growth curve of Callerya speciosa hairy roots, the effect of biomass in different media and the utilization of carbon source（sucrose）, N （NH4^＋ and NO3^-） and P （PO4^3-） source in 1/2 MS medium. Methods Fresh weight and dry weight were measured every 5 days to draw the growth curve when hairy roots were cultured in 1/2 MS medium, and the content of carbon source, NH4^＋, NO3^- and PO4^3- was detected by ion chromatography method. The biomass was also measured in day 30 when hairy roots were cultured with five kinds 0f media 1/2 1/4 MS, Ds, WPM）. Results The 8rowth trends of hairy roots was ＂slow-fast- slow＂, as ＂S＂ type curve, and the stable period was after cultured 30 days. The accumulation of biomass had a significant effect in different kinds media and 1/2 MS medium was the most suitable one because of the abundant nutritional components and appropriate proportion. Carbon source, NH4^＋, NO3^- and PO4^3- in 1/2 MS medium were utilized fully by hairy roots, displaying ＂fast-slow＂ or ＂slow-fast-slow＂ trend. No sucrose, NO3^- and PO4^3- and only a small content of glucose, fluctose and NH4^＋ were detected in day 30 of culture. Conclusion The growth cycle of Callerya speciosa hairy roots is 30 days. 1/2 MS medium is the suitable one for the accumulation of biomass, in which carbon source, N （NH4^＋ and NO3^-） and P （PO4^3-） source are utilized fully in the process of hairy roots growth.