培养基主成分优化提高大花金挖耳胞内黄酮产量

李玉平; 龚宁

doi:10.19303/j.issn.1008-0384.2019.06.005

培养基主成分优化提高大花金挖耳胞内黄酮产量

李玉平,
龚宁

Optimizing Medium for Flavonoid-producing Carpesium Macrocephalum Culture

摘要

摘要:
目的优化组合培养基中大量、微量元素及附加激素等培养条件，筛选出大花金挖耳细胞培养黄酮的生产培养基。
方法以NT+1.0 mg·L^-1 NAA+0.2 mg·L^-1 6-BA为基本培养基，先优化组合其大量元素NH₄⁺、NO₃^-、K⁺，再优化组合微量元素MoO₄^2-、Zn²⁺、BO₃^3-、Co²⁺、Cu²⁺、I^-、Mn²⁺，最后分别附加2，4-D、KT、GA₃，测定大花金挖耳悬浮培养细胞生长、黄酮含量及产量。
结果大量元素优化、微量元素优化及附加激素KT、GA₃，大花金挖耳细胞培养物中黄酮产量均得到了显著提升（P < 0.05），添加2，4-D降低了黄酮产量。本研究筛选得到大花金挖耳细胞培养生产黄酮类化合物的最佳NT培养基为：大量元素NH₄⁺、NO₃^-、K⁺分别为15.46 mmol·L^-1、14.10 mmol·L^-1、19.10 mmol·L^-1，微量元素MoO₄^2-、Zn²⁺、BO₃^3-、Co²⁺、Cu²⁺、I^-、Mn²⁺分别为3.0 μmol·L^-1、0.06 mmol·L^-1、0.4 mmol·L^-1、0.2 μmol·L^-1、0.4 μmol·L^-1、10 μmol·L^-1、0.2 mmol·L^-1，附加KT 0.2 mg·L^-1。在此培养条件下，大花金挖耳细胞培养物中黄酮含量达到2.05%，是基本培养基的1.68倍；黄酮产量为517.87 mg·L^-1，是基本培养基的1.80倍。
结论在前期研究的基础上，通过培养基主成分优化，提高了大花金挖耳细胞培养生产黄酮的产量，初步得到了黄酮生产培养基。

Abstract:
Objective Formulation of the medium for Carpesium macrocephalum culture to produce flavonoids was optimized from the one obtained by a previous study.
Method Using NT+NAA 1.0 mg·L^-1+0.2 mg·L^-1 6-BA as the base, the medium optimizations were firstly carried out on the macro-elements, NH₄⁺, NO₃^- and K⁺, then the micro-elements, MoO₄^2-, Zn²⁺, BO₃^3-, Co²⁺, Cu²⁺, I^-, and Mn²⁺ and followed by the added 2, 4-D, KT and GA₃. The cellular growth as well as the flavonoids yield in the culture suspension were monitored for evaluation.
Result With the optimized addition of the macro- and micro-elements plus KT or GA₃, the flavonoids produced by C. macrocephalum culture significantly increased (P < 0.05), but reduced by the presence of 2, 4-D. The finalized medium consisted of the base with added 15.46 NH₄⁺ mmol·L^-1, NO₃^- 14.10 mmol·L^-1, K⁺ 19.10 mmol·L^-1, MoO₄^2- 3.0 μmol·L^-1, Zn²⁺ 0.06 mmol·L^-1, BO₃^3- 0.4 mmol·L^-1, Co²⁺ 0.2 μmol·L^-1, Cu²⁺ 0.4 μmol·L^-1, I^- 10 μmol·L^-1, Mn²⁺ 0.2 mmol·L^-1, and KT 0.2 mg·L^-1. The resulting flavonoids content in the culture suspension was 2.05%, which was 1.68-fold of control, and the flavonoids yield 517.87 mg·L^-1, which was 1.80-fold of control.
Conclusion By further modifying the previously obtained medium, the flavonoids produced from the C. macrocephalum culture significantly increased.

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