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

doi: 10.19303/j.issn.1008-0384.2019.06.005

李玉平^{1, 2, 3,},
龚宁^4, ,

1.
西北农林科技大学生命科学学院, 陕西杨凌 712100
2.
青海大学研究生院, 青海西宁 810016
3.
西北农林科技大学陕西省生物农药工程技术研究中心, 陕西杨凌 712100
4.
西北农林科技大学化学与药学院, 陕西杨陵 712100

基金项目:

国家公益性行业（农业）科研专项 200903052

国家重点研发计划项目 2016YFC0402904

详细信息

作者简介:
李玉平(1970-), 男, 博士, 副教授, 主要从事植物资源保护与利用的教学与研究(E-mail:liyuping1970@nwafu.edu.cn)

通讯作者:
龚宁(1970-), 女, 博士, 副教授, 主要从事环境与食品安全研究(E-mail:Gongningcn@163.com)

中图分类号: Q813.1
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- 被引次数: 0
出版历程
- 收稿日期: 2019-04-18
- 修回日期: 2019-05-24
- 刊出日期: 2019-06-28

Optimizing Medium for Flavonoid-producing Carpesium Macrocephalum Culture

LI Yu-ping^{1, 2, 3
,},
GONG Ning^{4
, ,}

1.
College of Life Sciences, Northwest A & F University, Yangling, Shaanxi 712100, China
2.
Graduate School, Qinghai University, Xining, Qinghai 810016, China
3.
Technology and Engineering Center of Biopesticide of Shaanxi Province, Northwest A & F University, Yangling, Shaanxi 712100, China
4.
College of Chemistry & Pharmacy, Northwest A & F University, Yangling, Shaanxi 712100, China

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。
- 大量元素 /
- 微量元素 /
- 植物激素 /
- 黄酮 /
- 大花金挖耳
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.
- macroelements /
- microelements /
- phytohormone /
- flavonoids /
- Carpesium macrocephalum

HTML全文

图 1 大量元素优化组合对悬浮培养细胞生长、黄酮含量和产量的影响

注：图中不同字母表示不同处理方法在0.05水平存在显著差异(P < 0.05)，图 3~5同。

Figure 1. Effects of macro-elements on cell growth, flavonoids content and yield in culture suspension

Note:Different lowercase letters within column indicate significant difference between different treatment methods at the 0.05 level; the same below.The same as Fig. 3-5.

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图 2 基本培养基NT0和生产培养基NT1中的悬浮培养细胞(7 d)

Figure 2. Cells in suspension culture with basic medium NT0 or production medium NT1 (7 d)

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图 3 微量元素优化组合对悬浮培养细胞生长、黄酮含量和产量的影响

Figure 3. Effects of micro-elements on cell growth, flavonoids content and yield in culture suspension

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图 4 2, 4-D对悬浮培养细胞生长、黄酮含量和产量的影响

Figure 4. Effects of 2, 4-D in varied concentrations on cell growth, flavonoids content and yield in culture suspension

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图 5 KT对悬浮培养细胞生长、黄酮含量和产量的影响

Figure 5. Effects of KT in varied concentrations on cell growth, flavonoids content and yield in culture suspension

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图 6 GA₃对悬浮培养细胞生长、黄酮含量和产量的影响

Figure 6. Effects of GA3 in varied concentrations on cell growth, flavonoids content and yield in culture suspension

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表 1 大量元素氮、钾优化组合对大花金挖耳细胞培养生产黄酮类化合物试验

Table 1. Medium optimization on nitrogen and potassium for flavonoids-producing C. macrocephalum culture

试验组 Treatments	大量元素Macroelements/(mmol·L^-1)
试验组 Treatments	NH₄⁺	NO₃^-	K⁺
CK	10.31	9.40	14.40
Ⅰ	15.46	9.40	14.40
Ⅱ	15.46	14.10	19.10
Ⅲ	15.46	18.79	23.79
Ⅳ	20.61	9.40	14.40
Ⅴ	20.61	14.10	19.10
Ⅵ	20.61	18.79	23.79

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表 2 微量元素钼、锌优化组合对大花金挖耳培养细胞生产黄酮类化合物试验

Table 2. Medium optimization on molybdenum and zinc for flavonoids-producing C. macrocephalum culture

试验组 Treatments	微量元素Microelements
试验组 Treatments	MoO₄^2-/(μmol·L^-1)	Zn²⁺/(mmol·L^-1)
CK	1.0	0.03
A	3.0	0.06
B	3.0	0.09
C	2.0	0.06
D	2.0	0.09

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