培养基主成分优化提高大花金挖耳胞内黄酮产量  

Optimizing Medium for Flavonoid-producing Carpesium Macrocephalum Culture

作　　者：李玉平[1,2,3] 龚宁 LI Yu-ping;GONG Ning(College of Life Sciences,Northwest A&F University,Yangling,Shaanxi 712100,China;Graduate School,Qinghai University,Xining,Qinghai 810016,China;Technology andEngineering Center of Biopesticide of Shaanxi Province,Northwest A&F University,Yangling,Shaanxi 712100,China;College of Chemistry&Pharmacy,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[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

出　　处：《福建农业学报》2019年第6期652-659,共8页Fujian Journal of Agricultural Sciences

基　　金：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)

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+1.0mg·L^-1NAA+0.2mg·L^-16-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-、Zn^2+、BO3^3-、Co^2+、Cu^2+、I^-、Mn^2+,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15.46mmol·L^-1、14.10mmol·L^-1、19.10mmol·L^-1,微量元素MoO4^2-、Zn^2+、BO3^3-、Co^2+、Cu^2+、I^-、Mn^2+分别为3.0μmol·L^-1、0.06mmol·L^-1、0.4mmol·L^-1、0.2μmol·L^-1、0.4μmol·L^-1、10μmol·L^-1、0.2mmol·L^-1,附加KT0.2mg·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517.87mg·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。【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-,Zn^2+,BO3^3-,Co^2+,Cu^2+,I^-,and Mn^2+ and followed by the added 2,4-D,KT and GA 3.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 GA 3,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,Zn^2+ 0.06 mmol·L^-1,BO3^3- 0.4 mmol·L^-1,Co^2+ 0.2 μmol·L^-1,Cu^2+ 0.4 μmol·L^-1,I^- 10 μmol·L^-1,Mn^2+ 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.

关 键 词：大量元素 微量元素 植物激素 黄酮 大花金挖耳 

分 类 号：Q813.1[生物学—生物工程]