红花组织培养体系的建立及悬浮细胞中次生代谢产物的分析  

Tissue culture of safflower and analysis of secondary metabolites in suspension cells

作　　者：刘楠 吴晓毅[1] 宋雅迪[1] 高伟[1,3,4] 黄璐琦 LIU Nan;WU Xiao-yi;SONG Ya-di;GAO Wei;HUANG Lu-qi(School of Traditional Chinese Mediciney Capital Medical University,Beijing 100069,China;State Key Laboratory Breeding Base of Dao-di Herbs,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Pharmaceutical Sciences,Capital Medical University,Beijing 100069,China;Advanced Innovation Center for Human Brain Protection,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学中医药学院,北京100069 [2]中国中医科学院道地药材国家重点实验室培育基地,北京100700 [3]首都医科大学药学院,北京100069 [4]首都医科大学人脑保护高精尖创新中心,北京100069

出　　处：《中国中药杂志》2021年第17期4380-4388,共9页China Journal of Chinese Materia Medica

基　　金：中央本级重大增减支项目(2060302-1806-03);“万人计划”青年拔尖人才项目;横向课题项目(DYTKY180725)。

摘　　要：红花作为一种常见的价值丰富的药用植物,近年来被广泛关注。为了建立更加稳定的红花组织培养体系,为红花中次生代谢产物的深入研究提供更加稳定、优质的实验材料。该研究以红花种子为基础,进行了愈伤组织的培养,然后分化成无菌苗,进一步通过悬浮培养得到了稳定的悬浮细胞系。并以此悬浮细胞系为实验材料,通过UPLC-Q-TOF-MS等技术手段定性分析了其中所含的次生代谢产物,及其在4种不同的诱导子(茉莉酸甲酯、硝酸银、水杨酸和酵母提取物)作用下的变化规律。该研究利用Progenesis QI软件在红花悬浮细胞中预测了黄酮、苯丙素、生物碱、有机酸和芳香苷类等23种同样存在于红花植物体内的次生代谢产物,并进行了PCA分析发现其中11种化合物的相对含量在诱导子的作用下发生了改变,不同诱导子对红花悬浮细胞中次生代谢产物的积累有着不同的影响,其中酵母提取物表现出相对明显的正向诱导作用。这些都为进一步研究红花的功能基因搭建了平台,也为将来寻找靶向诱导剂和实现一些有价值的次生代谢产物的大规模生产提供了可能性。Safflower(Carthamus tinctorius),a valuable traditional Chinese medicinal plant,has attracted much attention in recent years.This study established a stable tissue culture system of safflower and analyzed the chromatogram of its secondary metabolites,providing high-quality experimental materials for further research on natural products in safflower.The calluses were established from the safflower seeds germinated in a sterile environment,and then they were differentiated into the aseptic seedlings,or cultured to obtain suspension cells in liquid medium.The ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS),Progenesis QI,and principal component analysis(PCA)were used to detect and analyze the secondary metabolites in the suspension cells before and after induction with different elicitors(methyl jasmonate,silver nitrate,salicylic acid and yeast extract).A total of 23 secondary metabolites including flavonoids,phenylpropanoids,alkaloids,fatty acids and aromatic glycosides were detected in safflower suspension cells.In response to the four elicitors,11 compounds showed increased or decreased relative content.The results indicate that different elicitors have various effects on the accumulation of secondary metabolites in safflower suspension cells,and yeast extract shows more obvious positive induction.Therefore,different elicitors may play a role in the expression of related genes in the biosynthetic pathway of specific secondary metabolites.The results facilitate the discovery of targeted elicitors and the large-scale production of valuable secondary metabolites in the future.

关 键 词：红花 组织培养 悬浮细胞 诱导子 UPLC-Q-TOF-MS 

分 类 号：S567.2[农业科学—中草药栽培]