麻花艽产黄酮类物质内生真菌的分离与鉴定及抗氧化活性分析  

Isolation,Identification and Antioxidant Activity of Endophytic Fungi Producing Flavonoids from Gentiana straminea Maxim.

作　　者：程庭峰 吕东晋[2] 王环[3] 陈世龙[1,4,5] 张辉 林鹏程[6] 沈建伟 邢睿[1] 周党卫[1,4,5,6] Cheng Tingfeng;Lü Dongjin;Wang Huan;Chen Shilong;Zhang Hui;Lin Pengcheng;Shen Jianwei;Xing Rui;Zhou Dangwei(Key Laboratory of Adaptation and Evolution of Plateau Biota,Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining,810008;Qinghai Province Center for Disease Prevention and Control,Xining,810007;Analysis and Testing Center,Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining,810008;Key Laboratory of Crop Molecular Breeding of Qinghai Province,Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining,810008;College of Life Sciences,University of Chinese Academy of Sciences,Beijing,100049;College of Pharmacy,Qinghai Nationalities University,Xining,810007)

机构地区：[1]中国科学院西北高原生物研究所,高原生物适应与进化重点实验室,西宁810008 [2]青海省疾病预防控制中心,西宁810007 [3]中国科学院西北高原生物研究所,分析测试中心,西宁810008 [4]中国科学院西北高原生物研究所,青海省作物分子育种重点实验室,西宁810008 [5]中国科学院大学生命科学学院,北京100049 [6]青海民族大学药学院,西宁810007

出　　处：《分子植物育种》2022年第19期6541-6549,共9页Molecular Plant Breeding

基　　金：青海省科技厅基础研究项目(2021-ZJ-734,2019-ZJ-7016);青海省自然科学基金项目(2019-ZJ-976Q);青海省青藏高原植物化学重点实验室项目(2020-ZJ-Y20)共同资助。

摘　　要：为了解高山植物麻花艽中产黄酮内生真菌的特征,寻找可替代产黄酮的微生物资源,从而更好地保护和利用野生麻花艽内生真菌资源。本研究采用组织块培养法,对青海省海东市互助土族自治县南门峡国家湿地公园采集到的一株野生麻花艽不同组织(根,茎和叶)的内生真菌进行分离培养,根据菌株的形态学特征,结合真菌ITS的系统进化分析对其进行鉴定,并通过液相色谱-串联质谱法(LC-MS/MS)对内生真菌的发酵产物进行分析,筛选产黄酮类物质的内生真菌。结果显示,从麻花艽的根、茎、叶中共分离得到78株内生真菌,将其划分为22个形态学种,隶属3门,8纲,5目,12个属,多样性丰富。从总体分离频率看,小球腔菌属(Leptosphaeria)(23.1%)和镰孢霉属(Fusarium)(16.3%)为优势菌群。通过LC-MS/MS分析,发现其中1株内生真菌Gs-6发酵液的甲醇提取液中存在3种黄酮类成分,其中异牡荆黄素含量为0.824 mg/L,槲皮素含量为1.110 mg/L,异荭草素为1.569 mg/L。形态学和分子鉴定,确认内生真菌Gs-6为Cadophora sp.,体外自由基活性分析表明,Gs-6对DPPH自由基清除具有较高活性,其IC_(50)为39.9μg/m L。To know about culture-endophytic fungi profiling of Gentiana straminea Maxim.,explore culture-endophytic fungi resource that produce flavonoids,and protect the wild G.straminea resource.Here,the endophytic fungi were isolated from root,leaf and stem of G.straminea using the tissue separation method.The fermentation products of the endophytic fungi were detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The screened endophytic fungi were then analyzed by the morphological traits and the flanking internal transcribed spacers sequencing(ITS).78 endophytic fungi were isolated from the roots,stems and leaves of G.straminea and they were divided into 22 morphological species,which belong to 3 phyla,5 classes,8 orders and 12 genera.On the whole,endophytic fungi were diversity and Leptosphaeria(23.1%)and Fusarium(16.3%)were comparatively advantageous taxa according to the colonization rate and isolation rate.Among them,the strain Gs-6 was found to produce quercetin,isoorientin,and isovitexin.The content of fermentation were 1.110,1.159 and 0.824(mg/L),respectively.Isoorientin was the highest of the three.Antioxidant effect showed that Gs-6 had high ability to scavenge DPPH and its IC_(50)was 39.9μg/mL.Further,through morphological and molecular biological identification,it was confirmed that the endophytic fungi Gs-6 was Cadophora sp..

关 键 词：麻花艽 内生真菌 LC-MS/MS 黄酮 抗氧化活性 

分 类 号：S567.239[农业科学—中草药栽培]