冬季油樟产黄酮内生真菌的分离鉴定及其抗氧化和挥发性成分分析
Isolation and identification of flavonoid-producing endophytic fungi with antioxidant and volatile components from Cinnamomum longepaniculatum in winter
刘雯雯 1刘晓凤 2沈爱军 3冯瑞章 4胡连清 3陈露 3魏琴 4周万海1
作者信息
- 1. 宜宾学院四川省油樟工程技术研究中心,宜宾 644007;宜宾学院农林与食品工程学部,宜宾 644007
- 2. 四川天一学院健康产业学院,德阳 618000
- 3. 宜宾学院农林与食品工程学部,宜宾 644007
- 4. 宜宾学院四川省油樟工程技术研究中心,宜宾 644007
- 折叠
摘要
目的 深入挖掘油樟内生真菌资源,从冬季油樟根、茎、叶中筛选产黄酮菌株并对其抗氧化和挥发性成分进行分析.方法 采用组织培养、形态学观察和分子生物学方法分离鉴定油樟内生真菌,借助显色反应和NaNO2-Al(NO3)3比色法筛选具有产黄酮能力菌株,利用清除自由基初步评估产黄酮最优菌株的抗氧化活性,并通过气相色谱-质谱法(GC-MS)分析菌株的发酵产物成分.结果 共获得61株内生真菌,隶属于3门23科29属,子囊菌门(Ascomycota)为优势门,肉座菌科(Hypocreaceae)和小丛壳科(Glomerellaceae)为优势科,木霉属(Trichoderma)和炭疽菌属(Colletotrichum)为优势属.不同组织间内生真菌种类和数量存在明显差异,其中茎部菌株多样性、均匀度和丰富度指数均最高,且茎部和叶部相似性系数最高.通过产黄酮筛选到WG5(Trichoderma)和WG15(青霉属,Penicilium)2株内生真菌,其中WG15总黄酮含量较高且表现出较好的抗氧化活性,DPPH和ABTS自由基清除率的IC5.分别为0.52和0.49 mg/L.挥发性物质分析显示,2株菌株代谢产物存在共性和差异,主要包括酯类、酸类、醇类和酚类等物质,共有苯乙醇、2,6-二叔丁基苯酚和棕榈酸成分.结论 WG15具有较好的产黄酮和抗氧化能力,可用于后续其他功能的开发,为内生真菌次生代谢产物的活性研究提供了菌种资源.
Abstract
Objective To explore the endophytic fungi resources of C.longepaniculatum,flavonoids-producing strains were screened from the roots,stems and leaves of C.longepaniculatum in winter,and their antioxidant and volatile components were analyzed.Methods The endophytic fungi of C.longepaniculatum were isolated and identified by tissue culture,morphological observation and molecular biology.The strains with flavonoid production capacity were screened by color reaction and the NaNO2-Al(NO3)3 colorimetric method.The antioxidant activity of the strains with optimal flavonoid production was preliminary evaluated by scavenging free radicals,and the fermentation products were analyzed by GC-MS.Results A total of 61 strains of endophytic fungi were obtained,belonging to 3 phyla,23 families and 29 genera.Ascomycota was the dominant phyla;Hypocreaceae and Glomerellaceae were the dominant families.Trichoderma and Colletotrichum were the dominant genera.There were significant differences in the number and species of endophytic fungi among different tissues,among which the diversity,evenness and richness indices of stem strains were the highest and the similarity coefficients of stem and leaf strains were the highest.Two endophytic fungi strains,WG5(Trichoderma)and WG15(Penicilium),were screened by flavonoids production,and the total flavonoids content of WG15 was high and showed good antioxidant activity.The IC50 of DPPH and ABTS free radical scavenging were 0.52 and 0.49 mg/L,respectively.Volatile substances analysis showed that the metabolites of the two strains had similarities and differences,mainly including esters,acids,alcohols and phenols,including phenylethyl alcohol,2,6-di-tert-butylphenol and palmitic acid.Conclusion WG15 has good flavonoid-producing and antioxidant capacity,which can be used for further development of other functions and provides a strain resource for the study of the activity of secondary metabolites in endophytic fungi.
关键词
油樟/内生真菌/物种鉴定/功能探索/多样性Key words
Cinnamomum longepaniculatum/Endophytic fungi/Functional verification/Functional exploration/Diversity引用本文复制引用
基金项目
四川省油樟工程技术研究中心开放基金项目(21YZY02)
四川省自然科学基金项目(2022NSFSC0176)
宜宾市双城协议保障科研经费科技项目(XNDX2022020010)
出版年
2024
NETL
NSTL
万方数据