Traditional theory holds that “drugs with long aging time have better effect” regarding Citrus reticulata pericarpium. It has been reported that microorganisms can change the pharmacodynamic material base during aging. High-throughput sequencing was used to systematically study the structure and characteristics of bacterial and fungal community on the surface of C. reticulata ‘Chachi’ with different aging time. The 16S rDNA or ITS1 fragments of the community isolated and sequenced. At genus level, the main bacteria were unidentified Rickettsiales, Phyllobacterium, Methylobacterium, unidentified Cyanobacteria, Pseudomonas, Sphingomonas, Stenotrophomonas, Lactococcus and Bacillus, and the main fungi were Xeromyces, Cladosporium, Zasmidium, Symmetrospora, Aureobasidium, Fusarium and Aspergillus. With the increasing of aging time, the dominant bacterial genera changed from Methylobacterium, Sphingomonas to Bacillus, Pseudomonas, Stenotrophomonas and Lactococcus, and the dominant fungal genera changed from Zasmidium, Cladosporium, Symmetrospora, Fusarium to Cladosporium, Aureobasidium, Aspergillus and Xeromyces. The results provide a theoretical basis for the subsequent isolation and screening of dominant functional microorganisms, in-depth study of the aging mechanism of C. reticulata pericarpium during aging.
[1] 国家药典委员会. 中华人民共和国药典:一部[M].北京:中国医药科技出版社,2020:199-200.
China Pharmacopoeia Committee.Pharmacopoeia of the People’s Republic of China (Edition one)[M].Beijing:Chinese Medical Science and Technology Press,2020:199-200.
[2] 鲍倩, 夏荃,潘超美,等.基于Delphi法对广陈皮商品规格等级划分[J].中国实验方剂学杂志,2017,23(22):48-54.
BAO Q,XIA Q,PAN C M,et al.Commercial specification of Citrus reticulata ‘Chachi’ based on delphi method[J].Chinese Journal of Experimental Traditional Medical Formulae,2017,23(22):48-54.
[3] 席秀利, 黄海波,楼步青,等.SCoT分子标记对茶枝柑及近缘种遗传多态性分析[J].中草药,2018,49(10):2 426-2 431.
XI X L,HUANG H B,LOU B Q,et al.Genetic polymorphism analysis of Citrus reticulata Blanco cv.Chachiensis Tanaka and its relatives by SCoT molecular marker[J].Chinese Traditional and Herbal Drugs,2018,49(10):2 426-2 431.
[4] 陶弘景. 本草经集注[M].上海:群联出版社,1955:360-362.
TAO H J.Collective commentaries on the classic of materia medica[M].Shanghai:Group Press,1955:360-362.
[5] 王智磊, 张鑫,刘素娟,等.陈皮“陈久者良”历史沿革和研究现状[J].中华中医药学刊,2017,35(10):2 580-2 584.
WANG Z L,ZHANG X,LIU S J,et al.Historical evolution and research status of Citri reticulatae pericarpium[J].Chinese Archives of Traditional Chinese Medicine,2017,35(10):2 580-2 584.
[6] 杨放晴, 何丽英,杨丹,等.不同陈化时间广陈皮中黄酮类成分的UPLC-Q-Orbitrap HRMS分析[J].中国实验方剂学杂志,2021,27(12):125-132.
YANG F Q,HE L Y,YANG D,et al.Analysis and identification of flavonoids in Citri reticulatae pericarpium with different aging time by UPLC-Q-Orbitrap HRMS[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(12):125-132.
[7] 王福, 张鑫,卢俊宇,等.陈皮“陈久者良”之黄酮类成分增加原因探究[J].中国中药杂志,2015,40(24):4 890-4 896.
WANG F,ZHANG X,LU J Y,et al.Study of reason of “the older,the better” of pericarpium Citri reticulatae (PCR) which contributed increase of flavonoids[J].China Journal of Chinese Materia Medica,2015,40(24):4 890-4 896.
[8] 刘丽娜. 基于微生物代谢对陈皮陈化活性物质转化机制的研究[D].湛江:广东海洋大学,2019.
LIU L N.Study on the mechanism of the active substance transformation of Citri reticulatae pericarpium aging based on microbial metabolism[D].Zhanjiang:Guangdong Ocean University,2019.
[9] 叶雷, 闫亚丽,陈庆森,等.高通量测序技术在肠道微生物宏基因组学研究中的应用[J].中国食品学报,2016,16(7):216-223.
YE L,YAN Y L,CHEN Q S,et al.Application of high-throughput sequencing technology in studying matagenomics of intestinal microbiota[J].Journal of Chinese Institute of Food Science and Technology,2016,16(7):216-223.
[10] TURCATTI G,ROMIEU A,FEDURCO M,et al.A new class of cleavable fluorescent nucleotides:Synthesis and optimization as reversible terminators for DNA sequencing by synthesis[J].Nucleic Acids Research,2008,36(4):25.
[11] 陈聪聪. 广陈皮陈化过程中微生物群落多样性解析及代谢物成分变化分析[D].广州:华南理工大学,2017.
CHEN C C.Characterization of microbial community diversity and metabolites changes of pericarpium Citri reticulatae ‘Chachiensis’ during aging process[D].Guangzhou:South China University of Technology,2017.
[12] 何静. 广陈皮细菌群落和植物化学物独特性及其相关性分析[D].广州:华南理工大学,2019.
HE J.The correlation of bacterial community and phytochemical peculiarities of Citri reticulatae pericarpium[D].Guangzhou:South China University of Technology,2019.
[13] 张鑫, 刘素娟,王智磊,等.橘皮表面真菌群落结构多样性分析[J].微生物学通报,2017,44(5):1 089-1 098.
ZHANG X,LIU S J,WANG Z L,et al.Community structure analysis of fungi isolated from citrus surfaces[J].Microbiology China,2017,44(5):1 089-1 098.
[14] WANG X Y,ZHU X G,BI Y L,et al.Dynamics of microbial community and changes of metabolites during production of type Ι sourdough steamed bread made by retarded sponge-dough method[J].Food Chemistry,2020,330:127 316.
[15] AWASTHI M K,CHEN H,WANG Q,et al.Succession of bacteria diversity in the poultry manure composted mixed with clay:studies upon its dynamics and associations with physicochemical and gaseous parameters[J].Bioresource Technology,2018,267:618-625.
[16] BOKULICH N A,SUBRAMANIAN S,FAITH J J,et al.Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing[J].Nature Methods,2013,10(1):57-59.
[17] CAPORASO J G,KUCZYNSKI J,STOMBAUGH J,et al.QIIME allows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336.
[18] HAAS B J,GEVERS D,EARL A M,et al.Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons[J].Genome Research,2011,21(3):494-504.
[19] EDGAR R C.UPARSE:Highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[20] 刘素娟. 陈皮陈化过程“环境因子—真菌—药效物质基础”相关性研究[D].成都:成都中医药大学,2018.
LIU S J.Correlation research of environmental factors,fungi and effective substance in the aging process of Citri reticulatae pericarpium[D].Chengdu:Chengdu University of Traditional Chinese Medicine,2018.
[21] 黎瑶依, 胡小霞,黄永光.茅台镇酱香型白酒酿造环境中真菌菌群多样性分析[J/OL].食品科学:1-11[2021-04-19].http://kns.cnki.net/kcms/detail/11.2206.TS.20200831.0939.012.html.
LI Y Y,HU X X,HUANG Y G.Analysis on the diversity of fungi flora in Maotai-flavor Liquor brewing environment in Maotai Town[J].Food Science:1-11[2021-04-19].http://kns.cnki.net/kcms/detail/11.2206.TS.20200831.0939.012.html.
[22] 伍雪莹. 陈化烤烟烟叶细菌群落鉴定及应用研究[D].广州:华南理工大学,2014.
WU X Y.Identification and application study of bacterial communities on flue-cured tobacco leaves[D].Guangzhou:South China University of Technology,2014.
[23] 周家喜, 喻理飞,张健,等.烟叶陈化过程细菌群落演替特征[J].生态学报,2018,38(21):7 739-7 748.
ZHOU J X,YU L F,ZHANG J,et al.Study on the characteristics of bacterial community succession in tobacco aging[J].Acta Ecologica Sinica,2018,38(21):7 739-7 748.
[24] WANG F,MEN X,ZHANG G,et al.Assessment of 16S rRNA gene primers for studying bacterial community structure and function of aging flue-cured tobaccos[J].AMB Express,2018,8(1):182.
[25] 赵铭钦, 刘云,李芳芳,等.陈化烤烟叶面优势菌的筛选鉴定与其增香效应[J].微生物学报,2009,49(5):625-631.
ZHAO M Q,LIU Y,LI F F,et al.Identification of dominant and fragrance-enhancing microorganisms of tobacco leaves during ripening[J].Acta Microbiologica Sinica,2009,49(5):625-631.
[26] 张鑫, 贺仪,刘素娟,等.陈皮“陈久者良”原因探究[J].食品科技,2017,42(1):90-95.
ZHANG X,HE Y,LIU S J,et al.The reason of “the older,the better” of Citri reticulatae pericarpium (PCR)[J].Food Science and Technology,2017,42(1):90-95.
[27] LI K K,ZHUO C,TENG C Y,et al.Effects of Ganoderma lucidum polysaccharides on chronic pancreatitis and intestinal microbiota in mice[J].International Journal of Biological Macromolecules,2016,93:904-912.
