杨娟;董醇波;陈万浩;梁建东;韩燕峰;梁宗琦;
YANG Juan;DONG Chun-bo;CHEN Wan-hao;LIANG Jian-dong;HAN Yan-feng;LIANG Zong-qi;Institute of Fungus Resources,College of Life Science,Guizhou University;Department of Microbiology,Guiyang College of Traditional Chinese Medicine;

• 1:贵州大学生命科学学院生态系真菌资源研究所
• 2:贵阳中医学院微生物教研室

摘要(Abstract)：

基于高通量测序及宏基因组分析法对采自贵州遵义县枫香镇、湖南慈利县零阳镇、四川旺苍县木门镇3个产地的杜仲树皮内生真菌的群落组成及生态功能结构进行分析。从所试样本中共获得内生真菌有效序列110 865条,聚类后OTUs对应的真菌类群涉及子囊菌门Ascomycota、担子菌门Basidiomycota、接合菌门Zygomycota的10纲、25目、41科、57个属的74种真菌。其中,四川旺苍样本(EWP)中优势属为丛赤壳科Nectriaceae中的未定属,相对丰度为55. 03%。湖南慈利样本(ECP)中优势属为子囊菌门Ascomycota中的未定属,相对丰度为40. 23%。贵州遵义样本(EZP)中优势属为扁孔腔菌属Lophiostoma,其相对丰度为47. 15%。α多样性分析表明,不同产地杜仲树皮内生真菌Shannon多样性指数从高到低依次为ECP(1. 340 2)>EZP(1. 380 4)>EWP(1. 168 3); Simpson多样性指数从高到低依次为,EWP (0. 427 3)> EZP (0. 332 5)> ECP (0. 313 6)。经FUNGuild软件平台解析,3个产地的杜仲树皮内生真菌包含14个生态功能群,即植物病原菌群、动物病原菌群和内生真菌群等,3产地杜仲样本的共有功能群数达到8个,超过了总数的二分之一。对杜仲皮内生真菌群落与4种活性成分进行了相关性分析,发现在四川旺苍样本(EWP)中,松脂醇二葡萄糖苷及绿原酸含量与内生真菌中大卫氏菌科Davidiellaceae中的未定属、被孢霉属Mortierella、毛壳菌属Chaetomium和拟盘多毛孢属Pestalotiopsis有不同程度的正关联。
Based on high-throughput sequencing and metagenomic technology,the community composition and ecological functions of endophytic fungi in the bark of Eucommia ulmoides from three producing areas,Fengxiang town in Zunyi county of Guizhou province,Lingyang town of Cili county of Hunan province and Mumen town of Wangcang county of Sichuan province,were analyzed. A total of110 865 effective sequences of endophytic fungi were obtained in the study. The corresponding fungal group of OTUs after clustering belonged to 3 phyla( Ascomycota,Basidiomycota,Zygomycota),10 classes,25 orders,41 families,57 genera and 74 species. Among them,the dominant genera of Sichuan Wangcang Bark of E. ulmoides( EWP) was an unclassified genus in the Nectriaceae of the Crimsonaceae,with a relative abundance of 54. 79%; The dominant species of Hunan Cili Bark of E. ulmoides( ECP) was the unclassified genus of Ascomycota,with a relative abundance of 39. 97% and the dominant species of Guizhou Zunyi bark( EZP) was Lophiostoma,and its relative abundance was 47. 07%. The analysis of α diversity indicated that the shannon diversity index of endophytic fungi from different places was as follows: ECP: 1. 340 2>EZP: 1. 380 4 > EWP: 1. 168 3. The simpson diversity index was: EWP( 0. 427 3) >EZP( 0. 332 5) > ECP( 0. 313 6). FUNGuild software platform analysis displayed that endophytic fungi of E. ulmoides bark from three producing areas contained the following 14 functional groups: plant pathogen,animal pathogen and endophyte et al.,the number of functional groups in the 3 samples of E. ulmoides reached up 8 genera and exceeded one half of the total number. Correlation analysis of Canonical correspondence analysis( CCA) between endophytic fungal community diversity and four active compounds of E. ulmoides were analyzed,the results showed that the contents of pinoresinol diglucoside and chlorogenic acid,to a certain extend,had a positive correlation with an unclassified genus of Davidiellaceae,Mortierella,Chaetomium and Pestalotiopsis from the endophytic fungi in EWP sample.

关键词(KeyWords)： 杜仲;宏基因组技术;内生真菌;真菌多样性
Eucommia ulmoides;metabacroding;endophytic fungi;fungal diversity

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31460010);; 科技部基础工作专项(2013FY110400);; 贵州省千层次人才项目(2014);; 贵州省自然科学基金项目(黔科合基础[2018]1014);; 贵州省生物学一流学科建设项目(GNYL[2017]009)

作者(Author): 杨娟;董醇波;陈万浩;梁建东;韩燕峰;梁宗琦;
YANG Juan;DONG Chun-bo;CHEN Wan-hao;LIANG Jian-dong;HAN Yan-feng;LIANG Zong-qi;Institute of Fungus Resources,College of Life Science,Guizhou University;Department of Microbiology,Guiyang College of Traditional Chinese Medicine;

Email:

DOI: 10.19540/j.cnki.cjcmm.20181226.005

参考文献(References)：

• [1]周政贤,郭光典.我国杜仲类型、分布及引种[J].林业科学,1980(S1):84.
• [2]神农本草经[M].吴普等述.孙星衍,孙冯翼编撰.北京:人民卫生出版社,1982.
• [3]Huang L,Guo L,Ma C,et al.Top-geoherbs of traditional Chinese medicine:common traits,quality characteristics and formation[J].Front Med,2011,5(2):185.
• [4]Huang L H,Yuan M Q,Ao X J,et al.Endophytic fungi specifically introduce novel metabolites into grape flesh cells in vitro[J].PLoS ONE,2018,13(5):e0196996.
• [5]崔晋龙,郭顺星,肖培根.内生菌与植物的互作关系及对药用植物的影响[J].药学学报,2017,52(2):214.
• [6]宁祎,李艳玲,周国英,等.青海上北山林场野生桃儿七根部内生真菌群落组成及多样性研究[J].中国中药杂志,2016,41(7):1227.
• [7]郑欢,张芝元,韩燕峰,等.刺槐内生真菌群落组成及其生态功能结构分析[J].菌物学报,2018,37(2):256.
• [8]Cho H,Kim M,Tripathi B,et al.Changes in soil fungal community structure with increasing disturbance frequency[J].Microb Ecol,2017,74(1):62.
• [9]Widder S,Allen R J,Pfeiffer T,et al.Challenges in microbial ecology:building predictive understanding of community function and dynamics[J].ISME J,2016,10(11):2557.
• [10]Root R B.The niche exploitation pattern of the blue-gray gnatcatcher[J].Ecol Monogr,1967,37(4):317.
• [11]Hamady M,Knight R.Microbial community profiling for human microbiome projects:tools,techniques,and challenges[J].Genome Res,2009,19:1141.
• [12]Wilson J B.Guilds,functional types and ecological groups[J].Oikos,1999,86(3):507.
• [13]Puri S C,Nazir A,Chawla R,et al.The endophytic fungus Trametes hirsuta as a novel alternative source of podophyllotoxin and related aryl Tetralin lignans[J].J Biotechnol,2006,122(4):494.
• [14]Nguyen N H,Song Z,Bates S T,et al.FUNGuild:an open annotation tool for parsing fungal community datasets by ecological guild[J].Fungal Ecol,2016,20:241.
• [15]梁雪娟,张水寒,张平,等.不同产地杜仲皮内生真菌种群结构的比较分析[J].中国中药杂志,2014,39(2):204.
• [16]Gamboa M A,Laureano S,Bayman P.Measuring diversity of endophytic fungi in leaf fragments:does size matter?[J].Mycopathologia,2002,156(1):41.
• [17]Carini P,Marsden P J,Leff J W,et al.Relic DNA is abundant in soil and obscures estimates of soil microbial diversity[J].Nat Microbiol,2017,2(3):16242.
• [18]Johnston P R,Park D,Smissen R D.Comparing diversity of fungi from living leaves using culturing and high-throughput environmental sequencing[J].Mycologia,2017,109(4):643.
• [19]Siddique A B,Unterseher M.A cost-effective and efficient strategy for Illumina sequencing of fungal communities:a case study of beech endophytes identified elevation as main explanatory factor for diversity and community composition[J].Fungal Ecol,2016,20:175.
• [20]Arnold A E,Lutzoni F.Diversity and host range of foliar fungal endophytes:are tropical leaves biodiversity hotspots?[J].Ecology,2007,88(3):541.
• [21]Stierle A,Storbel G,Stierle D.Taxol and taxane production by Taxomyces andrenae,an endophytic fungus of Paciifc Yew[J].Seienee,1993,260(1):154.
• [22]于维虹,刘艳,杨君杰,等.产松脂醇二葡萄糖苷杜仲内生菌的筛选与培养条件优化[J].曲阜师范大学学报,2017,43(3):69.
• [23]Shade A,Handelsman J.Beyond the Venn diagram:the hunt for a core microbiome[J].Environ Microbiol,2012,14(1):4.
• [24]Agler M T,Ruhe J,Kroll S,et al.Microbial hub taxa link host and abiotic factors to plant microbiome variation[J].PLoS Biol,2016,14(1):e1002352.
• [25]Turnbaugh P J,Ley R E,Hamady M,et al.The human microbiome project[J].Indian J Microbiol,2012,52(3):315.
• [26]Hawkins C P,Mac Mahon J A.Guilds:the multiple meanings of a concept[J].Annu Rev Entomol,1989,34(1):423.
• [27]糜亚男,张水寒,蔡媛,等.杜仲SSR-PCR反应体系的优化[J].中国实验方剂学杂志,2015,21(2):1.