檀香内生真菌多样性及其抗菌与促生特性的研究
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摘要
为了揭示檀香内生真菌分布特点及其在抗菌物质开发和檀香快速栽培研究中的潜在应用,该研究对檀香内生真菌的生物多样性及其抗菌与促生特性进行了研究。檀香内生真菌分离及类群分析结果显示:分离到的325株檀香内生真菌(其中来自根86株,茎105株,叶134株)隶属于16个属;不同部位内生真菌的分离率与定殖率呈现相同的变化规律,从高到低依次为叶、茎、根;根部内生真菌的多样性指数明显高于茎和叶,不同部位优势内生真菌类群存在较大的差异:根部为镰刀菌属(Fusarium,占50.00%)和链格孢属(Alternaria,占10.47%),茎部为链格孢属(占58.11%)和枝顶孢霉属(Acremonium,占20.00%),而叶部是盘长孢属(Pantoea),占74.63%。40株代表性檀香内生真菌菌株发酵液的抗菌活性分析结果显示:90%内生真菌至少对其中1种测试菌表现出抑制活性,其对大肠杆菌、产气肠杆菌、伤寒沙门菌、痢疾杆菌、金黄色葡萄球菌、枯草芽胞杆菌具有抑制活性的菌株分别占总数的45.0%,30%,47.5%,55%,72.5%,62.5%。对它们的促生特性进行分析,筛选到5株具溶磷活性的内生真菌,8株可分泌IAA的内生真菌,以及4株可产铁载体的内生真菌,其中内生真菌Monilia sp.TXRF45既具溶磷活性、又能产生IAA和铁载体。以上研究结果表明檀香内生真菌具有丰富的物种多样性且其分布具有一定的组织特异性,部分菌株具有较好的抗菌和促生活性,它们在抗菌活性物质的开发和快速栽培研究中具有潜在的应用前景。
The aim of this study was to investigated the biological diversity,antibacterial activites and the plant growth-promoting traits of endophytic fungi of sandal( Santalum album),and to assess their potential in the development of antibacterial substances and rapid cultivation of sandal. The results of isolation and taxa analysis of endophytic fungi from sandal showed that 325 strains of endophytic fungi belonging to 16 genera of endophytic fungi were isolated from sandal( of which 86 from roots,105 from stems and 134 from leaves). The isolation rate and colonization rate of endophytic fungi in different sandal parts showed the same pattern of change:leave > stems > roots. The diversity index of endophytic fungi in sandal roots was significantly higher than that of stems and leaves. The dominant endophytic fungi of sandal roots,stems and leaves showed significant differences. The dominant endophytic fungi of roots were Fusarium( 50. 00%) and Alternaria( 10. 47%),Alternaria( 58. 11%) and Acremonium( 20. 00%) for stems,and Pantoea( 74. 63%) for leaves. The antibacterial activity of 40 representative strains of sandal endophytic fungi were analyzed and the results showed that 90% of endophytic fungi exhibited inhibitory activity against at least one of the tested bacteria strains,and the strains with inhibitory activity to Escherichia coli,Enterobacter aerogenes,Shigella dysenteriae,Salmonella typhimurium,Staphylococcus aureus,and Bacillus subtilis accounted for 45. 0%,30%,47. 5%,55%,72. 5%,and 62. 5%,respectively. The sandal fungal endophytes with plant growth-promoting characteristics were screened,and 5 strains of endophytic fungi with phosphorus-solubilizing activity,8 strains of endophytic fungi producing IAA,and 4 strains of endophytic fungi producing siderophores were found. Among them,endophytic fungus Monilia sp TXRF45 clould produced IAA and siderophores,and also show phosphate-solubilizing activity. The results indicated that the endophytic fungi of Sandal were rich in species diversity and their distribution had a certain tissue specificity. Some strains showed good antibacterial activity and growth-promoting properties,which could potentially applicable for the development of antibacterial substances and rapid cultivation of sandal.
The aim of this study was to investigated the biological diversity,antibacterial activites and the plant growth-promoting traits of endophytic fungi of sandal( Santalum album),and to assess their potential in the development of antibacterial substances and rapid cultivation of sandal. The results of isolation and taxa analysis of endophytic fungi from sandal showed that 325 strains of endophytic fungi belonging to 16 genera of endophytic fungi were isolated from sandal( of which 86 from roots,105 from stems and 134 from leaves). The isolation rate and colonization rate of endophytic fungi in different sandal parts showed the same pattern of change:leave > stems > roots. The diversity index of endophytic fungi in sandal roots was significantly higher than that of stems and leaves. The dominant endophytic fungi of sandal roots,stems and leaves showed significant differences. The dominant endophytic fungi of roots were Fusarium( 50. 00%) and Alternaria( 10. 47%),Alternaria( 58. 11%) and Acremonium( 20. 00%) for stems,and Pantoea( 74. 63%) for leaves. The antibacterial activity of 40 representative strains of sandal endophytic fungi were analyzed and the results showed that 90% of endophytic fungi exhibited inhibitory activity against at least one of the tested bacteria strains,and the strains with inhibitory activity to Escherichia coli,Enterobacter aerogenes,Shigella dysenteriae,Salmonella typhimurium,Staphylococcus aureus,and Bacillus subtilis accounted for 45. 0%,30%,47. 5%,55%,72. 5%,and 62. 5%,respectively. The sandal fungal endophytes with plant growth-promoting characteristics were screened,and 5 strains of endophytic fungi with phosphorus-solubilizing activity,8 strains of endophytic fungi producing IAA,and 4 strains of endophytic fungi producing siderophores were found. Among them,endophytic fungus Monilia sp TXRF45 clould produced IAA and siderophores,and also show phosphate-solubilizing activity. The results indicated that the endophytic fungi of Sandal were rich in species diversity and their distribution had a certain tissue specificity. Some strains showed good antibacterial activity and growth-promoting properties,which could potentially applicable for the development of antibacterial substances and rapid cultivation of sandal.
引文
[1]刘小金,徐大平,杨曾奖,等.广东三地幼龄檀香生长和结香的早期评价[J].林业科学,2012,48(5):108.
[2]孙思胜,张大为,孟志霞,等.檀香形成部位内生真菌多样性研究[J].中国医药生物技术,2015,10(2):119.
[3]中国药典.一部[S].2010:357.
[4]李应兰,陈福莲.人工促成檀香结香的研究[J].热带亚热带植物学报,1994,2(3):39.
[5]莫小路,曾庆钱,邱蔚芬,等.檀香体细胞胚胎的发生及植株再生的研究[J].食品与药品,2008,10(1)35.
[6]Deng Z,Zhang R,Shi Y,et al.Characterization of Cd-,Pb-,Zn-resistant endophytic Lasiodiplodia sp.MXSF31 from metal accumulating Portulaca oleracea and its potential in promoting the growth of rape in metal-contaminated soils[J].Environ Sci Pollut Res,2014,21(3):2346.
[7]康冀川,靳瑞,文庭池,等.内生真菌产紫杉醇研究的回顾与展望[J].菌物学报,2011,30(2):168.
[8]Sun J Q,Guo L D,Zang W,et al.Research advances in the diversities of endophytic fungi in medicinal plants and their bioactive ingredients[J].Acta Bot Bor Occi Sin,2006,26(7):1505.
[9]Mayerhofer M S,Kernaghan G,Harper K A.The effects of fungal root endophytes on plant growth:a meta-analysis[J].Mycorrhiza,2013,23:119.
[10]颜仁梁,林励.檀香的研究进展[J].中药新药与临床药理,2003,14(3):218.
[11]Kim T H,Hatano T,Okamoto K,et al.Antifungal and ichthyotoxic sesquiterpenoids from Santalum album heartwood[J].Molecules,2017,22:1139.
[12]黄娟娟,杨艳,林奕云,等.檀香叶提取物抗氧化及抗菌活性初步研究[J].食品工业科学,2012,9:123.
[13]Sun S,Chen X,Guo S.Analysis of endophytic fungi in roots of Santalum album Linn.and its host plant Kuhnia rosmarinifolia Vent[J].J Zhejiang Univ Sci B,2014,15(2):109.
[14]Cao L X,You J L,Zhou S N.Endophytic fungi from Musa acuminate leaves and roots in South China[J].World J Microbiol Biotechnol,2002,18:169.
[15]魏景超.真菌鉴定手册[M].上海:科学技术出版社,1979.
[16]邓祖军,曹理想,Vrijmoed Lilian L P,等.红树林植物桐花树内生真菌类群分布的研究[J].热带海洋学报,2010,29(3):77.
[17]刘小莉,周剑忠,黄开红,等.古银杏内生真菌的分离及其抑菌活性[J].微生物学通报,2009,36(10):1513.
[18]You J L,Cao L X,Liu G F,et al.Isolation and characterization of actinomycetes antagonistic to pathogenic Vibrio spp.from nearshore marine sediments[J].World J Microbiol Biotechnol,2005,21,679.
[19]Xin G,Glawe D,Doty S L.Characterization of three endophytic,indole-3-acetic acid-producing yeasts occurring in Populus trees[J].Mycol Res,2009,113,973.
[20]Petrini O,Stone J,Carroll F E.Endophytic fungi in evergreen shrubs in western Oregon:a preliminary study[J].Can J Bot,1982,60:789.
[21]谢华蓉,徐在超,刘军,等.檀香内生真菌多样性及其对青枯菌的拮抗活性[J].微生物学通报,2017,44(5):1171.
[22]王亚妮,王丽琨,苗宗保,等.流苏石斛根部内生真菌多样性研究[J].生态科学,2014,33(4):672.
[23]李松岗.实用生物统计[M].2版.北京:北京大学出版社,2007.
[24]Camargo J A.Can dominance influence stability in competitive interactions?[J].Oikos,1992,64:605.
[25]臧威,孙翔,孙剑秋,等.南方红豆杉内生真菌的多样性与群落结构[J].应用生态学报,2014,25(7):2071.
[26]孙剑秋,郭良栋,臧威,等.药用植物内生真菌多样性及生态分布[J].中国科学C辑:生命科学,2008,38(5):475.
[27]朱军,李晓瑾,孙丽,等.新疆药用阿魏属植物内生真菌的生态分布与多样性[J].中国中药杂志,2015,40(2):356.
[28]Schulz B,Boyle C.The endophytic continuum[J].Mycol Res,2005,109(6):661.
[29]吴婧波,詹儒林,柳凤,等.杧果内生真菌分离鉴定及多样性分析[J].果树学报,2013,30(6):1030.
[30]Thongsandee W,Matsyda Y,Ito S.Temporal variations in endophytic fungal assemblages of Ginkgo biloba L.[J].J For Res,2012,17(2):213.
[31]钱一鑫,康冀川,耿坤,等.青蒿内生真菌的抗肿瘤抗氧化活性[J].菌物研究,2014,12(1):44.
[32]罗利琼,谢华蓉,邓祖军,等.蚌花内生真菌类群分布及抗菌、抗肿瘤活性的初筛[J].广东药科大学学报,2016,32(6):269.
[33]Wang L W,Zhang Y L,Lin F C,et al.Natural products with antitumor activity from endophytic fungi[J].Mini Rev Med Chem,2011,11(12):1056.
[34]张弘弛,刘瑞,李慧,等.恒山黄芪内生真菌Rhizopus sp.次生代谢产物的分离和鉴定[J].中国实验方剂学杂志,2017,23(4):71.
[35]曼琼,邓毅,杨志军,等.甘草内生菌菌株对痰浊阻肺型大鼠干预作用的对比研究[J].中国现代应用药学,2017,34(2):161.
[36]Strobel G A.Endophytes as sources of bioactive products[J].Microbes Infect,2003,5(6):535.
[37]Santoyo G,Moreno-Hagelsieb G,del Carmen Orozco-Mosqueda M,et al.Plant growth-promoting bacterial endophytes[J].Microbiol Res,2016,183:92.
[38]Kumar A,Guleria S,Mehta P,et al.Plant growth-promoting traits of phosphate solubilizing bacteria isolated from Hippophae rhamnoides L.(Sea-buckthorn)growing in cold desert Trans-Himalayan Lahul and Spiti regions of India[J].Acta Physiol Plant,2015,37:48.
[39]Khana A L,Halo B A,Elyassi A,et al.Indole acetic acid and ACC deaminase from endophytic bacteria improves the growth of Solanum lycopersicum[J].Electron J Biotechnol,2016,21:58.
[40]Rungin S,Indananda C,Suttiviriya P,et al.Plant growth enhancing effects by a siderophore-producing endophytic streptomycete isolated from a Thai jasmine rice plant(Oryza sativa L.cv.Kdml 105)[J].Anton Van Leeuw,2012,102:463.
[2]孙思胜,张大为,孟志霞,等.檀香形成部位内生真菌多样性研究[J].中国医药生物技术,2015,10(2):119.
[3]中国药典.一部[S].2010:357.
[4]李应兰,陈福莲.人工促成檀香结香的研究[J].热带亚热带植物学报,1994,2(3):39.
[5]莫小路,曾庆钱,邱蔚芬,等.檀香体细胞胚胎的发生及植株再生的研究[J].食品与药品,2008,10(1)35.
[6]Deng Z,Zhang R,Shi Y,et al.Characterization of Cd-,Pb-,Zn-resistant endophytic Lasiodiplodia sp.MXSF31 from metal accumulating Portulaca oleracea and its potential in promoting the growth of rape in metal-contaminated soils[J].Environ Sci Pollut Res,2014,21(3):2346.
[7]康冀川,靳瑞,文庭池,等.内生真菌产紫杉醇研究的回顾与展望[J].菌物学报,2011,30(2):168.
[8]Sun J Q,Guo L D,Zang W,et al.Research advances in the diversities of endophytic fungi in medicinal plants and their bioactive ingredients[J].Acta Bot Bor Occi Sin,2006,26(7):1505.
[9]Mayerhofer M S,Kernaghan G,Harper K A.The effects of fungal root endophytes on plant growth:a meta-analysis[J].Mycorrhiza,2013,23:119.
[10]颜仁梁,林励.檀香的研究进展[J].中药新药与临床药理,2003,14(3):218.
[11]Kim T H,Hatano T,Okamoto K,et al.Antifungal and ichthyotoxic sesquiterpenoids from Santalum album heartwood[J].Molecules,2017,22:1139.
[12]黄娟娟,杨艳,林奕云,等.檀香叶提取物抗氧化及抗菌活性初步研究[J].食品工业科学,2012,9:123.
[13]Sun S,Chen X,Guo S.Analysis of endophytic fungi in roots of Santalum album Linn.and its host plant Kuhnia rosmarinifolia Vent[J].J Zhejiang Univ Sci B,2014,15(2):109.
[14]Cao L X,You J L,Zhou S N.Endophytic fungi from Musa acuminate leaves and roots in South China[J].World J Microbiol Biotechnol,2002,18:169.
[15]魏景超.真菌鉴定手册[M].上海:科学技术出版社,1979.
[16]邓祖军,曹理想,Vrijmoed Lilian L P,等.红树林植物桐花树内生真菌类群分布的研究[J].热带海洋学报,2010,29(3):77.
[17]刘小莉,周剑忠,黄开红,等.古银杏内生真菌的分离及其抑菌活性[J].微生物学通报,2009,36(10):1513.
[18]You J L,Cao L X,Liu G F,et al.Isolation and characterization of actinomycetes antagonistic to pathogenic Vibrio spp.from nearshore marine sediments[J].World J Microbiol Biotechnol,2005,21,679.
[19]Xin G,Glawe D,Doty S L.Characterization of three endophytic,indole-3-acetic acid-producing yeasts occurring in Populus trees[J].Mycol Res,2009,113,973.
[20]Petrini O,Stone J,Carroll F E.Endophytic fungi in evergreen shrubs in western Oregon:a preliminary study[J].Can J Bot,1982,60:789.
[21]谢华蓉,徐在超,刘军,等.檀香内生真菌多样性及其对青枯菌的拮抗活性[J].微生物学通报,2017,44(5):1171.
[22]王亚妮,王丽琨,苗宗保,等.流苏石斛根部内生真菌多样性研究[J].生态科学,2014,33(4):672.
[23]李松岗.实用生物统计[M].2版.北京:北京大学出版社,2007.
[24]Camargo J A.Can dominance influence stability in competitive interactions?[J].Oikos,1992,64:605.
[25]臧威,孙翔,孙剑秋,等.南方红豆杉内生真菌的多样性与群落结构[J].应用生态学报,2014,25(7):2071.
[26]孙剑秋,郭良栋,臧威,等.药用植物内生真菌多样性及生态分布[J].中国科学C辑:生命科学,2008,38(5):475.
[27]朱军,李晓瑾,孙丽,等.新疆药用阿魏属植物内生真菌的生态分布与多样性[J].中国中药杂志,2015,40(2):356.
[28]Schulz B,Boyle C.The endophytic continuum[J].Mycol Res,2005,109(6):661.
[29]吴婧波,詹儒林,柳凤,等.杧果内生真菌分离鉴定及多样性分析[J].果树学报,2013,30(6):1030.
[30]Thongsandee W,Matsyda Y,Ito S.Temporal variations in endophytic fungal assemblages of Ginkgo biloba L.[J].J For Res,2012,17(2):213.
[31]钱一鑫,康冀川,耿坤,等.青蒿内生真菌的抗肿瘤抗氧化活性[J].菌物研究,2014,12(1):44.
[32]罗利琼,谢华蓉,邓祖军,等.蚌花内生真菌类群分布及抗菌、抗肿瘤活性的初筛[J].广东药科大学学报,2016,32(6):269.
[33]Wang L W,Zhang Y L,Lin F C,et al.Natural products with antitumor activity from endophytic fungi[J].Mini Rev Med Chem,2011,11(12):1056.
[34]张弘弛,刘瑞,李慧,等.恒山黄芪内生真菌Rhizopus sp.次生代谢产物的分离和鉴定[J].中国实验方剂学杂志,2017,23(4):71.
[35]曼琼,邓毅,杨志军,等.甘草内生菌菌株对痰浊阻肺型大鼠干预作用的对比研究[J].中国现代应用药学,2017,34(2):161.
[36]Strobel G A.Endophytes as sources of bioactive products[J].Microbes Infect,2003,5(6):535.
[37]Santoyo G,Moreno-Hagelsieb G,del Carmen Orozco-Mosqueda M,et al.Plant growth-promoting bacterial endophytes[J].Microbiol Res,2016,183:92.
[38]Kumar A,Guleria S,Mehta P,et al.Plant growth-promoting traits of phosphate solubilizing bacteria isolated from Hippophae rhamnoides L.(Sea-buckthorn)growing in cold desert Trans-Himalayan Lahul and Spiti regions of India[J].Acta Physiol Plant,2015,37:48.
[39]Khana A L,Halo B A,Elyassi A,et al.Indole acetic acid and ACC deaminase from endophytic bacteria improves the growth of Solanum lycopersicum[J].Electron J Biotechnol,2016,21:58.
[40]Rungin S,Indananda C,Suttiviriya P,et al.Plant growth enhancing effects by a siderophore-producing endophytic streptomycete isolated from a Thai jasmine rice plant(Oryza sativa L.cv.Kdml 105)[J].Anton Van Leeuw,2012,102:463.