云南白木香根内丛枝菌根真菌群落组成
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摘要
目的本实验对云南省西双版纳地区野生与种植园内栽培的白木香(Aquilaria sinensis)根内丛枝菌根真菌(AMF)进行群落结构多样性调查。方法本实验使用Nested-PCR和DNA克隆测序技术。结果共获得546个含有丛枝菌根真菌18S r DNA片段的克隆子,经限制性片段长度多态性(restriction fragment length polymorphism,RFLP)分析后,得到101个RFLP类型,将其划分为22个序列类型(OTUs),分属于4个科。在Maarj AM数据库中进行比对后,21个序列类型可鉴定为17个虚拟分类分子种,分布于11种不同的生境。经统计分析野生与种植白木香根内丛枝菌根真菌群落组成差异极显著(P <0. 001)。结论本实验比较野生与种植白木香根内丛枝菌根真菌群落差异,结合丛枝菌根真菌在生态系统中的分布特点,为种植白木香人工林的选址提供有力地环境指标数据。
OBJECTIVE The characterization of arbuscular mycorrhizal fungi( AMF) in both wild and cultivated Aquilaria sinensis roots was investigated. METHODS The nested-PCR,DNA cloning and sequencing techniques were used. RESULTS A total of546 clones containing 18 S r DNA genes of AMF from roots were screened by restriction fragment length polymorphism,and 100 different RFLP types were obtained which were further sequenced and divided into 22 operational taxonomic units belonging to four families.Furthermore,assessed for similarities against the MaarjAM database,21 OTUs divided into 17 corresponding molecular virtual taxon which isolated from eleven different habitats. By statistical analysis,AMF communities were significantly different between wild and cultivated Aquilaria sinensis roots( P < 0. 001). CONCLUSION This study compares the AMF community in wild and cultivated Aquilaria sinensis roots,combining with the distributed data of this AMF species in ecosystem,providing valuable environmental indicator data to choose a site for Aquilaria sinensis plantation.
OBJECTIVE The characterization of arbuscular mycorrhizal fungi( AMF) in both wild and cultivated Aquilaria sinensis roots was investigated. METHODS The nested-PCR,DNA cloning and sequencing techniques were used. RESULTS A total of546 clones containing 18 S r DNA genes of AMF from roots were screened by restriction fragment length polymorphism,and 100 different RFLP types were obtained which were further sequenced and divided into 22 operational taxonomic units belonging to four families.Furthermore,assessed for similarities against the MaarjAM database,21 OTUs divided into 17 corresponding molecular virtual taxon which isolated from eleven different habitats. By statistical analysis,AMF communities were significantly different between wild and cultivated Aquilaria sinensis roots( P < 0. 001). CONCLUSION This study compares the AMF community in wild and cultivated Aquilaria sinensis roots,combining with the distributed data of this AMF species in ecosystem,providing valuable environmental indicator data to choose a site for Aquilaria sinensis plantation.
引文
[1]Ch.P(2015)VolⅠ(中国药典2015年版.一部)[S].2015:185-186.
[2]CUI J L,XIAO P G,GUO S X,et al.Fungus inducing the formation of agilawood from Aquilaria sinensis[J].Chin Pharm J(中国药学杂志),2012,47(5):335-337.
[3]CUI J L,XIAO P G,GUO S X,et al.Basic biological characteristics of three fungi inducing the formation of agilawood from Aquilaria sinensis[J].Chin Pharm J(中国药学杂志),2013,48(20):1711-1714.
[4]CUI J L,GUO S X,FU S B,et al.Effects of inoculating fungi on agilawood formation in Aquilaria sinensis[J].Chin Sci Bull,2013,58(26):3280-3287.
[5]HE M L,QI S Y,HU L J.Induction of chromones in excised lateral roots culture of Aquilaria sinensis[J].Chin Tradit Herb Drugs(中草药),2010,41(2):281-284.
[6]Flora of China editorial committee of Chinese academy of sciences.Flora of China(中国植物志)[M].Beijing:Science Press,1994:114.
[7]GUO L P,HUANG L Q,JIANG Y X,et al.Soil deterioration during cultivation of medicinal plants and ways to prevent it[J].China J Chin Mater Med(中国中药杂志),2006,31(9):714-717.
[8]JIANG P,WANG M Y.Colonization rate and diversity of AMfungi in the rhizosphere of seven medicinal plants in Xiamen[J].Acta Ecol Sin(生态学报),2012,32(13):4043-4051.
[9]OPIK M,MOORA M,LIIRA J,et al.Composition of root-colonizing arbuscular mycorrhizal fungi communities in different ecosystems around the globe[J].J Ecol,2006,94(4):778-790.
[10]ZHAO J,HE X L.Resource and distribution of AM fungi in the rhizosphere of medicinal plants in Anguo city of Hebei province[J].J Agric Univ Hebei(河北农业大学学报),2010,33(1):39-44.
[11]LU H Z,SHA L Q,WANG J.Seasonal variation of soil respiration and its components in tropical rain forest and rubber plantation in Xishuangbanna,Yunnan[J].Chin J Appl Ecol(应用生态学报),2009,20(10):2315-2322.
[12]PHILLIPS J M,HAYMAN D S.Improved procedures for clearing roots and staining parasitic and vesicular mycorrhizal fungi for rapid assessment of infection[J].Transact Br Mycol Soc,1970,55(1):158-161.
[13]ZHOU L S,GUO S X.Molecular diversity of arbuscular mycorrhizal fungi in wild and cultured gynostemma pentaphyllum roots in Xishuangbanna,Xouthwest China[J].Chin J Appl Ecol(应用生态学报),2013,24(9):2503-2510.
[14]JAKOBSEN I,ABBOTT L K,ROBOSEN A D.External hyphae of vesicular-arbuscular mycorrhizal fungi associated with Trifolium subterraneum L.1.Spread of hyphae and phosphorus inflow into roots[J].New Phyto,1992,120(4):371-380.
[15]LEE J,LEE S,YOUNG J P W.Improved PCR primers for the detection and identification of arbuscular mycorrhizal fungi[J].FEMS Micro Ecol,2008,65(2):339-349.
[16]SIMON L,LEVESQUE R C,LALONDE M.Identification of endomycorrhizal fungi colonizing roots by fluorescent single-strand conformation polymorphism-polymerase chain reaction[J].App Enviro Micro,1993,59(12):4211-4215.
[17]HELGASON T,DANIEL T J,HUSBAND R,et al.Ploughing up the wood-wide web[J].Nature,1998,394(6692):431.
[18]SENES-GUERRERO C,TORRES-CORTES G,PFEIFFER S,et al.Potato-associated arbuscular mycorrhizal fungal communities in the Peruvian Andes[J].Mycorrhiza,2014,24(6):405-417.
[19]OPIK M,VANSTOA A,VANSTOA E,et al.The online database Maarj AM reveals global andecosystemic distribution patterns in arbuscular mycorrhizal fungi(Glomeromycota)[J].New Phyto,2010,188(1):223-241.
[20]TOLJANDER J F,SANTOS-GONZALEZ J C,TEHLER A,et al.Community analysis of arbuscular mycorrhizal fungi and bacteria in the maize mycorrhizosphere in a long-term fertilization trial[J].FEMS Micro Ecol,2008,65(2):323-338.
[21]HEUER H,SMALLA K.Evaluation of community-level catabolic profiling using BIOLOG GN microplates to study microbial community changes in potato phyllosphere[J].J Micro Methods,1997,30(1):49-61.
[22]CHAIYASEN A,YOUNG J P,TEAUMROONG N,et al.Characterization of arbuscular mycorrhizal fungus communities of aquilariacrassna and tectonagrandis roots and soils in thailand plantations[J].PLoS One,2014,9(11):e112591.
[23]TAMULI P,BORUAH P.Vesicular-abuscular mycorrhizal(VAM)associationof agarwood tree in Jorhat District of the Brahmatputra Valley[J].Indian For,2002,128(9):991-994.
[24]PIETIKAINEN A,KYTOVIITA M M,HUSBAND R,et al.Diversity and persistence of arbuscular mycorrhizas in a low-arctic meadow habitat[J].New Phyto,2007,176(3):691-698.
[25]BRUNDRETT M,ABBOTT L K,JASPER D A.Glomalean mycorrhizal fungi fromtropical Australia.I.Comparison of the effectiveness and specificity of different isolation procedures[J].Mycorrhiza,1999,8(6):305-314.
[26]HART M M,READER J R.Host plant benefit from association with arbuscular mycorrhizal fungi:variation due to differences in size of mycelium[J].Biol Fert Soils,2002,36(5):357-366.
[27]WU L S,WANG Y,LI M,et al.A survey of arbuscular mycorrhizal fungi in the rhizosphere of camellia sinensis in Laoshan[J].J Qingdao Agric Univ(青岛农业大学学报:自然科学版),2009,26(3):171-173.
[28]JIE W G,CAI B Y,GE J P,et al.Identification of arbuscular mycorrhizal fungi of phellodendren amurense rupr[J].Biotechnology(生物技术),2007,17(6):32-34.
[29]WANG J G.Study on the AMF Diversity and AM Improved the Drought Resistance of Amygdalus mongolica[D].Inner Mongolia:Inner Mongolia University,2011.
[30]HUANG W L,FAN X J,YAN Z Y,et al.Diversity study of arbuscular mycorrhizal fungi of Coptis deltoidea[J].Chin Tradit Herb Drugs(中草药),2012,35(5):689-693.
[31]DUAN X Y.Study on the ecology and diversity of AM fungi in the rhizospheres of hedysarum scoparium[D].Xi'an:Northwest Agriculture and Forestry University,2009.
[32]JIANG P,WANG M Y,LU J C.Arbuscular mycorrhizal fungi associated with medicinal plants in Zhangzhou,Fujian[J].Mycosystema(菌物学报),2012,31(5):676-689.
[33]ZHAO J L,CHENG X Q,GU X Y,et al.Relationship between am fungi of new eight medicinal plants and soil factors in Anguo of Hebei[J].J Henan Agric Sci(河南农业科学),2012,41(6):87-91.
[34]ZHAO J.Ecological Distribution of Arbuscular Mycorrhizal Fungi the Rhizospere of Ten Species of Medicinal Plants in Anguo City Hebei Provinc[D].Baoding:Hebei University,2010.
[35]GAO A X.Ecologicaldistribution of Arbuscular Mycorrhizal Fungi in the rhizospere of Medicinal plants in Anguo[D].Baoding:Hebei University,2007.
[36]MA Y F,YANG X H,LI M Q,et al.Investigation of the diversity of arbuscular mycorrhizal structure of medicinal plants in Chongqing[J].J Southwest Agricu Univ(西南农业大学学报:自然科学版),2005,27(3):406-409.
[37]CHEN L Q,HAN N L.Identification of ginkgo VA mycorrhizal fungi in Zhejiang Province[J].Forest Res(林业科学研究),1999,12(6):581-584.
[38]OPIK M,METSIS M,DANIELL T J,et al.Large-scale parallel454 sequencing reveals host ecological group specificity of arbuscular mycorrhizal fungi in a boreonemoral forest[J].New Phyto,2009,184(2):424-437.
[39]TRESEDER K K,CROSS A.Global distributions of arbuscular mycorrhizal fungi[J].Ecosystems,2006,9(2):305-316.
[2]CUI J L,XIAO P G,GUO S X,et al.Fungus inducing the formation of agilawood from Aquilaria sinensis[J].Chin Pharm J(中国药学杂志),2012,47(5):335-337.
[3]CUI J L,XIAO P G,GUO S X,et al.Basic biological characteristics of three fungi inducing the formation of agilawood from Aquilaria sinensis[J].Chin Pharm J(中国药学杂志),2013,48(20):1711-1714.
[4]CUI J L,GUO S X,FU S B,et al.Effects of inoculating fungi on agilawood formation in Aquilaria sinensis[J].Chin Sci Bull,2013,58(26):3280-3287.
[5]HE M L,QI S Y,HU L J.Induction of chromones in excised lateral roots culture of Aquilaria sinensis[J].Chin Tradit Herb Drugs(中草药),2010,41(2):281-284.
[6]Flora of China editorial committee of Chinese academy of sciences.Flora of China(中国植物志)[M].Beijing:Science Press,1994:114.
[7]GUO L P,HUANG L Q,JIANG Y X,et al.Soil deterioration during cultivation of medicinal plants and ways to prevent it[J].China J Chin Mater Med(中国中药杂志),2006,31(9):714-717.
[8]JIANG P,WANG M Y.Colonization rate and diversity of AMfungi in the rhizosphere of seven medicinal plants in Xiamen[J].Acta Ecol Sin(生态学报),2012,32(13):4043-4051.
[9]OPIK M,MOORA M,LIIRA J,et al.Composition of root-colonizing arbuscular mycorrhizal fungi communities in different ecosystems around the globe[J].J Ecol,2006,94(4):778-790.
[10]ZHAO J,HE X L.Resource and distribution of AM fungi in the rhizosphere of medicinal plants in Anguo city of Hebei province[J].J Agric Univ Hebei(河北农业大学学报),2010,33(1):39-44.
[11]LU H Z,SHA L Q,WANG J.Seasonal variation of soil respiration and its components in tropical rain forest and rubber plantation in Xishuangbanna,Yunnan[J].Chin J Appl Ecol(应用生态学报),2009,20(10):2315-2322.
[12]PHILLIPS J M,HAYMAN D S.Improved procedures for clearing roots and staining parasitic and vesicular mycorrhizal fungi for rapid assessment of infection[J].Transact Br Mycol Soc,1970,55(1):158-161.
[13]ZHOU L S,GUO S X.Molecular diversity of arbuscular mycorrhizal fungi in wild and cultured gynostemma pentaphyllum roots in Xishuangbanna,Xouthwest China[J].Chin J Appl Ecol(应用生态学报),2013,24(9):2503-2510.
[14]JAKOBSEN I,ABBOTT L K,ROBOSEN A D.External hyphae of vesicular-arbuscular mycorrhizal fungi associated with Trifolium subterraneum L.1.Spread of hyphae and phosphorus inflow into roots[J].New Phyto,1992,120(4):371-380.
[15]LEE J,LEE S,YOUNG J P W.Improved PCR primers for the detection and identification of arbuscular mycorrhizal fungi[J].FEMS Micro Ecol,2008,65(2):339-349.
[16]SIMON L,LEVESQUE R C,LALONDE M.Identification of endomycorrhizal fungi colonizing roots by fluorescent single-strand conformation polymorphism-polymerase chain reaction[J].App Enviro Micro,1993,59(12):4211-4215.
[17]HELGASON T,DANIEL T J,HUSBAND R,et al.Ploughing up the wood-wide web[J].Nature,1998,394(6692):431.
[18]SENES-GUERRERO C,TORRES-CORTES G,PFEIFFER S,et al.Potato-associated arbuscular mycorrhizal fungal communities in the Peruvian Andes[J].Mycorrhiza,2014,24(6):405-417.
[19]OPIK M,VANSTOA A,VANSTOA E,et al.The online database Maarj AM reveals global andecosystemic distribution patterns in arbuscular mycorrhizal fungi(Glomeromycota)[J].New Phyto,2010,188(1):223-241.
[20]TOLJANDER J F,SANTOS-GONZALEZ J C,TEHLER A,et al.Community analysis of arbuscular mycorrhizal fungi and bacteria in the maize mycorrhizosphere in a long-term fertilization trial[J].FEMS Micro Ecol,2008,65(2):323-338.
[21]HEUER H,SMALLA K.Evaluation of community-level catabolic profiling using BIOLOG GN microplates to study microbial community changes in potato phyllosphere[J].J Micro Methods,1997,30(1):49-61.
[22]CHAIYASEN A,YOUNG J P,TEAUMROONG N,et al.Characterization of arbuscular mycorrhizal fungus communities of aquilariacrassna and tectonagrandis roots and soils in thailand plantations[J].PLoS One,2014,9(11):e112591.
[23]TAMULI P,BORUAH P.Vesicular-abuscular mycorrhizal(VAM)associationof agarwood tree in Jorhat District of the Brahmatputra Valley[J].Indian For,2002,128(9):991-994.
[24]PIETIKAINEN A,KYTOVIITA M M,HUSBAND R,et al.Diversity and persistence of arbuscular mycorrhizas in a low-arctic meadow habitat[J].New Phyto,2007,176(3):691-698.
[25]BRUNDRETT M,ABBOTT L K,JASPER D A.Glomalean mycorrhizal fungi fromtropical Australia.I.Comparison of the effectiveness and specificity of different isolation procedures[J].Mycorrhiza,1999,8(6):305-314.
[26]HART M M,READER J R.Host plant benefit from association with arbuscular mycorrhizal fungi:variation due to differences in size of mycelium[J].Biol Fert Soils,2002,36(5):357-366.
[27]WU L S,WANG Y,LI M,et al.A survey of arbuscular mycorrhizal fungi in the rhizosphere of camellia sinensis in Laoshan[J].J Qingdao Agric Univ(青岛农业大学学报:自然科学版),2009,26(3):171-173.
[28]JIE W G,CAI B Y,GE J P,et al.Identification of arbuscular mycorrhizal fungi of phellodendren amurense rupr[J].Biotechnology(生物技术),2007,17(6):32-34.
[29]WANG J G.Study on the AMF Diversity and AM Improved the Drought Resistance of Amygdalus mongolica[D].Inner Mongolia:Inner Mongolia University,2011.
[30]HUANG W L,FAN X J,YAN Z Y,et al.Diversity study of arbuscular mycorrhizal fungi of Coptis deltoidea[J].Chin Tradit Herb Drugs(中草药),2012,35(5):689-693.
[31]DUAN X Y.Study on the ecology and diversity of AM fungi in the rhizospheres of hedysarum scoparium[D].Xi'an:Northwest Agriculture and Forestry University,2009.
[32]JIANG P,WANG M Y,LU J C.Arbuscular mycorrhizal fungi associated with medicinal plants in Zhangzhou,Fujian[J].Mycosystema(菌物学报),2012,31(5):676-689.
[33]ZHAO J L,CHENG X Q,GU X Y,et al.Relationship between am fungi of new eight medicinal plants and soil factors in Anguo of Hebei[J].J Henan Agric Sci(河南农业科学),2012,41(6):87-91.
[34]ZHAO J.Ecological Distribution of Arbuscular Mycorrhizal Fungi the Rhizospere of Ten Species of Medicinal Plants in Anguo City Hebei Provinc[D].Baoding:Hebei University,2010.
[35]GAO A X.Ecologicaldistribution of Arbuscular Mycorrhizal Fungi in the rhizospere of Medicinal plants in Anguo[D].Baoding:Hebei University,2007.
[36]MA Y F,YANG X H,LI M Q,et al.Investigation of the diversity of arbuscular mycorrhizal structure of medicinal plants in Chongqing[J].J Southwest Agricu Univ(西南农业大学学报:自然科学版),2005,27(3):406-409.
[37]CHEN L Q,HAN N L.Identification of ginkgo VA mycorrhizal fungi in Zhejiang Province[J].Forest Res(林业科学研究),1999,12(6):581-584.
[38]OPIK M,METSIS M,DANIELL T J,et al.Large-scale parallel454 sequencing reveals host ecological group specificity of arbuscular mycorrhizal fungi in a boreonemoral forest[J].New Phyto,2009,184(2):424-437.
[39]TRESEDER K K,CROSS A.Global distributions of arbuscular mycorrhizal fungi[J].Ecosystems,2006,9(2):305-316.