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丛枝菌根真菌可以通过与植物建立共生关系促进植物吸收矿质营养,在植物抵御逆境过程中起到重要的作用。新一代测序技术的快速发展极大地推动了微生物多样性研究,扩增子测序技术对植物和土壤样本中丛枝菌根真菌的检测起到了重要的作用。本研究报道一种基于Illumina MiSeq扩增子测序的丛枝菌根真菌检测方法,通过设计专门针对丛枝菌根真菌的引物,对植物和土壤样本中的丛枝菌根真菌进行扩增子测序检测。研究结果表明,设计的引物相较于通用引物f ITS7/ITS4在属水平上对丛枝菌根真菌有更高的检出率。建立了一种将测序数据与丛枝菌根真菌代表序列直接比对注释的方法,与OTU代表序列注释的方法相比具有较高的准确度。对丛枝菌根真菌代表序列和检出序列的系统发育分析显示出较好的系统发育关系。本研究有助于更高效检测植物和土壤样本中的丛枝菌根真菌。
Abstract：Arbuscular mycorrhizal fungi can promote plants to absorb mineral nutrients by establishing symbiotic relationship with plants,and play an important role in plant resistance to stress.The rapid development of next generation sequencing technology has greatly promoted the study of microbial diversity.Amplicon sequencing technology plays an important role in the detection of arbuscular mycorrhizal fungi in plant and soil samples.This study reported a method for detecting arbuscular mycorrhizal fungi based on Illumina MiSeq amplicon sequencing.Specific primers for arbuscular mycorrhizal fungi were designed,amplicon sequencing was performed for arbuscular mycorrhizal fungi detection in plant and soil samples.The results showed that the designed primers had higher detection rate of arbuscular mycorrhizal fungi than the universal primers fITS7/ITS4 at genus level.Method of direct comparison between sequencing data and representative sequences of arbuscular mycorrhizal fungi for annotation was established.Compared with the method of OTU representative sequence annotation,it achieves higher accuracy.Phylogenetic analysis of the representative and detected sequences of arbuscular mycorrhizal fungi showed a good phylogenetic relationship.This study is helpful to detect arbuscular mycorrhizal fungi in plant and soil samples more efficiently.
Blaxter M.,Mann J.,Chapman T.,Thomas F.,Whitton C.,Floyd R.,and Abebe E.,2005,Defining operational taxonomic units using DNA barcode data,Philos.Trans.R.Soc.Lond.B.Biol.Sci.,360(1462):1935-1943
Bokulich N.A.,Subramanian S.,Faith J.J.,Gevers D.,Gordon J.I.,Knight R.,Mills D.A.,and Caporaso J.G.,2013,Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing,Nat.Methods,10(1):57-59
Chen B.D.,Yu M.,Hao Z.P.,Xie W.,and Zhang X.,2019,Research progress in arbuscular mycorrhizal technology,Yingyong Shengtai Xuebao (Chinese Journal of Applied Ecology),30(3):1035-1046 (陈保冬,于萌,郝志鹏,谢伟,张莘,2019,丛枝菌根真菌应用技术研究进展,应用生态学报,30(3):1035-1046)
Dumbrell A.J.,Ashton P.D.,Aziz N.,Feng G.,Nelson M.,Dytham C.,Fitter A.H.,and Helgason T.,2011,Distinct seasonal assemblages of arbuscular mycorrhizal fungi revealed by massively parallel pyrosequencing,New Phytol.,190(3):794-804
Evelin H.,Kapoor R.,and Giri B.,2009,Arbuscular mycorrhizal fungi in alleviation of salt stress:a review,Ann.Bot.,104(7):1263-1280
Gollotte A.,van Tuinen D.,and Atkinson D.,2004,Diversity of arbuscular mycorrhizal fungi colonising roots of the grass species Agrostis capillaris and Lolium perenne in a field experiment,Mycorrhiza,14(2):111-117
Gosling P.,Hodge A.,Goodlass G.,and Bending G.D.,2006,Arbuscular mycorrhizal fungi and organic farming,Agr.Ecosyst.Environ.,113(1-4):17-35
Hildebrandt U.,Regvar M.,and Bothe H.,2007,Arbuscular mycorrhiza and heavy metal tolerance,Phytochemistry,68 (1):139-146
Huang C.L.,Jian F.Y.,Huang H.J.,Chang W.C.,Wu W.L.,Hwang C.C.,Lee R.H.,and Chiang T.Y.,2014,Deciphering mycorrhizal fungi in cultivated Phalaenopsis microbiome with next-generation sequencing of multiple barcodes,Fungal Divers.,66(1):77-88
Jeffries P.,Gianinazzi S.,Perotto S.,Turnau K.,and Barea J.M.,2003,The contribution of arbuscular mycorrhizal fungi in sustainable maintenance of plant health and soil fertility,Biol.Fert.Soils,37(1):1-16
Karlsson I.,Friberg H.,Steinberg C.,and Persson P.,2014,Fungicide effects on fungal community composition in the wheat phyllosphere,PLoS ONE,9(11):e111786
Krüger M.,Krüger C.,Walker C.,Stockinger H.,and Schüssler A.,2012,Phylogenetic reference data for systematics and phylotaxonomy of arbuscular mycorrhizal fungi from phylum to species level,New Phytol.,193(4):970-984
Kumar S.,Stecher G.,and Tamura K.,2016,MEGA7:molecular evolutionary genetics analysis version 7.0 for bigger datasets,Mol.Biol.Evol.,33(7):1870-1874
Morgan B.,and Egerton-Warburton L.M.,2017,Barcoded NS31/AML2 primers for sequencing of arbuscular mycorrhizal communities in environmental samples,Appl.Plant Sci.,5(8):1700017
pik M.,Vanatoa A.,Vanatoa E.,Moora M.,Davison J.,Kalwij J.M.,Reier U.,and Zobel M.,2010,The online database MaarjAM reveals global and ecosystemic distribution patterns in arbuscular mycorrhizal fungi (Glomeromycota),New Phytol.,188(1):223-241
pik M.,Zobel M.,Cantero J.J.,Davison J.,Facelli J.M.,Hiiesalu I.,Jairus T.,Kalwij J.M.,Koorem K.,Leal M.E.,Liira J.,Metsis M.,Neshataeva V.,Paal J.,Phosri C.,Plme S.,Reier譈.,Saks譈.,Schimann H.,Thiéry O.,Vasar M.,and Moora M.,2013,Global sampling of plant roots expands the described molecular diversity of arbuscular mycorrhizal fungi,Mycorrhiza,3(5):411-430
Sato K.,Suyama Y.,Saito M.,and Sugawara K.,2005,A new primer for discrimination of arbuscular mycorrhizal fungi with polymerase chain reaction-denature gradient gel electrophoresis,Grassl.Sci.,51(2):179-181
Vasar M.,Andreson R.,Davison J.,Jairus T.,Moora M.,Remm M.,Young J.P.W.,Zobel M.,and魻pik M.,2017,Increased sequencing depth does not increase captured diversity of arbuscular mycorrhizal fungi,Mycorrhiza,27(8):761-773
Wang Q.,Bao Y.Y.,Liu X.W.,and Du G.X.,2014,Spatio-temporal dynamics of arbuscular mycorrhizal fungi associated with glomalin-related soil protein and soil enzymes in different managed semiarid steppes,Mycorrhiza,24 (7):525-538
Wang Y.S.,and Liu R.J.,2017,A checklist of arbuscular mycorrhizal fungi in the recent taxonomic system of Glomeromycota,Junwu Xuebao (Mycosystema),36(7):820-850 (王幼珊,刘润进,2017,球囊菌门丛枝菌根真菌最新分类系统菌种名录,菌物学报,36(7):820-850)
Xu X.H.,Chen C.,Zhang Z.,Sun Z.H.,Chen Y.H.,Jiang J.D.,and Shen Z.G.,2017,The influence of environmental factors on communities of arbuscular mycorrhizal fungi associated with Chenopodium ambrosioides revealed by Mi Seq sequencing investigation,Sci.Rep.,7:45134
基本信息:
DOI：10.13417/j.gab.039.005617
中图分类号:Q945;Q93
引用信息:
[1]王启,刘广达.一种基于扩增子测序检测丛枝菌根真菌的方法[J].基因组学与应用生物学,2020,39(12):5617-5624.DOI:10.13417/j.gab.039.005617.
基金信息:
内蒙古自治区卫生和计划生育委员会科研计划项目(201703129);; 包头市青年创新人才项目;; 包头医学院青苗计划项目(BYJJ-QM201780);包头医学院秦文斌科技教育基金(BYJJ-QW201603)共同资助
2020-12-25
2020-12-25