马缨杜鹃根系微生物群落结构及其变化

【目的】为了解野生马缨杜鹃根系微生物的群落结构特征,比较百里杜鹃内马缨杜鹃根际土壤、根表及根内3个生态位微生物群落组成差异,探究杜鹃根系生态位之间、变化菌属间的相互关系,以期为今后杜鹃根系微生物研究提供具有参考价值的数据。【方法】对百里杜鹃内野生马缨杜鹃根系微生物16S rRNA V4区和ITS1区进行高通量测序,比较分析根际土壤、根表及根内微生物群落多样性及组成差异;并基于共变化网络分析进一步探究差异菌属间的相互关系。【结果】马缨杜鹃根系不同的生态位之间,微生物群落结构存在差异,其中以根际土壤与根表、根内差异更为显著,且细菌群落差异小于真菌群落。且从根际-根表-根内,马缨杜鹃根系细菌α-多样性显著下降。马缨杜鹃根系微生物分布于41细菌门和6个真菌门。优势细菌门为变形菌门(Proteobacteria,39.64%±0.08,69.47%±0.12,77.34%±0.07)、酸杆菌门(Acidobacteria,34.10%±0.11,11.03%±0.04,9.18%±0.04)以及放线菌门(Actinobacteria,10.19%±0.03,8.70%±0.02,7.08%±0.01),它们占整体细菌群落总丰度的80%以上。同时8个细菌菌门的相对丰度在根际土壤、根表和根内间显著变化,且它们的相对丰度占细菌群落总丰度的75%以上。真菌主要分布于接合菌门(Zygomycota)、担子菌门(Basidiomycota)和子囊菌门(Ascomycota),它们占整体菌群的99%以上。马缨杜鹃根系存在589个属的细菌,390个属的真菌,从根际-根表-根内,其中25个细菌属和10个真菌属的相对丰度发生显著变化。马缨杜鹃根系微生物群落共变化网络分析表明:在马缨杜鹃根系不同生态位间,除Waitea外,包括枝孢菌属(Cladosporium)、拟盘多毛孢属(Pestalotiopsis)等在内的8个差异真菌菌属均与细菌菌属显著相关,它们相互作用调控微生物群落结构的变化。Bryobacter、Nocardia、Rhizomicrobium和Telmatobacter等核心菌属对马缨杜鹃根系微生物群落共变化网络的变化具有十分重要的调控作用。【结论】百里杜鹃地区马缨杜鹃根际土壤、根表以及根内3个生态位间,微生物群落组成存在差异;而造成微生物组成存在差异这一结果,可能与马缨杜鹃根系密切相关。同时,共变化网络分析揭示出马缨杜鹃根系生态位之间,细菌和真菌彼此间互作。     

短花针茅荒漠草原土壤微生物群落组成及结构

为详细了解内蒙古短花针茅荒漠草原生态系统中土壤微生物群落组成与结构。对其土壤中微生物的总DNA提取后,采用高通量测序技术对土壤中细菌的16Sr DNA和真菌的ITS基因进行了序列测定,分析了短花针茅荒漠草原土壤中微生物群落结构特征。结果表明,共获得细菌OTUs13711个,真菌OTUs 5929个。物种分类显示,细菌种类隶属于29门57纲111目191科485属,其中优势类群为Gammaproteobacteria和Thermoleophilia,它们的相对丰度分别为32.68%和26.83%。真菌隶属于4门16纲45目78科105属,优势类群为Ascomycota和Basidiomycota,它们的相对丰度分别为35.76%和25.90%。土壤中细菌群落的多样性和丰富度均高于真菌。     

根腐病对茅苍术根际微生物群落多样性的影响

研究茅苍术根腐病与根际土壤微生物群落之间的关系,为茅苍术根腐病微生物调控防治技术提供依据。比较了未种植茅苍术土壤、茅苍术健康植株及根腐病病株根际土壤的土壤养分和酶活性变化,并结合Illumina MiSeq高通量测序,研究了茅苍术根腐病土壤中细菌和真菌的群落结构变化。茅苍术感染根腐病后,植株根际土壤pH降低,有机质、速效氮和速效磷显著增加;蔗糖酶活性增加,过氧化氢酶活性降低;细菌和真菌的多样性和丰度均显著降低。病原菌如镰刀菌属(Fusarium)和阿太菌属(Athelia)的丰度显著增加,而被孢霉属(Mortierella)、青霉属(Penicillium)和假单胞菌属(Pseudomonas)等病原微生物拮抗菌的丰度降低,这可能是诱发茅苍术根腐病的主要原因。通过分析细菌、真菌多样性及群落构成变化,表明多样性指数显著降低和群落结构失衡的打破可能和土壤pH、养分和酶活性的变化密切相关。     

高通量测序分析环保肥料增效剂对马铃薯根际土壤真菌多样性变化影响

【目的】高通量测序技术对研究环境样品中微生物群落组成具有很大的应用价值。土壤微生物群落结构和多样性及其变化在一定程度上反映了土壤的质量。旨在从微生物群落结构的角度阐述环保肥料增效剂对马铃薯根际土壤主要真菌类群结构的影响。【方法】通过高通量测序结果对比分析应用增效剂前后马铃薯根际真菌宏基因组ITS1区,并依据RDP中设置的分类阈值对序列进行物种分类。【结果】测序结果经过质量控制,共获得有效条带437 375条,依据97%的序列相似性做聚类分析,获得全部样品的可分类操作单元(OTUs)共633个。子囊菌的总体数量在所有样品中最多(相对丰度在56.95%-97.23%之间),且处理后呈增加趋势(HY除外),而担子菌门数量在处理后呈下降的趋势。基于真菌ITS1高通量测序结果获得的α指数显示,样品内部处理和对照之间真菌物种多样性有差别。基于真菌ITS1高通量测序获得的β指数显示,处理组与对照组的真菌多样性之间没有差别,这表明真菌多样性之间的差异更多地取决于样品采集地点。【结论】土壤特性是影响真菌种群的重要因素之一,环保肥料增效剂显著改善了土壤真菌的种群结构。     

两种混合样品策略对揭示杜鹃花根部真菌多样性的影响

由于土壤微生物群落物种组成的高度空间异质性,混合样品(sample pooling)被广泛应用于微生物多样性与群落结构研究。在根部真菌的分子检测中,样品混合策略以及测序的克隆数或序列数均对揭示真菌群落结构的准确性有影响。【目的】为建立一套能快速准确地反映杜鹃花属植物根部真菌的物种组成与群落结构的分子检测技术平台,【方法】本研究采集锈红杜鹃和亮鳞杜鹃多份根系样品分别提取DNA,比较PCR扩增前和扩增后混合策略构建的克隆文库中真菌物种组成的差异。【结果】在2种宿主植物根系中,多份样品在PCR扩增后混合构建的克隆文库检测到的根部真菌物种丰富度、真菌群落的Shannon-Wiener多样性指数均高于扩增前混合的克隆文库。高频度的根部真菌在2种克隆文库中均检测到,但低频度的真菌物种组成在2种克隆文库中完全不同。更重要的是,当采用广泛应用的真菌通用引物ITS1f和ITS4扩增根部真菌ITS序列时,PCR扩增后混合的方法能有效地减轻杜鹃花属植物ITS序列被优先扩增的现象。真菌物种累积曲线显示,当测序的真菌ITS片段克隆数达到50个左右,即能较全面地反映2种杜鹃花根部真菌物种组成。【结论】独立扩增多份根系样品DNA,再将PCR产物混合构建克隆文库的方法能更全面地揭示杜鹃花属植物根部真菌物种丰富度与物种组成。     

植被退化对滇西北高寒草地土壤微生物群落的影响

【目的】在同尺度下比较我国滇西北高寒草地土壤(GS)及其退化土壤(DGS)中细菌和真菌群落,研究植被退化对高寒草地土壤微生物群落的影响,并探索其环境驱动因子。【方法】分别以16SrRNA基因和ITS基因作为细菌和真菌分子生态学分析的靶标基因,采用定量PCR法测定基因数量来表征微生物群落丰度,采用Illumina Hiseq测序及生物信息学分析研究土壤微生物群落组成和群落结构。【结果】草地退化后,土壤pH值显著上升0.65个单位,土壤水分、总有机碳、可溶性氮含量和C/N比分别显著下降了18.4%、67.5%、47.2%和71.2%;草地退化显著降低了土壤细菌和真菌群落丰度,降低幅度分别为92.4%和94.9%;草地退化没有影响土壤细菌和真菌群落α-多样性,但显著改变了细菌和真菌群落β-多样性(群落结构);草地退化改变了土壤细菌和真菌在OTU水平上的物种组成,土壤真菌OTU种类变化更为显著;草地退化没有影响土壤细菌在门水平上的群落组成,但改变了细菌在纲水平上的群落组成(如Acidimicrobiia、Betaproteobacteria、Chloroplast等);草地退化没有影响土壤真菌在门水平和纲水平上的群落组成。【结论】本研究发现植被退化后滇西北高寒草地土壤质量显著降低,寄居在土壤中的微生物群落丰度也显著降低、微生物群落结构明显改变。     

两种麻山药典型病害根际土壤微生物多样性的研究

为了解两种典型病害土壤根际微生物群落结构,明确土壤病害与微生物多样性变化的关系,采用高通量测序对两种典型病害根际微生物多样性进行对比分析,结果表明,细菌多样性和丰度值表现为糊头病的土壤对照组根茎腐病组;而真菌变化更明显,两种病害土壤真菌多样性和丰富度均高于对照组。物种分类注释表明,不同处理土壤细菌种类与对照组基本一致,只是不同类别所占的比例各不相同,真菌中含有一些与病害发生有关的特有属,且比例变化较大,说明土壤微生物群落结构变化在一定程度上影响病害的发生。     

利用小亚基核糖体RNA 技术分析温室黄瓜近根土壤古菌和真菌多样性

土壤古菌和真菌在温室生态系统是仅次于细菌的微生物,具有类似于细菌的重要生态功能。通过构建古菌16S rRNA和真菌18S rRNA基因克隆文库,分析温室黄瓜近根土壤古菌和真菌群落结构组成,为开发利用温室这一特殊的生态环境中丰富的微生物资源以及理解微生物与植物间的互作提供参考依据。采用研磨-冻融-溶菌酶-蛋白酶K-SDS热处理以及CTAB处理等理化方法,提取和纯化微生物总DNA,构建古菌16S rRNA和真菌18S rRNA基因克隆文库。利用DOTUR软件将古菌和真菌序列按照相似性97％的标准分成若干个可操作分类单元 (OTUs)。土壤古菌克隆文库主要包括泉古菌门和未分类的古菌两大类,并有少部分广域古菌类群,所有泉古菌均属于热变形菌纲,共45个OTUs;真菌克隆文库包括真菌门的大多数亚门真菌,共24个OTUs,未发现担子菌亚门真菌。古菌多样性比较丰富,且发现少量的广域古菌 (甲烷菌),这一情况可能与温室长期高温高湿,高有机质含量,土壤处于缺氧环境有关;土壤真菌的优势种群为子囊菌,占到土壤真菌的80％以上,这可能与绝大多数植物真菌性病害属于土传病害,通过菌丝体、菌核或子囊壳在土壤病残体中越冬有一定的关系。     

黄精根际及药用部位内生真菌群落组成和生态功能分析

为寻找促进药用植物活性代谢产物合成的微生物,该文以黄精为研究对象,利用高通量测序技术和生态功能预测平台,测定根际土真菌、根茎和根内生真菌的ITS序列,分析其真菌多样性和群落组成,并预测根茎内生真菌的生态功能.结果表明:(1)测序得到1023个可操作分类单元(OTUs),根际、根茎和根真菌OTU数分别为703、128和1...     

羊肚菌烂柄病发生对土壤真菌群落结构的影响

为了解羊肚菌(Morel)烂柄病的发生对土壤真菌群落结构的影响,采用Illumina MiSeq高通量测序技术,对健康羊肚菌根际土、烂柄病发病子实体根际土及相同环境下未栽培羊肚菌土壤的真菌群落结构进行研究。结果表明,测序样品共获得344 163条序列,归为7个真菌门。各样品真菌多样性分析结果表明,未栽培土壤真菌群落多样性较高,栽培羊肚菌根际土壤真菌多样性显著降低,烂柄病发生后土壤真菌多样性增加。群落结构分析表明,烂柄病根际优势真菌类群为拟青霉属(Paecilomyces)、木霉属(Trichoderma)、葡萄穗霉属(Stachybotrys)、枝顶孢属(Acremonium)、Paratritirachium、Zopfiella、被孢霉属(Mortierella)和柄孢壳属(Podospora)。烂柄病的发生改变了土壤真菌群落结构,促进了根际真菌的繁殖。为了解羊肚菌烂柄病的发生、传播机理及防治提供了参考。     

Bacterial diversity in the bulk soil and rhizosphere fractions of Lolium perenne and Trifolium repens as revealed by PCR restriction analysis of 16S rDNA

The rhizosphere of Trifolium repens and Lolium perenne was divided into three fractions: the bulk soil, the soil adhering to the roots and the washed roots (rhizoplane and endorhizosphere). After isolation and purification of DNA from these fractions, 16S rDNA was amplified by PCR and cloned to obtain a collection of 16S rRNA genes representative of the bacterial communities of these three fractions. The genes were then characterized by PCR restriction analysis. Each different profile was used to define an operational taxonomic unit (OTU). The numbers of OTUs and the numbers of clones among these OTUs allowed to calculate a diversity index. The number of OTUs decreased as root proximity increased and a few OTUs became dominant, resulting in a lower diversity index. In the root fraction of T. repens, the restriction profile of the dominant OTU matched the theoretical profile of the 16S rRNA gene of Rhizobium leguminosarum. This study showed that plant roots create a selective environment for microbial populations.     

黑老虎内生真菌及根际土壤真菌的群落结构与生态功能分析

为探讨黑老虎(Kadsura coccinea)根际土壤和组织内生真菌菌群的组成及其生态功能，该研究采用ITS高通量测序技术对成熟黑老虎(根、茎、叶)内生真菌及根际土壤真菌群落结构、多样性和生态功能进行了分析。结果表明：(1)从12个样品中共获得2 241个可操作分类单元(OTU),涉及10门、41纲、95目、212科、367属，内生真菌(根、茎、叶)和根际土壤真菌OTU数分别为386、536、258、1 435个，其中共有的OTU为18个。在门水平上，黑老虎内生真菌及根际土壤真菌优势群落均为子囊菌门和担子菌门，其中子囊菌门在叶和茎中占比分别高达96.99%和95.37%;在属水平上，黑老虎根际土壤真菌中腐生真菌被孢霉属占比较高(为13.5%),叶和茎等生长旺盛的组织中子囊菌门未分类属和痂囊腔菌属占比较高。(2)α多样性分析结果显示，黑老虎根际土壤真菌群落的丰度和多样性明显高于内生真菌，茎中内生真菌丰度显著高于根和叶，而根、茎和叶组织间内生真菌多样性差异不显著；PCoA分析结果显示，叶和茎的真菌群落结构相似性更高。(3)利用FUNGuild数据库进行的功能预测分析结果显示，黑老虎根际土...     

High richness of ectomycorrhizal fungi and low host specificity in a coastal sand dune ecosystem revealed by network analysis

Ectomycorrhizal (EM) fungi are ubiquitous in temperate and boreal forests, comprising over 20,000 species forming root symbiotic associations with Pinaceae and woody angiosperms. As much as 100 different EM fungal species can coexist and interact with the same tree species, forming complex multispecies networks in soils. The degree of host specificity and structural properties of these interaction networks (e.g., nestedness and modularity) may influence plant and fungal community assembly and species coexistence, yet their structure has been little studied in northern coniferous forests, where trees depend on EM fungi for nutrient acquisition. We used high‐throughput sequencing to characterize the composition and diversity of bulk soil and root‐associated fungal communities in four co‐occurring Pinaceae in a relic foredune plain located at Îles de la Madeleine, Québec, Canada. We found high EM fungal richness across the four hosts, with a total of 200 EM operational taxonomic units (OTUs), mainly belonging to the Agaricomycetes. Network analysis revealed an antinested pattern in both bulk soil and roots EM fungal communities. However, there was no detectable modularity (i.e., subgroups of interacting species) in the interaction networks, indicating a low level of specificity in these EM associations. In addition, there were no differences in EM fungal OTU richness or community structure among the four tree species. Limited shared resources and competitive exclusion typically restrict the number of taxa coexisting within the same niche. As such, our finding of high EM fungal richness and low host specificity highlights the need for further studies to determine the mechanisms enabling such a large number of EM fungal species to coexist locally on the same hosts.     

Analogous wheat root rhizosphere microbial successions in field and greenhouse trials in the presence of biocontrol agents Paenibacillus peoriae SP9 and Streptomyces fulvissimus FU14

Two Pythium-infested soils were used to compare the wheat root and rhizosphere soil microbial communities from plants grown in the field or in greenhouse trials and their stability in the presence of biocontrol agents. Bacteria showed the highest diversity at early stages of wheat growth in both field and greenhouse trials, while fungal diversity increased later on, at 12 weeks of the crop cycle. The microbial communities were stable in roots and rhizosphere samples across both soil types used in this study. Such stability was also observed irrespective of the cultivation system (field or greenhouse) or addition of biocontrol coatings to wheat seeds to control Pythium disease (in this study soil infected with Pythium sp. clade F was tested). In greenhouse plant roots, Archaeorhizomyces, Debaryomyces, Delftia, and unclassified Pseudeurotiaceae were significantly reduced when compared to plant roots obtained from the field trials. Some operational taxonomic units (OTUs) represented genetic determinants clearly transmitted vertically by seed endophytes (specific OTUs were found in plant roots) and the plant microbiota was enriched over time by OTUs from the rhizosphere soil. This study provided key information regarding the microbial communities associated with wheat roots and rhizosphere soils at different stages of plant growth and the role that Paenibacillus and Streptomyces strains play as biocontrol agents in supporting plant growth in infested soils.     

Fungal soil communities in a young transgenic poplar plantation form a rich reservoir for fungal root communities

Fungal communities play a key role in ecosystem functioning. However, only little is known about their composition in plant roots and the soil of biomass plantations. The goal of this study was to analyze fungal biodiversity in their belowground habitats and to gain information on the strategies by which ectomycorrhizal (ECM) fungi form colonies. In a 2-year-old plantation, fungal communities in the soil and roots of three different poplar genotypes (Populus × canescens, wildtype and two transgenic lines with suppressed cinnamyl alcohol dehydrogenase activity) were analyzed by 454 pyrosequencing targeting the rDNA internal transcribed spacer 1 (ITS) region. The results were compared with the dynamics of the root-associated ECM community studied by morphotyping/Sanger sequencing in two subsequent years. Fungal species and family richness in the soil were surprisingly high in this simple plantation ecosystem, with 5944 operational taxonomic units (OTUs) and 186 described fungal families. These findings indicate the importance that fungal species are already available for colonization of plant roots (2399 OTUs and 115 families). The transgenic modification of poplar plants had no influence on fungal root or soil communities. Fungal families and OTUs were more evenly distributed in the soil than in roots, probably as a result of soil plowing before the establishment of the plantation. Saprophytic, pathogenic, and endophytic fungi were the dominating groups in soil, whereas ECMs were dominant in roots (87%). Arbuscular mycorrhizal diversity was higher in soil than in roots. Species richness of the root-associated ECM community, which was low compared with ECM fungi detected by 454 analyses, increased after 1 year. This increase was mainly caused by ECM fungal species already traced in the preceding year in roots. This result supports the priority concept that ECMs present on roots have a competitive advantage over soil-localized ECM fungi.     

Reiteration in the Monodominant Tropical Tree Dicymbe corymbosa (Caesalpiniaceae) and its Potential Adaptive Significance

The tropical monodominant tree Dicymbe corymbosa reiterates via epicormic shoots and roots, resulting in multistemmed trees with complex pseudotrunks and root mounds. In 2 ha of primary forest on the Guiana Shield, we quantified the reiterative structure and aboveground soil development of 307 D. corymbosa individuals ≥ 10 cm dbh and investigated the potential adaptive significance of reiteration in terms of genet persistence and root exploitation of aboveground soil accumulations. We also investigated the incidence of the heart rot fungus Phellinus robustus in D. corymbosa and examined its relationship to the reiteration process. Large trees contained more and larger reiterations, greater trunk, root mound, and organic soil volumes, and a higher incidence of Phellinus than smaller trees. Roots and ectomycorrhizas were abundant in aboveground soils on the trees, occurred at higher densities than those of the surrounding forest floor, and may be important in recycling mineral nutrients. Stem turnover and reiteration were associated with Phellinus heart rot and appeared to be cumulative over time, resulting in persistent, structurally complex trees of indeterminate lifespan. Dicymbe corymbosa provides a rare example of a tree species that exploits both persistence and recruitment niches, as it successfully recruits through mast fruiting.     

Structure and species composition of ectomycorrhizal fungal communities colonizing seedlings and adult trees of Pinus montezumae in Mexican neotropical forests

Mexico is a center of diversity for pines, but few studies have examined the ectomycorrhizal (ECM) fungal communities associated with pines in this country. We investigated the ECM communities associated with Pinus montezumae seedlings and mature trees in neotropical forests of central Mexico and compared their structure and species composition. Root tips were sampled on both planted seedlings and naturally occurring adult trees. A total of 42 ECM operational taxonomic units (OTUs) was found on P.?montezumae. Diversity and similarity indices showed that community structure was similar for both plant growth stages, but phylogenetic diversity and Chao-estimated richness were higher for seedlings. Species composition differed between communities. The dominant OTUs belonged to the families Atheliaceae, Cortinariaceae, and Sebacinaceae, although different taxa appeared to colonize seedlings and adults. Only 12 OTUs were shared between seedlings and adults, which suggests that ECM fungi which colonize seedlings are still not fully incorporated into mycelial networks and that ECM taxa colonizing young individuals of P.?montezumae are likely to come from fungal propagules. Intra-generic diversity could be an insurance mechanism to maintain forest productivity under stressed conditions. This is the first report describing the abundance of Atheliaceae in tree roots in neotropical ecosystems.     

Medicago species affect the community composition of arbuscular mycorrhizal fungi associated with roots

Sequencing of the 5' end of the large ribosomal subunit (LSU rDNA) and quantitative polymerase chain reaction (qPCR) were combined to assess the impact of four annual Medicago species (Medicago laciniata, Medicago murex, Medicago polymorpha and Medicago truncatula) on the genetic diversity of arbuscular mycorrhizal (AM) fungi, and on the relative abundance of representative AM fungal genotypes, in a silty-thin clay soil (Mas d'Imbert, France). Two hundred and forty-six Glomeromycete LSU rDNA sequences from the four plant species and the bulk soil were analysed. The high bootstrap values of the phylogenetic tree obtained allowed the delineation of 12 operational taxonomic units (OTUs), all belonging to Glomus. Specific primers targeting Glomeromycetes and major OTUs were applied to quantify their abundance by qPCR. Glomeromycetes and targeted OTUs were significantly more abundant in the root tissues than in the bulk soil, and the frequencies of three of them differed significantly in the root tissues of the different plant species. These differences indicate that, despite the absence of strict host specificity in mycorrhizal symbiosis, there was a preferential association between some AM fungal and plant genotypes.     

Diversity of root-associated fungal endophytes in Rhododendron fortunei in subtropical forests of China

To investigate the diversity of root endophytes in Rhododendron fortunei, fungal strains were isolated from the hair roots of plants from four habitats in subtropical forests of China. In total, 220 slow-growing fungal isolates were isolated from the hair roots of R. fortunei. The isolates were initially grouped into 17 types based on the results of internal transcribed spacer-restriction fragment length polymorphism (ITS-RFLP) analysis. ITS sequences were obtained for representative isolates from each RFLP type and compared phylogenetically with known sequences of ericoid mycorrhizal endophytes and selected ascomycetes or basidiomycetes. Based on phylogenetic analysis of the ITS sequences in GenBank, 15 RFLP types were confirmed as ascomycetes, and two as basidiomycetes; nine of these were shown to be ericoid mycorrhizal endophytes in experimental cultures. The only common endophytes of R. fortunei were identified as Oidiodendron maius at four sites, although the isolation frequency (3–65%) differed sharply according to habitat. Phialocephala fortinii strains were isolated most abundantly from two habitats which related to the more acidic soil and pine mixed forests. A number of less common mycorrhizal RFLP types were isolated from R. fortunei at three, two, or one of the sites. Most of these appeared to have strong affinities for some unidentified root endophytes from Ericaceae hosts in Australian forests. We concluded that the endophyte population isolated from R. fortunei is composed mainly of ascomycete, as well as a few basidiomycete strains. In addition, one basidiomycete strain was confirmed as a putative ericoid mycorrhizal fungus.     

High specificity of a rare terrestrial orchid toward a rare fungus within the North American tallgrass prairie

The Orchidaceae are globally distributed and represent a diverse lineage of obligate mycotrophic plants. Given their dependence on symbiotic fungi for germination and/or plant development, fungal community structure in substrates is expected to influence the distribution and persistence of orchid species. Yet, simultaneous characterization of orchid mycorrhizal fungal (OMF) communities in roots and in soil is rarely reported. To explain the co-distributions of OMF in roots, orchid-occupied, and bulk soil, we characterized mycorrhizal fungi associated with Platanthera praeclara over multiple years across its entire natural distribution within the North American tallgrass prairie. Root derived OMF communities included 24 Ceratobasidiaceae and 7 Tulasnellaceae operational taxonomic units (OTUs) though the orchid exhibited high spatio-temporal specificity toward a single Ceratobasidiaceae OTU, which was strongly stable across population sizes and phenological stages of the sampled individuals. The preferred OMF OTUs were primarily restricted to orchid-occupied locations while infrequent or absent in bulk soil. Variation in soil OMF assemblies was explained most by soil moisture, magnesium, manganese, and clay. In this first study of coupled root and soil OMF communities across a threatened grassland ecosystem, we report a strong relationship, further nuanced by soil chemistry, between a rare fungus and a rare orchid.