Identifying airborne fungi in Seoul,Korea using metagenomics

Fungal spores are widespread and common in the atmosphere. In this study, we use a metagenomic approach to study the fungal diversity in six total air samples collected from April to May 2012 in Seoul, Korea. This springtime period is important in Korea because of the peak in fungal spore concentration and Asian dust storms, although the year of this study (2012) was unique in that were no major Asian dust events. Clustering sequences for operational taxonomic unit (OTU) identification recovered 1,266 unique OTUs in the combined dataset, with between 223?96 OTUs present in individual samples. OTUs from three fungal phyla were identified. For Ascomycota, Davidiella (anamorph: Cladosporium) was the most common genus in all samples, often accounting for more than 50% of all sequences in a sample. Other common Ascomycota genera identified were Alternaria, Didymella, Khuskia, Geosmitha, Penicillium, and Aspergillus. While several Basidiomycota genera were observed, Chytridiomycota OTUs were only present in one sample. Consistency was observed within sampling days, but there was a large shift in species composition from Ascomycota dominant to Basidiomycota dominant in the middle of the sampling period. This marked change may have been caused by meteorological events. A potential set of 40 allergyinducing genera were identified, accounting for a large proportion of the diversity present (22.5?7.2%). Our study identifies high fungal diversity and potentially high levels of fungal allergens in springtime air of Korea, and provides a good baseline for future comparisons with Asian dust storms.     

竹叶兰根系共生真菌群落结构及生物学功能初探

为研究竹叶兰Arundina graminifolia根系共生真菌的群落结构及对寄主的营养作用,采用高通量测序技术对野生竹叶兰根围土壤、根表、根内3个生态位真菌的种类及生物学功能进行鉴定与预测。所测竹叶兰根系共生真菌的平均辛普森指数为根围0.972、根 表0.905、根内0.703,说明3个位点都具有丰富的共生真菌;根围土壤真菌香农指数7.393显著高于根表和根内香农指数4.728和2.872,根表真菌多样性显著高于根内（P<0.05）。根表、根内有子囊菌门、担子菌门、罗兹壶菌门、壶菌门、毛霉菌门和被孢霉门,较根围土壤缺少球囊菌门和梳霉菌门;分子方差分析（AMOVA）证实3个生态位的真菌群落结构差异极显著（P=0.004）。各生态位丰度最高的20个真菌仅属于子囊菌和担子菌的40个属,其中子囊菌有30个属,种类数量占优势,而担子菌的单个种类丰度高于子囊菌。根围土壤腐生型真菌占优势（44.07%）;根表主要有腐生、病原、共生型真菌;根内真菌营养类型差异大,样品M1的共生型真菌为优势类型（97.21%）,M2、M3以腐生型占优（90.96%）。优势共生型真菌有革菌属Threlephora、根须腹菌属Rhizopogon和Dentiscutata,腐生型真菌主要有长毛孔菌属Funalia和Vuilleminia,均为担子菌。研究结果为揭示竹叶兰与根系真菌的营养关系以及共生真菌的开发提供了依据。     

黄山典型植被类型土壤真菌群落特征及其影响因素

黄山地势高差明显,植被类型多样,生态系统保存完整,是研究森林生态系统土壤真菌群落的天然实验室。本研究采集黄山典型植被下土壤样本,利用Illumina NovaSeq高通量测序技术分析土壤真菌群落结构,结合土壤理化性质探讨不同植被类型影响真菌群落组成的潜在因素。结果共检测到13个真菌门,优势真菌门依次为：担子菌门Basidiomycota,获得38目,202属,相对丰度介于7.30%-90.71%,在常绿落叶阔叶混交林、山地矮林及落叶阔叶林中出现高值,局部呈现先增后减的单峰变化格局;子囊菌门Ascomycota有56目,393属,相对丰度介于4.69%-53.07%,随着典型植被类型变化无明显变化规律;被孢霉门Mortierellomycota获得1目和2属,相对丰度介于2.88%-29.92%,随着典型植被类型变化呈现U型变化模式;5种植被类型土壤中共检测到34个不同分类单元的真菌指示类群,落叶阔叶林土壤真菌指示类群最为丰富,占67%;pH显著影响土壤真菌α多样性（Pearson,P<0.001）,是黄山土壤真菌群落变异的主控因子（Monte Corlo 检验,P<0.01）。     

Diversity of microorganisms in decaying maize stalks revealed by a molecular method

Microbial diversity in decaying maize stalk was characterized by constructing and analyzing rRNA gene clone library. Total 47 OTUs were obtained from 82 bacterial clones, including Proteobacteria (64.6%), Actinobacteria (30.5%), Bacteroidetes (2.4%) and Firmicutes (2.4%). Most proteobacterial clones were members of Rhizobium, Pseudomonas and Stenotrophomonas. Eighty-four percent of Actinobacteria was related to Microbacterium. Only 14 OTUs were identified from 124 fungal clones, including Ascomycota (88%) and Basidiomycota (12%). Sixty percent of Ascomycota were members of Eupenicillium and Paecilomyces but all Basidiomycota were close to Kurtzmanomyces nectairei.     

Manganese superoxide dismutase in pathogenic fungi: an issue with pathophysiological and phylogenetic involvements

Manganese-containing superoxide dismutases (MnSODs) are ubiquitous metalloenzymes involved in cell defence against endogenous and exogenous reactive oxygen species. In fungi, using this essential enzyme for phylogenetic analysis of Pneumocystis and Ganoderma genera, and of species selected among Ascomycota, Basidiomycota and Zygomycota, provided interesting results in taxonomy and evolution. The role of mitochondrial and cytosolic MnSODs was explored in some pathogenic Basidiomycota yeasts (Cryptococcus neoformans var. grubii, Cryptococcus neoformans var. gattii, Malassezia sympodialis), Ascomycota filamentous fungi (Aspergillus fumigatus), and Ascomycota yeasts (Candida albicans). MnSOD-based phylogenetic and pathogenic data are confronted in order to evaluate the roles of fungal MnSODs in pathophysiological mechanisms.     

黑胫病感染对烟草茎秆及根际土壤真菌群落结构的影响

烟草黑胫病是由寄生疫霉烟草变种引起的一种对烟草生产造成巨大经济损失的土传病害。本文以健康烟株与感染黑胫病烟株茎秆和根际土壤为研究对象,通过PCR技术扩增样本中真菌转录间隔区的ITS1区域,采用Illumina Miseq高通量测序技术对扩增片段进行测序,旨在了解黑胫病感染对于烟草茎秆和根际土壤真菌群落结构与多样性的影响。本研究20个测序样品,共获得755 599条高质量序列片段,最短序列为200bp,最长序列为356bp,平均序列长度248bp。结果表明,健康烟株和发病烟株根际土壤的优势真菌为子囊菌门Ascomycota、接合菌门Zygomycota和担子菌门Basidiomycota,所有茎秆样品的优势真菌为子囊菌门、担子菌门。健康烟株与感染黑胫病烟株根际土壤样品相对丰度大于1%的属有镰刀菌属Fusarium、被孢霉属Mortierella、隐球菌属Cryptococcus、链格孢属Alternaria和赤霉菌属Gibberella等,其中,健康烟株根际土壤优势属为镰刀菌属（39.35%）和被孢霉属（14.19%）,发病烟株根际土壤镰刀菌属和被孢霉属相对丰度分别为40.26%和20.77%;健康茎秆样品优势属为隐球菌属（31.12%）、链格孢属（18.28%）、镰刀菌属（15.67%）和红酵母属Rhodotorula（13.34%）;病健交界茎秆样品优势属为镰刀菌属（41.36%）、隐球菌属（28.15%）和链格孢属（22.32%）;发病茎秆优势属为隐球菌属（62.14%）和链格孢属（27.75%）。烟株感染黑胫病后,其根际土壤与茎秆样品真菌优势属种类与健康烟株无明显变化,但属水平的相对丰度变化显著。发病烟株茎秆与根际土壤样品真菌群落Sobs、Chao1、Shannon指数较健康烟株降低,Simpson指数上升,表明烟株发病后根际土壤真菌与茎秆内生真菌群落丰富度与多样性降低。该结果对于研究烟草黑胫病发生的微生态机制及其生物防治具有一定的理论指导意义。     

免耕覆盖及有机肥施用对土壤真菌群落组成及多样性的影响

为揭示土壤真菌群落对免耕、覆盖及有机肥施用的响应机制,在宁夏南部山区冬小麦农田连续3年采用免耕覆盖+施有机肥(NF)、免耕覆盖+不施有机肥(NC)、传统耕作不覆盖+施有机肥(TF)和传统耕作不覆盖+不施有机肥(TC)4个处理,依托Illumina MiSeq 高通量测序平台,研究了土壤真菌群落组成、多样性及其与土壤环境因子间的关系。结果表明: 4个处理共获得3490个可操作分类单元(OTUs),其中含有一些未知真菌。在已知的真菌群落中,子囊菌门和担子菌门为优势菌门,其相对丰度占总丰度的82.1%～94.2%,且TF处理下子囊菌门座囊菌纲的相对丰度最高,NF处理下担子菌门银耳菌纲的相对丰度最高。4个处理真菌群落多样性指数(Shannon和Simpson指数)依次为NC>TC>NF>TF。多元分析结果表明,微生物生物量碳是影响担子菌门和接合菌门相对丰度的主要因素,而土壤全磷、速效钾和速效磷含量高低是驱动子囊菌门真菌相对丰度变化的关键因素。在宁夏南部山区推广以免耕覆盖并施用有机肥的保护性耕作措施有利于提高土壤真菌群落多样性。     

Fungal Community Analyses of Endophytic Fungi from Two Oak Species,Quercus mongolica and Quercus serrata,in Korea

Fungal endophytes have been recorded in various plant species with a richness of diversity, and their presence plays an essential role in host plant protection against biotic and abiotic stresses. This study applied the Illumina MiSeq sequencing platform based on the amplification of fungal ribosomal ITS2 region to analyze fungal endophytic communities of two oak species (Quercus mongolica and Q. serrata) with different oak wilt disease susceptibilities in Korea. The results showed a total of 230,768 sequencing reads were obtained and clustered at a 97% similarity threshold into 709 operational taxonomic units (OTUs). The OTUs of Q. serrata were higher than that of Q. mongolica with the number of 617 OTUs and 512 OTUs, respectively. Shannon index also showed that Q. serrata had a significantly higher level of fungal diversity than Q. mongolica. Total of OTUs were assigned into 5 fungal phyla, 17 classes, 60 orders, 133 families, 195 genera, and 280 species. Ascomycota was the dominant phylum with 75.11% relative abundance, followed by Basidiomycota with 5.28%. Leptosillia, Aureobasidium and Acanthostigma were the most abundant genera detected in Q. serrata with the average relative abundance of 2.85, 2.76, and 2.19%, respectively. On the other hand, Peltaster, Cladosporium and Monochaetia were the most common genera detected in Q. mongolica with the average relative abundance of 4.83, 3.03, and 2.87%, respectively. Our results indicated that fungal endophytic communities were significantly different between two oak species and these differences could influence responses of host trees to oak wilt disease caused by Raffaelea quercus-mongolicae.     

Manganese superoxide dismutase in pathogenic fungi: An issue with pathophysiological and phylogenetic involvements

Manganese-containing superoxide dismutases (MnSODs) are ubiquitous metalloenzymes involved in cell defence against endogenous and exogenous reactive oxygen species. In fungi, using this essential enzyme for phylogenetic analysis of Pneumocystis and Ganoderma genera, and of species selected among Ascomycota, Basidiomycota and Zygomycota, provided interesting results in taxonomy and evolution. The role of mitochondrial and cytosolic MnSODs was explored in some pathogenic Basidiomycota yeasts (Cryptococcus neoformans var. grubii, Cryptococcus neoformans var. gattii, Malassezia sympodialis), Ascomycota filamentous fungi (Aspergillus fumigatus), and Ascomycota yeasts (Candida albicans). MnSOD-based phylogenetic and pathogenic data are confronted in order to evaluate the roles of fungal MnSODs in pathophysiological mechanisms.     

Cryopreservation of filamentous micromycetes and yeasts using perlite

The viability, growth and morphology of 48 strains of Ascomycota (including 17 yeasts) and 20 strains of Zygomycota were determined after a 2-d and then after 1-year storage in liquid nitrogen using a new cryopreservation method with perlite as a particulate solid carrier. In case of Ascomycota, 45 strains (94 %) out of 48 survived both 2-d and 1-year storage in liquid nitrogen, respectively. In case of Zygomycota, all 20 strains survived both storage. In addition, 3 strains of Basidiomycota counted among yeasts were tested and all survived the 1 year storage. In all surviving cultures no negative effects of cryopreservation by this method have been observed after 1-year of storage in liquid nitrogen. The results indicate that the perlite protocol can be successfully used for cryopreservation of taxonomically different groups of fungi and also for fungi which failed to survive other routinely used preservation procedures.     

Contrasting Diversity and Host Association of Ectomycorrhizal Basidiomycetes versus Root-Associated Ascomycetes in a Dipterocarp Rainforest

Root-associated fungi, including ectomycorrhizal and root-endophytic fungi, are among the most diverse and important belowground plant symbionts in dipterocarp rainforests. Our study aimed to reveal the biodiversity, host association, and community structure of ectomycorrhizal Basidiomycota and root-associated Ascomycota (including root-endophytic Ascomycota) in a lowland dipterocarp rainforest in Southeast Asia. The host plant chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) region and fungal internal transcribed spacer 2 (ITS2) region were sequenced using tag-encoded, massively parallel 454 pyrosequencing to identify host plant and root-associated fungal taxa in root samples. In total, 1245 ascomycetous and 127 putative ectomycorrhizal basidiomycetous taxa were detected from 442 root samples. The putative ectomycorrhizal Basidiomycota were likely to be associated with closely related dipterocarp taxa to greater or lesser extents, whereas host association patterns of the root-associated Ascomycota were much less distinct. The community structure of the putative ectomycorrhizal Basidiomycota was possibly more influenced by host genetic distances than was that of the root-associated Ascomycota. This study also indicated that in dipterocarp rainforests, root-associated Ascomycota were characterized by high biodiversity and indistinct host association patterns, whereas ectomycorrhizal Basidiomycota showed less biodiversity and a strong host phylogenetic preference for dipterocarp trees. Our findings lead to the working hypothesis that root-associated Ascomycota, which might be mainly represented by root-endophytic fungi, have biodiversity hotspots in the tropics, whereas biodiversity of ectomycorrhizal Basidiomycota increases with host genetic diversity.     

绿背山雀人工巢箱真菌多样性差异分析

真菌是巢穴微生物的重要组成部分,与鸟类的生存、繁殖和环境适应息息相关。本研究通过悬挂人工巢箱招引绿背山雀(Parus monticolus)入住,基于内转录间隔区(ITS)测序技术,对绿背山雀繁殖成功巢箱与筑巢未产卵巢箱真菌群落的组成差异进行探究。结果显示,在门水平上,子囊菌门(Ascomycota,98.81%)是繁殖成功巢箱内微生物的主要菌门;子囊菌门(85.59%)和担子菌门(Basidiomycota,8.33%)是筑巢未产卵巢箱内微生物的主要菌门。在属水平上,繁殖成功巢箱的优势属为拟单宽皿菌属(Phialemoniopsis,83.04%)、曲霉属(Aspergillus,4.75%)、子囊菌属(Arthroderma,4.29%)和柄帚霉属(Scopulariopsis,1.78%);筑巢未产卵巢箱的优势属为拟单宽皿菌属(36.06%)、曲霉属(14.53%)、青霉属(Penicilliu,6.22%)、单端孢霉属(Trichothecium,5.80%)、德巴利酵母菌属(Debaryomyces,1.67%)和蝶孔耳属(Papiliotrema,1.09%)。Alpha多样性分析表明,筑巢未产卵巢箱中真菌的多样性和丰富度均显著高于繁殖成功巢箱(P < 0.05);Beta多样性分析表明,繁殖成功巢和筑巢未产卵巢箱之间的真菌群落存在显著差异;LEfSe分析共检测到19个具有统计学差异的生物标记物,繁殖成功巢箱和筑巢未产卵巢箱的显著生物标志物分布在子囊菌门和担子菌门中,两种巢箱的标记物种存在显著差异。整体来说,与繁殖成功巢相比,筑巢未产卵巢内分布有更多的潜在病原菌。     

不同种植年限薰衣草根际土壤真菌群落结构的演变

土壤真菌群落结构和多样性对植物生长起着重要作用,了解种植年限对薰衣草根际土壤真菌群落结构的影响对薰衣草病害防治和增产的研究具有重要意义。探究不同种植年限薰衣草根际土壤真菌群落结构和多样性特征,及其随种植年限的演变规律。采集新疆伊犁霍城县种植年限1、3、5 a,以及未种植薰衣草土壤,对ITS序列进行Illumina高通量测序。对测序结果进行分析,比较各样品组真菌多样性和群落分布规律及与种植年限的关联。结果表明,Alpha多样性分析显示随着种植年限的增加Shannon指数逐渐降低,而Chao1指数先降低后增高。在种植薰衣草土壤中共检测到12个门,28个纲,72个目,146个科,236个属。在门水平上,子囊菌门(Ascomycota)、担子菌门(Basidiomycota)、球囊菌门(Glomeromycota)为优势菌门。随着种植年限的增加,子囊菌门相对丰度逐渐降低,担子菌门逐渐增高,而球囊菌门先增加后降低。在属水平上优势菌属为Xylodon、锐孔菌属(Oxyporus)、镰刀菌属(Fusarium)、枝孢属(Cladosporium),均属于植物致病菌。随着种植年限的增加,Xylodon...     

天山一号冰川前沿生态系统真菌群落结构演替及分布格局

【目的】分析真菌群落结构和多样性随着一号冰川退缩前沿年代序列的变化,揭示真菌群落的演替轨迹及环境因子对群落组成的影响。【方法】采用宏基因组学研究方法,结合生物信息学和统计学分析技术,对取自一号冰川末端表面冰尘,底部和前沿14个样品进行总DNA的提取,ITS基因的扩增并使用Illumina Miseq平台测序,通过相关生物地理化学特性综合分析在不同年代序列下真菌群落结构及其演替规律。【结果】经测序,筛选和质控分析获得185103条rawreads,占78.3%的非单序列在97%的相似度聚类分析共得到300个操作分类单元(OTU),共划分为6个门:子囊菌门(Ascomycota,52.7%)、担子菌门(Basidiomycota,16.9%)、壶菌门(Chytridiomycota,15.1%)、接合菌门(Zygomycota,2.4%)和球囊菌门(Glomeromycota,1.2%)。从演替初期到后期阶段虽然子囊菌的序列数逐渐下降而担子菌出现缓慢上升趋势,但子囊菌随着土壤年代序列的增加始终为优势类群,壶菌在冰川底部和前沿基层普遍存在且丰度仅次于子囊菌和担子菌。我们在缺乏植被的最新退缩基层发现依靠自养型宿主存活的活体营养菌,如Taphrinomycetes、Urediniomycetes和Ustilaginomycetes。从冰川底部和前沿基层检测到丰度较高的酵母菌,而粪生真菌(coprophilous fungi)仅仅出现在冰川前沿基层,共23个操作分类单元。球囊菌仅在前沿部分样品中存在,有着十分狭小的生态位分布。【结论】一号冰川前沿随着年代序列的增加真菌群落存在明显的演替轨迹和多样性的显著变化,不同生态位真菌类群组成的相似性较低且都存在明显的指示性真菌类群。     

Molecular analysis of fungal diversity associated with three bryophyte species in the Fildes Region, King George Island, maritime Antarctica

The fungal communities associated with three bryophytes species (the liverwort Barbilophozia hatcheri, the mosses Chorisodontium aciphyllum and Sanionia uncinata) in the Fildes Region, King George Island, maritime Antarctica, were studied using clone library analysis. Fungal communities showed low diversity; the 680 clones belonged to 93 OTUs. Of these, 78 belonged to the phylum Ascomycota, 13 to the phylum Basidiomycota, 1 to the phylum Zygomycota, and 1 to an unknown phylum. Among the OTUs, the most common orders in the Ascomycota were Helotiales (42 OTUs) and Chaetothyriales (14 OTUs) and the most common orders in the Basidiomycota were Sebacinales (3 OTUs) and Platygloeales (3 OTUs). Most OTUs clustered within clades that contained phylotypes identified from samples in Antarctic or Arctic ecosystems or from bryophytes in other ecosystems. In addition, we found that host-related factor may shape the fungal communities associated with bryophytes in this region. This is the first systematic study of the fungal community in Antarctic bryophytes to be performed using culture-independent method and the results may improve understanding of the endophytic fungal evolution and ecology in the Antarctic ecosystem.     

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。     

Analyses of soil fungal communities in adjacent natural forest and hoop pine plantation ecosystems of subtropical Australia using molecular approaches based on 18S rRNA genes

Soil fungal communities were studied using 18S rDNA-based molecular techniques. Soil DNA was analyzed using temperature gradient gel electrophoresis (TGGE), single-stranded conformational polymorphism (SSCP), cloning and sequencing methods, following community DNA extraction and polymerase chain reaction (PCR). The extracted community DNA was successfully amplified using the primer pair of EF4f-Fung5r which produced ca. 550bp 18S rDNA fragments. TGGE screening of the PCR products showed some differences in band position and intensity between two soil samples in adjacent natural forest (YNF) and hoop pine plantation (YHP) ecosystems at Yarraman in subtropical Australia. TGGE and SSCP could be used for screening PCR products. However, care must be exercised when interpreting the TGGE and SSCP results with respect to microbial diversity, because one band may not necessarily represent one species. It is recommended that the PCR products should be purified before TGGE or SSCP screening. SSCP screening of the clone sequences revealed differences among the clones. Sequence and phylogenetic analyses revealed that all obtained clones were affiliated to the kingdom Fungi, including three phyla, i.e., Zygomycota, Ascomycota and Basidiomycota. Our results suggested that community DNA extraction, PCR, cloning, SSCP screening of clones, sequencing of selected clones and phylogentic analyses could be a good strategy in investigation of soil fungal community and diversity.     

Assessment of soil fungal communities using pyrosequencing

Pyrosequencing, a non-electrophoretic method of DNA sequencing, was used to investigate the extensive fungal community in soils of three islands in the Yellow Sea of Korea, between Korea and China. Pyrosequencing was carried out on amplicons derived from the 5′ region of 18S rDNA. A total of 10,166 reads were obtained, with an average length of 103 bp. The maximum number of fungal phylotypes in soil predicted at 99% similarity was 3,334. The maximum numbers of phylotypes predicted at 97% and 95% similarities were 736 and 286, respectively. Through phylogenetic assignment using BLASTN, a total of 372 tentative taxa were identified. The majority of true fungal sequences recovered in this study belonged to the Ascomycota (182 tentative taxa in 2,708 reads) and Basidiomycota (172 tentative taxa in 6,837 reads). The predominant species of Ascomycota detected have been described as lichen-forming fungi, litter/wood decomposers, plant parasites, endophytes, and saprotrophs: Peltigera neopolydactyla (Lecanoromycetes), Paecilomyces sp. (Sordariomycetes), Phacopsis huuskonenii (Lecanoromycetes), and Raffaelea hennebertii (mitosporicAscomycota). The majority of sequences in the Basidiomycota matched ectomycorrhizal and wood rotting fungi, including species of the Agaricales and Aphyllophorales, respectively. A high number of sequences in the Thelephorales, Boletales, Stereales, Hymenochaetales, and Ceratobasidiomycetes were also detected. By applying high-throughput pyrosequencing, we observed a high diversity of soil fungi and found evidence that pyrosequencing is a reliable technique for investigating fungal communities in soils.     

Fungal spore transport by omnivorous mycophagous slug in temperate forest

Slugs are important consumers of fungal fruiting bodies and expected to carry their spores. In this study, we examined whether slugs (Meghimatium fruhstorferi) can act as effective dispersers of spores of basidiomycetes. The microscopic observation confirmed the presence of basidiospores in feces of field‐collected slugs, and the DNA metabarcoding study revealed that Ascomycota and Basidiomycota were major fungal taxa found in the feces. In Basidiomycota, the dominant order was Agaricales followed by Trichosporonales and Hymenochaetales. The laboratory experiments using Tylopilus vinosobrunneus showed that slugs carried a large number of spores in their digestive tracts. It was also observed that Pleurotus, Armillaria, and Gymnopilus spores excreted by slugs had a higher germination capacity than control spores collected from spore prints. The field experiments showed that slugs traveled 10.3 m in 5 h at most by wandering on the ground, litter layers, wood debris, and tree trunks. These results suggest that slugs could carry spores of ectomycorrhizal, saprophytic, and wood‐decaying fungi to appropriate sites for these fungi to establish colonies.