Mycorrhiza-plant colonization patterns on a subalpine glacier forefront as a model system of primary succession

Lyman glacier in the North Cascades Mountains of Washington has a subalpine forefront characterized by a well-developed terminal moraine, inconspicuous successional moraines, fluting, and outwash. These deposits were depleted of symbiotic fungi when first exposed but colonized by them over time after exposure. Four major groups of plant species in this system are (1) mycorrhiza-independent or facultative mycotrophic, (2) dependent on arbuscular mycorrhizae (AM) (3) dependent on ericoid mycorrhiza (ERM) or ectomycorrhizae (EM), and (4) colonized by dark-septate (DS) endophytes. We hypothesized that availability of mycorrhizal propagules was related to the success of mycorrhiza-dependent plants in colonizing new substrates in naturally evolved ecosystems. To test this hypothesis roots samples of 66 plant species were examined for mycorrhizal colonization. The plants were sampled from communities at increasing distances from the glacier terminus to compare the newest communities with successively older ones. Long established, secondary successional dry meadow communities adjacent to the glacier forefront, and nearby high alpine communities were sampled for comparison. DS were common on most plant species on the forefront. Nonmycorrhizal plants predominated in the earlier successional sites, whereas the proportion of mycorrhizal plants generally increased with age of community. AM were present, mostly at low levels, and nearly absent in two sites of the forefront. ERM were present in all species of Ericaceae sampled, and EM in all species of Pinaceae and Salicaceae. Roots of plants in the long established meadow and heath communities adjacent to the forefront and the high alpine community all had one or another of the colonization types, with DS and AM predominating.     

Ericoid mycorrhizal fungi are common root inhabitants of non-Ericaceae plants in a south-eastern Australian sclerophyll forest

Fungi were isolated from the roots of 17 plant species from the families Apiaceae, Cunoniaceae, Cyperaceae, Droseraceae, Fabaceae-Mimosoideae, Lomandraceae, Myrtaceae, Pittosporaceae, Proteaceae and Stylidiaceae at a sclerophyll forest site in New South Wales, Australia. Internal transcribed spacer (ITS) restriction fragment length polymorphism (RFLP) and sequence comparisons indicated that the isolated fungi had affinities to a range of ascomycetes, basidiomycetes and zygomycetes. Four RFLP types had closest affinities to previously identified Helotiales ericoid mycorrhizal (ERM) or Oidiodendron spp. Isolates representing six RFLP types, which were variously isolated from all 17 plant species, formed ERM coils in hair root epidermal cells of Woollsia pungens (Ericaceae) under gnotobiotic conditions. Three of these isolates formed intercellular hyphae, intracellular hyphae and/or microsclerotia, which are typical of dark septate endophyte infection, in roots of Stylidium productum (Stylidiaceae), indicating an ability to form different types of association with roots of different hosts. Overall the data indicate that a broad range of plant taxa may act as repositories for ERM fungi in sclerophyll forest soil.     

Dark septate endophytes: a review of facultative biotrophic root-colonizing fungi

Dark septate root endophytes (DSE) are conidial or sterile fungi (Deuteromycotina, Fungi Imperfecti) likely to be ascomycetous and colonizing plant roots. They have been reported for nearly 600 plant species representing about 320 genera and 100 families. DSE fungi occur from the tropics to arctic and alpine habitats and comprise a heterogeneous group that functionally and ecologically overlaps with soil fungi, saprotrophic rhizoplane-inhabiting fungi, obligately and facultatively pathogenic fungi and mycorrhizal fungi. Numerous species of undescribed sterile and anamorphic taxa may also await discovery. Although DSE are abundant in washed root and soil samples from various habitats, and are easily isolated from surface-sterilized roots of ecto-, ectendo-, endo- and non-mycorrhizal host species, their ecological functions are little understood. Studies of DSE thus far have yielded inconsistent results and only poorly illustrate the role of DSE in their natural habitats. These inconsistencies are largely due to the uncertain taxonomic affinities of the strains of DSE used. In addition, because different strains of a single anamorph taxon seem to vary greatly in function, no clear generalizations on their ecological role have been drawn. This paper reviews the current literature on DSE and the ecology and discusses the need for and direction of future research.     

Seasonal and temporal dynamics of arbuscular mycorrhizal and dark septate endophytic fungi in a tallgrass prairie ecosystem are minimally affected by nitrogen enrichment

Root colonization by arbuscular mycorrhizae (AM) and dark septate endophytic (DSE) fungi in nitrogen amended and unamended mixed tallgrass prairie communities were analyzed monthly over two growing seasons. Roots were stained with Trypan blue and Sudan IV and fungal structures quantified using the modified magnified intersections method. Root length colonized (RLC) by DSE exceeded AM colonization during early part of the growing season. Fungal colonization varied among the years and was greater in 2003 than in 2002. Seasonal variation among the months within a growing season was observed in 2002 but not in 2003 for both AM and DSE. AM fungi were most abundant during the peak growing season of dominant C₄ vegetation while DSE were most abundant during the early part of the growing season. Hyperparasitism of AM hyphal coils by melanized septate fungi was frequently observed and increased with AM coil frequency. Although nitrogen amendment had altered the plant community composition, it had no impact on the colonization by AM or DSE fungi.     

Identical genotypes of an ericoid mycorrhiza-forming fungus occur in roots of Epacris pulchella (Ericaceae) and Leptospermum polygalifolium (Myrtaceae) in an Australian sclerophyll forest

Assemblages of fungi associated with roots of cooccurring Epacris pulchella ( Ericaceae ) and Leptospermum polygalifolium ( Myrtaceae ) seedlings at a sclerophyll forest site in New South Wales, Australia, were investigated by direct DNA extraction and analysis of rRNA gene internal transcribed spacer (ITS) products by denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) analyses. While ordination of the DGGE data suggested that the assemblages did not differ significantly between the two plant taxa, T-RFLP data provided marginal statistical support for the presence of different assemblages. Fungi isolated from roots of both plants were identified by ITS sequence comparisons largely as ascomycetes, several of which had close sequence identity to Helotiales ericoid mycorrhizal (ERM) fungi. One isolate morphotype from E. pulchella had close sequence similarity to ectomycorrhizal fungi in the Cenococcum geophilum complex, and neighbour-joining analysis grouped this strongly with other Australian C. geophilum- like sequences. Distribution of genotypes of an ERM Helotiales ascomycete in root systems of the two plant taxa was also investigated using inter-simple sequence repeat (ISSR)-PCR. Nineteen ISSR genotypes were identified, two of which were present in roots of both plant taxa. The results are discussed in the context of potential mycelial connections between Ericaceae and non- Ericaceae plants.     

The cultivable endophytic community of Norway spruce ectomycorrhizas from microhabitats lacking ericaceous hosts is dominated by ericoid mycorrhizal Meliniomyces variabilis

Most of the temperate conifers associate with ectomycorrhizal fungi, but their roots also harbour a wide range of endophytes. We focused on ascomycetes associating with basidiomycetous ectomycorrhizas of Norway spruce in a temperate montane forest in central Europe and found that the majority of the co-associated fungi belonged to the Rhizoscyphus ericae aggregate (REA), being dominated by Meliniomyces variabilis. We further tested the ability of representative isolates to colonize spruce root tips and European blueberry (Vaccinium myrtillus) hair roots in an agar system as well as their effect on blueberry growth in a peat-agar system. M. variabilis intracellularly colonized spruce (Picea abies) root tip cortex, formed ericoid mycorrhizas in blueberry and enhanced blueberry shoot and root growth in comparison with non-inoculated plants. Our findings suggest that spruce ectomycorrhizas may represent selective niches for ericoid mycorrhizal fungi in habitats lacking suitable ericaceous hosts.     

海南霸王岭热带雨林常见植物丛枝菌根真菌调查

对海南霸王岭热带雨林的12科16种常见植物的丛枝菌根状况进行了调查,用碱解离-酸性品红染色法进行了真菌鉴定。结果表明,13种植物形成典型的丛枝菌根,占所调查植物的81%;3种植物没有形成丛枝菌根,占所调查植物的19%。用湿筛沉淀法从这些植物根际土壤中共分离鉴定出了3属11种丛枝菌根真菌(AMF),即无梗囊霉属(Acaulos-pora)3种,球囊霉属(Glomus)7种,巨孢囊霉属(Gigaspora)1种;其中,球囊霉属是样地的优势属。在AMF中,孔窝无梗囊霉(A.foveata)分离频率最高,在14种植物的根际土中都有发现;此外,大果球囊霉(G.macrocarpum)的相对多度最大,为59%,具有最强的产孢能力。同时,在11种植物的根中发现了深色有隔内生真菌(DSE),占调查植物的69%;其中,11种植物同时被DSE和AMF感染。     

宁夏枸杞深色有隔内生真菌的侵染特征及多样性

【背景】枸杞是享誉中外的名贵药材,深色有隔内生真菌(Dark septate endophytes,DSE)是枸杞内生真菌的重要组成部分。【目的】从宁夏枸杞栽培品种和野生品种的根系分离获得DSE菌株,研究枸杞DSE的群落组成、物种多样性及在宿主植物内的侵染定殖情况。【方法】从宁夏枸杞栽培园采集栽培品种宁杞1号、宁杞3号、宁杞5号、宁杞6号、宁杞7号、宁杞8号以及野生品种黄果枸杞和黑果枸杞共8个枸杞品种的根系,分离DSE菌株,运用形态学特征、r DNA-ITS序列分析进行菌株鉴定,采用回接试验方法确定DSE真菌。【结果】DSE在枸杞根系能形成大量"微菌核"典型结构。从8个枸杞品种根系中共分离获得DSE菌株279株,分属于18个属,具有丰富的物种多样性。镰刀菌属(Fusarium)为各品种的共有属和优势属,相对频率最高达85%。Monosporascus、蓝状菌属(Talaromyces)和俄氏孔菌属(Earliella)为枸杞内首次报道的DSE。枸杞不同品种中DSE群落物种多样性指数、均匀度指数和Simpson指数差异显著。【结论】DSE在枸杞栽培品种及野生品种中具有丰富的生物多样性,能够与枸杞根系形成良好的共生关系,增强了枸杞对生态环境的适应性。     

Diversity and antimicrobial activity of endophytic fungi isolated from Nyctanthes arbor-tristis, a well-known medicinal plant of India

Endophytic fungi from Nyctanthes arbor-tristis were isolated and evaluated for their antimicrobial activity. A total of 19 endophytic fungi were isolated from 400 segments of healthy leaf and stem tissues of N. arbor-tristis. Eighteen endophytic fungi were obtained from leaf, while only ten from stem. Alternaria alternata had the highest colonization frequency (15.0%) in leaf, whereas Cladosporium cladosporioides ranked first in stem with a colonization frequency of 12%. The diversity and species richness were found higher in leaf tissues than in stem. The similarity indices between leaf and stem were 0.473 for Jaccard’s and 0.642 for the Sorenson index, respectively. Of 16, 12 (75%) endophytic fungal extracts showed antibacterial activity against either one or more pathogenic bacteria. The endophytic Nigrospora oryzae showed maximum inhibition against Shigella sp. and Pseudomonas aeruginosa. The leaf endophytes Colletotrichum dematium and Chaetomium globosum exhibited a broad range of anibacterial activity and were active against Shigella flexnii, Shigella boydii, Salmonella enteritidis, Salmonella paratyphi, and P. aeruginosa. Nine out of 16 (56.25%) endophytic fungi exhibited antifungal activity to one or more fungal pathogens. Colletotrichum dematium inhibited 55.87% of the radial growth of the phytopathogen Curvularia lunata. The antimicrobial activity of these endophytic microorganisms could be exploited in the biotechnological, medicinal, and agricultural industries.     

Red list plants: colonization by arbuscular mycorrhizal fungi and dark septate endophytes

Since information concerning the mycorrhization of endangered plants is of major importance for their potential re-establishment, we determined the mycorrhizal status of Serratula tinctoria (Asteraceae), Betonica officinalis (Lamiaceae), Drosera intermedia (Droseraceae) and Lycopodiella inundata (Lycopodiaceae), occurring at one of two wetland sites (fen meadow and peat bog), which differed in soil pH and available P levels. Root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) was quantified. Colonization by AMF appeared to be more frequent in the fen meadow than in the peat bog, and depended on the host plant. Roots of S. tinctoria and B. officinalis were well colonized by AMF in the fen meadow (35–55% root length) and both arbuscules and vesicles were observed to occur in spring as well as in autumn. In the peat bog, L. inundata showed a low level of root colonization in spring, when vesicles were found frequently but no arbuscules. In roots of D. intermedia from the peat bog, arbuscules and vesicles were observed, but AMF colonization was lower than in L. inundata. In contrast, the amount of AMF spores extracted from soil at the peat bog site was higher than from the fen meadow soil. Spore numbers did not differ between spring and autumn in the fen meadow, but they were higher in spring than in autumn in the peat bog. Acaulospora laevis or A. colossica and Glomus etunicatum were identified amongst the AMF spores extracted from soil at the two sites. S. tinctoria and B. officinalis roots were also regularly colonized by DSE (18–40% root length), while L. inundata was only rarely colonized and D. intermedia did not seem to be colonized by DSE at all.     

金沙江支流普渡河、小江干热河谷的丛枝菌根

调查具有不同植物群落的金沙江支流普渡河、小江干热河谷中91种常见植物的丛枝菌根真菌的侵染率及孢子密度。普渡河样地调查了56种植物,其中54种(96%)植物能形成典型的丛枝菌根,其平均孢子密度为1423±175/100g土;小江样地35种植物中有34种(97%)植物能形成典型的丛枝菌根,其平均孢子密度为601±103/100g土。单因素方差分析表明两个样地植物的AMF总感染率差异不显著,但其根际土壤中AMF孢子密度却存在显著差异,小江样地的AMF孢子密度明显低于普渡河样地。相关性分析表明,干热河谷植物的AMF感染率与其根际土壤中的AMF孢子密度之间不存在相关性。此外,调查还发现91种植物中,有61种植物(67%)在形成AM的同时,也被黑色有隔内生菌感染。     

Species composition and colonization of dark septate endophytes are affected by host plant species and soil depth in the Mu Us sandland,northwest China

Investigations into the edaphic associations, host affiliations and soil depth of dark septate endophytes (DSE) in arid desert environments can help explain their spatial distribution and the response mechanisms in desert ecosystems. Soils were sampled to a depth of 50 cm in the rhizospheres of Psammochloa villosa, Hedysarum laeve and Artemisia ordosica in the Mu Us sandland of northwest China in July 2015. The plant species and soil depth significantly influenced the distribution and colonization of DSE. Hyphal and total root colonization were significantly higher under P. villosa than the others in the 0–20 cm layer. The maximum colonization of P. villosa and H. laeve occurred in the 10–20 cm and 20–30 cm soil layers, respectively, while 30–40 cm soil layer under A. ordosica. Of twelve DSE species isolated from the roots of these plants, Phoma radicina and Bipolaris zeae were reported in desert ecosystems for the first time. Hyphal colonization was significantly and positively correlated with soil total nitrogen (TN) and significantly and negatively correlated with the soil carbon/nitrogen (C/N) ratio. Microsclerotial colonization was significantly and positively correlated with soil organic carbon (SOC), and total colonization was significantly and positively correlated with soil TN and total phosphorus (TP) and significantly and negatively correlated with soil C/N. Variation of DSE colonization was mostly attributed to effects of plant species. We concluded that the species composition and colonization of the DSE fungi were influenced by the plant species, soil depth and soil nutrient availability in this desert ecosystem. This research provides a basis for further understanding the ecological adaptability of DSE and their roles in promoting vegetation restoration and reducing desertification in arid ecosystems.     

Diversity of endophytic enterobacteria associated with different host plants

Fifty-three endophytic enterobacteria isolates from citrus, cocoa, eucalyptus, soybean, and sugar cane were evaluated for susceptibility to the antibiotics ampicillin and kanamycin, and cellulase production. Susceptibility was found on both tested antibiotics. However, in the case of ampicillin susceptibility changed according to the host plant, while all isolates were susceptible to kanamycin. Cellulase production also changed according to host plants. The diversity of these isolates was estimated by employing BOX-PCR genomic fingerprints and 16S rDNA sequencing. In total, twenty-three distinct operational taxonomic units (OTUs) were identified by employing a criterion of 60% fingerprint similarity as a surrogate for an OTU. The 23 OTUs belong to the Pantoea and Enterobacter genera, while their high diversity could be an indication of paraphyletic classification. Isolates representing nine different OTUs belong to Pantoea agglomerans, P. ananatis, P. stewartii, Enterobacter sp., and E. homaechei. The results of this study suggest that plant species may select endophytic bacterial genotypes. It has also become apparent that a review of the Pantoea/Enterobacter genera may be necessary.     

Wetland dicots and monocots differ in colonization by arbuscular mycorrhizal fungi and dark septate endophytes

As an initial step towards evaluating whether mycorrhizas influence composition and diversity in calcareous fen plant communities, we surveyed root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytic fungi (DSE) in 67 plant species in three different fens in central New York State (USA). We found colonization by AMF and DSE in most plant species at all three sites, with the type and extent of colonization differing between monocots and dicots. On average, AMF colonization was higher in dicots (58±3%, mean±SE) than in monocots (13±4%) but DSE colonization followed the opposite trend (24±3% in monocots and 9±1% in dicots). In sedges and cattails, two monocot families that are often abundant in fens and other wetlands, AMF colonization was usually very low (<10%) in five species and completely absent in seven others. However, DSE colonization in these species was frequently observed. Responses of wetland plants to AMF and DSE are poorly understood, but in the fen communities surveyed, dicots appear to be in a better position to respond to AMF than many of these more abundant monocots (e.g., sedges and cattails). In contrast, these monocots may be more likely to respond to DSE. Future work directed towards understanding the response of these wetland plants to AMF and DSE should provide insight into the roles these fungal symbionts play in influencing diversity in fen plant communities.     

Community composition of root‐associated fungi in a Quercus‐dominated temperate forest: “codominance” of mycorrhizal and root‐endophytic fungi

In terrestrial ecosystems, plant roots are colonized by various clades of mycorrhizal and endophytic fungi. Focused on the root systems of an oak‐dominated temperate forest in Japan, we used 454 pyrosequencing to explore how phylogenetically diverse fungi constitute an ecological community of multiple ecotypes. In total, 345 operational taxonomic units (OTUs) of fungi were found from 159 terminal‐root samples from 12 plant species occurring in the forest. Due to the dominance of an oak species (Quercus serrata), diverse ectomycorrhizal clades such as Russula, Lactarius, Cortinarius, Tomentella, Amanita, Boletus, and Cenococcum were observed. Unexpectedly, the root‐associated fungal community was dominated by root‐endophytic ascomycetes in Helotiales, Chaetothyriales, and Rhytismatales. Overall, 55.3% of root samples were colonized by both the commonly observed ascomycetes and ectomycorrhizal fungi; 75.0% of the root samples of the dominant Q. serrata were so cocolonized. Overall, this study revealed that root‐associated fungal communities of oak‐dominated temperate forests were dominated not only by ectomycorrhizal fungi but also by diverse root endophytes and that potential ecological interactions between the two ecotypes may be important to understand the complex assembly processes of belowground fungal communities.     

Occurrence and coexistence of arbuscular mycorrhizal fungi and dark septate fungi in aquatic macrophytes in a tropical river-floodplain system

In this study, carried out in four water bodies in the Upper Paraná River floodplain, we assessed the occurrence of root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate fungi (DSF), as well as the AMF species richness associated with 24 species of aquatic macrophytes belonging to different life forms. AMF were found in nine species of macrophytes and DSF in 16 species among the 24 investigated. AM colonization occurred mainly in eudicotiledons (five of the six species evaluated) and the Paris morphology was the most common type. Co-occurrence of AMF and DSF was observed in seven species of macrophytes (Commelinaceae sp. 1, Limnobium laevigatum (H.B.K. ex Willd) Heine, Hygrophila cf. costata, Myriophyllum brasiliense (Camb), Polygonum acuminatum Kunth, P. ferrugineum Wedd and P. stelligerum Cham). Four species of macrophytes (Pistia stratiotes L., Eichhornia crassipes (Mart.) Solms, Egeria najas Planch and Nymphaea amazonum Mart. & Zucc) were not colonized by any type of fungi. In total, 27 morphotypes of AMF were recorded, and spores occurred both in the rhizosphere of macrophytes whose roots were internally colonized by AMF and in non-colonized macrophytes. Acaulospora delicata, Acaulospora aff. laevis, Acaulospora longula, Glomus lamellosum, Glomus luteum and NID 1 (a non-identified species) were the most frequent species. Samples collected close to the roots of N. amazonum had the highest AMF richness (20 species), but this plant was not colonized by fungi. A species richness curve indicated that more root-associated fungi than reported here are likely present in this floodplain.     

Strategies of carbon and nitrogen acquisition by saprotrophic and ectomycorrhizal fungi in Finnish boreal Picea abies-dominated forests

We compared the δ¹³C and δ¹⁵N of forest material with an extensive sporocarp collection to elucidate the role of litter, wood and soil as fungal carbon and nitrogen sources in Finnish boreal Picea abies-dominated forests. Ectomycorrhizal Hydnum and Cortinarius had higher δ¹⁵N than other ectomycorrhizal fungi, suggesting use of ¹⁵N-enriched, deeper nitrogen. Russula had lower δ¹⁵N than other ectomycorrhizal fungi and resembled some litter decay genera, suggesting use of litter-derived nitrogen. There was little variation in δ¹⁵N among other genera of ectomycorrhizal fungi, indicating limited functional diversity in nitrogen use. Saprotrophic Leotia, Gymnopus, Hypholoma, Pholiota, Rhodocollybia and Calocera had δ¹⁵N values similar to ectomycorrhizal fungi, indicating overlap in use of older nitrogen from soil or roots or use of newly fixed nitrogen. Genera of litter and wood decay fungi varied up to 6‰ in δ¹³C and 10‰ in δ¹⁵N, suggesting large differences in carbon and nitrogen sources and processing. Similar δ¹³C between white and brown rot wood decay fungi also suggest that white rot fungi do not use lignin-derived carbon. Together, these δ¹³C and δ¹⁵N patterns of fungi from Finnish boreal forests enhance our knowledge of fungal functional diversity and indicate broad use of litter, wood and soil resources.     

Spatial dynamics of dark septate endophytes in the roots and rhizospheres of Hedysarum scoparium in northwest China and the influence of edaphic variables

Spatial dynamics of DSE in the roots and rhizospheres of Hedysarum scoparium Fisch. et Mey. and soil factors were investigated at seven arid and semi-arid locations in northwest China in July 2015. Sampling sites were found to have a significant influence on the morphology, components, distribution, and infection of DSE in the roots of H. scoparium. Of nine DSE species isolated from H. scoparium roots, five are reported here for the first time from desert ecosystems. Hyphal infection in roots was positively correlated with soil urease and phosphatase activity. The presence of microsclerotia in roots was positively correlated with soil ammonium, and negatively correlated with soil organic matter and pH. We conclude that DSE infection is spatially predictable, and is influenced by nutrient availability and enzymatic activity. This research provides a basis for further understanding the ecological functions of DSE, and their roles in the promotion of vegetation restoration and in reducing erosion and desertification in arid ecosystems.     

Richness of cultivable endophytic fungi along an altitudinal gradient in wet forests of Costa Rica

We collected various plant species along an altitudinal gradient ranging from 400 to 2900 m in tropical wet forests of Costa Rica, isolated the associated endophytic fungi, and performed bioinformatic analyses to determine whether changes in altitude are related to changes in their richness and community structure. We showed that the richness of endophytic fungi varied along the altitudinal gradient, being higher in the lowest stratum and decreasing as elevation increases. Each stratum presented a particular composition and diversity of endophytes, although the whole population was characterized by the presence of a few dominant and apparently ubiquitous species, coexisting with a number of less abundant species that presented a more limited host range. These results have important implications for better understanding the role of altitude on the distribution and composition of endophytic fungal populations in tropical forests, but also for maximizing the number and diversity of endophytic isolates in bioprospecting campaigns.