Soil-environment interaction with arbuscular mycorrhizal fungi in three cold desert plants of Himalaya, India

SummaryThe mycorrhizal symbiosis formed between plant roots and the arbuscular mycorrhizal (AM) fungi or Glomales is of great interest to ecologists because of its potential influence on ecosystem processes, its role in determining plant diversity in natural communities and the ability of the fungi to induce a wide variety of growth responses in coexisting plant species. Little attention, however, has been paid to the ecological role of diversity of AM fungi. Difficulties in identification, the inability to grow the fungi in pure culture, problems of taxonomic classification, and a lack of basic information on the life histories of AM fungi hinder studies of the ecological significance of diversity of AM fungi. Nucleic acid based techniques have the potential to fill this gap in our knowledge by offering better means of identification and the opportunity to study links between the genetic diversity of AM fungi and functional and morphological diversity. The application of genus‐specific molecular markers has shown that different genera of AM fungi coexist in plant roots and that this is a common occurrence. Molecular techniques have also shown that natural AM fungal populations exhibit unexpectedly high genetic diversity, despite the assumption that diversity in these seemingly asexual fungi should be low. The high diversity occurs in multicopy ribosomal genes and their internal transcribed spacers, which are normally well conserved and homogeneous within an individual organism. The results show that sequence heterogeneity of the ribosomal genes can occur even in single spores of AM fungi, and we discuss how genetic diversity may be promoted and maintained. Contrasting results, indicating that genetic diversity among replicate spores from pot‐cultured material is low (even though they contain within spore sequence heterogeneity), suggest that there are mechanisms which promote high genetic diversity of AM fungi in natural ecosystems.We propose that AM fungi could be heterokaryotic as a result of the exchange of nuclei following hyphal fusion with other individuals but that other mechanisms, such as gene turnover and molecular drive, might also explain the generation of high genetic diversity without any exchange of genetic material among individuals. The high diversity in ribosomal gene sequences in AM fungi might cause problems in their use as molecular markers in field studies. A better understanding of the levels of genetic diversity of ribosomal genes within spores, among spores of the same morphology, and among spores of differing morphology is essential to the development of sound molecular markers for field studies and to the development of a phylogenetic classification.We conclude that an understanding of the mechanisms which promote and maintain genetic diversity in the AM fungi is crucial, not only to further advances in ecological and evolutionary studies but also to studies of the molecular basis of the regulation of the symbiosis. Moreover, we predict that while observational investigations on AM fungal ecology and diversity using molecular techniques are of high value they will not give an understanding of the role of AM fungi in natural ecosystems and that further studies should also aim to fill the gaps in current knowledge of links between genetic diversity and distribution of AM fungi in natural ecosystems, and their functional diversity.

James Hutton Institute, Dundee DD2 5DA, UKBelowground organisms, such as arbuscular mycorrhizal(AM) fungi, have long been credited with altering plant fit-ness. More recently, research on belowground organismshas revealed that AM fungi also influence a wide variety ofaboveground organisms via plants (reviewed in Van Dam H Bennett 2010). Schausberger et al. (2012) demon-strate that the presence of an AM fungus in the roots of ahost plant alters volatile emissions and host plant attractive-ness to parasitoids in the presence of herbivores. Thisextends previous studies that have focused on direct inter-actions of AM with plants (e.g. mycorrhizal fungal–plant–herbivore interactions; reviewed in Gehring & Bennett2009), but have not conclusively demonstrated how below-ground organisms, and AM fungi in particular, influencethird trophic level organisms such as parasitoids (Gange,Brown & Aplin 2003; Guerrieri et al. 2004; Hempel et al.2009; Leitner et al. 2010; Hoffmann, Vierheilig & Schaus-berger 2011a,b; Wooley & Paine 2011) via the release ofplant volatiles that attract parasitoids that attack herbivoreson host plants. Until recently, these studies failed to conclu-sively document the effects of AM fungi on both volatilerelease and attraction of parasitoids. For example, Wooley& Paine (2011) and Gange, Brown & Aplin (2003) haveshown variation in parasitoid attraction to plants hostingdifferent strains and species of Glomus as compared to non-mycorrhizal plants. Hoffmann, Vierheilig & Schausberger(2011a) also showed greater preference by parasitoids foreggs oviposited on plants associated with a single AMfungus. In addition, a single AM fungus in the roots of ahost plant has been shown to positively influence parasitoidlife-history characteristics (Hempel et al. 2009; Hoffmann,Vierheilig & Schausberger 2011b). However, none of thesestudies measured volatile profiles for host plants, so parasit-oid attraction could not be directly attributed to volatiles.A study on AM fungal influenced volatile release revealeddifferences but did not test whether changes in volatilesinfluenced parasitoids (Leitner et al. 2010). One study com-bined both parasitoid attractiveness and measurement ofvolatiles, but they primarily tested effects of attraction toplants in the absence of herbivory and never made compari-sons between mycorrhizal and non-mycorrhizal plantsexperiencing herbivory (Guerrieri et al. 2004). Unlike theseprevious experiments, Schausberger et al. measured bothchanges in volatile chemistry as well as parasitoid attractionin a fully factorial design.The results presented by Shausberger et al. open up multi-plefutureopportunitiesinabove–belowgroundresearch.Thefirst of these opportunities involves identifying the mecha-nisms by which AM fungi alter parasitoid attraction. Forexample,whatarethe biochemicalortranscriptionalchangesthat occur following AM fungal colonization that result inaltered volatile profiles? Are the mechanisms suggested forAM fungal alteration of direct chemical defences the samemechanisms that alter volatile profiles? Colonization by AMfungi has been shown to turn on the salicylic acid pathwaytemporarily,aprocessthatmayprimethejasmonicacidpath-way for herbivore attack (reviewed in Pozo & Azcon-Aguilar2007).Theinductionofvolatilesislinkedtothejasmonicacidpathway (reviewed in Heil 2008), and therefore, plants maybeprimedforafasterorgreaterreleaseofvolatileswhencolo-nizedbyAMfungi.However, there may be other mechanisms by which AMfungi influence volatile release. For example, given that AMfungi increase plant biomass and fitness in the PhaseolusvulgarissystemstudiedbyShausbergeret al.(aswellasmanyothersystems),itcouldsimplybethattheincreasedresourcesprovided by the mutualism allow plants to allocate moreresourcestoplantdefensivecharacteristics(e.g.directconstit-utiveandinduceddefencesaswellasindirectdefencesviavol-atile attraction; Bennett, Alers-Garcia & Bever 2006) or thatchanges in plant size or structure in association with AMfungi benefit or hinder parasitoid searching capabilities(Gange,Brown&Aplin2003).What characteristics of the volatile blends produced in thepresence of AM fungi are attractive for parasitoids? Shaus-bergeret al.showedtherewerefewerchemicalspresentinthevolatileblendsofAMfungalplantsbeforeherbivory(relativeto plants not hosting AM fungi), but this difference disap-peared after herbivory. However, different volatile chemicalswere released from plants experiencing herbivory and colo-nizedornotbyAMfungi(seealsoLeitneret al.2010).Shaus-berger et al. did not address whether increased attraction toplants hosting AM fungi is associated with a particular vola-

The aim of this work was to evaluate arbuscular mycorrhizal (AM) fungi as soil indicators and the mycorrhization of native and exotic tree species planted in the Acarau basin, a transition area from the coast to the Brazilian semiarid region. Plots with 6-year-old trees of four native and three non-native species as well as one non-forested area were evaluated in terms of the diversity of AM fungi in the mycorrhizosphere and the root colonization by AM and ectomycorrhizal (EcM) fungi. Twenty-four AM fungi were identified; Claroideoglomus etunicatum, Glomus sinuosum, Paraglomus albidum, Acaulospora laevis, and Acaulospora brasiliensis were abundant in the forest soil. Diversity, dominance, evenness and richness indices of AM fungi were higher in plots with native trees. All root samples were colonized by AM fungi and only Anadenanthera colubrina, Acacia mangium, Casuarina equisetifolia and Eucalyptus urophylla formed associations with EcM fungi. Acaulospora morphotypes served as soil indicators for coverings with the native species Astronium fraxinifolium and Colubrina glandulosa. Exotic species may favor the proliferation of rarer AM fungi. These fungi–plant relationships may be important in the management of forest systems, and the evidence with mycorrhizal associations allows the inclusion of Brazilian species in tropical reforestation.

The association between terrestrial plants and arbuscular mycorrhizal (AM) fungi is one of the most widespread mutualistic plant-fungus interactions in natural and cropping systems. We studied the effect of different plant densities (70,000 and 100,000 plant ha-1) on species diversity and community structure of AM fungi associated with maize (Zea mays L.) in a long-term crop production experiment established in Martonvásár, Hungary. Based on the differences in small subunit (SSU)/18S ribosomal genes nested-PCR procedure was used to identify groups of AM fungi that are active in the colonization of maize roots. Shannon-Wiener diversity index of AM fungi were 1.43 ± 0.37 and 1.31 ± 0.50 at 70,000 plant ha-1 and 100,000 plant ha-1 respectively. All of sequence types we found belonging to the Glomus clade. Besides Glomus A fungi, only the members of the Glomus B group occurred, however at significantly lower frequency. There were differences in the phylogenetic group composition of AM fungi demonstrating the effect of different plant densities on the diversity of arbuscular mycorrhizal fungi.

Recent studies have shown that continuous cropping in soybean causes substantial changes to the microbial community in rhizosphere soil. In this study, we investigated the effects of continuous cropping for various time periods on the diversity of rhizosphere soil arbuscular mycorrhizal (AM) fungi in various soybean cultivars at the branching stage. The soybean cultivars Heinong 37 (an intermediate cultivar), Heinong 44 (a high-fat cultivar) and Heinong 48 (a high-protein cultivar) were seeded in a field and continuously cropped for two or three years. We analyzed the diversity of rhizosphere soil AM fungi of these soybean plants at the branching stage using morphological and denaturing gradient gel electrophoresis (DGGE) techniques. The clustering analysis of unweighted pair-group method with arithmetic averages (UPGMA) was then used to investigate the AM fungal community shifts. The results showed that increasing the number of years of continuous cropping can improve the colonization rate of AM fungi in different soybean cultivars at the branching stage. The dominant AM fungi in the experimental fields were Funneliformismosseae and Glomus spp. The number of years of continuous cropping and the soybean cultivar both had obvious effects on the diversity of AM fungi, which was consistent with the results of colonization rate analysis. This study establishes a basis for screening dominant AM fungi of soybean. In addition, the results of this study may be useful for the development of AM fungal inoculants.

Arbuscular mycorrhizal fungi (AMF) have the potential to increase crop productivity and play a key role in the functioning and sustainability of most agroecosystems. However, limited information is available on the divervisity of AMF associated with upland rice varieties in Southwest Nigeria. Field survey was conducted to investigate colonization and diversity of AMF in 13 upland rice varieties commonly grown in Southwest Nigeria. Root and soil samples were collected from rice fields in 2012. The results showed natural root colonization of all the rice varieties by AMF with highest root colonization in ITA 157and Ofada. The spore densities retrieved from the different rhizospheres were relatively high, varying from 13 spores in UORW 111 to 174 spores in Ofada with a mean of 67.6 spores per 20 g dry soil. Glomus was observed to be the most abundant AMF genus. Funneliformis mosseae was the most frequently occurring AMF species (96.2%) with relative density (RD) of 32.2%, followed by Glomus intraradices, Claroideoglomus etunicatum, and Glomus clareium. This study showed that AMF naturally colonized the roots of these rice varieties and diversity of different AMF genera in rice rhizosphere. This study will help draw attention to natural colonization of AMF in rice producing areas of Nigeria that can influence future possibility of using inocula of the dominant AMF species in upland rice cultivation.

The diversity of arbuscular mycorrhizal (AM) fungi in deforested (Mantoushan) and natural forest (Banruosi) land in the subtropical region of Dujiangyan was surveyed and compared. A total of 44 taxa of AM fungi were isolated, and the same number of AM fungus taxa (34 taxa) was found in both deforested and natural forest land. Acaulospora and Glomus were the dominant genera in the two sites. Glomus convolutum and G. versiforme were the dominant species in the natural forest land, while only G. versiforme was dominant in the deforested land. There was no significant difference in total spore density of AM fungi between the two sites, but the total species richness of AM fungi was significantly higher in the deforested land than in the natural forest land. The Shannon-Weiner index of AM fungus diversity was a higher in the natural forest land (2.67) than in the deforested land (2.15). There was high AM fungus composition similarity (Sorenson's coefficient CS=0.71) between the two sites. We suggest that there was little effect of deforestation on the diversity of AM fungi, and that annual herbaceous plants play a major role in maintaining and increasing AM fungus spore density and species richness in deforested land.

To understand the effects of agricultural management activities on soil arbuscular mycorrhizal (AM) fungi diversity, the high-throughput sequencing based on Illumina MiSeq platform, and the fatty acids fingerprints were used to examine the effects of maize straw returning on soil arbuscular mycorrhizal fungi. The relationships between AM fungal community composition, AM fungal biomass and soil factors after maize straw returning were examined for four continuous years. A total of 2430 operational taxonomic units (OTUs) of AM fungi were classified into 10 genera and 143 species, respectively, which belonged to 1 phylum, 3 classes, 4 orders, 8 families. There was no significant difference in AM fungal community richness (Chaoles index and ACE index) and diversity (Shannon, Simpson diversity indices) in different treatments. Paraglomus and Glomus were dominant genera among all AM fungal communities. With the increase of the maize straw returned amounts, the abundance of Glomus reduced. Under the treatments of 3000 and 9000 kg·hm-2 straw returned, the abundance of Glomus and Acaulospora had significant differences with the control (0 kg·hm-2). Compared with the control, there were significant differences between Archaeospora, Paraglomus and Glomus in the treatment of 3000 kg·hm-2 straw returned. Results from non-metric multi-dimensional scale (NMDS) analysis showed that under 9000 and 12000 kg·hm-2 straw returning treatments, the difference between the β diversity of soil AM fungi and the spatial distance of controls was farther apart than the other treatments. The effect of straw returning on the β diversity of AM fungi was significant. The multivariate analysis results revealed the relationship of the spatial variation between the soil physicochemical properties and AM fungi richness and diversity could be explained at 82.8% cumulative variables. The total nitrogen and available nitrogen were the most important factors driving soil microbial communities biomass marked by PLFAs and AM fungal biomass (NLFAs). The continuous maize straw returning to the field changed the genera composition of AM fungi. With the increases of straw returning amounts, the specific species of AM fungi decreased and the similarity between AM fungi community composition decreased. Straw returning increased soil AM fungi biomass and its contribution to soil total microbial biomass.

Arbuscular mycorrhizal (AM) fungi are crucial for ecosystem functioning and can contribute to the formation and maintenance of soil aggregates through the exudation of glomalin by extraradical hyphae. Monitoring fertilization effects on AM fungi may help us to develop sound management strategies. The objectives of this study were to investigate the impacts of long-term fertilization on AM fungal parameters and to find out the key factor that affects the diversity and function of AM fungi. A long-term fertilization experiment established in a sandy loam soil at northern China has received continuous fertilization treatments for 21 years, including control; mineral fertilizers of NK, PK, NP, and NPK; organic manure (OM); and half organic manure N plus half mineral fertilizer N (1/2 OMN). Top soil samples (0–15 cm) from three individual plots per treatment were collected for the analysis of chemical properties and fungal parameters. The population size of soil AM fungi was determined by real-time PCR, and the community composition was analyzed using PCR-denature gradient gel electrophoresis (DGGE), cloning, and sequencing techniques. The external mycelium of AM fungi was assessed using the grid-line intersect method, and the glomalin-related soil protein (GRSP) was extracted with citrate solution using bovine serum albumin as a standard. Long-term fertilization significantly increased (P < 0.05) soil organic C content, AM fungal population, species richness (R), Shannon–Wiener index (H), and GRSP content, except for the P-deficiency (NK) fertilization treatment. OM had a significantly greater (P < 0.05) impact on AM fungal population and GRSP content compared to mineral fertilizers but significantly decreased the length of external mycelium compared to the control (P < 0.05). Fertilization also changed the community composition of AM fungi, and the P-deficiency treatment again had the slightest influence. In addition, most species recovered from the DGGE profiles belonged to three genera, Glomus, Diversispora, and Archaeospora. Redundancy analysis showed that the population size and species richness of AM fungi and the GRSP content all significantly correlated to soil organic C content (P < 0.05). Long-term P-containing fertilization, especially the application of OM, greatly increased the population size, species richness, and species diversity of AM fungi, as well as the contents of GRSP and soil organic C, but tended to decrease the length of external mycelium, while the P-deficiency fertilization had no such effect, suggesting that P was the key factor to maintain soil fertility as well as soil AM fungal diversity in this sandy loam soil.

厦门市七种药用植物根围AM真菌的侵染率和多样性

PurposeGrapevine (Vitis vinifera L.) is a relevant crop, which is associated to arbuscular mycorrhizal fungi (AMF) that are influenced by agricultural practices. The hypothesis of this study is that organic/biodynamic management stimulates grapevine mycorrhizal colonisation and increases AMF diversity in Chilean vineyards. The aim of this study was to determine the influence of agricultural management on AMF association and AMF diversity in Chilean vineyards.MethodsMycorrhizal colonisation of grapevine roots from organic/biodynamic and conventional vineyards in Northern (Elqui Valley), Central (Casablanca and Cachapoal Valleys), and Southern Chile (Maule and Itata Valleys), was determined under a microscope. AMF diversity was analysed by morphological, and molecular characterisation of spores through SSU-ITS-LSU rRNA region sequence analyses.ResultsAMF colonisation of grapevine roots was influenced by vineyard management independent of the season. Higher mycorrhizal colonisation was detected in organic/biodynamic grapevine soils (20 − 35%), compared with conventional soils (6 − 31%). Twelve AMF species were identified in vineyards, belonging to five Glomeromycota families. Interestingly, organic/biodynamic vineyards showed higher AMF diversity. The three predominant morphotypes were Funneliformis verruculosum (GL1), Septoglomus sp. (GL4) and Septoglomus constrictum (GL5). Molecular analyses of AMF spores highlighted the occurrence of Septoglomus, Acaulospora, Pacispora and Cetraspora genera in vineyards.ConclusionsIn this study, AMF diversity in Chilean vineyards is described for the first time. The diversity of AMF in vineyards in Chile was higher than the diversity reported in other wine-producing ecosystems. The understanding of agricultural practices on AMF activity and diversity may be crucial to improve the vineyard management.

Knowledge about the distribution and local diversity patterns of arbuscular mycorrhizal (AM) fungi are limited for extreme environments such as the Arctic, where most studies have focused on spore morphology or root colonization. We here studied the joint effects of plant species identity and elevation on AM fungal distribution and diversity. We sampled roots of 19 plant species in 18 locations in Northeast Greenland, using next generation sequencing to identify AM fungi. We studied the joint effect of plant species, elevation and selected abiotic conditions on AM fungal presence, richness and composition. We identified 29 AM fungal virtual taxa (VT), of which six represent putatively new VT. Arbuscular mycorrhizal fungal presence increased with elevation, and as vegetation cover and the active soil layer decreased. Arbuscular mycorrhizal fungal composition was shaped jointly by elevation and plant species identity. We demonstrate that the Arctic harbours a relatively species-rich and nonrandomly distributed diversity of AM fungi. Given the high diversity and general lack of knowledge exposed herein, we encourage further research into the diversity, drivers and functional role of AM fungi in the Arctic. Such insight is urgently needed for an area with some of the globally highest rates of climate change.

BackgroundUnderstanding diversity of arbuscular mycorrhizal fungi (AMF) is important for optimizing their role for phosphorus (P) nutrition of soybeans (Glycine max (L.) Merr.) in P-limited soils. However, it is not clear how soybean growth and P nutrition is related to AMF colonization and diversity of AMF communities in a continuous P-unfertilized cover cropping system. Thus, we investigated the impact of P-application and cover cropping on the interaction among AMF colonization, AMF diversity in soybean roots, soybean growth and P nutrition under a five-year P-unfertilized crop rotation.MethodsIn this study, we established three cover crop systems (wheat, red clover and oilseed rape) or bare fallow in rotation with soybean. The P-application rates before the seeding of soybeans were 52.5 and 157.5 kg ha−1 in 2014 and 2015, respectively. We measured AMF colonization in soybean roots, soybean growth parameters such as aboveground plant biomass, P uptake at the flowering stage and grain yields at the maturity stage in both years. AMF community structure in soybean roots was characterized by specific amplification of small subunit rDNA.ResultsThe increase in the root colonization at the flowering stage was small as a result of P-application. Cover cropping did not affect the aboveground biomass and P uptake of soybean in both years, but the P-application had positive effects on the soybean performance such as plant P uptake, biomass and grain yield in 2015. AMF communities colonizing soybean roots were also significantly influenced by P-application throughout the two years. Moreover, the diversity of AMF communities in roots was significantly influenced by P-application and cover cropping in both years, and was positively correlated with the soybean biomass, P uptake and grain yield throughout the two years.DiscussionOur results indicated that P-application rather than cover cropping may be a key factor for improving soybean growth performance with respect to AMF diversity in P-limited cover cropping systems. Additionally, AMF diversity in roots can potentially contribute to soybean P nutrition even in the P-fertilized cover crop rotational system. Therefore, further investigation into the interaction of AMF diversity, P-application and cover cropping is required for the development of more effective P management practices on soybean growth performance.

Arbuscular mycorrhizal fungal (AMF) communities are fundamental in organic cropping systems where they provide essential agro-ecosystem services, improving soil fertility and sustaining crop production. They are affected by agronomic practices, but still, scanty information is available about the role of specific crops, crop rotations and the use of winter cover crops on the AMF community compositions at the field sites. A field experiment was conducted to elucidate the role of diversified cover crops and AMF inoculation on AMF diversity in organic tomato. Tomato, pre-inoculated at nursery with two AMF isolates, was grown following four cover crop treatments: Indian mustard, hairy vetch, a mixture of seven species and a fallow. Tomato root colonization at flowering was more affected by AMF pre-transplant inoculation than by the cover crop treatments. An enormous species richness was found by morphological spore identification: 58 AMF species belonging to 14 genera, with 46 and 53 species retrieved at the end of cover crop cycle and at tomato harvest, respectively. At both sampling times, AMF spore abundance was highest in hairy vetch, but after tomato harvest, AMF species richness and diversity were lower in hairy vetch than in the cover crop mixture and in the mustard treatments. A higher AMF diversity was found at tomato harvest, compared with the end of the cover crop cycle, independent of the cover crop and pre-transplant AMF inoculation. Our findings suggest that seasonal and environmental factors play a major role on AMF abundance and diversity than short-term agronomic practices, including AMF inoculation. The huge AMF diversity is explained by the field history and the Mediterranean environment, where species characteristic of temperate and sub-tropical climates co-occur.

Elevated nitrogen (N) deposition may stimulate a plant’s dependency on arbuscular mycorrhizal (AM) fungi in phosphorus (P)-deficient subtropical forests. However, the ecological assembly processes and the responses of AM fungal diversity and community structure to N deposition in both the roots and rhizosphere are still unclear. We collected root and soil samples from a Cunninghamia lanceolata plantation forest after four years of N addition and examined the community structure and assembly of AM fungi. Elevated N deposition decreased the AM fungal community diversity in both rhizosphere soil and roots. Glomeraceae was the dominant family of the AM fungal community in both soil and roots across all N addition treatments, followed by Gigasporaceae and Ambisporaceae. However, N addition induced differential variation in the community composition of AM fungi between soil and roots. For soil AM fungi, N addition decreased the Glomeraceae abundance and increased the Gigasporaceae and Ambisporaceae abundance. In contrast, the root AM fungal community was dominated by Glomeraceae under N addition treatments. Furthermore, N addition increased the deterministic community assembly that acted as an environmental filter for soil AM fungi. In contrast, N addition decreased the importance of determinism, implying that the selection of plants on root AM fungi decreased with increasing N addition. Altogether, our findings suggest that the community structure of AM fungi responds differently to N deposition in the soil and roots in subtropical forests and highlight the important role of soil AM fungi in helping host plants respond to N deposition.