Arbuscular Mycorrhizal Fungi Species Diversity Research Articles

Arbuscular mycorrhizal fungi and vascular plant species abundance and community structure in tallgrass prairies with varying agricultural disturbance histories

Tallgrass prairie is one of the most critically endangered habitats in Canada, with less than one percent of its original extent remaining. Information on arbuscular mycorrhizal fungi (AMF) in Canadian prairie, which may be beneficial to tallgrass prairie restoration, is lacking. We conducted a descriptive ecological study of the communities of arbuscular mycorrhizal fungi (AMF) and vascular plants at five former or extant tallgrass prairie sites on Walpole Island First Nation, Ontario: one undisturbed prairie, two naturally recovered prairies and two old fields abandoned from agriculture 19 and 3 years ago, respectively. The AMF were identified morphologically using spores from trap cultured soil samples, and the number of spores of each taxon was used as an estimate of their abundance. We performed ester linked extraction and measured fungal lipid biomarkers to assay the biomass of AMF. Sixteen species of AMF and 111 plant species were identified. Species composition, richness and diversity of both AMF and plants varied among sites. Principal component analysis distinguished fungal and plant communities at tallgrass prairie sites from those at more recently disturbed fields. However, AMF biomass, as assessed by the concentration of the fatty acid methyl esters C16:1cis11 and C18:1cis11, was not significantly different between old fields and tallgrass prairie. Community composition of AMF was a better indicator of agricultural disturbance than AMF abundance, species richness, or biomass. Therefore, if AMF play an important role in the reestablishment of tallgrass prairie plant species, it is most likely through plant host specificity with particular species or taxonomic groups in the AMF community.

The mosaic of habitats in the high-altitude Brazilian rupestrian fields is a hotspot for arbuscular mycorrhizal fungi

The high diversity in rupestrian field vegetation has been attributed to the mosaic of environments formed by several soil classes, rugged relief and microclimatic variation. Although advances in the knowledge of some biological areas in rupestrian fields have been made, little is known about the relevance of soil microorganisms and their relationships with the vegetation. Symbiosis with arbuscular mycorrhizal fungi (AMF) is one of the most studied interactions between microorganisms and plants, because they are ubiquitous and contribute to the sustainability of ecosystems. This study aimed to investigate the occurrence and diversity of AMF species and to evaluate their relationship with soil physicochemical attributes and plant diversity in different habitats of the rupestrian fields from the Cadeia do Espinhaço, Serra do Cipó, Brazil. These rupestrian fields were delimited into five distinct habitats: rock outcrop, quartz gravel fields, sandy bogs, peat bogs and the Cerrado. Forty-nine AMF species were identified as belonging to nine families and twelve genera. Among them, Acaulospora colossica and Pacispora dominikii were found for the first time in Brazil. The results of this study suggest that the diversity of AMF is related to the heterogeneity of habitats and that the soil texture (coarse sand, gravel and silt) is better related to the structure of these fungi communities than to the soil chemical attributes. Plant species richness was related to AMF richness only in the quartz gravel field, rocky outcrop, and sandy bog habitats. Considering these habitats constitute one of the most menaced ecosystems on the planet, our survey provides information to improve knowledge about rupestrian field biodiversity, thus supporting policy actions for its conservation and preservation.

Dosage-dependent shift in the spore community of arbuscular mycorrhizal fungi following application of tannery sludge

The controlled disposal of tannery sludge in agricultural soils is a viable alternative for recycling such waste; however, the impact of this practice on the arbuscular mycorrhizal fungi (AMF) communities is not well understood. We studied the effects of low-chromium tannery sludge amendment in soils on AMF spore density, species richness and diversity, and root colonization levels. Sludge was applied at four doses to an agricultural field in Rolândia, Paraná state, Brazil. The sludge was left undisturbed on the soil surface and then the area was harrowed and planted with corn. The soil was sampled at four intervals and corn roots once within a year (2007/2008). AMF spore density was low (1 to 49 spores per 50cm(3) of soil) and decreased as doses of tannery sludge increased. AMF root colonization was high (64%) and unaffected by tannery sludge. Eighteen AMF species belonging to six genera (Acaulospora, Glomus, Gigaspora, Scutellospora, Paraglomus, and Ambispora) were recorded. At the sludge doses of 9.0 and 22.6Mgha(-1), we observed a decrease in AMF species richness and diversity, and changes in their relative frequencies. Hierarchical grouping analysis showed that adding tannery waste to the soil altered AMF spore community in relation to the control, modifying the mycorrhizal status of soil and selectively favoring the sporulation of certain species.

Arbuscular mycorrhizal status and root phosphatase activities in vegetative Carica papaya L. varieties

The arbuscular mycorrhizal (AM) status and root phosphatase activities were studied in four vegetative Carica papaya L. varieties viz., CO-1, CO-2, Honey Dew and Washington. Standard techniques were used to ascertain information on spore density and species diversity of AM fungi. Although in case of estimation of root colonization and root phosphatase activities, the existing methods were slightly modified. Root colonization and spore density of AM fungi along with root phosphatase (acid and alkaline) activities varied significantly in four papaya varieties. The present study recorded higher acid root phosphatase activity when compared with alkaline root phosphatase activity under P-deficient, acidic soil conditions. The present study revealed that the root colonization of AM fungi influenced acid root phosphatase activity positively and significantly under P-deficient, acidic soil conditions. A total of 11 species of AM fungi belonging to five genera viz., Acaulospora, Dentiscutata, Gigaspora, Glomus and Racocetra were recovered from the rhizosphere of four papaya varieties.

Influence of Long-Term Fixed Fertilization on Diversity of Arbuscular Mycorrhizal Fungi

Diversity of arbuscular mycorrhizal fungi (AMF) was investigated in a field that had received long-term fixed fertilization (LFF) for 26 years. There were a total of 12 treatments in triplicates with different amounts of manure, urea, calcium phosphate, and potassium chloride. Rhizosphere soil samples of maize and wheat grown in the experimental field in Shandong Province, China, were collected in September 2003 and May 2004, respectively. Arbuscular mycorrhizal fungal spores were isolated and identified using morphological characters. Mycorrhizal colonization percentage, spore density (SD), species richness (SR), relative abundance (RA), and Shannon-Weiner index (SWI) were determined. Nineteen recognized species of AMF belonging to 5 genera were identified. Long-term fixed fertilization significantly influenced colonization percentage, SR, SD, and species diversity of AMF. The adaptability of AMF to soil fertility was different among species. Species richness and SD of AMF in maize and wheat rhizosphere soils were the highest in the nonfertilization treatment (control) and lowest in the high manure + high nitrogen treatment (M 2N 2). The SWI decreased as the fertilization level increased except in the low manure treatment (M 1) on maize. Compared with the other treatments, Treatment M 2N 2 significantly reduced SD of Glomus, and the high manure + low nitrogen treatment (M 2N 1) significantly retarded sporulation of Scutellospora. Manure treatments stimulated sporulation of Glomus mosseae. Spore density of G. mosseae was higher in the high nitrogen + phosphorus + potassium treatment (N 2PK) than in the high nitrogen + phosphorus treatment (N 2P) and the high nitrogen + potassium treatment (N 2K). The SD of S. pellucida was higher in Treatment N 2K than Treatments N 2PK and N 2P. In conclusion, long-term fixed fertilization, especially with high levels of manure and N, decreased SR, SD, and colonization and changed the species composition of AMF.

DNA‐based species level detection of Glomeromycota: one PCR primer set for all arbuscular mycorrhizal fungi

* At present, molecular ecological studies of arbuscular mycorrhizal fungi (AMF) are only possible above species level when targeting entire communities. To improve molecular species characterization and to allow species level community analyses in the field, a set of newly designed AMF specific PCR primers was successfully tested. * Nuclear rDNA fragments from diverse phylogenetic AMF lineages were sequenced and analysed to design four primer mixtures, each targeting one binding site in the small subunit (SSU) or large subunit (LSU) rDNA. To allow species resolution, they span a fragment covering the partial SSU, whole internal transcribed spacer (ITS) rDNA region and partial LSU. * The new primers are suitable for specifically amplifying AMF rDNA from material that may be contaminated by other organisms (e.g., samples from pot cultures or the field), characterizing the diversity of AMF species from field samples, and amplifying a SSU-ITS-LSU fragment that allows phylogenetic analyses with species level resolution. * The PCR primers can be used to monitor entire AMF field communities, based on a single rDNA marker region. Their application will improve the base for deep sequencing approaches; moreover, they can be efficiently used as DNA barcoding primers.

Effects of Organic Farming on Communities of Arbuscular Myeorrhizal Fungi

Red pepper (Capsicum annum L.) roots and soils representing different agricultural management practices such as conventional (CON), no-chemical (NOC), and organic farming systems (ORG) were collected from 32 farm field sites in Kyunggi, Korea to investigate the effects of these agricultural practices on arbuscular mycorrhizal (AM) symbiosis. ORG inoculum significantly increased plant growth compared to inoculum from CON and NOC. A community analysis of AM fungi (AMF) using morphological features of spores revealed that AMF spore abundance and species diversity were significantly higher in ORG than in CON. Additionally, a community analysis of AMF colonizing roots using a molecular technique revealed higher AMF diversity in ORG than in CON. These results suggest that agricultural practices significantly influence AM fungal community structure and mycorrhizal inoculum potential.

Diversity and infectivity of arbuscular mycorrhizal fungi in agricultural soils of the Sichuan Province of mainland China

Knowledge about the presence and diversity of arbuscular mycorrhizal fungi (AMF) in a specific area is an essential first step for utilizing these fungi in any application. The community composition of AMF in intensively managed agricultural soil in the Sichuan Province of southwest China currently is unknown. In one set of samples, AMF were trapped in pot cultures from 40 fields growing legumes in the Panxi region, southeast Sichuan. In a second set of samples, the MPN method with four-fold dilutions and maize as host was used to estimate infective propagules in soil from another 50 agricultural sites throughout the province. Soil types were heterogeneous and were classified as purple, yellow, paddy and red. Crops at each site were either maize, wheat or sweet orange. From this set of soil, AMF spores were also extracted and identified. Including all ninety soils, thirty glomeromycotan species in Glomus (20 species), Acaulospora (four species), Scutellospora (three species), Ambispora (one species), Archaeospora (one species) and Paraglomus (one species) were identified. Yellow, red and purple soils yielded similar numbers of AMF species, while AMF species diversity was clearly lower in paddy soil. In trap culture soils, the most frequent species were Glomus aggregatum or Glomus intraradices, Glomus claroideum and Glomus etunicatum. The species Acaulospora capsicula, Acaulospora delicata, G. aggregatum (or intraradices), G. claroideum, Glomus epigaeum, G. etunicatum, Glomus luteum, Glomus monosporum, Glomus mosseae and Glomus proliferum were successfully cultured as single-species pot cultures in Plantago lanceolata. The three most frequent species in field soils were G. mosseae, Glomus caledonium and Glomus constrictum. MPN values varied between 17 and 3334 propagules 100 g soil(-1) among the fifty field sites sampled. Regression analysis, including host&soil, log(P) and pH as explanatory variables explained 59% of the variation in log(MPN). The highest MPN estimates were found in purple soil cropped with maize and citrus, 324 and 278 propagules 100 g soil(-1), respectively. The lowest MPN value, 54 propagules 100 g soil(-1), was measured in wheat in purple and yellow soil. Despite intensive agricultural management that can include often repeated tillage, our examination of 90 agricultural sites revealed that soils of the Sichuan region have moderate to high numbers of infective AMF propagules as well as a high AMF species diversity. This opens possibilities for further studies and utilization of AMF in agriculture and horticulture in the Sichuan province, People's Republic of China.

MYCORRHIZAL COMMUNITY DYNAMICS FOLLOWING NITROGEN FERTILIZATION: A CROSS-SITE TEST IN FIVE GRASSLANDS

Arbuscular mycorrhizal fungi (AMF) are considered both ecologically and physiologically important to many plant communities. As a result, any alteration in AMF community structure following soil nitrogen (N) enrichment may impact plant community function and contribute to widespread changes in grassland productivity. We evaluated the responses of AMF communities to N fertilization (≥100 kg N·ha−1·yr−1) in five perennial grasslands within the Long-Term Ecological Research network to generate a broader understanding of the drivers contributing to AMF species richness and diversity with increasing soil N fertility, and subsequent effects to host-plant communities. AMF spore and hyphal community data at three mesic sites (Cedar Creek, Kellogg Biological Station, Konza Prairie) and two semiarid sites (Sevilleta, Shortgrass Steppe) were collected over two consecutive years and used to test four hypotheses about AMF responses to N fertilization. Under ambient soil N, plant annual net primary productivity and soil phosphorus (P) were strongly related to climatic differences in AMF communities (semiarid vs. mesic). Following N fertilization, the drivers of AMF community structure were soil N availability, N:P supply ratio, and host-plant photosynthetic strategy (C3 vs. C4) but not climate. In P-rich soils (low N:P), N fertilization reduced AMF productivity, species richness, and diversity and intensified AMF community convergence due to the loss of rare AMF species and the increased abundance of Glomus species. In P-limited soils (high N:P), AMF productivity, species richness, and diversity increased with N fertilization; the most responsive AMF taxa were Acaulospora, Scutellospora, and Gigaspora. Soil N or N:P × host-plant (C3, C4) interactions further modified these responses: AMF hyphae (primarily Gigasporaceae) associated with C3 plants increased in abundance with N fertilization, whereas C4 plants hosted nitrophilous Glomus species. Such responses were independent of the duration or quantity of N fertilization, or the time since cessation of N fertilization. This synthesis provides a new understanding of AMF community patterns and processes, and it identifies three key drivers (soil N, N:P, host plant) of AMF community structure that may be tested in other communities.

Community structure of arbuscular mycorrhizal fungi in a primary successional volcanic desert on the southeast slope of Mount Fuji

Community structure of arbuscular mycorrhizal fungi (AMF), evaluated as spore samples and mycorrhizal roots of four herbaceous plant species, was investigated at different altitudes in a primary successional volcanic desert on Mount Fuji using molecular methods (fragment and sequence analysis of the large ribosomal subunit RNA gene). In total, 17 different AMF clades were identified, and most were members of the Glomaceae, Acaulosporaceae, and Gigasporaceae. The AMF community structures detected by spore sampling were inconsistent with those from plant roots. Of all AMF clades, six (35.3%) were detected only on the basis of spores, six (35.3%) only in roots, and five corresponded to both spores and roots (29.4%). Although an Acaulospora species was the most dominant among spores (67.1%), it accounted for only 6.8% in root samples. A species analysis of AMF communities at different altitudes demonstrated that AMF species diversity increased as altitude decreased and that the species enrichment at lower altitudes resulted from the addition of new species rather than species replacement. The inconsistencies in the species composition of spore communities with those in roots and the change in species diversity with altitude are discussed.

Community structure of arbuscular mycorrhizal fungi at different soil depths in extensively and intensively managed agroecosystems

The vertical distribution of spores of arbuscular mycorrhizal fungi (AMF) was investigated in soil profiles of extensively and intensively managed agroecosystems, including two permanent grasslands, a vineyard and two continuously mono-cropped maize fields. The number of AMF spores decreased with increasing soil depth--most drastically in the grasslands and the vineyard--but there was a large diversity of AMF species even in the deepest soil layers (50-70 cm). This was particularly striking in the maize fields where the highest species numbers were found below ploughing depth. Some species sporulated mainly, or exclusively, in the deep soil layers, others mainly in the top layers. Soil samples were used to inoculate trap cultures. Up to 18 months after inoculation, there was no conspicuous difference in the species composition among the trap cultures representing different soil depths, and only a weak match to the species composition determined by analysis of field samples. Our results indicate that the AMF communities in deep soil layers are surprisingly diverse and different from the topsoil. Thus, deep soil layers should be included in studies to get a complete picture of AMF diversity.

Impact of long-term conventional and organic farming on the diversity of arbuscular mycorrhizal fungi.

Previous work has shown considerably enhanced soil fertility in agroecosystems managed by organic farming as compared to conventional farming. Arbuscular mycorrhizal fungi (AMF) play a crucial role in nutrient acquisition and soil fertility. The objective of this study was to investigate the diversity of AMF in the context of a long-term study in which replicated field plots, at a single site in Central Europe, had been cultivated for 22 years according to two "organic" and two "conventional" farming systems. In the 23rd year, the field plots, carrying an 18-month-old grass-clover stand, were examined in two ways with respect to AMF diversity. Firstly, AMF spores were isolated and morphologically identified from soil samples. The study revealed that the AMF spore abundance and species diversity was significantly higher in the organic than in the conventional systems. Furthermore, the AMF community differed in the conventional and organic systems: Glomus species were similarly abundant in all systems but spores of Acaulospora and Scutellospora species were more abundant in the organic systems. Secondly, the soils were used to establish AMF-trap cultures using a consortium of Plantago lanceolata, Trifolium pratense and Lolium perenne as host plants. The AMF spore community developing in the trap cultures differed: after 12 months, two species of the Acaulosporaceae (A. paulinae and A. longula) were consistently found to account for a large part of the spore community in the trap cultures from the organic systems but were found rarely in the ones from the conventional systems. The findings show that some AMF species present in natural ecosystems are maintained under organic farming but severely depressed under conventional farming, indicating a potentially severe loss of ecosystem function under conventional farming.

The history of Solidago canadensis invasion and the development of its mycorrhizal associations in newly-reclaimed land.

This study examined the associations between an invasive alien plant (Solidago canadensis L.) and arbuscular mycorrhizal fungi (AMF) in relation to the invasion history of S. canadensis on Chongming Island, China. A significant positive correlation was found between invasion time and AMF colonisation rate. The total number of AMF species increased with increasing invasion time and was positively related to the number of plant species occurring in plant communities, suggesting that the invasion time and plant diversity influence the diversity of AMF species. The relative abundance of two dominant AMF species, Glomus mosseae and G. constrictum, varied systematically with invasion time. In dry habitats, the relative abundance of G. mosseae increased with the invasion time of S. canadensis, while that of G. constrictum decreased with the invasion time. However, these relationships did not exist in the wet habitats.

Molecular diversity of arbuscular mycorrhizal fungi colonising Hyacinthoides non‐scripta (bluebell) in a seminatural woodland

Arbuscular mycorrhizal (AM) fungi form symbiotic associations with plant roots. Around 150 species have been described and it is becoming clear that many of these species have different functional properties. The species diversity of AM fungi actively growing in roots is therefore an important component of ecosystem diversity. However, it is difficult to identify AM fungi below the genus level from morphology in planta, as they possess few informative characters. We present here a molecular method for identifying infrageneric sequence types that estimate the taxonomic diversity of AM fungi present in actively growing roots. Bluebell roots were sampled from beneath two different canopy types, oak and sycamore, and DNA sequences were amplified from roots by the polymerase chain reaction with fungal‐specific primers for part of the small subunit ribosomal RNA gene. Restriction fragment length polymorphism among 141 clones was assessed and 62 clones were sequenced. When aligned, discrete sequence groups emerged that cluster into the three families of AM fungi: Acaulosporaceae, Gigasporaceae and Glomaceae. The sequence variation is consistent with rRNA secondary structure. The same sequence types were found at both sampling times. Frequencies of Scutellospora increased in December, and Acaulospora increased in abundance in July. Sites with a sycamore canopy show a reduced abundance of Acaulospora, and those with oak showed a reduced abundance of Glomus. These distribution patterns are consistent with previous morphological studies carried out in this woodland. The molecular method provides an alternative method of estimating the distribution and abundance of AM fungi, and has the potential to provide greater resolution at the infrageneric level.