Identification Of Arbuscular Mycorrhizal Fungi Research Articles

EUKARYOME: the rRNA gene reference database for identification of all eukaryotes.

Molecular identification of micro- and macroorganisms based on nuclear markers has revolutionized our understanding of their taxonomy, phylogeny and ecology. Today, research on the diversity of eukaryotes in global ecosystems heavily relies on nuclear ribosomal RNA (rRNA) markers. Here, we present the research community-curated reference database EUKARYOME for nuclear ribosomal 18S rRNA, internal transcribed spacer (ITS) and 28S rRNA markers for all eukaryotes, including metazoans (animals), protists, fungi and plants. It is particularly useful for the identification of arbuscular mycorrhizal fungi as it bridges the four commonly used molecular markers-ITS1, ITS2, 18S V4-V5 and 28S D1-D2 subregions. The key benefits of this database over other annotated reference sequence databases are that it is not restricted to certain taxonomic groups and it includes all rRNA markers. EUKARYOME also offers a number of reference long-read sequences that are derived from (meta)genomic and (meta)barcoding-a unique feature that can be used for taxonomic identification and chimera control of third-generation, long-read, high-throughput sequencing data. Taxonomic assignments of rRNA genes in the database are verified based on phylogenetic approaches. The reference datasets are available in multiple formats from the project homepage, http://www.eukaryome.org.

Keanekaragaman Fungi Mikoriza Arbuskula pada Tegakan Kelicung (Diospyros macrophylla Blume) di KHDTK Rarung, Lombok Tengah

Kelicung tree (Diospyros macrophylla) is a native tree species of West Nusa Tenggara. It is a slow-growing tree with very good wood ornament, strength, and durability properties. Symbiotic mutualism between kelicung and mycorrhiza has not been documented. Mycrorrhiza is a mutualism symbiosis between fungi and plant roots that could improve plant growth due to increase nutrient and water absorbtion. Therefore, the objective of this study is to investigate mycorrhizal symbiosis in kelicung roots. Soil and root samples were taken in the Rarung Special Purpose Forest Area at 0 – 20 cm depth. Isolation and identification of arbuscular mycorrhizal fungi used screening and centrifugation methods to identify spore density, diversity, abundance, and frequency. Furthermore, root staining and colonization of arbuscular mycorrhizal fungi were carried out by observing the structure of vesicles, arbuscules, hyphae, and spores on Kelicung roots. The research results showed that the average number of spores found was 846 spores/50 g of soil. Spores were identified in two genera: Glomus sp and Acaulospora sp. Glomus spores found were round, the colour of the spores was yellow, the spores did not react when Melzer's solution was dropped, and the spore walls of the Arbuscular mycorrhizal fungi of the genus Glomus consisted of 1-2 layers of cell walls. Observation of root colonization, no structures of root arbuscular mycorrhizal fungi were found in Kelicung.
 Keywords: Acaulospora sp., Arbuscular mycorrhizal fungi, Diospyros macrophylla, Glomus sp., Kelicung

Identification of Arbuscular Mycorrhizal Fungi associated with Arabica coffee root (Coffea arabica) in the Arfak Mountains region of West Papua, Indonesia

Abstract. Suparno A, Prabawardani S, Nisa DK, Ruimassa RR. 2023. Identification of Arbuscular Mycorrhizal Fungi associated with Arabica coffee root (Coffea arabica) in the Arfak Mountains region of West Papua, Indonesia. Biodiversitas 24: 3207-3213. Coffee is a global commodity widely consumed as a beverage globally and has a high economy. Currently, the trend of drinking coffee is no longer dominated by older people but has become part of the millennial and celebrity lifestyle; therefore, worldwide demand for coffee continues to increase. Efforts to develop area expansion and productivity continue to be pursued. The Arfak Mountains Region of West Papua, located 800 - 2,500 m above sea level (masl), is a potential area for coffee plantations. The local farmers in this area have grown coffee independently and with gradual government support for the last years. Given the benefits of Arbuscular Mycorrhizal Fungi (AMF), which can increase plant growth and productivity, this study aims to identify the types of AMF that are associated with coffee plants in 4 districts, Mokwam, Anggi Giji, Anggi Gida, and Membey in Arfak Mountains region, West Papua. Identification of AMF types was observed based on the morphology of mycorrhizal spores. The research was conducted using the observation method with a purposive sampling technique from November 2022 to March 2023. Based on the observation, the coffee plants in 4 districts were associated with AMF. The types of AMF associated with coffee in Mokwam were more numerous than in other locations, namely the Acaulosporaceae Genus with five species, Glomeaceae Genus with five species, one species of Simiglomus sensu stricto, one species of Funneliformis sensu stricto, and one species of Septoglomus sensu stricto. AMF in Anggi Giji District consisted of Acaulosporaceae Genus with two species, Glomeaceae Genus with four species, while in Anggi Gida District consisted of Acaulosporaceae Genus with one species, Glomeaceae Genus with three species, one species Septoglomus sensu stricto. AMF species in the coffee plants of the Membey District are the Acaulosporaceae Genus with three species, the Glomeaceae Genus with two species, and one species of Septoglomus sensu stricto. From the soil analysis results, the soil fertility level is low; conversely, the mycorrhizal presence level is higher because of the more infertile soil, the more active mycorrhiza.

Mycorrhizal status of Argania spinosa (L.) Skeels in northeastern of Morocco

The argan tree, an endemic species in Morocco, has been exhaustively studied in its southwestern range but neglected in its northeastern territory. Thus, the present study sets as objectives the identification of arbuscular mycorrhizal fungi and the evaluation of the argan tree’s roots’ mycorrhization level in two stations: one in the region of Béni Snassène (Berkane): Jebel Takermine with 7 prospected sites and the other at Jebel Aklim Alkabir with 2 sites. At the first station (Jebel Takermine), the mycorrhizal frequency of Argania spinosa roots varied between 64% and 100% with the root mycorrhizal intensity in the range of 30.77% and 66%. The arbuscular contents are higher at sites 2 (50.46%), 4 (50.33%), 7 (39.44%) and 1 (30.5%) against 18% and 20% at sites 6, 5 and 3. Argan trees from Jbel Aklim Alkbir, exhibited a high mycorrhization frequency and intensity ranging from 88% to 100% and between 39.4% and 73.4% respectively. Regarding arbuscular and vesicular rates, the highest values were associated to the roots of site 1 with 59.3% and 29.4% respectively compared to the lowest rates of 20% and 14% in those of site 2. Spore densities in the rhizosphere of the studied argan trees in the two stations were in the range of 78 and 697 spores/100 g of soil. The identification of isolated mycorrhizal spores revealed the presence of 28 species encompassing 7 genera: Acaulospora, Dentiscutata Claroideoglomus, Funneliformis, Glomus, Rhizophagus, Pacispora, and 5 Families: Glomeraceae (7 species), Acaulosporaceae (10 species), Pacisporaceae (2 species), Claroideoglomeraceae (2 species), Gigasporaceae (1 species).

Arbuscular Mycorrhizal Fungi as an Important Factor Enabling the Adaptation of Anthyllis vulneraria L. to Zn-Pb-Polluted Tailings.

The old Zn-Pb-contaminated (calamine) tailings in southern Poland are spontaneously colonized by metal-tolerant Anthyllis vulneraria L. (Fabaceae), which can form simultaneously symbiotic association with nitrogen-fixing rhizobia and phosphorus-acquiring arbuscular mycorrhizal fungi (AMF). So far, fungal colonization and the AMF diversity of calamine-inhabiting legumes have been poorly studied. Thus, we determined AMF spore density in the substratum and the mycorrhizal status of nodulated A. vulneraria plants occurring on calamine tailings (M) and on a reference non-metallicolous (NM) site. The results indicate the presence of the Arum-type of arbuscular mycorrhiza in the roots of both Anthyllis ecotypes. Despite the presence of AM fungi in M plant roots, the dark septate endophyte (DSE) fungi (hyphae and microsclerotia) were occasionally also detected. Metal ions were accumulated mainly in the nodules and intraradical fungal structures rather than thick plant cell walls. Mycorrhization parameters (frequency of mycorrhization and intensity of root cortex colonization) for M plants were markedly higher and differed in a statistically significant manner from the parameters for NM plants. Heavy metal excess had no negative effect on the number of AMF spores, the amounts of glomalin-related soil proteins and AMF species composition. Molecular identification of AMF using PCR-DGGE analysis based on the 18S rDNA ribosomal gene by nested-PCR with primers AM1/NS31 and NS31-GC/Glo1 revealed similar genera/species of AMF in the roots of both Anthyllis ecotypes: Rhizophagus sp., R. fasciculatus, and R. iranicus. The results of this work indicate the presence of unique fungal symbionts, which may enhance A. vulneraria tolerance to heavy metal stress and plant adaptation to extreme conditions on calamine tailings.

Isolation and identification of arbuscular mycorrhizal fungi from an abandoned uranium mine and their role in soil-to-plant transfer of radionuclides and metals

Arbuscular mycorrhizal fungi were recovered from soil samples from the naturally radioactive soil at the long-abandoned South Terras uranium mine in Cornwall, UK. Species of Rhizophagus, Claroideoglomus, Paraglomus, Septoglomus, and Ambispora were recovered, and pot cultures from all except Ambispora were established. Cultures were identified to species level using morphological observation and rRNA gene sequencing combined with phylogenetic analysis. These cultures were used in pot experiments designed with a compartmentalised system to assess the contribution of fungal hyphae to the accumulation of essential elements, such as copper and zinc, and non-essential elements, such as lead, arsenic, thorium, and uranium into root and shoot tissues of Plantago lanceolata. The results indicated that none of the treatments had any positive or negative impact on shoot and root biomass. However, Rhizophagus irregularis treatments showed higher accumulation of copper and zinc in shoots, while R. irregularis and Septoglomus constrictum enhanced arsenic accumulation in roots. Moreover, R. irregularis increased uranium concentration in roots and shoots of the P. lanceolata plant. This study provides useful insight into fungal-plant interactions that determine metal and radionuclide transfer from soil into the biosphere at contaminated sites such as mine workings.

Diversity of arbuscular mycorrhizal fungi in asphalt post-mining land in Buton Island, Indonesia

Abstract. Tuheteru FD, Husna, Albasri, Effendy HM, Arif A, Basrudin, Tuheteru EJ, Mulyono S, Irianto RSB. 2022. Diversity of arbuscular mycorrhizal fungi in asphalt post-mining land in Buton Island, Indonesia. Biodiversitas 23: 6327-6334. Studies on the presence and diversity of Arbuscular Mycorrhizal Fungi (AMF) on overburden (OB) and asphalt post-mining areas in Buton Island are still limited. The purpose of this study was to examine the presence and diversity of AMF on OB and asphalt post-mining stockpiles in Buton Island, Southeast Sulawesi Province. The sampling of soil and roots of successional plants was carried out at eight post-mining sites at the IUP Kabungka (Pasarwajo) (6 sites) PT. WIKA Aspal and IUP Lawele (2 sites), PT. WIKA Bitumen. The AMF spores were isolated using the wet filter pour method, followed by the identification of AMF which was done by observing the morphology of AMF spores. The results showed that all successional plants were in symbiosis with AMF, which was indicated by the existence of AMF structure in plant roots with the percentage of AMF colonization ranging from 5 - 95%. The number of spores per 50 g ranged from 1 to 9 spores. A total of 9 AMF species were found to belong to five genera, i.e., Claroideoglomus, Haloatospora, Glomus, Gigaspora, and Cetraspora from three families. Claroideoglomus claroideum was found to be the most dominant species isolated with the high occurrence and the highest Importance and Shannon-Wiener Diversity Indices followed by Haloatospora cf. pansihalos, Glomus sp. 5, and Glomus sp. 4. This study also showed that the AMF diversity index differed among locations. The presence of AMF can accelerate the emergence and development of adaptive plants on post-asphalt mining land in Buton Island.

Diversity of Arbuscular Mycorrhizal Fungi of Kalappia celebica: An endemic and endangered plant species in Sulawesi, Indonesia

Abstract. Husna, Tuheteru FD, Albasri, Arif A, Basrudin, Nurdin WR, Arman E, Agustin DI, Saribadu J, Rahmat, Dermawansyah A, Daliana, Lody LP, Deri AS, Safitri I. 2022. Diversity of Arbuscular Mycorrhizal Fungi of Kalappia celebica.): An endemic and endangered plant species in Sulawesi, Indonesia. Biodiversitas 23: 5290-5297. Arbuscular Mycorrhizal Fungi (AMF) are a group of fungi belonging to Phylum Glomeromycota. AMF has a symbiotic relationship with higher plants, including endangered species in various climatic regions and land types. Research on AMF diversity related to endemic and endangered tree species in the tropics is still limited. This study aimed to examine the diversity of AMF in the natural habitat of Kalappia celebica Kosterm. (kalapi) in Southeast Sulawesi Province, Indonesia. Samples of soil and roots of K. celebica were collected from six natural habitat locations, i.e., Konawe District (Anggatoa and Abuki Villages), East Kolaka District (Lalingato and Anggaloosi Villages) and Kolaka District (Lalonaha and Lapao Pao Villages). The AMF spores were isolated by using the pouring method, followed by wet filtration. AMF identification was carried out by observing the morphology of AMF spores. A total of eight AMF species belonging to three genera Viz. Glomus, Funneliformis and Rhizophagus were isolated. Among the species identified, five species belong to genus Glomus, two species belong to the genus Funneliformis and one species belongs to the genus Rhizophagus. Glomus sp.1 and 2 and F. mosseae had the highest spore density, relative density and frequency, and importance index values. The AMF highest spore density was obtained in Anggaloosi Village of East Kolaka District. Soil properties such as organic C, total N and clay were positively correlated with spore density and negatively correlated with AMF colonization. The Simpson’s Index ranged from 0.42 ± 0.117 to 0.86 ± 0.068. The range of Shannon-Wiener Index was 0.12 ± 0.208 to 0.99 ± 0.198, while the Evenness Index ranged from 0.17 ± 0.189 to 0.75 ± 0.106. These findings indicated that K. celebica species are rich in AMF diversity. These symbiont fungi are key components of the ecosystem. Further research is needed to develop their use as promoters of plant establishment in conservation and restoration of those sites.

Diversity of Arbuscular Mycorrhizal Fungi of Kalappia celebica: An endemic and endangered plant species in Sulawesi, Indonesia

Abstract. Husna, Tuheteru FD, Albasri, Arif A, Basrudin, Nurdin WR, Arman E, Agustin DI, Saribadu J, Rahmat, Dermawansyah A, Daliana, Lody LP, Deri AS, Safitri I. 2022. Diversity of Arbuscular Mycorrhizal Fungi of Kalappia celebica.): An endemic and endangered plant species in Sulawesi, Indonesia. Biodiversitas 23: 5290-5297. Arbuscular Mycorrhizal Fungi (AMF) are a group of fungi belonging to Phylum Glomeromycota. AMF has a symbiotic relationship with higher plants, including endangered species in various climatic regions and land types. Research on AMF diversity related to endemic and endangered tree species in the tropics is still limited. This study aimed to examine the diversity of AMF in the natural habitat of Kalappia celebica Kosterm. (kalapi) in Southeast Sulawesi Province, Indonesia. Samples of soil and roots of K. celebica were collected from six natural habitat locations, i.e., Konawe District (Anggatoa and Abuki Villages), East Kolaka District (Lalingato and Anggaloosi Villages) and Kolaka District (Lalonaha and Lapao Pao Villages). The AMF spores were isolated by using the pouring method, followed by wet filtration. AMF identification was carried out by observing the morphology of AMF spores. A total of eight AMF species belonging to three genera Viz. Glomus, Funneliformis and Rhizophagus were isolated. Among the species identified, five species belong to genus Glomus, two species belong to the genus Funneliformis and one species belongs to the genus Rhizophagus. Glomus sp.1 and 2 and F. mosseae had the highest spore density, relative density and frequency, and importance index values. The AMF highest spore density was obtained in Anggaloosi Village of East Kolaka District. Soil properties such as organic C, total N and clay were positively correlated with spore density and negatively correlated with AMF colonization. The Simpson’s Index ranged from 0.42 ± 0.117 to 0.86 ± 0.068. The range of Shannon-Wiener Index was 0.12 ± 0.208 to 0.99 ± 0.198, while the Evenness Index ranged from 0.17 ± 0.189 to 0.75 ± 0.106. These findings indicated that K. celebica species are rich in AMF diversity. These symbiont fungi are key components of the ecosystem. Further research is needed to develop their use as promoters of plant establishment in conservation and restoration of those sites.

English

The papaya tree is not very exploitable on the international market because of the post-harvest decline of the fruits. In this study, we identified arbuscular mycorrhizal fungi (AMF) from the rhizosphere of papaya in two agroecological zones of Cameroon (Njombe-Pendja: NP and Yaounde: Y). Morphological studies on the density, the rate of root colonization, the diversity of AMF species were evaluated. Molecular identification of AMF was performed by sequencing the small subunit (SSU) 18S rDNA gene region from soil samples. These genes were amplified from the AML and NS2 primers. The total number of spores per 100 g of dry soil was significantly different (P<0.05) depending on the sites, ranging from 391 in NP to 790 in Y sites. The frequency and intensity of root colonization was not significantly different in the two sites. Eleven (11) AMF belonging to seven (7) genera (Scutellospora, Gigaspora, Acaulospora, Entrophospora, Funneliformis, Glomus and Racocetra) were recorded. Two AMF species (T1: Scutellospora rosa and T6: Racocetra gregaria) were recorded at NP and absent at Y. The Shannon-Wiener index revealed a low diversity of AMF species. Homogeneity of species was recorded. Molecular analysis reveals that the AMF species obtained have sizes of 250 bp. Key words: Arbuscular mycorrhizal fungi, Carica papaya L., fungal, molecular and morphological identification, rhizosphere.

Seasonal variation of arbuscular mycorrhizal fungi in ecotone forests of the northern region of Brazilian Amazonia

The ecotone forests in the northern region of Brazilian Amazonia are important areas representing transition zones between forest and non-forest ecosystems. These areas have soils nutrient-poor that poorly drain. Under these environmental conditions, Peltogyne gracilipes (Leguminoseae), an endemic tree species, can form natural monodominant forests. Here, we assessed the arbuscular mycorrhizal fungi (AMF) community in three forest types on the eastern side of Maracá Island and the relationship of these microorganisms with the monodominance of P. gracilipes. In this study, soil samples were collected in two seasons (dry and rainy). The samples were collected in 9 plots, in rich areas, poor areas and areas without P. gracilipes. Soil samples were evaluated for chemical and particle size analysis, spore density and morphology, and identification of AMF. AMF species were identified using two approaches: spores collected in the field and trap cultures. Eighteen and 13 AMF species were identified in the dry and rainy seasons, respectively, for spores extracted from the field. Six total species were detected exclusively in trap cultures in the dry season. AMF communities were co-dominated by members of the Gigasporaceae, Acaulosporaceae and Glomeraceae families. Redundancy analyses indicated that several soil attributes, such as pH, Fe, Mg, and sand content associated with the AMF species richness in both seasons. We conclude that the ecotone forests in the eastern region of Maracá Island are home to important richness and diversity of AMF species and that various soil factors influence the composition of the AMF community in this ecosystem.

Environmental identification of arbuscular mycorrhizal fungi using the LSU rDNA gene region: an expanded database and improved pipeline

Arbuscular mycorrhizal fungi (AMF; Glomeromycota) are difficult to culture; therefore, establishing a robust amplicon-based approach to taxa identification is imperative to describe AMF diversity. Further, due to low and biased sampling of AMF taxa, molecular databases do not represent the breadth of AMF diversity, making database matching approaches suboptimal. Therefore, a full description of AMF diversity requires a tool to determine sequence-based placement in the Glomeromycota clade. Nonetheless, commonly used gene regions, including the SSU and ITS, do not enable reliable phylogenetic placement. Here, we present an improved database and pipeline for the phylogenetic determination of AMF using amplicons from the large subunit (LSU) rRNA gene. We improve our database and backbone tree by including additional outgroup sequences. We also improve an existing bioinformatics pipeline by aligning forward and reverse reads separately, using a universal alignment for all tree building, and implementing a BLAST screening prior to tree building to remove non-homologous sequences. Finally, we present a script to extract AMF belonging to 11 major families as well as an amplicon sequencing variant (ASV) version of our pipeline. We test the utility of the pipeline by testing the placement of known AMF, known non-AMF, and Acaulospora sp. spore sequences. This work represents the most comprehensive database and pipeline for phylogenetic placement of AMF LSU amplicon sequences within the Glomeromycota clade.

Exploration and Identification of Arbuscular Mycorrhizal Fungi (AMF) in the Rhizosphere of Cocoa (Theobroma cacao. L) in West Sumatra

West Sumatra was declared as a center for Indonesian cocoa in the western portion. Almost all areas in West Sumatra are suitable for cocoa cultivation, so the cocoa planting area covers almost all the districts in the province. However, West Sumatran Cocoa production decline occurs because cocoa plantations are still mostly cultivated with smallholder plantations. The occurrence of land conversion and reduced land carrying capacity, including soil fertility, is due to the continuous use of inorganic fertilizers, which results in a decrease in the ecological quality of the land. Efforts to improve land conditions biologically by utilizing soil biotechnology. One of the soil microbes that can be utilized is Arbuscular Mycorrhizal Fungi (AMF). AMF that live in a symbiotic mutualism with plant roots and helps the absorption of plant nutrients and live in various places. The study aimed to identify arbuscular mycorrhiza Cocoa rhizosphere three districts, West Sumatra province, from the month of November 2020 to May 2021. The results showed the number of spores and diversity of AMF. The number of spores was 67-218 spores per 20 g soil. The root colonization was in the range of 21.3 - 24.6%. While the AMF diversity before trapped found three types of sport Glomus sp., Acaulospora sp., dan Gigaspora sp

Isolation and identification of indigenous Arbuscular Mycorrhizal Fungi (AMF) of forest clove rhizosphere from Maluku, Indonesia

Abstract. Mahulette AS, Alfian A, Kilkoda KA, Lawalata IJ, Marasabessy DA, Tanasale VL, Makaruku MH. 2021. Isolation and identification of indigenous Arbuscular Mycorrhizal Fungi (AMF) of forest clove rhizosphere from Maluku, Indonesia. Biodiversitas 22: 3613-3619. Forest clove is classified as wild-type and endemic to the Maluku (Moluccas) Islands, Indonesia. The different condition of growing areas causes various types of Arbuscular Mycorrhizal Fungi (AMF) associated with forest clove. The study aimed to identify and obtain indigenous AMF inoculums from the forest clove rhizosphere from two distribution areas in Maluku. The results of AMF identification found two types of spores from the genus Glomus in the rhizosphere of forest cloves from Ambon Island with a spore density of 35/50 g of soil. In comparison, three spores were found in Seram Island, two from the genus Scutellospora and one from the Acaulospora. With an overall spore density of 5/50 g of soil. After culture trapping, there was a change in type and an increase in spore density in soil samples from the rhizosphere of the two forest clove distribution areas. Soil samples from Ambon after trapping culture obtained two new types of spores from the genus Acaulospora with a total spore number of 57/50 g soil while in soil samples from Seram found three new types of spores from the genus Glomus with a total spore count of 104/50 g of soil.

Arbuscular mycorrhizal fungal community assembly in agroforestry systems from the Southern Brazil

Soil ecosystem disturbances promoted by the biological invasion can promote soil food web degradation by disrupting arbuscular mycorrhizal fungi (AMF) community structure. We aimed to compare the benefit of native tree plantings (e.g., Araucaria angustifolia, Ilex paraguariensis, and Mimosa scabrella) in a 5-year agroforestry system to overcome the soil disturbance due to invasion by Pine trees on AMF community composition in the Southern Brazil to natural restoration (unassisted forest ecosystem). We used morphological AMF identification and the natural forest as reference ecosystem. AMF diversity and dominance in the native tree species and unassisted forest ecosystem were similar, but these environments showed higher values when compared to the natural ecosystem. The PCA analysis showed soil pH, microbial biomass carbon, AMF richness, and the presence of Claroideoglomus etunicatum, Dentiscutata cerradensis, Gigaspora species, and Glomus sp. as the main factors contributing to the variance of the samples. In general, the most abundant AMF species presented in all studied ecosystems was Funneliformis mosseae. Differences in AMF community structure and soil properties were associated with (1) ecosystem disturbance, (2) changes in soil properties such as soil pH, (3) rootability by fine root production, and (4) rhizodeposition by soil microbial carbon as observed into the PCA analysis. These results contribute to a deeper view of the AMF communities in agroforestry systems and open new perspective for the management of degraded lands by biological invasion involving AMF species and native tree species in the Southern Brazil.

Diversity and species composition of arbuscular mycorrhizal fungi across maize fields in the southern part of Belgium.

Arbuscular mycorrhizal fungi (AMF) are key actors among soil microbial inhabitants, forming beneficial associations with most horticultural plants and crops (e.g., maize). For maize, the world most cultivated cereal, data on AMF species diversity in fields is sparse and even totally nonexistent in the southern part of Belgium where maize represents 8% of the cultivated area. In the present study, 14 maize fields in South Belgium under conventional, conversion, or organic management were analyzed for AMF diversity and species composition using 454 pyrosequencing. A large part (54%) of the 49 AMF species observed were unknown or have not been described in the literature. AMF diversity highly varied among fields, with the number of species ranging between 1 and 37 according to the field. A statistically significant effect of management was measured on AMF diversity, with the highest Hill index values (diversity and richness) under the organic management system compared with conventional management or conversion. Our results suggest a positive effects of organic management on AMF diversity in maize. They also highlight the rather high diversity or richness of AMF and the large portion of sequences not yet ascribed to species, thereby emphasizing a need to intensify AMF identification in cropping systems.

Identification of arbuscular mycorrhizal fungi in soils of the North Caucasus based on Illumina MiSeq data for ITS1 and ITS2 regions

The objective of our research was to analyze the efficiency of identification of arbuscular mycorrhizal fungi (AMF) for 2 regions: ITS1 and ITS2 regions of AMF DNA isolated from the soils of the North Caucasus (Karachay-Cherkessia). For the first time the necessity of different AMF species identification using both ITS regions was revealed, but not one region. The research demonstrated: 1) the set of taxa is different using ITS1- and ITS2-based identification; 2) analysis of the ITS1 region reveals a greater number of operational taxonomic units; 3) ITS2 allows identification of AMF at the species level more often. Sample preparation for Illumina MiSeq analysis was optimized. Obligatory stages in the sample preparation were the purification of DNA in the agarose gel in Silica after isolation, as well as separate amplification of ITS1 and ITS2 followed by combining and joint sequencing for each sample. The results showed the highest AMF biodiversity for the 176Te sample from the ecosystem of the subalpine meadow of the southeastern slope of Malaya Hatipara mountain (43°25′48.0″N 41°42′31.0″E; 2401 m above sea level), in which 8 species of AMF were identified (Archaeospora spainiae, Claroideoglomus claroideum, Diversispora versiformis, Entrophpora infrequens, Funneliformis mosseae, Glomus indicum, Paraglomus laccatum, Rhizophagus irregularis).

Exploring plant root-fungal interactions in a neotropical freshwater wetland

Background: Wetlands in Neotropics harbor high fungal diversity, including arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE). This study describes the interaction of plant roots with AMF and DSE in a freshwater wetland belonging to a hotspot of biodiversity.
 Hypothesis: Differential root colonization between arbuscular mycorrhizal and dark septate endophyte fungi is influenced by plant species and abiotic conditions in a freshwater wetland. 
 Studied species: Plant species colonized by arbuscular mycorrhizal and dark septate endophyte fungi.
 Methods: Properties of soils and the water column, floristic composition, root colonization by AMF and DSE, and molecular identification of AMF inside roots were studied.
 Results: Soils were Gleysol and flooded during the rainy season. Most of identified plant species were herbaceous, with Cyperus articulatus and Mimosa pigra as the dominant species. Seven of 8 analyzed plant species exhibited differential co-colonization between AMF and DSE. Repeated sampling for one year under flooding/dry conditions demonstrated that C. articulatus and M. pigra were mainly associated with DSE and AMF, respectively. A positive correlation between dissolved O2 in the water column and fungal colonization was observed in C. articulatus . Glomerales and Archaeosporales were molecularly identified inside roots containing arbuscules of M. pigra .
 Conclusions: Findings highlight differential coexistence between AMF and DSE in plant roots; fungal colonization was influenced by flooding/dry conditions in a neotropical wetland; the community of AMF inside arbusculated roots of M. pigra includes at least four clades.

Occurrence of Arbuscular Mycorrhizal Fungi in Hemp (Cannabis sativa) Plants and Soil Fertilized with Sewage Sludge and Phosphogypsum

ABSTRACT The colonization of soil and roots by arbuscular mycorrhizal fungi and the formation of spores under the influence of fertilization with phosphogypsum and sewage sludge were examined. Identification of arbuscular mycorrhizal fungi was carried out in soil and in roots of outdoor experimental hemp plantation. Assessment of the colonization was carried out by molecular and microscopic methods. The material for the isolation of DNA consisted of soil samples taken from mycorrhizal soil and hemp roots of cv. Białobrzeskie, cv. Tygra, and cv. Beniko. In nested PCR reactions part of the small subunit (SSU) region, the region of the internal transcribed spacer (ITS) and part of large subunit (LSU) rDNA region were amplified. As a result of the cloning and sequencing of the DNA, it was found that the greatest similarity of the obtained sequences present in the samples of roots and soil was to the following species: Diversispora sp., Funneliformis sp., Glomus sp., Funneliformis mosseae, Glomus caledonium, Funneliformis geosporum, Glomus occultum. The colonization by the fungi was estimated and found to differ depending on the hemp cultivar and doses of fertilizers. Cultivar Tygra showed no negative impact on biomass production by intensive mycorrhization, proved its high tolerance to phosphogypsum and sewage sludge pollution, and responded positively regarding biomass production.

Morphological and molecular characterization of arbuscular mycorrhizal fungal communities inhabiting the roots and the soil of saffron (Crocus sativus L.) under different agricultural management practices

ABSTRACTCrocus sativus L. cultivation is expanding to areas with low soil fertility, where mycorrhizal fungi are supposed to be essential for plants growth and ecosystems functioning. Agricultural practices applied under these conditions should lead to good saffron productivity and quality. Our objective was to study the density and diversity of mycorrhizal fungi populations associated with saffron grown in Taliouine (Morocco) under different agricultural management practices (fertilization type, age and plantation method). Morpho-anatomical studies identified rhizospheric mycorrhizal spores and assessed root colonization by arbuscular mycorrhizal fungi (AMF). Molecular identification of AMF was realized by sequencing the Large Subunit (LSU) rDNA gene region. Among the eleven species of AMF spores identified, Funneliformis and Rhizoglomus species were the most abundant (> 35%). Modern saffron plantation showed higher roots colonization rates (mycorrhization intensity (100%) and frequency (51.6%)), while in traditional plantations lower mycorrhization frequency values were found (17.4%). LSU sequencing identified five AMF genera and three unknown genomic groups, whereas Shannon diversity index indicated that AMF community composition changed significantly according to plantation age and fertilization type. Our results contribute to a better knowledge of saffron AMF communities and open new perspectives for a rational utilization of the agricultural practices for organic saffron production.