Distribution and diversity of arbuscular mycorrhizal fungi in grapevines from production vineyards along the eastern Adriatic coast

The colonisation and diversity of arbuscular mycorrhizal fungi (AMF) on roots of grapevines were investigated in production vineyards located along a 500-km-long stretch of karst along the coast of the Adriatic Sea. AMF communities on roots of grapevines were analysed using temporal temperature gel electrophoresis and sequencing of the 18S and internal transcribed spacer segments of the rDNA operon. The AMF colonisation of these grapevines roots was consistent along the whole of this east Adriatic karst region, at 64 to 82 % of fine roots. The comparison of the AMF communities on the roots of these grapevines showed that the fungal community associated with grapevine roots seems to be relatively stable, with inter-vineyard variability comparable to intra-vineyard variability. Some of the changes in the fungal communities were attributed to environmental factors (plant-available P) and location of the vineyard, although the latter could also have been influenced by an unmeasured environmental factor. A total of 27 taxa of fungi were identified, including taxa from Glomus group B, based on the sequencing of 18S rDNA. Sequencing of the internal transcribed spacer rDNA yielded 30 different fungal taxa, which comprised eight different Glomeromycota taxa, including Glomus sinuosum and Glomus indicum. To our knowledge, this is the first report of grapevine colonisation by G. indicum.     

Influence of Habitat and Climate Variables on Arbuscular Mycorrhizal Fungus Community Distribution,as Revealed by a Case Study of Facultative Plant Epiphytism under Semiarid Conditions

In semiarid Mediterranean ecosystems, epiphytic plant species are practically absent, and only some species of palm trees can support epiphytes growing in their lower crown area, such as Phoenix dactylifera L. (date palm). In this study, we focused on Sonchus tenerrimus L. plants growing as facultative epiphytes in P. dactylifera and its terrestrial forms growing in adjacent soils. Our aim was to determine the possible presence of arbuscular mycorrhizal fungi (AMF) in these peculiar habitats and to relate AMF communities with climatic variations. We investigated the AMF community composition of epiphytic and terrestrial S. tenerrimus plants along a temperature and precipitation gradient across 12 localities. Epiphytic roots were colonized by AMF, as determined by microscopic observation; all of the epiphytic and terrestrial samples analyzed showed AMF sequences from taxa belonging to the phylum Glomeromycota, which were grouped in 30 AMF operational taxonomic units. The AMF community composition was clearly different between epiphytic and terrestrial root samples, and this could be attributable to dispersal constraints and/or the contrasting environmental and ecophysiological conditions prevailing in each habitat. Across sites, the richness and diversity of terrestrial AMF communities was positively correlated with rainfall amount during the most recent growing season. In contrast, there was no significant correlation between climate variables and AMF richness and diversity for epiphytic AMF communities, which suggests that the composition of AMF communities in epiphytic habitats appears to be largely determined by the availability and dispersion of fungal propagules from adjacent terrestrial habitats.     

Effects of long-term NP-fertilization on abundance and diversity of arbuscular mycorrhizal fungi under a maize cropping system

Diversity of arbuscular mycorrhizal fungi (AMF) in 27-year long-term NP-fertilization plots under a maize cropping system in Thailand was studied through spore morphological characterization. The plots received 0–0, 60–60, 120–120 and 180–180 kg N-P₂O₅ ha^–1 year^–1 as ammonium sulfate and triple superphosphate. The plots were sampled monthly for one year, the AMF spores were counted and morphotyped, and taxa were identified after morphotyping and monospecific pot culture. Spore number g^–1 soil, relative spore abundance and Shannon-Wiener indexes were calculated. Sixteen putative taxa were recorded from the field of which nine sporulated on maize roots in pot culture. The long-term fertilization caused decreases in AMF total spore numbers and variation in species diversity depended on sampling time. Effects of fertilization on spore number and also relative spore abundance varied with species and sampling time. Among the nine species sporulating under maize, only Acaulospora sp.1 showed no change (P > 0.003 after Bonferroni correction) in spore number with fertilization in the field; and was therefore classified as an AMF species insensitive to fertilization. Spores of Entrophospora schenckii, Glomus mosseae, Glomus sp.1, Glomus geosporum-like and Scutellospora fulgida, though they decreased in absolute numbers in response to fertilization, showed no change (P > 0.003 after Bonferroni correction) in relative abundance; these species were classified as AMF species slightly sensitive to fertilization. Three unidentified species of Glomus, though they decreased in absolute numbers in response to fertilization, showed decreases (P < 0.003 after Bonferroni correction) in relative abundance; these species were classified as AMF species highly sensitive to fertilization.     

Molecular Community Analysis of Arbuscular Mycorrhizal Fungi in Roots of Geothermal Soils in Yellowstone National Park (USA)

To better understand adaptation of plants and their mycorrhizae to extreme environmental conditions, we analyzed the composition of communities of arbuscular mycorrhizal fungi (AMF) in roots from geothermal sites in Yellowstone National Park (YNP), USA. Arbuscular mycorrhizal fungi were identified using molecular methods including seven specific primer pairs for regions of the ribosomal DNA that amplify different subgroups of AMF. Roots of Dichanthelium lanuginosum, a grass only occurring in geothermal areas, were sampled along with thermal and nonthermal Agrostis scabra and control plants growing outside the thermally influenced sites. In addition, root samples of Agrostis stolonifera from geothermal areas of Iceland were analyzed to identify possible common mycosymbionts between these geographically isolated locations. In YNP, 16 ribosomal DNA phylotypes belonging to the genera Archaeospora, Glomus, Paraglomus, Scutellospora, and Acaulospora were detected. Eight of these phylotypes could be assigned to known morphospecies, two others have been reported previously in molecular studies from different environments, and six were new to science. The most diverse and abundant lineage was Glomus group A, with the most frequent phylotype corresponding to Glomus intraradices. Five of the seven phylotypes detected in a preliminary sampling in a geothermal area in Iceland were also found in YNP. Nonthermal vegetation was dominated by a high diversity of Glomus group A phylotypes while nonthermal plants were not. Using multivariate analyses, a subset of three phylotypes were determined to be associated with geothermal conditions in the field sites analyzed. In conclusion, AMF communities in geothermal soils are distinct in their composition, including both unique phylotypes and generalist fungi that occur across a broad range of environmental conditions. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Role of native and exotic mycorrhizal symbiosis to develop morphological,physiological and biochemical responses coping with water drought of date palm,Phoenix dactylifera

Key Message

Arbuscular mycorrhizal (AM) symbiosis can improve date palm growth and alleviate drought-related impacts than non-mycorrhizal plants due to the ability of AMF for modifying plant metabolism and physiology.

Abstract

Date palm (Phoenix dactylifera L.) is an important agricultural and commercial crop in the North of Africa and Middle Eastern countries. During the last decade, date palm plantations were subjected to degradation due to an extensive exploitation and to drastic environmental conditions such as drought. Currently, there is a growing interest in the valorization of water due to environmental problems and economic aspects. The use of arbuscular mycorrhizal fungi (AMF) can offer a possibility to overcome these problems. The objective of this study was to study the influence of different Glomus species—Glomus intraradices, G. mosseae and Complex Aoufous (native AMF)—on the development of date palm grown under two water regimes (optimal irrigation, 75 % of field capacity or water deficit, 25 % of field capacity). Our results revealed that the beneficial effect of mycorrhizal symbiosis on plant growth depended on the fungal species and the water regime applied to the palm date seedling. While the native Complex Aoufous was the most effective in increasing the shoot height and biomass under well-watered conditions, G. intraradices was the most beneficial fungus for improving growth of plants that undergo restricted water supply. This positive effect of G. intraradices under drought conditions was not related to an enhancement of the antioxidant enzymatic activities in leaves; the association of palm date with G. intradices caused an increase in the elasticity of cell walls in leaves and allowed maintaining high water content in leaves without lowering leaf water potential under stressful conditions. The adequate selection of the AMF species is crucial for improving growth of palm date seedlings, and it must be in accordance with the water regime that will be applied to plants.     

Differences of arbuscular mycorrhizal fungal diversity and community between a cultivated land,an old field,and a never-cultivated field in a hot and arid ecosystem of southwest China

We investigated the spore density, species composition, and diversity of arbuscular mycorrhizal fungi (AMF) in a cultivated land (CL), an old field (OF), and a never-cultivated field (NCF), which are located adjacently in a slope in the hot and arid ecosystem of southwest China. AMF spores in the rhizosphere soils of representative plants in the three habitats were extracted by wet-sieving and decanting. A total of 47 taxa of AMF including 31 taxa from the genus Glomus, 8 from Acaulospora, 6 from Scutellospora, 1 from Entrophospora, and 1 from Gigaspora were extracted and identified morphologically. The highest spore density occurred in NCF, slightly lower in OF and lowest in CL, and the Shannon–Wiener index of species diversity was reversed. The dominant species of AMF were different in the three habitats. OF resembled NCF more than CL in AMF spore density, species richness, and community composition, which means that AMF community in the OF has been developing from cultivated land to natural habitat. Cluster analysis based on the similarity in AMF community composition indicated that the distribution of AMF was not random over space and that AMF community composition associated with a given plant species was greatly habitat-convergence. Following the cluster analysis, we hypothesized that the effect of habitats on AMF communities were greater than that of the host preference to AMF. L-F. Li and T. Li contributed equally to this work.     

The composition of arbuscular mycorrhizal fungal communities in the roots of a ruderal forb is not related to the forest fragmentation process

Land‐use changes and forest fragmentation have strong impact on biodiversity. However, little is known about the influence of new landscape configurations on arbuscular mycorrhizal fungal (AMF) community composition. We used 454 pyrosequencing to assess AMF diversity in plant roots from a fragmented forest. We detected 59 virtual taxa (VT; phylogenetically defined operational taxonomic units) of AMF – including 10 new VT – in the roots of Euphorbia acerensis. AMF communities were mainly composed of members of family Glomeraceae and were similar throughout the fragmented landscape, despite variation in forest fragment size (i.e. small, medium and large) and isolation (i.e. varying pairwise distances). AMF communities in forest fragments were phylogenetically clustered compared with the global, but not regional and local AMF taxon pools. This indicates that non‐random community assembly processes possibly related to dispersal limitation at a large scale, rather than habitat filtering or biotic interactions, may be important in structuring the AMF communities. In this system, forest fragmentation did not appear to influence AMF community composition in the roots of the ruderal plant. Whether this is true for AMF communities in soil and the roots of other ecological groups of host plants or in other habitats deserves further study.     

Seasonal Dynamics of Arbuscular Mycorrhizal Fungal Communities in Roots in a Seminatural Grassland

Symbiotic arbuscular mycorrhizal fungi (AMF) have been shown to influence both the diversity and productivity of grassland plant communities. These effects have been postulated to depend on the differential effects of individual mycorrhizal taxa on different plant species; however, so far there are few detailed studies of the dynamics of AMF colonization of different plant species. In this study, we characterized the communities of AMF colonizing the roots of two plant species, Prunella vulgaris and Antennaria dioica, in a Swedish seminatural grassland at different times of the year. The AMF small subunit rRNA genes were subjected to PCR, cloning, sequencing, and phylogenetic analysis. Nineteen discrete sequence types belonging to Glomus groups A and B and to the genus Acaulospora were distinguished. No significant seasonal changes in the species compositions of the AMF communities as a whole were observed. However, the two plant species hosted significantly different AMF communities. P. vulgaris hosted a rich AMF community throughout the entire growing season. The presence of AMF in A. dioica decreased dramatically in autumn, while an increased presence of Ascomycetes species was detected.     

Effects of Alliaria petiolata (garlic mustard; Brassicaceae) on mycorrhizal colonization and community structure in three herbaceous plants in a mixed deciduous forest

Herbaceous plant species are important components of forest ecosystems, and their persistence in forests may be affected by invasive plant species that reduce mycorrhizal colonization of plant roots. I examined the effect of the invasive plant Alliaria petiolata on arbuscular mycorrhizal fungi (AMF) colonizing the roots of three forest plant species. AMF root colonization and community structure was examined from plants that were growing either in the absence or presence of Alliaria under natural forest conditions. AMF root colonization varied among the plant species but was not significantly affected by Alliaria. With molecular methods, ～12 different taxa of AMF could be distinguished among the root samples, and these taxa belonged to the genera Acaulospora and Glomus, with Glomus dominating AMF communities. There were significant differences between the community of AMF colonizing roots of Maianthemum racemosum and Trillium grandiflorum, but only AMF communities of Maianthemum roots were significantly affected by Alliaria. Indicator species analysis found that an Acaulospora species type was a significant indicator of Maianthemum plants grown in the absence of Alliaria. These results suggest invasive plants like Alliaria may selectively suppress AMF fungi, and this suppression can affect AMF communities colonizing the roots of some native plant species.     

A method for assessing arbuscular mycorrhizal fungi group distribution in tree roots by intergenic transcribed sequence variation

We identified five taxonomic groups of arbuscular mycorrhizal fungi (AMF) inside roots of young trees of six species of legumes and six species of non-legumes from a field site in southern Costa Rica using an AMF group-specific PCR assay of the intergenic transcribed sequence and 18S rRNA gene fragment. Assay specificity was verified by cloning and sequencing representatives from four of the five AMF groups. We found no difference in overall AMF diversity levels between legumes and non-legumes or between plant species. Some groups of AMF may associate more frequently with legumes than others, as Glomus Group A (Glomus mosseae/intradices group) representatives were detected more frequently in legumes than non-legumes relative to Glomus Group B (Glomus etunicatum/claroideum) representatives.     

Effects of arbuscular mycorrhizal fungi on zinnia and the different colonization between Gigaspora and Glomus

Zinnia (Zinnia elegans) was inoculated with four arbuscular mycorrhizal fungi (AMF) i.e. Gigaspora margarita, Gigaspora rosea, Glomus intraradices, and Glomus mosseae, either singly or mixture of two species of Gigaspora and Glomus. Results indicated that Glomus significantly enhanced the leaf size and the shoot biomass. G. mosseae was more effective than G. intraradices. Only G. mosseae increased number and size of flowers. Mixed inoculations were not much effective in the growth-promotion than the corresponding singly inoculation with Glomus. Comparison of colonization percent demonstrated that the highest colonization by G. mosseae, and followed by G. intraradices and Gigaspora species. In semi-quantitative PCR amplifications, Glomus was dominant in the roots. Our results suggest that G. mosseae is good for inoculation to zinnia and the interaction between different AMF species should be given full consideration in the application.     

Diversity of AMF associated with Ammophila arenaria ssp. arundinacea in Portuguese sand dunes

Dune vegetation is essential for the formation and preservation of sand dunes and the protection of the coast line. Coastal sand dunes are harsh environments where arbuscular mycorrhizal fungi (AMF) play an important role in promoting plant establishment and growth. We present a study of the diversity of AMF associated with A. arenaria ssp. arundinacea in two locations of the Portuguese coast under a Mediterranean climate. These two locations were selected to compare a well-preserved dune system from a protected area with a degraded dune system from a public beach. AMF diversity was assessed mainly by cloning and sequencing of a fragment of the ribosomal SSU using the primer NS31 and AM1. Most of the 89 AMF clones obtained from the rhizosphere and roots of A. arenaria belonged to the genus Glomus, the largest clade within the Glomeromycota. Higher AMF diversity was found in the least disturbed site, in which spores of Scutellospora persica, Glomus constrictum and Glomus globiferum were found in the rhizosphere of A. arenaria.     

Effect of arbuscular mycorrhizal fungi on the acclimatization of micropropagated banana plantlets

 The effect of arbuscular mycorrhizal fungi (AMF) on micropropagated banana plantlets was evaluated during the acclimatization period. Plants were inoculated with Acaulospora scrobiculata, Glomus clarum or Glomus etunicatum. After cultivation in a greenhouse for 3 months, height, leaf area, fresh weight and dry matter of root and shoots, level of AMF colonization, nutrient level, photosynthesis and transpiration rate, water potential and stomatal conductance were measured. The number of AMF spores produced in each treatment was also determined. Plantlets inoculated with AMF had greater height, leaf area and fresh weight of shoots and roots, as well as higher rates of photosynthesis and transpiration than controls. Plants inoculated with Glomus were superior in most of the evaluated parameters. Accepted: 24 May 1999     

Species richness and spore abundance of arbuscular mycorrhizal fungi across distinct land uses in Western Brazilian Amazon

Arbuscular mycorrhizal fungi (AMF) were surveyed for species richness and abundance in sporulation in six distinct land uses in the western Amazon region of Brazil. Areas included mature pristine forest and sites converted to pasture, crops, agroforestry, young and old secondary forest. A total of 61 AMF morphotypes were recovered and 30% of them could not be identified to known species. Fungal communities were dominated by Glomus species but Acaulospora species produced the most abundant sporulation. Acaulospora gedanensis cf., Acaulospora foveata, Acaulospora spinosa, Acaulospora tuberculata, Glomus corymbiforme, Glomus sp15, Scutellospora pellucida, and Archaeospora trappei sporulated in all land use areas. Total spore numbers were highly variable among land uses. Mean species richness in crop, agroforestry, young and old secondary forest sites was twice that in pristine forest and pasture. fungal communities were dominated in all land use areas except young secondary forest by two or three species which accounted for 48% to 63% of all sporulation. Land uses influenced AMF community in (1) frequency of occurrence of sporulating AMF species, (2) mean species diversity, and (3) relative spore abundance. Conversion of pristine forest into distinct land uses does not appear to reduce AMF diversity. Cultural practices adopted in this region maintain a high diversity of arbuscular mycorrhizal fungi.     

Local and landscape drivers of arthropod diversity and decomposition processes in oil palm leaf axils

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典型农田土壤中丛枝菌根真菌-根际细菌互作及与氮磷利用的关系

在农业生态系统中,土壤微生物是土壤-作物系统养分循环的重要驱动力,其中丛枝菌根真菌(Arbuscularmycorrhizalfungi,AMF)能够促进作物对养分的吸收,适应逆境胁迫。【目的】进一步揭示AMF和根际细菌群落的跨界网络互作,挖掘与作物氮磷利用显著相关的关键微生物类群,揭示关键类群的生态网络特征。【方法】利用Illumina测序技术对3种典型农田旱地土壤(黑土、潮土和红壤)中AMF和根际细菌群落结构进行分析;构建互作网络并利用偏冗余分析、相关性分析探究了与氮磷利用相关的潜在关键类群。【结果】3种土壤中AMF与根际细菌均以正相互作用为主。不同土壤中AMF与根际细菌互作关系差异明显,在红壤中跨界互作最为密切,其中球囊霉属真菌(Glomus)与根际细菌中的放线菌(Actinobacteria)和变形菌(Proteobacteria)之间的交互作用最多。而在黑土中主要体现为根际细菌的界内互作。与氮磷利用率显著相关的关键微生物类群主要属于球囊霉属真菌、放线菌和α变形菌。【结论】典型旱地土壤中AMF与根际细菌的正相互关系对作物氮磷利用有潜在促进作用,关键类群在有机质和养分贫乏的红壤中可能起到更重要的作用。     

Fern-associated arbuscular mycorrhizal fungi are represented by multiple Glomus spp.: do environmental factors influence partner identity?

Symbioses involving arbuscular mycorrhizal fungi (AMF) are among the most important ecological associations for many plant species. The diversity of AMF associated with ferns, however, remains poorly studied. Using recently designed Glomus-specific primers, we surveyed the AMF community associated with ferns from deciduous, broad-leaved second-growth forest habitats at the eastern edge of the piedmont region of central Virginia, USA. Results indicate that this molecular approach may be a useful tool for detecting AMF in ferns compared to traditional techniques based on morphology. Over 30 potential fungal ribotypes were identified from eight fern species using denaturing gradient gel electrophoresis. Fungal ribotypes were found to differ widely in terms of (1) the number of fern partners with which they interact and (2) their relative frequency within each fern. Sequence analysis of fungal isolates from three species of fern indicated that the primers were generally highly specific for Glomus species but some non-target DNA was also amplified. Cloned polymerase chain reaction (PCR) products from Polystichum acrostichoides and Osmunda regalis revealed several phylogenetically distinct Glomus species. A single Glomus species was identified in the cloned PCR products from Botrychium virginianum. These findings challenge the hypothesis that the extent or degree of fern–fungal symbiosis is somehow tied to root complexity. Environmental factors appear to influence the suite of AMF that form partnerships with ferns. Some species of fern from similar habitats associated with dissimilar fungal partners (e.g., P. acrostichoides and Athyrium filix-femina var. asplenioides), whereas others harbored uniform fungal communities (e.g., Asplenium platyneuron). The significance of these data in terms of ecological and evolutionary dynamics of the AMF–fern symbiosis is discussed. Brittany West, Jessica Brandt, and Kay Holstien contributed equally to this work.     

Production and specificity of polyclonal antibodies against soluble proteins from the arbuscular mycorrhizal fungus Glomus intraradices

Rabbit polyclonal antibodies were produced against a soluble protein fraction from a vesicle and spore mixture of the arbuscular mycorrhizal fungus (AMF) Glomus intraradices. The protocol for isolation of vesicles and spores from plant roots was optimized to minimize debris contamination. Protein extract purification and preparation for immunization was adapted to increase protein content and immunogenicity. Active antisera were produced starting from the second boost immunization. Antibodies obtained were specific for surface antigens of AMF and revealed different patterns of soluble protein antigens in G. intraradices, G. constrictum and an unidentified Glomus species. Accepted: 6 December 2000     

Assessing the diversity of arbuscular mycorrhizal fungi in semiarid shrublands dominated by Artemisia tridentata ssp. wyomingensis

Variation in the abiotic environment and host plant preferences can affect the composition of arbuscular mycorrhizal (AMF) assemblages. This study analyzed the AMF taxa present in soil and seedlings of Artemisia tridentata ssp. wyomingensis collected from sagebrush steppe communities in southwestern Idaho, USA. Our aims were to determine the AMF diversity within and among these communities and the extent to which preferential AMF–plant associations develop during seedling establishment. Mycorrhizae were identified using molecular methods following DNA extraction from field and pot culture samples. The extracted DNA was amplified using Glomeromycota specific primers, and identification of AMF was based on phylogenetic analysis of sequences from the large subunit-D2 rDNA region. The phylogenetic analyses revealed seven phylotypes, two within the Claroideoglomeraceae and five within the Glomeraceae. Four phylotypes clustered with known species including Claroideoglomus claroideum, Rhizophagus irregularis, Glomus microaggregatum, and Funneliformis mosseae. The other three phylotypes were similar to several published sequences not included in the phylogenetic analysis, but all of these were from uncultured and unnamed glomeromycetes. Pairwise distance analysis revealed some phylotypes with high genetic variation. The most diverse was the phylotype that included R. irregularis, which contained sequences showing pairwise differences up to 12 %. Most of the diversity in AMF sequences occurred within sites. The smaller genetic differentiation detected among sites was correlated with differences in soil texture. In addition, multiplication in pot cultures led to differentiation of AMF communities. Comparison of sequences obtained from the soil with those from A. tridentata roots revealed no significant differences between the AMF present in these samples. Overall, the sites sampled were dominated by cosmopolitan AMF taxa, and young seedlings of A. tridentata ssp. wyomingensis were colonized in relation to the abundance of these taxa in the soil.     

Seasonal dynamics of arbuscular mycorrhizal fungal communities in roots in a seminatural grassland

Symbiotic arbuscular mycorrhizal fungi (AMF) have been shown to influence both the diversity and productivity of grassland plant communities. These effects have been postulated to depend on the differential effects of individual mycorrhizal taxa on different plant species; however, so far there are few detailed studies of the dynamics of AMF colonization of different plant species. In this study, we characterized the communities of AMF colonizing the roots of two plant species, Prunella vulgaris and Antennaria dioica, in a Swedish seminatural grassland at different times of the year. The AMF small subunit rRNA genes were subjected to PCR, cloning, sequencing, and phylogenetic analysis. Nineteen discrete sequence types belonging to Glomus groups A and B and to the genus Acaulospora were distinguished. No significant seasonal changes in the species compositions of the AMF communities as a whole were observed. However, the two plant species hosted significantly different AMF communities. P. vulgaris hosted a rich AMF community throughout the entire growing season. The presence of AMF in A. dioica decreased dramatically in autumn, while an increased presence of Ascomycetes species was detected.