Warming and rainfall reduction alter soil microbial diversity and co-occurrence networks and enhance pathogenic fungi in dryland soils.

In this 9-year manipulative field experiment, we examined the impacts of experimental warming (2 °C, W), rainfall reduction (30 % decrease in annual rainfall, RR), and their combination (W + RR) on soil microbial communities and native vegetation in a semi-arid shrubland in south-eastern Spain. Warming had strong negative effects on plant performance across five coexisting native shrub species, consistently reducing their aboveground biomass growth and long-term survival. The impacts of rainfall reduction on plant growth and survival were species-specific and more variable. Warming strongly altered the soil microbial community alpha-diversity and changed the co-occurrence network structure. The relative abundance of symbiotic arbuscular mycorrhizal fungi (AMF) increased under W and W + RR, which could help buffer the direct negative impacts of climate change on their host plants nutrition and enhance their resistance to heat and drought stress. Indicator microbial taxa analyses evidenced that the marked sequence abundance of many plant pathogenic fungi, such as Phaeoacremonium, Cyberlindnera, Acremonium, Occultifur, Neodevriesia and Stagonosporopsis, increased significantly in the W and W + RR treatments. Moreover, the relative abundance of fungal animal pathogens and mycoparasites in soil also increased significantly under climate warming. Our findings indicate that warmer and drier conditions sustained over several years can alter the soil microbial community structure, composition, and network topology. The projected warmer and drier climate favours pathogenic fungi, which could offset the benefits of increased AMF abundance under warming and further aggravate the severe detrimental impacts of increased abiotic stress on native vegetation performance and ecosystem services in drylands.

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.

Host preferences of arbuscular mycorrhizal fungi colonizing annual herbaceous plant species in semiarid Mediterranean prairies.

In this study, we have analyzed and compared the diversities of the arbuscular mycorrhizal fungi (AMF) colonizing the roots of five annual herbaceous species (Hieracium vulgare, Stipa capensis, Anagallis arvensis, Carduus tenuiflorus, and Avena barbata) and a perennial herbaceous species (Brachypodium retusum). Our goal was to determine the differences in the communities of the AMF among these six plant species belonging to different families, using B. retusum as a reference. The AMF small-subunit rRNA genes (SSU) were subjected to nested PCR, cloning, sequencing, and phylogenetic analysis. Thirty-six AMF phylotypes, belonging to Glomus group A, Glomus group B, Diversispora, Paraglomus, and Ambispora, were identified. Five sequence groups identified in this study clustered to known glomalean species or isolates: group Glomus G27 to Glomus intraradices, group Glomus G19 to Glomus iranicum, group Glomus G10 to Glomus mosseae, group Glomus G1 to Glomus lamellosum/etunicatum/luteum, and group Ambispora 1 to Ambispora fennica. The six plant species studied hosted different AMF communities. A certain trend of AMF specificity was observed when grouping plant species by taxonomic families, highlighting the importance of protecting and even promoting the native annual vegetation in order to maintain the biodiversity and productivity of these extreme ecosystems.

Assessing the diversity of AM fungi in arid gypsophilous plant communities.

In the present study, we used PCR-Single-Stranded Conformation Polymorphism (SSCP) techniques to analyse arbuscular mycorrhizal fungi (AMF) communities in four sites within a 10 km(2) gypsum area in Southern Spain. Four common plant species from these ecosystems were selected. The AM fungal small-subunit (SSU) rRNA genes were subjected to PCR, cloning, SSCP analysis, sequencing and phylogenetic analyses. A total of 1443 SSU rRNA sequences were analysed, for 21 AM fungal types: 19 belonged to the genus Glomus, 1 to the genus Diversispora and 1 to the Scutellospora. Four sequence groups were identified, which showed high similarity to sequences of known glomalean species or isolates: Glo G18 to Glomus constrictum, Glo G1 to Glomus intraradices, Glo G16 to Glomus clarum, Scut to Scutellospora dipurpurescens and Div to one new genus in the family Diversisporaceae identified recently as Otospora bareai. There were three sequence groups that received strong support in the phylogenetic analysis, and did not seem to be related to any sequences of AM fungi in culture or previously found in the database; thus, they could be novel taxa within the genus Glomus: Glo G4, Glo G2 and Glo G14. We have detected the presence of both generalist and potential specialist AMF in gypsum ecosystems. The AMF communities were different in the plant studied suggesting some degree of preference in the interactions between these symbionts.

Complexity of semiarid gypsophilous shrub communities mediates the AMF biodiversity at the plant species level.

The community composition of arbuscular mycorrhizal fungi (AMF) was analyzed in roots of Gypsophila struthium growing in gypsum soils under semiarid conditions. In order to investigate the effect of plant community degradation on the AMF biodiversity at the single species level, on the basis of the plant community complexity level, we selected four areas affected by degradation and shrub species spatial heterogeneity. The AM fungal community colonizing G. struthium was investigated from the morphological and molecular points of view. All plants were well colonized and showed a high level of infective AM propagules. Roots were analyzed by polymerase chain reaction, restriction fragment length polymorphism screening, and sequence analyses of the ribosomal DNA small subunit region. Four AM fungal types were identified and clustered into the AM fungal family: Glomeraceae, Glomus being the only taxon present. One fungal type was present in all the selected areas. Two fungal types are distinct from any previously published sequences and could be specific to gypsum soils. The chemical-physical properties of the soil were not correlated with the AMF diversity in roots. Our data show vegetation cover complexity-dependent differences in the AM fungal community composition.

The cover crop determines the AMF community composition in soil and in roots of maize after a ten-year continuous crop rotation.

Intensive agricultural practices are responsible for soil biological degradation. By stimulating indigenous arbuscular mycorrhizal fungi (AMF), cover cropping enhances soil health and promotes agroecosystem sustainability. Still, the legacy effects of cover crops (CCs) and the major factors driving the AM fungal community are not well known; neither is the influence of the specific CC. This work describes a field experiment established in Central Spain to test the effect of replacing winter fallow by barley (Hordeum vulgare L.) or vetch (Vicia sativa L.) during the intercropping of maize (Zea mays L.). We examined the community composition of the AMF in the roots and rhizosphere soil associated with the subsequent cash crop after 10 years of cover cropping, using Illumina technology. The multivariate analysis showed that the AMF communities under the barley treatment differed significantly from those under fallow, whereas no legacy effect of the vetch CC was detected. Soil organic carbon, electrical conductivity, pH, Ca and microbial biomass carbon were identified as major factors shaping soil AMF communities. Specific AMF taxa were found to play a role in plant uptake of P, Fe, Zn, Mn, and Cd, which may shed light on the functionality of these taxa. In our conditions, the use of barley as a winter CC appears to be an appropriate choice with respect to promotion of AMF populations and biological activity in agricultural soils with intercropping systems. However, more research on CC species and their legacy effect on the microbial community composition and functionality are needed to guide decisions in knowledge-based agriculture.

The impact of tillage practices on arbuscular mycorrhizal fungal diversity in subtropical crops.

Arbuscular mycorrhizal fungi (AMF) are a main component of soil microbiota in most agrosystems. As obligately mutualistic symbionts, they colonize the roots of the majority of plants, including crop plants. We used molecular techniques to investigate how different tillage systems (moldboard, shred-bedding, subsoil-bedding, and no tillage) can influence the AM fungal community colonizing maize, bean, and sorghum roots in an experimental site located in northern Tamaulipas, Mexico. Roots from 36 plants were analyzed using AM fungal-specific primers to partially amplify the small subunit (SSU) of the ribosomal DNA genes. More than 880 clones were screened for restriction fragment length polymorphism (RFLP) variation, and 173 of these were sequenced. Ten AM fungal types were identified and clustered into three AM fungal families: Gigasporaceae, Glomaceae, and Paraglomaceae. Glomus was the dominating taxon in all the samples. Four of the 10 identified types were distinct from any previously published sequences and could correspond to either known unsequenced species or unknown species. The fungal diversity was low in the four agriculture management systems, but the multidimensional scaling (MDS) analysis and log-linear-saturated model indicated that the composition of the AMF community was significantly affected by the tillage system. In conclusion, since some fungal types were treatment specific, agricultural practices could directly or indirectly influence AM biodiversity.

Arbuscular mycorrhizal fungi inoculation mediated changes in rhizosphere bacterial community structure while promoting revegetation in a semiarid ecosystem.

The main goal of this study was to assess the effect of the inoculation of four autochthonous shrub species with the arbuscular mycorrhizal (AM) fungus Rhizophagus intraradices on the rhizosphere bacterial community and to ascertain whether such an effect is dependent on the host plant species. Additionally, analysis of rhizosphere soil chemical and biochemical properties was performed to find relationships between them and the rhizosphere bacterial communities. Non-metric multidimensional scaling analysis and subsequent permutational multivariate analysis of variance revealed differences in bacterial community composition and structure between non-inoculated and inoculated rhizospheres. Moreover, an influence of the plant species was observed. Different bacterial groups were found to be indicator taxonomic groups of non-inoculated and inoculated rhizospheres, Gemmatimonadetes and Anaerolineaceae, respectively, being the most notable indicators. As shown by distance based redundancy analysis, the shifts in bacterial community composition and structure mediated by the inoculation with the AM fungus were mainly related to changes in plant nutrients and growth parameters, such as the shoot phosphorus content. Our findings suggest that the AM fungal inoculum was able to modify the rhizosphere bacterial community assemblage while improving the host plant performance.

Arbuscular mycorrhizal fungal assemblages in biological crusts from a Neotropical savanna are not related to the dominant perennial Trachypogon.

Knowledge of the arbuscular mycorrhizal fungal assemblages in the Trachypogon savanna ecosystems is very important to a better understanding of the ecological processes mediated by this soil microbial group that affects multiple ecosystem functions. Considering the hypothesis that the biocrusts can be linked to vegetation through the arbuscular fungi mycelial network, the objectives proposed in this study were to determine (i) whether there are arbuscular mycorrhizal fungi (AMF) in the biocrusts (ii) whether arbuscular mycorrhizal fungal assemblages are linked to the Trachypogon patches, and (iii) whether the composition of the assemblages is related to soil properties affected by microbiological activity. The community structure of the AMF was investigated in three habitats: rhizospheric soil and roots of Trachypogon vestitus, biological soil crusts, and bare soil. The canonical correspondence analysis showed that two soil properties related to enzymatic activity (protease and ß-glucosidase) significantly affected the community composition of the AMF. The biocrusts in the Venezuelan savanna are colonized by an AM fungal community linked to that of the bare soil and significantly different from that hosted by the roots of the surrounding T. vestitus, suggesting that assemblages of AMF in biocrusts might be related more closely to those of annual plant species appearing in favorable conditions.

Arbuscular mycorrhizal fungi communities in a coral cay system (Morrocoy, Venezuela) and their relationships with environmental variables.

Knowledge of the natural diversity of arbuscular mycorrhizal fungi (AMF) and understanding of their biogeographical patterns and what drive them might help to the maintenance and preservation of ecosystems under a changing environment. The objective of this study was to evaluate the contribution of different environmental factors to the determination of the composition of AMF assemblages in representative sites within the Morrocoy National Park (Venezuela). The community structure of the AMF under the canopy of Coccoloba uvifera was investigated in four cays (Borracho, Muerto, Peraza, and Paiclás) and one mainland location (Las Luisas). Based on partial sequences of the nuclear small subunit ribosomal DNA gene, the AM fungi in soil samples were divided into 31 operational taxonomic units, grouped in eight families. The canonical correspondence analysis showed that environmental factors related to insularity (the mean annual rainfall, the distance to the mainland coast, and the cay land area) and a soil property related to biological activity (the total carbohydrate content) were significantly related to the distribution of the AMF communities.

Subcellular localization of nerve growth factor receptors in identified cells of the rat nucleus basalis magnocellularis: an immunocytochemical study.

The subcellular location of nerve growth factor receptor in the ventromedial portion of rat globus pallidus was investigated with affinity-purified monoclonal 192-IgG following the unlabelled antibody peroxidase-antiperoxidase immunocytochemical procedure. At the light microscopic level, punctate immunoreaction product was observed in the perinuclear region and in the plasma membrane of large, probably cholinergic neurons. Examination in the electron microscope of these neurons confirmed that nerve growth factor receptor-stained cells were basal forebrain cholinergic neurons. Within these cells, immunostaining occurred in the Golgi apparatus, in multivesicular bodies and, occasionally, in rough endoplasmic reticulum cisternae and the nuclear envelope. Moreover, patches of immunoreactivity were observed associated with the outer surface of the plasma membrane of the soma and their proximal dendrites and also with the plasma membrane of distal dendrites showing scarcity of synaptic input. Positive immunostaining was never observed in synaptic clefts, but filled the space between the plasma membranes of immunoreactive neurons and those of thin glial processes in their vicinity. The location of membrane nerve growth factor receptor in close apposition to membranes of neighbouring astrocytes rather than near synaptic complexes, suggests that glial cells may be a physiological source of nerve growth factor.

Application of composted urban residue enhanced the performance of afforested shrub species in a degraded semiarid land.

Improvement of physical-chemical soil quality is a key step for carrying out revegetation programs of degraded lands in Mediterranean semiarid areas. Organic residue addition may restore the quality of these areas. A field experiment was conducted in a silt-loam soil (Typic Petrocalcid) from a degraded semiarid Mediterranean area to evaluate the effect of the addition of a composted urban residue on soil aggregate stability, bulk density and chemical properties and on the establishment of Pistacia lentiscus and Retama sphaerocarpa seedlings. The composted residue was applied at a rate of 6.7 kg m(-2) before planting. The nutrient content (NPK), total organic C and water soluble C were increased and bulk density was decreased, in the rhizosphere soil of both shrub species, by the composted residue. The addition of composted residue significantly increased the soil aggregate stability by about 22% for both shrub species. The beneficial effect of the composted residue on soil quality still persisted 18 months after addition. Eighteen months after planting, the addition of composted residue to soil had increased significantly the production of shoot biomass by P. lentiscus and R. sphaerocarpa, by about 160% and 320% respectively, compared to control values. Composted residue addition to soil can be considered an effective preparation method of a degraded area for carrying out successful revegetation programs with Mediterranean shrubs under semiarid conditions.

Establishment of two ectomycorrhizal shrub species in a semiarid site after in situ amendment with sugar beet, rock phosphate, and Aspergillus niger.

A field experiment was carried out to assess the effectiveness of the addition of sugar beet, rock phosphate, and Aspergillus niger directly into the planting hole, and the mycorrhizal inoculation of seedlings with Scleroderma verrucosum, for promotion of plant growth of Cistus albidus L. and Quercus coccifera L. and enhancement of soil physicochemical, biochemical, and biological properties, in a degraded semiarid Mediterranean area. One year after planting, the available phosphorus content in the amended soils of both species was about fourfold higher than in the nonamended soil. Amendment addition increased the aggregate stability of the rhizosphere of C. albidus (by 56% with respect to control soil) while the mycorrhizal inoculation increased only the aggregate stability of the rhizosphere of Q. coccifera (by 13% with respect to control soil). Biomass C content and enzyme activities (dehydrogenase, urease, protease-BAA, acid phosphatase, and beta-glucosidase) of the rhizosphere of C. albidus were increased by amendment addition but not by mycorrhizal inoculation. Both treatments increased enzyme activities of the rhizosphere of Q. coccifera. The mycorrhizal inoculation of the seedlings with S. verrucosum was the most effective treatment for stimulating the growth of C. albidus (by 469% with respect to control plants) and Q. coccifera (by 74% with respect to control plants). The combined treatment, involving mycorrhizal inoculation of seedlings and addition of the amendment directly into soil, had no additive effect on the growth of either shrub species.

Changes in physical and biological soil quality indicators in a tropical crop system (Havana, Cuba) in response to different agroecological management practices.

The objective of our study was to assess the response of physical (aggregate stability and bulk density) and biological (enzyme activities and microbial biomass) soil quality indicators to the adoption of agroecological management practices, such as the planting of forage species (forage area) and the rotation of local crops (polycrop area), carried out in a representative tropical pasture on an integrated livestock-crop farm. The pasture system was used as control (pasture area). In all three areas, the values of water-soluble C were higher in the rainy season compared to the dry season. Pasture and forage areas had the highest percentage of stable aggregates in the rainy season, while polycrops developed soils with less stable aggregates. Soil bulk density was lower in the pasture and forage areas than in the polycrop area. In the pasture area, the microbial biomass C values, dehydrogenase, urease, protease-BAA, acid phosphatase, and beta-glucosidase activities were higher than in the forage and polycrop areas, particularly in the dry season. The highest increase in the microbial biomass C in the rainy season, with respect to the dry season, was recorded in the pasture area (about 1.2-fold). In conclusion, the planting of forage species can be considered an effective practice for carrying out sustainable, integrated livestock-crop systems, due to its general maintenance of soil quality, while the adoption of polycrop rotations appears to be less favorable because it decreases soil quality.