Fermentation of sugar beet waste by Aspergillus niger facilitates growth and P uptake of external mycelium of mixed populations of arbuscular mycorrhizal fungi

Sugar beet waste has potential value as a soil amendment and this work studied whether fermentation of the waste by Aspergillus niger would influence the growth and P uptake of arbuscular mycorrhizal (AM) fungi. Plants were grown in compartmentalised growth units, each with a root compartment (RC) and two lateral root-free compartments (RFC). One RFC contained untreated soil while the other RFC contained soil, which was uniformly mixed with sugar beet waste, either untreated (SB) or degraded by A. niger (ASB) in a rock phosphate (RP)-supplied medium. The soil in each pair of RFC was labelled with ³³P and ³²P in order to measure P uptake by the AM fungal mycelium, of which length density was also measured. Whole cell fatty acid (WCFA) signatures were used as biomarkers of the AM fungal mycelium and other soil microorganisms. The amount of biomarkers of saprotrophic fungi and both Gram-positive and Gram-negative bacteria was higher in SB than in ASB treatments. Whilst ASB increased growth and activity of AM mycelium, SB had the opposite effect. Moreover, shoot P content was increased by the addition of ASB, and by inoculation with AM fungi. Modification of soil microbial structure and production of exudates by A. niger, as a consequence of fermentation process of sugar beet waste, could possibly explain the increase of AM growth in ASB treatments. On the other hand, the highest P uptake was a result of the solubilisation of rock phosphate by A. niger during the fermentation.     

Pre-symbiotic and symbiotic interactions between Glomus intraradices and two Paenibacillus species isolated from AM propagules. In vitro and in vivo assays with soybean (AG043RG) as plant host

Two indole-producing Paenibacillus species, known to be associated with propagules of arbuscular mycorrhizal (AM) fungi, were examined for their mycorrhization helper bacteria activity at pre-symbiotic and symbiotic stages of the AM association. The effects were tested under in vitro and in vivo conditions using an axenically propagated strain of the AM fungus Glomus intraradices and Glycine max (soybean) as the plant host. The rates of spore germination and re-growth of intraradical mycelium were not affected by inoculation with Paenibacillus strains in spite of the variation of indole production measured in the bacterial supernatants. However, a significant promotion in pre-symbiotic mycelium development occurred after inoculation of both bacteria under in vitro conditions. The Paenibacillus rhizosphaerae strain TGX5E significantly increased the extraradical mycelium network, the rates of sporulation, and root colonization in the in vitro symbiotic association. These results were also observed in the rhizosphere of soybean plants grown under greenhouse conditions, when P. rhizosphaerae was co-inoculated with G. intraradices. However, soybean dry biomass production was not associated with the increased development and infectivity values of G. intraradices. Paenibacillus favisporus strain TG1R2 caused suppression of the parameters evaluated for G. intraradices during in vitro symbiotic stages, but not under in vivo conditions. The extraradical mycelium network produced and the colonization of soybean roots by G. intraradices were promoted compared to the control treatments. In addition, dual inoculation had a promoting effect on soybean biomass production. In summary, species of Paenibacillus associated with AM fungus structures in the soil, may have a promoting effect on short term pre-symbiotic mycelium development, and little impact on AM propagule germination. These findings could explain the associations found between some bacterial strains and AM fungus propagules.     

Mycelium of arbuscular mycorrhizal fungi increases soil water repellency and is sufficient to maintain water-stable soil aggregates

Using an in vitro bioreactor system in which the arbuscular mycorrhizal (AM) fungus Glomus intraradices was grown in a soil devoid of detectable living microbes, we could show that the mycelium of this fungus contributed to the maintenance of water-stable soil aggregates and increased soil water repellency, as measured by water drop penetration time. This is to our knowledge the first demonstration of a causal link between AM fungal growth and water repellency of soil aggregates. Our results also place AM fungal contributions to soil aggregation on a firm mechanistic footing by showing that hyphae are sufficient to produce effects, in the absence of other soil biota, which have always been included in previous studies.     

Interactions between the external mycelium of the mycorrhizal fungus Glomus intraradices and other soil microorganisms as affected by organic matter

The influence of organic matter on the interactions between external mycelium of the arbuscular mycorrhizal (AM) fungus Glomus intraradices, the bacterium Burkholderia cepacia and other soil microorganisms was studied in a root-free sand environment. Organic matter amendment, in terms of ground barley leaves, markedly increased the growth of the external mycelium of G. intraradices as estimated both with the fatty acid biomarker 16:1ω5 and hyphal length measurements. Mycelial proliferation of G. intraradices in sand with organic matter was unaffected by both inoculation with B. cepacia and a soil filtrate containing a mixed population of indigenous microorganisms. On the other hand, in the absence of organic matter, both inoculation with B. cepacia and the soil filtrate reduced the growth of G. intraradices, as estimated with measurements of 16:1ω5. In contrast, B. cepacia inoculation increased hyphal length density of G. intraradices in the absence of organic matter. Overall, the presence of external mycelium of G. intraradices increased the bacterial biomass and counteracted a suppressive effect of B. cepacia on the growth of saprotrophic fungi.     

The mycorrhizal fungus Glomus intraradices and rock phosphate amendment influence plant growth and microbial activity in the rhizosphere of Acacia holosericea

Plants inoculated with arbuscular mycorrhizal (AM) fungi utilize more soluble phosphorus from soil mineral phosphate than non-inoculated plants. However, there is no information on the response of soil microflora to mineral phosphate weathering by AM fungi and, in particular, on the catabolic diversity of soil microbial communities.The AM fungus, Glomus intraradices was examined for (i) its effect on the growth of Acacia holosericea, (ii) plant-available phosphate and (iii) soil microbial activity with and without added rock phosphate.After 4-months culture, AM fungal inoculation significantly increased the plant biomasses (by 1.78× and 2.23× for shoot and root biomasses, respectively), while mineral phosphate amendment had no effect in a sterilized soil. After 12-months culture, the biomasses of A. holosericea plants growing in a non-sterilized soil amended with mineral phosphate were significantly higher than those recorded in the control treatment (by 2.5× and 5× for shoot and root biomasses, respectively). The fungal inoculation also significantly stimulated plant growth, which was significantly higher than that measured in the mineral phosphate treatment. When G. intraradices and mineral phosphate were added together to the soil, shoot growth were significantly stimulated over the single treatments (inoculation or amendment) (1.45×). The P leaf mineral content was also higher in the G. intraradices+mineral phosphate treatment than in G. intraradices or rock phosphate amendment. Moreover, the number of fluorescent pseudomonads has been significantly increased when G. intraradices and/or mineral phosphate were added to the soil. By using a specific type of multivariate analysis (co-inertia analysis), it has been shown that plant growth was positively correlated to the metabolization of ketoglutaric acid, and negatively linked to the metabolisation of phenylalanine and other substrates, which shows that microbial activity is also affected.G. intraradices inoculation is highly beneficial to the growth of A. holosericea plants in controlled conditions. This AM symbiosis optimises the P solubilization from the mineral phosphate and affects microbial activity in the hyphosphere of A. holosericea plants.     

Differential effects of soil disturbance and plant residue retention on function of arbuscular mycorrhizal (AM) symbiosis are not reflected in colonization of roots or hyphal development in soil

The effects of soil disturbance and residue retention on the functionality of the symbiosis between medic (Medicago truncatula L.) and arbuscular mycorrhizal fungi (AMF) were assessed in a two-stage experiment simulating a crop rotation of wheat (Triticum aestivum L.) followed by medic. Plants were inoculated or not with the AMF, Glomus intraradices and Gigaspora margarita, separately or together. The contribution of the arbuscular mycorrhizal (AM) pathway for P uptake was determined using ³²P-labeled soil in a small hyphal compartment accessible only to hyphae of AMF. In general AM colonization was not affected by soil disturbance or residue application and disturbance did not affect hyphal length densities (HLDs) in soil. At 4 weeks disturbance had a negative effect on growth and phosphorus (P) uptake of plants inoculated with G. margarita, but not G. intraradices. By 7 weeks disturbance reduced growth of plants inoculated with G. margarita or AMF mix and total P uptake in all inoculated plants. With the exception of plants inoculated with G. margarita in disturbed soil at 4 weeks, the AM pathway made a significant contribution to P uptake in all AM plants at both harvests. Inoculation with both AMF together eliminated the negative effects of disturbance on AM P uptake and growth, showing that a fungus insensitive to disturbance can compensate for loss of contribution of a sensitive one. Application of residue increased growth and total P uptake of plants but decreased ³²P in plants inoculated with the AMF mix in disturbed soil, compared with plants receiving no residue. The AMF responded differently to disturbance and G. intraradices, which was insensitive to disturbance, compensated for lack of contribution by the sensitive G. margarita when they were inoculated together. Colonization of roots and HLDs in soil were not good predictors of the outcomes of AM symbioses on plant growth, P uptake or P delivery via the AM pathway.     

Suppression of other soil microorganisms by mycelium of arbuscular mycorrhizal fungi in root-free soil

The influence of mycelium of two arbuscular mycorrhizal (AM) fungi, Glomus intraradices and Glomus mosseae, on other soil microorganisms, was examined in root-free soil with and without organic substrate amendment in terms of cellulose. The AM fungi were grown in symbiosis with cucumber in a compartmented growth system, which allowed AM fungal external mycelium to grow into root-free compartments. The fungicide Benomyl was applied to the root-free compartments to create an alternative non-mycorrhizal control treatment. Whole cell biomarker fatty acids were employed to quantify different groups of soil microorganisms including the two AM fungi. Abundance of most microbial groups were reduced by external mycelium of both AM fungi, though differential effects on the microbial community composition were observed between the two AM fungi as revealed from principal component analysis. Inhibition of other soil microorganisms was more pronounced in root-free soil with mycelium of G. mosseae than with mycelium of G. intraradices. In general, cellulose increased the amount of biomarker fatty acids of most groups of soil microorganisms, but cellulose did not affect the influence of AM fungi on other soil microorganisms. Benomyl suppressed growth of the external mycelium of the two AM fungi and had limited non-target effects on other microbial groups. In conclusion, our results show differential effects of external mycelium of AM fungi on other soil microbial communities, though both AM fungi included in the study overall inhibited most microbial groups as examined using whole cell biomarker fatty acids.     

Fraxinus-Glomus-Pisolithus symbiosis: Plant growth and soil aggregation effects

It has been established that arbuscular mycorrhizal (AM) fungi are involved in the conservation of soil structure. However, the effect of ectomycorrhizal (EM) fungi alone or in interaction with AM fungi in soil structure has been much less studied. This experiment evaluated EM and AM fungi effects on soil aggregation and plant growth. Ash plants (Fraxinus uhdei) were grown in pots, and were inoculated with Glomus intraradices and Pisolithus tinctorius separately but also in combination. Our results showed that F. uhdei established a symbiotic association with EM and AM fungi, and that these organisms, when interacting, showed synergistic and additive effects on plant growth compared to singly inoculated treatments. EM and AM fungi prompted changes in root morphology and increased water-stable aggregates. AM fungi affect mainly small-sized macroaggregates, while EM and EM-AM fungi interaction mainly affected aggregates bigger than 0.5 mm diameter. These results suggest that ectomyccorrhizal as well as arbuscular mycorrhizal fungi should be considered in restoration programs with Fraxinus plants.     

Interactions between the arbuscular mycorrhizal fungus Glomus intraradices and the plant growth promoting rhizobacteria Paenibacillus polymyxa and P. macerans in the mycorrhizosphere of Cucumis sativus

Interactions between the arbuscular mycorrhizal (AM) fungus Glomus intraradices and bacteria from the genus Paenibacillus (P. macerans and P. polymyxa) were examined in a greenhouse pot experiment with Cucumis sativus with and without organic matter amendment (wheat bran). P. polymyxa markedly suppressed AM fungus root colonization irrespective of wheat bran amendment, whereas P. macerans only suppressed AM fungus root colonization in combination with wheat bran amendment. Dual inoculation with P. macerans and G. intraradices in combination with wheat bran amendment also caused severe plant growth suppression. Inoculation with G. intraradices was associated with increased levels of dehydrogenase activity and available P in the growth substrate suggesting that mycorrhiza formation accelerated the decomposition of organic matter resulting in mobilization of phosphorus. Inoculation with both Paenibacillus species increased all measured microbial fatty acid biomarkers in the cucumber rhizosphere, except for the AM fungus biomarker 16:1ω5, which was reduced, though not significantly. Similarly, inoculation with G. intraradices increased all measured microbial fatty acid biomarkers in the cucumber rhizosphere, except for the Gram-positive bacteria biomarker 15:0 anteiso, which was overall decreased by G. intraradices inoculation. In combination with wheat bran amendment G. intraradices inoculation caused a 39% reduction in the amount of 15:0 anteiso in the treatment with P. polymyxa, suggesting that G. intraradices suppressed P. polymyxa in this treatment. In conclusion, plant growth promoting species of Paenibacillus may have suppressive effects of AM fungi and plant growth, especially in combination with organic matter amendment. The use of an inert plant growth media in the present study allowed us to study rhizosphere microbial interactions in a relative simple substrate with limited interference from other soil biota. However, the results obtained in the present work mainly show potential interactions and should not be directly extrapolated to a soil situation.     

Elevated CO2 increases the effect of an arbuscular mycorrhizal fungus and a plant-growth-promoting rhizobacterium on structural stability of a semiarid agricultural soil under drought conditions

In arid and semiarid Mediterranean regions, an increase in the severity of drought events could be caused by rising atmospheric CO₂ concentrations. We studied the effects of the interaction of CO₂, water supply and inoculation with a plant-growth-promoting rhizobacterium (PGPR), Pseudomonas mendocina Palleroni, or inoculation with an arbuscular mycorrhizal (AM) fungus, Glomus intraradices (Schenk & Smith), on aggregate stabilisation of the rhizosphere soil of Lactuca sativa L. cv. Tafalla. The influence of such structural improvements on the growth of lettuce was evaluated. We hypothesised that elevated atmospheric CO₂ concentration would increase the beneficial effects of inoculation with a PGPR or an AM fungus on the aggregate stability of the rhizosphere soil of lettuce plants. Leaf hydration, shoot dry biomass and mycorrhizal colonisation were decreased significantly under water-stress conditions, but this decrease was more pronounced under ambient vs elevated CO₂. The root biomass decreased under elevated CO₂ but only in non-stressed plants. Under elevated CO₂, the microbial biomass C of the rhizosphere of the G. intraradices-colonised plants increased with water stress. Bacterial and mycorrhizal inoculation and CO₂ had no significant effect on the easily-extractable glomalin concentration. Plants grown under elevated CO₂ had a significantly higher percentage of stable aggregates under drought stress than under well-watered conditions, particularly the plants inoculated with either of the assayed microbial inocula (about 20% higher than the control soil). We conclude that the contribution of mycorrhizal fungi and PGPR to soil aggregate stability under elevated atmospheric CO₂ is largely enhanced by soil drying.     

Increases in growth and nutrient uptake of alfalfa grown in soil amended with microbially-treated sugar beet waste

Sugar beet waste (SB), treated by Aspergillus niger under the conditions of 10-, 20-, and 30-day-solid state fermentation, supplemented or not with rock phosphate (RP), was added to a soil-plant system. Plant growth responses depended on the time period of preincubation of the agrowaste characterized by different lignocellulosic composition and N and P contents before introduction into soil. Maximum growth and nutrient uptake of alfalfa during three crop cycles were recorded in a soil amended with microbially-treated SB waste+RP. This effect was more pronounced in treatments with arbuscular mycorrhizal (AM) fungus grown in soil enriched with 10- and 20-day-microbially-treated SB+RP, when the respective average total plant growth increased 233% and 343% over the non-mycorrhizal control containing untreated SB. Compared to other treatments, plant mycorrhization was ineffective when 30-day-treated agrowaste was used. Similarly, plant nodule numbers and uptake of metal ions depended on both the time period of waste preincubation and mycorrhization.     

Growth promotion of plants by plant growth-promoting rhizobacteria under greenhouse and two different field soil conditions

This study was conducted with sugar beet in greenhouse and field at two soil type with different organic matter (containing 2.4 and 15.9% OM, referred as the low- and high-OM soil) conditions in order to investigate seed inoculation of sugar beet, with five N₂-fixing and two phosphate solubilizing bacteria in comparison to control and mineral fertilizers (N and P) application. Three bacterial strains dissolved P; all bacterial strains fixed N₂ and significantly increased growth of sugar beet. In the greenhouse, inoculations with PGPR increased sugar beet root weight by 2.8-46.7% depending on the species. Leaf, root and sugar yield were increased by the bacterial inoculation by 15.5-20.8, 12.3-16.1, and 9.8-14.7%, respectively, in the experiment of low- and high-OM soil. Plant growth responses were variable and dependent on the inoculants strain, soil organic matter content, growing stage, harvest date and growth parameter evaluated. The effect of PGPR was greater at early growth stages than at the later. Effective Bacillus species, such as OSU-142, RC07 and M-13, Paenibacillus polymyxa RC05, Pseudomonas putida RC06 and Rhodobacter capsulatus RC04 may be used in organic and sustainable agriculture.     

Distribution of the arbuscular mycorrhizal biomarker C16:1cis11 among neutral, glyco and phospholipids extracted from soil during the reproductive growth of corn

Arbuscular mycorrhiza (AM) fungi form symbiotic relationships with the majority of land plants and are known for their positive effects on plant P acquisition and soil quality. The extramatrical growth of the mycelium is a key factor in nutrient acquisition by the symbiont. Soil grinding and extraction/fractionation of lipids were used in a field experiment to identify probable sources of the AM biomarker C16:1cis11 and its functional significance during reproductive growth of corn (Zea mays L.). Chambers, enclosed with a 1 mm mesh fabric to allow roots and hyphae to pass into the enclosed soil volume, were installed in two field sites cropped to continuous corn in central Nebraska. The chambers were installed at tasselling and removed after 3, 6 and 9 weeks. Soil from the chambers was analyzed by ester-linked fatty acid (EL-FAME) and chloroform-methanol fatty acid (CM-FAME) analysis. We also separated and analyzed the neutral lipid (NLFA), glycolipid (GLFA) and phospholipid (PLFA) fatty acid fractions. Roller milling the soil gave up to two-fold increases in the recovery of EL- and CM-FAMEs common to saprophytic fungi (C16:0, C18:1cis9, C18:2cis9,12) and AM fungi (C16:0, C16:1cis11, C18:1cis11) but not those specific to bacteria or fauna. Resistant AM fungal structures were enriched in NLFA and GLFA C16:1cis11, but not PLFA, indicating that storage lipids and possibly cell-wall lipids are released by roller milling. Similar proportional increases in C16:1cis11 on roller milling indicates that mild alkaline hydrolysis (EL-FAME) is as inefficient as chloroform-methanol (CM-FAME) in extracting lipids from AM spores. EL- and CM-FAME C16:1cis11 increased by one-third between R5 and R6, indicating C allocation from the plant to the AM fungus during the reproductive stages of corn. This increase was attributed to accumulation of NLFA and GLFA in lipid-containing structures of the extramatrical mycelium and AM structures within roots, not increased sporulation. We propose EL-FAME C16:1cis11 as a simple measure of AM biomass in soils that largely reflects the AM hyphal network important to nutrient acquisition by the plant.     

The application of an organic amendment modifies the arbuscular mycorrhizal fungal communities colonizing native seedlings grown in a heavy-metal-polluted soil

A mesocosm experiment was conducted to investigate whether communities of arbuscular mycorrhizal (AM) fungi associated with roots of native (Piptatherum miliaceum, Retama sphaerocarpa, Psoralea bituminosa, Coronilla juncea, and Anthyllis cytisoides) and for comparison (Lolium perenne) seedlings in a heavy-metal-contaminated, semiarid soil were affected by the application of composted sugar beet waste. We also investigated whether there were relation between AMF diversity and metal concentration (Al, Cd, Cu, Fe, Mn, Pb and Zn) and total P in shoot as well as some soil parameters (total organic carbon and total N) when the SB waste was added to the soil. We analyzed a portion of approximately 795 base pairs of the small-subunit (SSU) rRNA gene by nested PCR, cloning, sequencing, and phylogenetic analyses. Twelve different AMF sequence types were distinguished: seven of these belonged to Glomus group A, one to Glomus group B, one to Diversispora, one to Archaeospora, and two to Paraglomus. The AM fungal populations colonizing roots in a heavy-metal-polluted soil were quite dependent on the host plant, the highest diversity values being obtained in authochtonous plants recognized as metallophytes, such as P. bituminosa, and in an allochtonous, invasive species (L. perenne). No significant correlation was found between AMF diversity and plant metal concentration and soil parameters. Excepting P. bituminosa, when sugar beet waste was added to soil, the populations of AM fungi in roots increased and the shoot metal concentrations decreased in all host plant species studied. Therefore, the addition of sugar beet waste can be considered a good strategy for the remediation and/or phytostabilization of mine tailing sites.     

Physiological characteristics (SDH and ALP activities) of arbuscular mycorrhizal colonization as affected by Bacillus thuringiensis inoculation under two phosphorus levels

The effect of Bacillus thuringiensis (B.t.) inoculation on plant growth and on the intra- and extraradical mycorrhizal development of lettuce roots colonized by Glomus mosseae or Glomus intraradices was examined in an inert, soil-less substrate. Histochemical determination of succinate dehydrogenase (SDH) and alkaline phosphatase (ALP) activities which indicate active fungal metabolism was carried out at two phosphorus (P) levels. The presence of B.t. increased extra- and intraradical colonization [measured as frequency (%F), intensity (%I) and percentage of arbuscules (%A)] for both arbuscular mycorrhizal fungi (AMF) rather than plant growth or nutrition regardless P level. Under the lowest level of P fertilization, B.t. enhanced to a similar extent the extra- and intraradical development of both endophytes, but the proportion of fungal tissue showing SDH or ALP was increased in G. intraradices-colonized plants. [SDH: 458% (M) and 512% (A); ALP: 358% (M) and 300% (A)]. P supply decreased G. intraradices colonization to a higher extent than G. mosseae. Nevertheless, the totality of G. intraradices structures developed in P-amended medium showed intraradical o extraradical activity, while in G. mosseae-colonized roots, SDH and ALP activities highly decreased relative to fungal tissue determined by TB staining as affected by P. Our results show that bacterial inoculation compensates the negative effect of P on the intraradical fungal growth and vitality. P amendment reduced in a higher extent G. intraradices infection intensity (non-vital and vital staining) and G. mosseae activity (ALP staining). Thus, big differences in the proportion of SDH-active infection showing ALP activity in mycelium developed by each endophyte were noted at the highest P level. Physiological plant parameters such as photosynthetic activity did not explain specific changes on each arbuscular-mycorrhizal fungus as affected by P or B.t. inoculation. The increased extraradical mycelium development and metabolic fungal activity as a result of B.t. inoculation positively affected N and P plant content and photosynthetic rate in G. intraradices-colonized plants under the lowest P conditions. In general, the increased metabolically active fungal biomass in co-inoculated plants was irrespective of P level and was not related to the P plant uptake from the inert soil-less substrate. These results show the bacterial effect increasing the physiological and metabolic status of AM endophytes, which not only confirms but also extends previous findings on arbuscular mycorrhizae-bacteria interactions. The present study emphasizes the ecological and practical importance of rhizosphere free-living bacteria as mycorrhizae-helper microorganisms.     

Spatial distribution of phosphatase activity associated with ectomycorrhizal plants is related to soil type

Several ectomycorrhizal fungi, including Hebeloma cylindrosporum, actively release large quantities of phosphatase enzymes into their growth medium. We fractionated the phosphatase activity of the ectomycorrhizal association between H. cylindrosporum and its host plant, Pinus pinaster, with the aim to quantify its spatial and temporal variation in response to contrasting soil phosphorus conditions. Seedlings were grown in mini-rhizoboxes and the phosphomonoesterase activity of rhizosphere soil, released by roots, surface-bound to roots or mycelium was determined spectrophotometrically with the p-nitrophenyl phosphate method or microscopically with the ELF-method as a function of culture time. We showed that acid phosphatase activity of the soil and the root increased with mycorrhizal association. We also observed that the phosphatase activity associated with ectomycorrhizal plants was related to soil type. All phosphatase fractions decreased over culture time, except the proportion of hyphae exhibiting phosphatase activity in the extramatrical mycelium, which increased over time. The specific fractions of phosphatase activity associated with the mycorrhizal plants were clearly related to the soil phosphorus type and content. Soils showed an increase in acid phosphomonoesterase activity with mycorrhizal association, supporting a role for this enzyme in the degradation of soil bound phosphorus. The gradually increasing proportion of hyphae in the extramatrical mycelium exhibiting alkaline phosphatase activity, particularly under low phosphorus conditions, indicates an induction of alkaline phosphatase activity by phosphorus limitation.     

Organic acid exudation by mycorrhizal Andropogon virginicus L. (broomsedge) roots in response to aluminum

Elevated aluminum (Al) availability limits plant growth on acidic soils. Although this element is found naturally in soils, acidic conditions create an environment where Al solubility increases and toxic forms of Al impact plant function. Plant resistance to Al is often attributed to organic acid exudation from plant roots and the chelation of cationic Al in the rhizosphere. The association of arbuscular mycorrhizal (AM) fungi with the roots of plants may alleviate Al toxicity by altering soil Al availability or plant exposure through the binding of Al to fungal structures or through the influence of fungi on exudation from roots. Diverse communities of AM fungi are found in soil ecosystems and research suggests that AM fungi exhibit functional diversity that may influence plant performance under varying edaphic environments. In the present study, we evaluated acidic isolates of six AM species in their responses to Al. Andropogon virginicus (broomsedge), a warm-season grass that commonly grows in a range of stressful environments including acidic soils, was used as a plant host for Acaulospora morrowiae, Glomus claroideum, Glomus clarum, Glomus etunicatum, Paraglomus brasilianum, and Scutellospora heterogama. Fungal spores were germinated and exposed to 0 or 100 μM Al on filter paper in sand culture or were grown and exposed to Al in sand culture in association with A. virginicus. Short- and long-term responses to Al were evaluated using direct measurements of fungal spore germination, hyphal elongation, and measurements of A. virginicus colonization and plant growth as a phytometer of AM function in symbio. Spore germination and hyphal elongation varied among AM species in response to Al, but patterns were not consistent with the influences of these AM species on A. virginicus under Al exposure. Exposure to Al did not influence colonization of roots, although large differences existed in colonization among fungal species. Plants colonized by G. clarum and S. heterogama exhibited the least reduction in growth when exposed to Al, produced the highest concentrations of Al-chelating organic acids, and had the lowest concentrations of free Al in their root zones. This pattern provides evidence that variation among AM fungi in Al resistance conferred to their plant hosts is associated with the exudation of Al-binding organic acids from roots and highlights the role that AM fungal diversity may play in plant performance in acidic soil environments.     

Stability of desiccated rhizosphere soil aggregates of mycorrhizal Juniperus oxycedrus grown in a desertified soil amended with a composted organic residue

Adequate soil structural stability favours the establishment and viability of a stable plant cover, protecting the soil against water erosion in desertified Mediterranean environments. We studied the effect of soil drying-rewetting, inoculation with a mixture of three exotic arbuscular mycorrhizal (AM) fungi (Glomus intraradices Schenck & Smith, Glomus deserticola (Trappe, Bloss. & Menge) and Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe) and addition of a composted organic residue on aggregate stabilisation of the rhizosphere soil of Juniperus oxycedrus. The AM fungi and composted residue produced similar increases in plant growth, independently of the water conditions. Under well-watered conditions, the highest percentages of stable aggregates were recorded in the amended soil, followed by the soil inoculated with AM fungi. Excepting microbial biomass C, the soil drying increased labile C fractions (water soluble C, water soluble and total carbohydrates), whereas the rewetting decreased significantly such C fractions. Desiccation caused a significant increase in aggregate stability of the rhizosphere soil of all plants, particularly in the amended and inoculated plants. In all treatments, the aggregates formed after soil drying were unstable, since, in the rewetting, they disappear, reaching the initial levels before soil drying. Our results suggest that the aggregation mechanisms developed by rhizosphere microbial community of the amended and inoculated plants under water stress can be particularly relevant in desertified soils exposed to long desiccation periods.     

Soil inoculation with the biocontrol agent Clonostachys rosea and the mycorrhizal fungus Glomus intraradices results in mutual inhibition, plant growth promotion and alteration of soil microbial communities

Interactions between the biocontrol fungus Clonostachys rosea IK 726 and a tomato/Glomus intraradices BEG87 symbiosis were examined with and without wheat bran, which served as a food base for C. rosea. In soil without wheat bran amendment, inoculation with C. rosea increased plant growth and altered shoot nutrient content resulting in an increase and decrease in P and N content, respectively. Inoculation with G. intraradices had no effect on plant growth, but increased the shoot P content. Dual inoculation with G. intraradices and C. rosea followed the pattern of C. rosea in terms of plant growth and nutrient content. Wheat bran amendment resulted in marked plant growth depressions, which were counteracted by both inoculants and dual inoculation increased plant growth synergistically. Amendment with wheat bran increased the population density of C. rosea and reduced mycorrhizal fungus colonisation of roots. The inoculants were mutually inhibitory, which was shown by a reduction in root colonisation with G. intraradices in treatments with C. rosea and a reduction in colony-forming units (cfu) of C. rosea in treatments with G. intraradices, irrespective of wheat bran amendment. Moreover, both inoculants markedly influenced soil microbial communities examined with biomarker fatty acids. Inoculation with G. intraradices increased most groups of microorganisms irrespective of wheat bran amendment, whereas the influence of C. rosea on other soil microorganisms was affected by wheat bran amendment. Overall, inoculation with C. rosea increased and decreased most groups of microorganisms without and with wheat bran amendment, respectively. In conclusion, despite mutual inhibition between the two inoculants this interaction did not impair their observed plant growth promotion. Both inoculants also markedly influenced other soil microorganisms, which should be further studied in relation to their plant growth-promoting features.     

The bacterial community of tomato rhizosphere is modified by inoculation with arbuscular mycorrhizal fungi but unaffected by soil enrichment with mycorrhizal root exudates or inoculation with Phytophthora nicotianae

Arbuscular mycorrhizal (AM) fungi have been shown to induce the biocontrol of soilborne diseases, to change the composition of root exudates and to modify the bacterial community structure of the rhizosphere, leading to the formation of the mycorrhizosphere. Tomato plants were grown in a compartmentalized soil system and were either submitted to direct mycorrhizal colonization or to enrichment of the soil with exudates collected from mycorrhizal tomato plants, with the corresponding negative controls. Three weeks after planting, the plants were inoculated or not with the soilborne pathogen Phytophthora nicotianae growing through a membrane from an adjacent infected compartment. At harvest, a PCR-Denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments amplified from the total DNA extracted from each plant rhizosphere was performed. Root colonization with the AM fungi Glomus intraradices or Glomus mosseae induced significant changes in the bacterial community structure of tomato rhizosphere, compared to non-mycorrhizal plants, while enrichment with root exudates collected from mycorrhizal or non-mycorrhizal plants had no effect. Our results support that the effect of AM fungi on rhizosphere bacteria would not be mediated by compounds present in root exudates of mycorrhizal plants but rather by physical or chemical factors associated with the mycelium, volatiles and/or root surface bound substrates. Moreover, infection of mycorrhizal or non-mycorrhizal plants with P. nicotianae did not significantly affect the bacterial community structure suggesting that rhizosphere bacteria would be less sensitive to the pathogen invasion than to mycorrhizal colonization. Of 96 unique sequences detected in the tomato rhizosphere, eight were specific to mycorrhizal fungi, including two Pseudomonas, a Bacillus simplex, an Herbaspirilium and an Acidobacterium. One Verrucomicrobium was common to rhizospheres of mycorrhizal plants and of plants watered with mycorrhizal root exudates.