Nickel-tolerant Brevibacillus brevis and arbuscular mycorrhizal fungus can reduce metal acquisition and nickel toxicity effects in plant growing in nickel supplemented soil

The growth of clover (Trifolium repens ) and its uptake of N, P and Ni were studied following inoculation of soil with Rhizobium trifolii, and combinations of two Ni-adapted indigenous bacterial isolates (one of them was Brevibacillus brevis) and an arbuscular mycorrhizal (AM) fungus (Glomus mosseae). Plant growth was measured in a pot experiment containing soil spiked with 30 (Ni I), 90 (Ni II) or 270 (Ni III) mg kg⁻¹ Ni-sulphate (corresponding to 11.7, 27.6 and 65.8 mg kg⁻¹ available Ni on a dry soil basis). Single inoculation with the most Ni-tolerant bacterial isolate (Brevibacillus brevis) was particularly effective in increasing shoot and root biomass at the three levels of Ni contamination in comparison with the other indigenous bacterial inoculated or control plants. Single colonisation of G. mosseae enhanced by 3 fold (Ni I), by 2.4 fold (Ni II) and by 2.2 fold (Ni III) T. repens dry weight and P-content of the shoots increased by 9.8 fold (Ni I), by 9.9 fold (Ni II) and by 5.1 fold (Ni III) concomitantly with a reduction in Ni concentration in the shoot compared with non-treated plants. Coinoculation of G. mosseae and the Ni-tolerant bacterial strain (B. brevis) achieved the highest plant dry biomass (shoot and root) and N and P content and the lowest Ni shoot concentration. Dual inoculation with the most Ni-tolerant autochthonous microorganisms (B. brevis and G. mosseae) increased shoot and root plant biomass and subtantially reduced the specific absorption rate (defined as the amount of metal absorbed per unit of root biomass) for nickel in comparison with plants grown in soil inoculated only with G. mosseae. B. brevis increased nodule number that was highly depressed in Ni I added soil or supressed in Ni II and Ni III supplemented soil. These results suggest that selected bacterial inoculation improved the mycorrhizal benefit in nutrients uptake and in decreasing Ni toxicity. Inoculation of adapted beneficial microorganisms (as autochthonous B. brevis and G. mosseae) may be used as a tool to enhance plant performance in soil contaminated with Ni.     

Exudates of dark septate endophyte (DSE) modulate the development of the arbuscular mycorrhizal fungus (AMF) Gigaspora rosea

Exudates of a dark septate endophyte (DSE) identified as Dreschlera sp., a common endophyte isolated by the inner cortical cells of the grass Lolium multiflorum, were put in contact with the arbuscular mycorrhizal fungus (AMF) Gigaspora rosea. These exudates stimulated the hyphal length and the hyphal branching of the AMF. A negative effect on the extramatrical phase of the AMF was detected. This is the first report to show how exudates of DSE can affect the development of AMF. These results show that DSE could be modifying the mycorrhizal status of the plants, modulating a different symbiosis in the rhizosphere.     

Interaction between the soil yeast Rhodotorula mucilaginosa and the arbuscular mycorrhizal fungi Glomus mosseae and Gigaspora rosea

The effect of the soil yeast, Rhodotorula mucilaginosa LBA, on Glomus mosseae (BEG n°12) and Gigaspora rosea (BEG n°9) was studied in vitro and in greenhouse trials. Hyphal length of G. mosseae and G. rosea spores increased significantly in the presence of R. mucilaginosa. Exudates from R. mucilaginosa stimulated hyphal growth of G. mosseae and G. rosea spores. Increase in hyphal length of G. mosseae coincided with an increase in R. mucilaginosa exudates. No stimulation of G. rosea hyphal growth was detected when 0.3 and 0.5 ml per petri dish of yeast exudates was applied. Percentage root length colonization by G. mosseae in soybean (Glycine max L. Merill) and by G. rosea in red clover (Trifolium pratense L. cv. Huia) was increased only when the soil yeast was inoculated before G. mosseae or G. rosea was introduced. Beneficial effects of R. mucilaginosa on arbuscular mycorrhizal (AM) colonization were found when the soil yeast was inoculated either as a thin agar slice or as a volume of 5 and 10 ml of an aqueous solution. R. mucilaginosa exudates (20 ml per pots) applied to soil increased significantly the percentage of AM colonization of soybean and red clover.     

Use of specific phospholipid fatty acids for identifying and quantifying the external hyphae of the arbuscular mycorrhizal fungus Gigaspora rosea

The external hypha of arbuscular mycorrhizal (AM) fungi, extending from roots out into soil, is an important structure in the uptake of phosphate from the depletion zone around each root. In this paper, we analysed some phospholipid fatty acids (PLFAs) derived from external hyphae of four AM fungi (Glomus etunicatum, Glomus clarum, Gigaspora margarita and Gigaspora rosea) to find fatty acids which may be useful as specific markers for identifying and quantify the external hyphae of Gigaspora species. Leek (Allium porrum L.) seedlings inoculated with each AM fungus were grown in river sand. Sand samples were collected and four PLFAs (16:1ω5, 18:1ω9, 20:1ω9 and 20:4) in the sand were analysed. In addition, the hyphal biomass in the sand was determined by the direct microscopic method. PLFAs 18:1ω9 and 20:4 were found in all the AM-inoculated and non-inoculated sand samples. PLFA 16:1ω5 was detected in the sand inoculated with G. etunicatum, G. clarum and Gi. rosea. PLFA 20:1ω9 was detected only in the sand inoculated with Gi. rosea. PLFAs 16:1ω5 and 20:1ω9 were not found in the sand inoculated with Gi. margarita. The amount of PLFA 20:1ω9 was closely correlated with the amount of biomass of external hyphae of Gi. rosea (r=0.937, P<0.001), whereas no correlation was observed for PLFA 16:1ω5. The 20:1ω9 content of Gi. rosea was approximately 6.56 nmol mg⁻¹ hyphal biomass. We suggest that PLFA 20:1ω9 can be used as a specific marker for identifying and quantifying the external hyphae of Gi. rosea, at least in controlled experimental systems.     

Short-chain polyphosphate in arbuscular mycorrhizal roots colonized by Glomus spp.: A possible phosphate pool for host plants

We investigated the relationship between plant growth and polyphosphate (poly P) content in Allium fistulosum roots colonized by Glomus sp. R10 or Glomus etunicatum with different levels of phosphate addition. Poly P was quantified by two enzymatic assays: long-chain poly P (>∼20 inorganic phosphate [P_i] residues) with polyphosphate kinase, and total poly P with polyphosphate exopolyphosphatase. The difference between these values was designated as short-chain poly P (<∼20 P_i residues). The content of long-chain poly P in mycorrhizal roots and plant growth was poorly correlated or not, depending in fungal species. However, the content of short-chain poly P and plant growth was highly correlated in both species. This difference suggests that short-chain poly P in mycorrhizal roots colonized by Glomus spp. functions as a pool of P_i for the plant.     

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.     

Changes in agricultural management drive the diversity of Burkholderia species isolated from soil on PCAT medium

In order to assess the diversity of culturable Burkholderia populations in rhizosphere and bulk soil and to evaluate how different agricultural management regimes and land use history affect this diversity, four treatments were evaluated: permanent grassland; grassland converted into maize monoculture; arable land and arable land converted into grassland. Burkholderia isolates obtained on PCAT medium were grouped in 47 clusters using 16S ribosomal RNA gene based PCR-DGGE combined with BOX genomic fingerprinting (DGGE-BOX). The distribution of the isolates in the DGGE-BOX clusters was used to calculate the Shannon diversity index per treatment. Interestingly, we observed that the Burkholderia diversity was affected by changes in the agricultural management, since the highest diversity was observed in permanent grassland and in continuous arable land. In addition, the diversity tended to be higher in the rhizosphere than in the corresponding bulk soil. The use of species abundance models indicated that rhizosphere communities had more even distributions than communities collected from the bulk soil. Identification of isolates revealed that only 2% of these belonged to the B. cepacia complex and that the majority was assigned to either (1) new Burkholderia species or (2) Burkholderia species that had originally been isolated from soil. Isolates classified as B. hospita, B. caledonica and Burkholderia sp. ‘LMG 22934’ and ‘LMG 22936’ were found mainly in the arable land, while isolates belonging to Burkholderia sp. ‘LMG 22929’ and B. phytofirmans were associated with the grassland area. Another potentially new Burkholderia species, ‘LMG 22932’, was found in both areas, in close association with the maize rhizosphere.     

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.     

Root exudate-stimulated RNA accumulation in the arbuscular mycorrhizal fungus Gigaspora rosea

Plant root exudates induce the transition from asymbiosis to presymbiosis in arbuscular mycorrhizal fungi. In order to get an insight into this developmental switch, two libraries of Gigaspora rosea and one library of Gigaspora gigantea were screened for fragments of genes that show enhanced RNA accumulation 1 h after addition of a semi-purified exudate fraction of carrot roots. Among 150 clones, 40 seemed to contain inserts of root exudate-induced genes. One of the genes, GrosRbp1, putatively encoding an RNA binding protein involved in developmental control showed RNA accumulation which correlates to the extent of stimulation of presymbiotic hyphal branching.     

Effects of soil application of olive mill wastewaters on the structure and function of the community of arbuscular mycorrhizal fungi

Recycling of olive mill wastewaters (OMW) into agricultural soils is a controversial issue since benefits to soil fertility should counterbalance potential short-term toxicity effects. We investigated the short-term effects of OMW on the soil-plant system, regarding the diversity, structure and root colonization capacity of arbuscular mycorrhizal (AM) fungi and the respective growth response of Vicia faba L, commonly used as green manure in olive-tree plantations. A compartmentalized pot system was used that allowed the establishment of an AM fungal community in one compartment (feeder) and the application of three OMW dose levels in an adjacent second compartment (receiver). At 0, 10, and 30 days after OMW treatment (DAT), V. faba pre-germinated seeds were seeded in the receiver compartment. At harvest, shoot and root dry weights, AM fungal root colonization, soil hyphal length and P availability were recorded in the receiver compartment. In addition, OMW effects on AM fungal diversity in plant roots were studied by DGGE. A transient effect of OMW application was observed; plant growth and AM fungal colonization were initially inhibited, whereas soil hyphal length was stimulated, but in most cases differences were absent when seeding was performed 30 DAT. Similarly, changes induced in the structure of the root AM fungal community were of transient nature. Cloning and sequencing of all the major DGGE bands showed that roots were colonized by Glomus spp. The transient effects of OMW on the structure and function of AM fungi could be attributed to OMW-derived phytoxicity to V. faba plants or to an indirect effect via alteration of soil nutritional status. The high OMW dose significantly increased soil P availability in the presence of AM fungi, suggesting efficient involvement of AM fungi in organic-P minerilization. Overall our results indicate that soil application of OMW would cause transient changes in the AM fungal colonization of V. faba plants, which, would not impair their long-term plant growth promoting ability.     

Changes in soil aggregation and glomalin-related soil protein content as affected by the arbuscular mycorrhizal fungal species Glomus mosseae and Glomus intraradices

Arbuscular mycorrhizal (AM) fungi are key organisms of the soil/plant system, influencing soil fertility and plant nutrition, and contributing to soil aggregation and soil structure stability by the combined action of extraradical hyphae and of an insoluble, hydrophobic proteinaceous substance named glomalin-related soil protein (GRSP). Since the GRSP extraction procedures have recently revealed problems related to co-extracting substances, the relationship between GRSP and AM fungi still remains to be verified. In this work the hypothesis that GRSP concentration is positively correlated with the occurrence of AM fungi was tested by using Medicago sativa plants inoculated with different isolates of Glomus mosseae and Glomus intraradices in a microcosm experiment. Our results show that (i) mycorrhizal establishment produced an increase in GRSP concentration - compared to initial values - in contrast with non-mycorrhizal plants, which did not produce any change; (ii) aggregate stability, evaluated as mean weight diameter (MWD) of macroaggregates of 1-2 mm diameter, was significantly higher in mycorrhizal soils compared to non-mycorrhizal soil; (iii) GRSP concentration and soil aggregate stability were positively correlated with mycorrhizal root volume and weakly correlated with total root volume; (iv) MWD values of soil aggregates were positively correlated with values of total hyphal length and hyphal density of the AM fungi utilized.The different ability of AM fungal isolates to affect GRSP concentration and to form extensive and dense mycelial networks, which may directly affect soil aggregates stability by hyphal enmeshment of soil particles, suggests the possibility of selecting the most efficient isolates to be utilized for soil quality improvement and land restoration programs.     

Can the arbuscular mycorrhizal fungus Glomus intraradices actively mobilize P from rock phosphates?

Plants colonized by arbuscular mycorrhizal (AM) fungi have been shown to respond positively to the application of insoluble forms of inorganic phosphorus (P) such as rock phosphates (RPs). The mechanism(s) underlying such responses remain(s) unknown and although it has been hypothesized, there is no experimental support for the production of chelating agents by AM fungal hyphae. Here we investigate whether AM fungi can solubilize P from RPs and transfer it to plant roots. Using root-organ cultures of Daucus carrota L. inoculated or not with Glomus intraradices Schenk & Smith and containing P from different RP sources, we predicted that: (1) roots inoculated with G. intraradices would take up more P than those uninoculated; that (2) the amount of P taken up by roots through G. intraradices would be positively correlated with the RP reactivity; and that (3) G. intraradices would have access to RP through localized alterations of pH and/or by the production of organic acid anions that may act as chelating agents. The RP reactivity was positively correlated with P uptake. However, mycorrhizal roots grew initially slower and did not respond differently to any P treatment than those uninoculated. There was no evidence of localized changes in pH in proximity of G. intraradices hyphae, indicating that responses to RP by mycorrhizal plants observed in previous studies do not appear to result from the release of H⁺ ions alone or in combination with organic acid anions.     

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.     

Impacts of earthworms and arbuscular mycorrhizal fungi (Glomus intraradices) on plant performance are not interrelated

Earthworms and arbuscular mycorrhizal fungi (AMF) might interactively impact plant productivity; however, previous studies reported inconsistent results. We set up a three-factorial greenhouse experiment to study the effects of earthworms (Aporrectodea caliginosa Savigny and Lumbricus terrestris L.) and AMF (Glomus intraradices N.C. Schenck & G.S. Sm.) on the performance (productivity and shoot nutrient content) of plant species (Lolium perenne L., Trifolium pratense L. and Plantago lanceolata L.) belonging to the three functional groups grasses, legumes and herbs, respectively. Further, we investigated earthworm performance and plant root mycorrhization as affected by the treatments. Our results accentuate the importance of root derived resources for earthworm performance since earthworm weight (A. caliginosa and L. terrestris) and survival (L. terrestris) were significantly lower in microcosms containing P. lanceolata than in those containing T. pratense. However, earthworm performance was not affected by AMF, and plant root mycorrhization was not modified by earthworms. Although AMF effectively competed with T. pratense for soil N (as indicated by δ¹⁵N analysis), AMF enhanced the productivity of T. pratense considerably by improving P availability. Remarkably, we found no evidence for interactive effects of earthworms and AMF on the performance of the plant species studied. This suggests that interactions between earthworms and AMF likely are of minor importance.     

Novel strains of nodulating Burkholderia have a role in nitrogen fixation with papilionoid herbaceous legumes adapted to acid, infertile soils

We investigated the taxonomic position and symbiotic capabilities of two root-nodule bacterial strains isolated from the South African herbaceous, papilionoid legume Rhynchosia ferulifolia. The 16S rRNA gene sequence of the two strains was determined along with intragenic sequences of nodA and nifH, together with their symbiotic capabilities when inoculated onto the papilionoid legumes R. ferulifolia, Rhynchosia caribaea, Rhynchosia minima and Macroptilium atropurpureum (Siratro). Burkholderia phymatum STM815^T, Cupriavidus taiwanensis LMG 19424^T and root-nodule bacteria isolated from R. minima and Rhynchosia totta were included in the study. Root-nodule bacteria isolated from R. ferulifolia, WSM3937 and WSM3930, belong to the genus Burkholderia and are most closely related to Burkholderia terricola (98.8% similarity). The phylogenetic analysis of nodA and nifH revealed substantial similarity of the novel strains with Burkholderia tuberum STM678^T, a β-rhizobium also originated from South Africa, and only a distant relationship with South American Mimosa-nodulating β-rhizobia. R. ferulifolia was effectively nodulated only by Burkholderia sp. WSM3937 and WSM3930 and not by bradyrhizobia isolated from Rhynchosia minima and Rhynchosia totta or STM815 and LMG 19924. Nodules induced by the novel strains were determinate and hosted well organized symbiosomes within infected cells. In this study we describe a new symbiotic N-fixing relationship between Burkholderia sp. and the South African legume R. ferulifolia. This is the first report of N-fixation between β-rhizobia and an herbaceous, papilionoid legume from which the strains were originally isolated. The level of N-fixation in this symbiosis approached that achieved by effectively nodulated Medicago sativa and suggests that the β-rhizobia may have a role in N-fixation in agricultural systems.     

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.     

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.     

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.     

Sugar beet waste and its component ferulic acid inhibits external mycelium of arbuscular mycorrhizal fungus

External arbuscular mycorrhiza (AM) mycelium plays an important role in soil while interacting with a range of biotic and abiotic factors. One example is the soil organic amendment sugar beet waste. The fermented Aspergillus niger-sugar beet waste (ASB) increases growth and P uptake by the AM mycelium in soil whereas non-fermented waste (SB) had a strong inhibitory effect. The underlying mechanisms are not understood.We used gas chromatography-mass spectrometry to identify differences in composition of water extracts of ASB and SB. The chromatograms showed that ferulic acid was present in SB and absent in ASB. We compared the effects of the water extracts of SB and ASB and ferulic acid upon the growth of Glomus intraradices in in vitro monoxenic cultures.Hyphal growth of the AM fungus G. intraradices was extremely reduced in ferulic acid and SB treatments. Moreover, AM hyphae appeared disorganized, undulated and tangled. In contrast, ASB increased hyphal length and numbers of branched absorbing structures and of spores. We conclude that ferulic acid is one compound in SB which is responsible for its inhibition of AM extraradical growth. The relevance of these findings is discussed.